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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016) 5–7 February 2016 Venue: Yashada Auditorium, Pune, India

Welcome Message from the Organizers Dear Delegates, On behalf of the organizing committee, I am really happy to welcome you all at the “3rd International Conference on Biotechnology and Bioinformatics”, being organized from 5-7 February 2016, at Yashada Auditorium, Pune, India. This is a platform where scientists, professors and students from various fields of biotechnology and bioinformatics will gather and exchange the existing knowledge and generate new ideas for future research. There will be more than 150 poster presentations on various topics within the conference themes spread over three days and you should take advantage of this gathering to build your network of scientist within your own area of research interest. Our mission and slogan is “Spreading the Knowledge & Bringing People Together” and every year this conference is getting bigger. We hope that this gathering of young students, scientists and faculty members will fulfill our objectives and help in your career advancements. We look forward to welcome you again next year at the 4th ICBB-2017. The city and venue will be announced soon at our website www.icbb.in. We wish you all a very happy stay in Pune and enjoy the conference!

Yours sincerely,

Dr. Sheo Mohan Singh, Organizer, MSc(UK), PhD(Germany), PDF(USA,UK)

Co-organizers: Dr. Christoph Steinbeck, Head of Cheminformatics & Metabolism, European Bioinformatics Institute (EMBL-EBI), Wellcome Trust Genome Campus, Cambridge, UK Dr. Stefan Bohlander, Marijanna Kumerich Chair in Leukemia & Lymphoma Research, Department of Molecular Medicine and Pathology, University of Auckland, Auckland, New Zealand Dr. Laxman Gangwani, Associate Professor, Department of Biomedical Sciences, Texas Tech University Health Sciences Center, El Paso, USA Dr. Arun Trivedi, Senior Scientist, Central Drug Research Institute (CDRI), Lucknow, India

Conference Secretariat: Dr. Sheo Mohan Singh, Director, ICSCCB, A-304, Orange Province, Vishal Nagar, Pimple Nilakh, Pune-411027, India Web: www.icscc.in, Email: [email protected], Tel: +91-9545089202

www.icsccb.org International Centre for Stem Cells,Cancer and Biotechnology (ICSCCB), Pune, India

Programs 6-Months Dissertation/Certificate in Stem Cell Biology & Molecular Medicine (PGC-SCBMM) 1-Year Post Graduate Program in Stem Cell Biology & Molecular Medicine (PGP-SCBMM) 1-Week Hands-on Workshop on Human/Cancer Cell Culture Techniques & MTT Assay 5 days Hands-on Workshop on Molecular Biotechnology and Bioinformatics Director:

Scientific Advisors:

Prof. Dr. Sheo Mohan Singh

Prof. Dr. Keith Humphries, Canada Prof. Dr. Christian Buske, Germany Prof. Dr. Laxman Gangwani, USA Prof. Dr. M. Buitenhuis, Netherlands Dr. S.N. Singh, Bangalore Dr. Sambuddha Ghosh, Pune

MSc(UK), PhD(Germany), PDF(USA,UK)

Please log on to www.icsccb.org for more information and application. Contact: Email: [email protected], [email protected] Tel: +91-9545089202

APS LABS KITS 1. BACTERIAL CULTURE 2. GEL EXTRACTION AND PCR CLEAN UP KIT 3. TISSUE AND BLOOD CULTURES 4. RNA PURIFICATION 5. FOOD EXTRATION 6. VIRAL NUCLEIC ACID 7. PLANT RESEARCH KITS 8. DNA RELATED PRODUCTS 9. PROTEIN RELATED PRODUCTS 10. PCR/RT PCR PRODUCTS 11. RT PCR KIT 12. PCR CLONING AND LIGATION 13. LUMIX-BOX –BLUE LIGHT (U V trans illuminator alternatives)

APS LABS SERVICES 1. 2. 3. 4. 5. 6. 7. 8. 9. 10.

MICROARRAY & NGS SERIVICES FLOWCYTOMERTY & GENOMIC SERVICES PRIMERS & GENE SYNTHESIS CELL CULTURE PRODUCTS & SERVICES PLANT GENOMIC SERVICES CUSTOMISED PROJECTS & TURNKEY SOLUTIONS FOOD SAFETY TESTINGS & CLINICAL DIAGNOSTICS ENVIRONMENTAL & BIOANALYTICS ATC & DRUG DISCOVERY PLATFORM ANIMAL HOUSE & CUTOM ANTIBODIES SOLUTIONS

Contact: 020-65607783 Web: www.apslifetech.com Email: [email protected]

 

3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

                                 

 

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Organized by: ICSCCB, www.icsccb.org, Pune, India

3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

1. Eco-friendly crop protection strategies using plant secondary metabolites Ashok P. Giri Senior Scientist, Plant Molecular Biology Unit, Division of Biochemical Sciences, CSIR-National Chemical Laboratory, Dr. Homi Bhabha Road, Pune 411 008 (MS), India

Available history manifests contemporary diversity that exists in plant-insect interactions. A radical thinking is necessary for developing strategies that can co-opt natural insect-plant mutualism, ecology and environmental safety for crop protection since current agricultural practices can reduce species richness and evenness. The global environmental changes, such as increased temperature, carbon dioxideand ozone levels, biological invasions, land use change and habitat fragmentation together play a significant role in reshaping the plant-insect multi-trophic interactions. Diverse natural products need to be studied and explored for their biological functions as insect pest control agents. Plant produces variety of secondary

metabolites

for

specific

application.

Here

I

focus

on

understanding the role of plant secondary metabolites in plant defense against insect pest.In our search for insecticidal or insect growth inhibitors,particularlysmall molecules, we identified a plant natural phenol, caffeic acid (CA);and terpenoids - camphor, limonene and β-caryophyllene, as potent inhibitors of Helicoverpa armigeragrowth and development. In silico and in vitro screening of multiple molecules supported by in vivo validations suggested that CAinhibitsH. armigera gut proteases. Protease activity and gene expression were altered in CA-fed larvae. The structureactivity relationship of CA highlighted that all the functional groups are crucial for inhibition of protease activity. In another experiments, H. armigera Page | 2   

larvae

fed

on

artificial

diet

supplemented

with

 

Organized by: ICSCCB, www.icsccb.org, Pune, India

(i)

3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Ocimumkilimandscharicum leaf extract, (ii) camphor, (iii) limonene, and (iv) β-caryophylleneshowed growth retardation, increased mortality rates and pupal deformities. Digestive enzymes of H. armigerafed on these dietsnamely, amylase, protease and lipase showed variable patterns after feeding on O. kilimandscharicum, which implies striving of the larvae to attain required nutrition for growth, development and metamorphosis. Altogetherourabove

findings

suggest

that

these

plant

secondary

metabolitescan be implicated as potent insecticidal molecules and could be explored for the development of effective dietary pesticides. Related publications: 1. Mishra M, Lomate PR, Joshi R, Punekar SA, Gupta VS, Giri AP(2015) Ecological turmoil in evolutionary dynamics of plant-insect interactions: Defense to offence. Planta 242, 761-771 2. Joshi RS, Wagh T, Sharma N, Fayaj M, Sonawane U, Thulasiram HV, Joshi R, Gupta VS, Giri AP(2014) Way towards “Dietary Pesticides”: Mechanistic insight into insecticidal action of natural phenols. Journal of Agriculture and Food Chemistry62, 1084710854 3. Singh PA, Ramesha HJ, Sarate PJ, Kulkarni MJ, Thulasiram HV, Giri AP (2014) Insecticidal potential of defense metabolites from Ocimum kilimandscharicum against Helicoverpa armigera.PLoS ONE9(8), e104377

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

2. Therapeutic Potential of Bone Marrow-derived, Allogeneic Mesenchymal Stromal Cells for Patients with Critical Limb Ischemia due to Buerger’s disease Gupta, PK1, Anoop CH, Balasubramanian, S1, Mathiazhagan, R1,Rajkumar M2, Krishna M3, Dutta S4, Sarkar, U5, Desai, S6, Radhakrishnan, R7, Dhar,A8,Prasanth KV1, Majumdar, AS1 1

Stempeutics Research, Bangalore, 2Dept. of Vascular Surgery, SRM Medical College Hospital

and Research Centre, Kattankulathur,Chennai, 3Dept. of Vascular Surgery, Sri Jayadeva Inst. of Cardiovascular Sciences, Bangalore, 4Dept. of Cardiovascular Surgery, Nightingale Hospital, Kolkata, 5Dept. of Cardiovascular Surgery, Health Point Hospital, Kolkata, 6Dept. of Vascular Surgery, M.S Ramaiah Medical College & Hospitals, Bangalore, 7Dept.of Vascular Surgery, Sri Ramchandra Medical College, Chennai,8Dept. Of Vascular Surgery, All India Institute of Medical Sciences, New Delhi.

Thromboangiitis obliterans (Buerger’s disease), is a non-atherosclerotic, segmental inflammatory disease that most commonly affects the small and medium-sized vessels. Although the exact etiology of Buerger’s disease is not clear, the condition is strongly associated with heavy tobacco use. Critical Limb Ischemia (CLI)is a severe form of the disease that results in rest

pain

and

non-healing

ulcers

and

gangrene

of

the

lower

extremity.Approximately 40% to 50% of “no-option” patients of CLI will undergo amputation within 6 – 12 months and approximately 15% will also require contralateral amputation within 2 years. Several important characteristics of multipotent mesenchymal stromal cells (MSC) suggest that these cells may offer therapeutic benefit to these “no option” patients of CLI. Bone marrow derived MSC (BMMSC) are known to possess strong immunomodulatotory and anti-inflammatory properties, and promote angiogenesis

and

tissue

regeneration

through

paracrine

activity.

Stempeucel® is manufactured from BMMSC obtained from healthy volunteers and the product is comprised of pooled allogeneic BMMSCs Page | 4   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

obtained from multiple donors. Stempeucel® express MSC-associated surface markers, differentiate into bone, cartilage and adipose cells, possess potent potent immunosuppressive activity and secrete various angiogenic factors including VEGF, angiopoietin, IL8 and HGF. Preclinical studies have demonstrated that stempeucel® is non-toxic and nontumorigenic. Administration of the product ameliorates limb necrosis, promote blood flow and prevent limb loss in a mouse model of hind limb ischemia.We have recently completed a phase II, clinical trial in patients with Buerger’s disease(NCT01484574) with two doses of stempeucel® (1 and 2 million cells/kg body weight).36 patients each were enrolled in the cell armsand 18 patients in the control arm who received standard protocol of care (SPOC). The cells were injected intramuscularly around the gastrocnemius muscle and around the ulcer. At six months follow updata revealedthat both the primary end points (relief of rest pain & healing of ulcers) were significantly better in 2M/kg group as compared to SPOC. Rest pain reduced 0.3 units (SE=0.13) per month, which was statistically significant (p = 0.0193), CI= (-0.55,-0.05) whereas, there was 11% (SE=0.05) significant decrease in ulcer size per month (p=0.0253) CI= (0.80, 0.99).

In addition, Ankle Brachial Pressure Index(p=0.0132)also

show significant improvement in 2M/kg dose group, suggesting improved blood flow in the affected limb. The improvement in quality of life, total walking distance has also been observed in patients in the stempeucel® treatment arms. None of the adverse events in the trial was found to be associated with stempeucel® administration. Hence, it is concluded that stempeucel® is safe and provides therapeutic benefits to “no-option” patients suffering from Buerger’s disease.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Keywords: Buerger’s disease, allogeneic mesenchymal stem cells, rest pain 3. Does your alcohol alter the mitochondrial structural (fusion/fission) genes? Subbiah, Ramasamy1,*, Sivakumar, Anusha1 , Muniswamy Madesh2

1. Cardiac Hypertrophy Laboratory, Department of Molecular Biology, School of Biological Sciences, Madurai Kamaraj University, Madurai- 625 021. 2. Centre for Translational Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania, PA, USA. *Correspondence: [email protected]

Mitochondria are indispensible, dynamic subcellular organelles for cellular homeostasis. Energy demand, mitochondrial genome integrity and copy number relies on their counter balanced and equipoised fusion-fission phenomenon, arbitrated by GTP-dependent fusion proteins (MFN1/2, OPA1) and fission factors (Drp1, Fis1). With several recent reports implicating deregulated mitochondrial dynamics in neurodegenerative diseases, cardiovascular diseases and diabetes, our study aims to elucidate the role of these machinery protein genes during chronic ethanolinduced liver damage. Confocal microscopic images of mitochondria, probed with mito-GFP, exposed to varying doses of ethanol display fragmented and punctate structures in a dose-dependent manner. Superoxide, generated during ethanol metabolism, also show a dosedependent increase when stained with mitoSOX-Red while mitochondrial membrane potential, oxygen consumption, ATP biosynthesis and NADH oxidation is significantly reduced. Fusion/fission machinery genes do not show significant alterations at the transcriptional level but western blot of MFN1 Page | 6   

depicts

a

processed/degraded

fragment

at

higher

 

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ethanol

3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

concentration. Taken together, these data reveal that ethanol induced mitochondrial fragmentation occurs via excessive superoxide generation and stimulating cytosolic or mitochondrial proteolytic factors that degrade/ process the fusion protein, MFN1. Identification and inhibition of these factor(s) could resuscitate mitochondrial structure and function, providing a therapeutic strategy for liver damage in habitual alcoholics.

4. Phenomics in anti-parasitic drug discovery Dhanasekaran Shanmugam

Biochemical Sciences Division, CSIR National Chemical Laboratory, Pune, India

Parasitic infections are a huge scourge to human and animal health and newer drugs are needed to effectively combat these infections. In CSIRNCL, we are conducting early stage drug discovery work on two important model parasitic organisms – the human malaria parasite Plasmodium falciparum and the agent for human and animal toxoplasmosis, Toxoplasma gondii. In contrast to classical whole cell live-dead inhibitor screening assays, our focus is to carryout phenotypic screens to gain insights into the mechanism of action of the inhibitory molecules. This talk will highlight the utility of different screens such as replication timing assays, cell cycle progression assays, delayed death assays and finally metabolic perturbation assays, for tracking the phenotypic effects of the test molecules on both Plasmodium and Toxoplasma. The talk will also cover the results from the phenotypic screens we have run on a collection of potent anti-parasitic compounds call ‘Malaria Box’, a likely source for future anti-malarial and other anti-parasitic drugs.   Page | 7   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Curcumin and extracellular matrix proteins synergistically act to inhibit the proliferation of breast cancer cells Bilen Lemma Regassa*, Ashwin Wadaskar** and Anuradha Vaidya*** *M.Sc. Student, **Ph.D. Student, ***Corresponding author, Deputy Director, Symbiosis School of Biomedical Sciences (SSBS), Symbiosis International University (SIU), Symbiosis Knowledge Village, Gram: Lavale, Taluka: Mulshi, District: Pune-412115

Natural substances have been gaining attention as they are accessible, cheap and associated with milder adverse effects as compared to conventional treatment. Curcumin is the active ingredient of turmeric plant and is known to possess anti-inflammatory, anti oxidant and anti metastatic effects. Here we have systematically studied the effects of curcumin individually and in combination with ECM (extracellular matrix) proteins on the MCF-7 cell line. MTT assay was performed to screen the effect of various concentrations of curcumin on MCF -7 cells. Flow cytometry and caspase-3 assays were done to detect apoptosis. Nitric oxide (NO) and reactive oxygen species (ROS) generation levels were also examined as both are known to play very important roles in cancer initiation and progression. It was observed that at low concentrations curcumin exhibited significant antiproliferative activity which was supported with increased apoptosis and reduced levels of ROS and NO. Moreover, the combined treatment

of

curcumin

and

ECM

proteins

further

boosted

the

antiproliferative effect of low concentrations of curcumin indicating that they act synergistically to inhibit the proliferation of breast cancer cells. This finding could be further investigated for development of novel anti cancer therapies.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Pka RII Sub Unit Phosphopentapeptide: A Candidate Substrate For Calcineurin 1,2 1

Devaraju K S, 1Nivedita Shireshi 2Harish B M, 2Saraswathi R, and 3Hemalatha N

Department of Biochemistry, Karnatak University, Pavate Nagar, Dharwad-580 003,

India. Department of 2Biotechnology, J B Campus, Bangalore University, Bangalore-560 056, India. 3

Department of Biochemistry and Nutrition, CFTRI, V V Moholla, Mysore-570 020, India. [email protected]

The measurement of Calcineurin (CN) activity plays an important role in understanding the mechanism underlying many pathological conditions like cancer, metabolic aberrations and neurodegenerative disorders. Till date contemporary methods to assay CN use specific protein phosphatase inhibitors or use of specific substrate protein kinase A regulatory sub unit type II.

Here we have used a small phosphopeptide substrate derived

from protein kinase A regulatory sub unit type II, being employed to assay CN. Before developing an assay, we have done the affinity studies for series of small phospho peptides with CN by using Molgro Virtual Docking software to find out the binding free energy. Further the candidate phosphopenta peptide was synthesized by employing F-moc chemistry (Devaraju K S et. al., 2007). MASS spectrometry.

Purified over HPLC and characterized by

This phospho pentapeptide was used for in vitro

assay of CN by Malachite Green Method and the results were compared with R II peptide.

The kinetics study indicates that the small

Phosphopentapeptide could be used as a candidate substrate for the assay of calcinuerin in place of RII peptide. Key words: Calcineurin, Protein Kinase-A (PKA), Docking, Peptide, Enzyme Assay,

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Comparison of Copper Surface Mediated Toxicity in Gram-Positive, Gram-Negative Bacteria and Saccharomyces cerevisiae. Nidhi Gadura. Biology Department, Queensborough Community College, City University of New York, Bayside, NY 11364

Copper alloy surfaces are known as antimicrobial sanitizing agents that have the ability to kill microorganisms. However, the mechanism by which cell death occurs still remains unclear. The aim of our project is to determine the relationship between exposure to copper alloy surfaces or copper ions, lipid peroxidation, and killing of Gram positive, Gram negative bacteria and yeast. We hypothesize that we might see differences in the mode of copper mediated cell death in different bacterial and fungal species. Using Gram positive (Staphylococcus aureus), Gram negative (Pseudomonas aeruginosa) bacteria and Saccharomyces cerevisiaestrain BY4741 quantitative dilutions series was performed to test for bacterial and yeast cell death. Our results indicate a biphasic killing curve for Gram positive, Gram negative and Saccharomyces cerevisiaewhen exposed to copper chips. TBARS assay was used to measure the amount of lipid peroxidation that occurred. The bacterial and yeast killing rate upon exposure to copper surfaces also correlates to increased levels of lipid peroxidation. Invitrogen Live/Dead Assay was done using Fluorescent Microscopy to confirm the cell death. Genomic DNA was extracted to study the mode of cell death, apoptotic vs. necrotic.There are some differences seen in the kinetics of cell death that correlates with the levels of lipid peroxidation between the two bacterial strains and yeast. Genomic DNA extraction indicates that the mode of cell death seems to be necrotic in all cases.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Spectroscopic Investigations on the interaction of natural alkaloid and ct-DNA and G-Quadruplex DNA: A Comparative Study Paulami Mandal University of Calcutta Department of Biochemistry Kolkata-700019 [email protected]

Present investigation intends to observe the interaction of natural alkaloid Harmine with ct-DNA and G-quadruplex DNA by using different spectroscopic techniques. G-quadruplex DNA arrangement is highly polymeric in nature and is weakly abundant as compared to duplex-DNA, hence targeting this DNA is quite challenging in itself. Till date most of the investigations were focused on targeting duplex DNA. Only recently Gquadruplex motifs are identified as biologically crucial structure and hence ligand-G quadruplex DNA interaction study is emerging as a new field of research. Formation of G-quadruplex structures can inhibit the activity of telomerase enzyme that elongates the G rich strand of telomeric DNA. This enzyme is active in about 85% of tumors, leading the cancer cells to infinite lifetime. Therefore the small molecules that induce and stabilize the formation of quadruplex are considered to be potential telomerase inhibitors. This study involves htel22 AGGGTTAGGGTTAGGGTTAGGG sequence. In the presence of metal ions such as K+ or Na+, telomeric DNA can form structures of the G-quadruplexes. In our case quadruplex structure is formed in the presence of K+ions. TE buffer solution (10mMol L1

Tris-HCl, 1mMol L-1 EDTA, pH 7.4) is used as the starting stock solution.

In this investigation, steady state and time resolved fluorescence along with circular dichroism (CD) spectroscopic and anisotropy investigations were made to reveal the nature of the interactions between Harmine and both Page | 12   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

DNAs. The goal of this project is to explore the critical differences in effects of that particular small molecule on the two forms of DNAs like, dissimilarities in the efficiency of binding and unwinding, dissociation. These could be useful in targeting tumor and/or cancer cells and designing more effective drugs. Phytochemicals as a source of herbal Antibiotics: A new horizons Priti Mathur 1*,Chandni Tandon1,Manodeep Sen2 and Sanjeev Kanojiya3,Mohini chaurasia1** 1*,1

Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow- 226028

1**

Amity Institute of Pharmacy, Amity University Uttar Pradesh, Lucknow- 226028

2

Department of Microbiology, Dr. Ram Manohar Lohia Institute of Medical Sciences, Lucknow-226010

3

Sophisticated Analytical Instrument Facility, CSIR-Central Drug Research Institute, Lucknow-226031 [email protected]; [email protected]

The emergence of antibiotic-resistant forms of pathogenic S. aureus e.g. Methicillin-resistant Staphylococcus aureus (MRSA) is a worldwide problem in clinical medicine. Although S.aureus is not always pathogenic, it is a common cause of skin infections, respiratory disease (e.g. sinusitis), and food poisoning. Keeping the problem in view, widely growing plants of tropical area like Andrographis Paniculata, Cannabis Sativa, Datura innoxiaet care investigated for its active ingredient. The isolated extract was tested for its antibacterial activity at crude and purified fraction and characterizes during different analytical technique and bioinformatics software. Antibacterial activity was tested upon clinically isolated different strains of Staphylococcus aureus. Bioassay done in terms of inhibition(ZOI),minimum

inhibition

concentration(MIC),

zone of minimum

Bacteriocidal Content (MBC) and for shelf life. Further, the purified extract Page | 13   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

was subjected to semisolid formulation and evaluated for pH, Loss on drying, Density, Appearance, Spreadibility and extrudibility. All the formulations were found to be smooth in nature. Weight before and after drying and density was measured to ensure batch to batch uniformity. Optimized formulation showed semistiff nature with spread diameter between 42-45mm. The formulation was found to have excellent extrudibility when allowed to extrude from lacquered aluminum collapsible tube. The formulations tested for its antibacterial activity and found to have moderate antibacterial properties. The present experiment showed that compound from plant source could be utilized as herbal antibiotics. Key words: Antibacterial phytocompound, ZOI, MIC, MBC, shelf life, analytical techniques, antibacterial formulation CUX2 Functions As an Accessory Factor in the Repair of Oxidative DNA Damage Ranjana Pal1, 8,§, Zubaidah M Ramdzan1,§, Simran Kaur1,2, Philippe M. Duquette5, Richard Marcotte6, Lam Leduy1, Sayeh Davoudi2, Nathalie Lamarche-Vane5, Angelo Iulianella7, Alain Nepveu1,2,3,4 Goodman Cancer Research Centre 1, and Departments of Biochemistry2, Medicine3 and Oncology4, Anatomy and Cell Biology5, McGill University, 1160 Pine Avenue West, Montreal, Quebec, Canada, H3A1A3. Princess Margaret Cancer Centre, University Health Network 6, Toronto, M5G 1L7, Canada. 7

Department of Medical Neuroscience , Dalhousie University, Life Science Research Institute, 1348 Summer Street, Halifax, B3H 4R2, Canada 8

Present address : Department of Biological Sciences, Presidency University, Kolkata, [email protected]

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India §

Both authors contribute equally to this work.

CUX1 and CUX2 proteins are characterized by the presence of three highly similar regions called Cut repeats 1, 2 and 3. While CUX1 is ubiquitously expressed, CUX2 plays an important role in the specification of neuronal cells and continues to be expressed in post mitotic neurons. Cut repeats from the CUX1protein were recently shown to stimulate 8-oxoguanine DNA glycosylase, (OGG1), an enzyme that removes oxidized purines from DNA and introduces a single-strand breakthrough its AP-lyase activity to initiate base excision repair. Here, we investigated whetherCUX2 plays a similar role in the repair of oxidative DNA damage. CUX2 knockdown in embryonic cortical neurons increased levels of oxidative DNA damage. In vitro, Cut repeats from CUX2 increased the binding of OGG1 to 8-oxoG containing DNA and stimulated both the glycosylase and AP-lyase activities of OGG1.Genetic inactivation in mouse embryo fibroblasts or CUX2 knockdown in HCC38 cells delayed DNA repair and increased DNA damage. Conversely, ectopic expression of Cut repeats from CUX2 accelerated DNA repair and reduced levels of oxidative DNA damage. These results demonstrate that CUX2 functions as an accessory factor that stimulates the repair of oxidative DNA damage. Neurons produce high level of reactive oxygen species because of their dependence on aerobic oxidation of glucose as their source of energy. Our results suggest that the persistent expression of CUX2 in post mitotic neurons contributes to the maintenance of genome integrity through its stimulation of oxidative DNA damage repair.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Identification, expression, purification, characterization and role in proinflammatory response of YghJ, a secreted metalloprotease from neonatal septicemic Escherichia coli Rima Tapader1, Sulagna Basu2, Amit Pal1 1

Division of Pathophysiology, National Institute of Cholera and Enteric Diseases,

Kolkata, West Bengal, India, 2Division of Bacteriology, National Institute of Cholera and Enteric Diseases, Kolkata, West Bengal, India. [email protected]

Among the Gram negative pathogens involved in sepsis of newborns, Escherichia coli occupies a pivotal position in developing countries and has emerged as the major pathogen for sepsis related mortality among preterm infants. In search for a novel virulence factor in neonatal septicemic E. coli (NSEC) we identified YghJ, a secreted metalloprotease. YghJ has recently been shown to be highly immunogenic and has been identified as one of the potential vaccine candidates for extraintestinal pathogenic E. coli. YghJ was cloned, expressed and purified from EB260, a clinical NSEC strain. YghJ showed optimal activity at pH 7-8 and at temperature of 3740°C. YghJ showed cytotoxicity by exhibiting clear changes in cellular morphologies of Int407, HT-29 and HEK293 cell lines. Sepsis is the systemic inflammatory response of host against an infection which involves the release of an array of endogenous mediators. Proinflammatory cytokines are one of the mediators attributed to play key role in pathogenesis of sepsis and are implicated in the mortality and morbidity associated with septic shock. The proinflammatory cytokines are considered as an important diagnostic marker for early diagnosis of neonatal sepsis. Bacterial LPS are known to activate proinflammatory response of host. However the role of virulence factors in NSEC isolates responsible for stimulating the proinflammatory response has not yet been Page | 16   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

elucidated. We report for the first time that YghJ can stimulate the production of proinflammatory cytokines such as IL-1α, IL-1β, TNF-α, IL-8 in murine macrophages and human intestinal epithelial cell lines. YghJ can also down-regulate the production of anti-inflammatory cytokines such as IL-10. Interestingly, in our study, yghJ is present in both septicemic (78%) and fecal E. colii solates (54%). However, expression of YghJ is significantly higher among the septicemic (80%) than the fecal isolates (33%). Our results demonstrate that YghJ essentially contributes to the pathogenesis of NSEC. Study of Cellulases from Extremophiles Sangeeta Shetty1,2, Vishwas Sarangdhar2, Sheela Kamat3, Dileep V. Kamat3 1

Department of Life Science and Biochemistry, 2Caius Research Laboratory, St. Xavier's College (Autonomous), Mumbai, and 3Department of Microbiology, Mithibai College, Mumbai. [email protected]

Developing sustainable technology to convert plant biomass into fuels is one of the core research areas of the Clean Energy Initiative's put forth by the Government of India. Microbial Cellulases have been used to convert the lingo cellulosic mass to bio-ethanol, but this technology is not commercially viable because of low conversion efficiency of the currently available enzymes. This problem can be addressed by harnessing the potential of enzymes from extremophiles as these enzymes are known to possess higher stability and efficiency in unusual conditions. The objective of this study was to isolate extremophiles with high cellulolytic activity. Soil samples were spread on Avicel-agar plates at extreme pH ranges and varying temperatures of incubation. The cellulosedegrading bacteria were detected by the Congo red staining method. This Page | 17   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

yielded many cellulase-positive strains, some of which had ability to grow at pH of 3.0, some preferred pH 10, whereas some could grow even at pH 12. Some of the isolates could grow and digest crystalline cellulose (Avicel) at 50ºC. The enzymes produced by these were assayed qualitatively using the supernatants and the pellets of the bacterial cultures. The cellulases produced by these isolates were extracellular and constitutive. Molecular phylogenetic characterization of some of these isolates was carried out using bio-informatic tools to analyze the 16s rRNA gene sequences. Some microbial strains that were isolated showed presence of two different enzymes, one which is active at acidic pH, and the other at both alkaline and acidic pH. Induction of these enzymes depends on the pH of the growth medium. Study on the unique cellulolytic strains capable of variable induction depending on pH will enhance our understanding of the biology of cellulose-degraders. These isolates could have application in biofuel production, as they can tolerate pH up to 12.0 and/ or temperature of 50°C.

Recent Developments in assessing the overall quality of ProteinProtein Interaction structural models: their scoring and ranking. Sankar Basu, Bjorn Wallner Bioinformatics Division Department of Physics, Chemistry and Biology, Linkoping University Sweden 58183.

Protein-Protein Interactions (PPI) serve as an indispensable component in understanding the basic molecular biology of life on earth and its associated disorders. Building of computational, structural models for PPI Page | 18   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

complexes in the absence of the experimental structure is an active field of research. These computationally built PPI atomic models must be associated with confidence estimates (scores) of their correctness with respect to the hypothetical experimental target. Again, the accurate design and training of such a 'knowledge-based' scoring function requires a suitable quality measure (e.g., Cα-RMS deviation) calculated in the presence of experimentally solved native structures (targets) in a database. The state-of-the-art for these quality measures is currently based on three related yet independent measures as proposed and standardized by the CAPRI1 community. These quality measures quantify different aspects of the quality of a particular docking pose and need to be viewed together to reveal the overall true quality. This evaluation protocol is integrated with applying various ad-hoc cutoffs on these measures to classify a model into different classes. This classification has been useful in CAPRI, but also rather limiting, making it difficult to rank models, correlate with scoring functions or use it as target function in machine learning algorithms. The current work presents the design and utility of a single quality measure (DockQ; http://bioinfo.ifm.liu.se/DockQ/) based on the CAPRI evaluation protocol and also the development of a scoring function (CompEn) trained by machine learning using DockQ as a target function. CompEn elegantly combines shape and electrostatic complementarity at the protein-protein interface with different energy terms and performs better than the currently available energy functions in scoring and ranking PPI models in CAPRI. In addition, a structure validation tool for PPI complexes, namely the Complementarity

Plot

(http://bioinfo.ifm.liu.se/SARAMAint/SARAMAint-

liupage.html)2 is also presented, to be used effectively in conjugation with a

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

similar tool developed earlier for single domain globular proteins (http://www.saha.ac.in/biop/www/sarama.html)3-5. Related Publications: 1. Lensink, M.F., and Wodak, S.J. 2013. Proteins 81, 2082–2095. 2.Basu et al., Journal of Bioinformatics and Intelegent Controll, 2014, 3:309-314. 3. Basu et al., Biophysical Journal 2012, 102 (11): 2605-2614. 4. Basu et al., Journal of Bioinformatics and Intelegent Controll 2013, 2 (4): 321-323. 5. Basu et al., Indian Journal of Biochemistry and Biophysics, 2014, 51 (June) : 188-200.

Molecular studies on Alpinia calcarata Silvy Mathew1, S John Britto2 The Rapinat herbarium and Centre for Molecular Systematics, St. Joseph’s college, Tiruchirappalli, 620002, Tamilnadu St. Dominic’s college, Kanjirappally, Kottayam (Dt), 686512, Kerala [email protected]

A. calcarata is a rare multipotent, medicinal plant. The isolation of good quality DNA is the prerequisite for molecular research. DNA barcoding based on cpDNA is a useful technique for species identification and assessing genetic diversity. The query sequences of A. calcarata trnL - trnF have 99% identical with previously reported A.zerumbet sequences. Chloroplast DNA sequencing resulted in an unambiguous 396bp for A. calcarata.

Also, In the present investigation 7 Alpinia species were

collected and identified from Western Ghats (A. malaccensis, A. purpurata, Page | 20   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

A. galanga, A. vittata, A. mutica, A.abundiflora) and multiple sequence alignment and the phylogeny studied by using the MEGA6 software. Six sequences of Alpinia from the Western Ghats were compared thoroughly with A.calcarata by molecular phylogenetic analysis like maximum likelihood method, neighbor - joining method, minimum evolution method, UPGMA method and maximum parsimony analysis to assess the performance in species discrimination. From the phylogenetic trees it is clear that A.calcarata shows high affinity to A.mutica based on trnL-trnF sequences.  

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India      

                                              Page | 22   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Photonic crystals made up of stimuli-responsive nanoparticles: Possible applications for color tunable biosensors Aashish Agarwal1,Madhusmita Palai2 ,Kriti Agarwal3 Department of Biotechnology KIIT University, Bhubaneswar [email protected]

In the current study, we have synthesized a novel type of nano-particle so called

stimuli-responsive

microgels.

These

are

poly

(Nisopropylacrylamide)(PNIPAm) co Poly(acrylic acid) based microgels having a cross-linked polymeric structures that can undergo reversible continuous or discontinuous volume transitions upon variations of external conditions such as temperature, pH or ionic strength. We have synthesized these microgel particles using precipitation polymerization and then these microgelparticles are characterized using dynamic light with respect to their size, swelling ratio. In a next step, we are able to crystallize these microgel particles at higher concentration. These crystals diffract light at a particular lattice constant which is of the order of wavelength in the visible region and follows Bragg’s law, 2μdsinθ=nλ (d: Average inter-planer spacing, μ : Refractive index, θ: Angle of diffraction, n: Order of diffraction, λ: Wave length of light). The average inter planner spacing depends on the volume fraction of the microgel particles.

As these microgels particles are pH and thermo-

responsive, the volume of the particle can be tuned simply by varying the pH and temperature of the solvent. Hence in a concentrated suspension, the wavelength of diffraction can be tuned by changing the volume of the particle which can be done by changing pH or temperature of the solvent. We believe that these unique properties (Stimuli-responsive swelling deswelling of the microgels) can be effectively used for a designing a color Page | 23   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

tunable biosensor. A biosensor is a device made up of a transducer and some biological elements. Biological elements can be proteins, enzymes, polymers etc. These elements interact with analytes which convert the biological response to an electrical signal. In our case of microgel particles, it is expected that the sensing molecules swell the microgel particle thus changing the interparticle distance that cause a shift in Bragg’s peak wavelength. A comparative review of PprA gene with respect to Deinococcus radiodurans using Bioinformatics tools Jayarama Reddy, Sushma I Hiremath, Adeline Rebecca Hubert and Chandra Mohana Centre for Molecular and Computational Biology, St.Joseph’s College, 36, Langford Road, Bangalore – 560027. [email protected]

Deinococcus radiodurans a member of the family Deinococcaceae, is an extremophile which shows remarkable resistance to a range of damages caused by ionizing radiations, dessication, ultraviolet radiations and oxidizing agents. The extent of radiation resistance of this bacterium is due to efficiency of mechanism by combination of proteins such as PprA, Rec A, SSB, IrrE, DdrA and DdrD. The PprA (Pleiotropic Protein Promoting DNA Repair A) protein repairs the DNA double stranded breaks. It plays a critical role in a non-homologous end joint (NHEJ) pathway for the repair of radiation induced DNA double strand breaks. This study involves a comparative analysis of the function and mechanism of PprA gene in Deinococcus

radiodurans

with

respect

to

other

organisms.

The

bioinformatic analysis of the PprA gene was done using tools such as BLAST, FASTA, UniProtKB and ClustalW server.

Page | 24   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Veterinary Sciences and Animal Biotechnology: Applications Of Various Techniques Aishwarya Iyer, Tejal Patil, Arpita Wagle SIES Graduate School of Technology Sri Chandrasekaendra Saraswathy Vidyapuram Navi Mumbai [email protected]; [email protected]; [email protected]

Veterinary Science and Animal Biotechnology is the science of diagnosing, treating and curing the diverse types of diseases in birds and animals. It involves not only taking care of animal health but also includes scientific breeding and handling of livestock. The major goals of veterinary vaccines are to improve the health and welfare of companion animals, increase production of livestock in a cost-effective manner, and prevent animal-tohuman transmission from both domestic animals and wildlife. Veterinary science in the diagnosis involves the emergence of microarray-based technologies and the design of DNA biochips allow high-throughput studies of RNA expression in cell and tissues. In treatment Nanotechnology has a tremendous potential to revolutionize agriculture and livestock sector. Nanotechnology tools like micro fluidics, nanomaterials, bio analytical nanosensors, etc. has the potential to solve many more puzzles related to animal health, production, reproduction and prevention and treatment of diseases. Also new technologies of assisted reproduction are being developed to apply the properties of spermatogonial stem cells to preserve endangered animal species. Artificial insemination (AI) is a technique which is widely used by veterinarians and veterinary specialists in most domestic as well as wild animal species. In production animals, it is a way to increase reproductive efficiency, production and has proven to be a very effective reproductive technology that selectively increases genetic gain Page | 25   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

through increased selection pressure on males. The above mentioned techniques have been proven to be effective in the field of veterinary sciences and animal biotechnology and are therefore gaining more importance. Keywords:

veterinary

sciences,

animal

biotechnology,

microarray,

nanotechnology, artificial insemination, stem cells.

Isolation and Characterization of xylanolytic bacteria from insect gut Aishwarya Puranik, Pallavi Purohit, Sona Umloti, Sowjanya K BVB College of Engg. & Tech., Hubballi, India. [email protected]

Bombyx moriL (Lepidoptera: Bombycidae) have been domesticated and widely used for silk production. It feeds on mulberry leaves. Mulberry leaves are mainly composed of pectin, xylan, cellulose and starch. Some of the digestive enzymes that degrade these carbohydrates might beproduced by gut bacteria. Three isolates were obtained from the digestive tract of B. mori, whichwere Gram-negative motile and spore forming. The gut of B. mori has an alkaline pH and all of the isolated bacterial strainswere found to grow and degrade polysaccharides at alkaline pH. Screening was done using Congo red method. Different characterization methods like gramstaining and catalase test were carried out to characterize the isolates. The isolates were found to be catalase positive. Submerged fermentation was carried out for three isolates of silkworm for the enzymeassay.Xylanase activity was determined by submerged fermentation.Potent xylanolytic bacteria were obtained from the gut.These bacteria provide digestive enzymes in a synergic manner and contribute to larval growth. Enzyme

Page | 26   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

production had begun from the second day of the submerged fermentation itself and it increased with the increase in the incubation period.Also the media for the growth of these xylanolytic gut bacteria was optimized. Nitrogen source was found to be very necessary for the growth of xylanolytic bacteria.The bacterial isolates obtained from 5th instar larvae were found to have the ability to digest cellulose, xylan, pectin and starch, all of which are found in mulberry leaves.This suggests that bacteria secrete enzymes important in digestion. Keywords: Bombyx mori, xylanolytic bacteria,xylan. Stem Cells And Breast Cancer Aishwarya S Patil Department of Biotechnology SIES graduate school of technology Nerul, Navi Mumbai. [email protected]

The concept of cancer stem cells responsible for tumour origin, maintenance and resistance to treatment has gained prominence in the field of breast cancer research. Here, we review the role of stem cells in the healthy breast, the role of breast cancer stem cells in disease, together with their implications in breast cancer progression, treatment. There is increasing evidence that this cancer is originated in and maintained by a small population of undifferentiated cells with self-renewal properties. The cancer stem cells are now considered as backbone in the development of the cancer. A number of agents capable of targeting breast cancer stem cells in preclinical models are currently entering clinical trials. Assessment of the efficacy of the agents will require development of innovative clinical trial designs with appropriate biologic and clinical end points. The effective Page | 27   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

targeting of breast cancer stem cells has the potential to significantly improve outcome for women with both early-stage and advanced breast cancer. Keywords:

Stem

cell,

Breast

cancer,

Tumour,

Self-renewal,

Undifferentiated cells RamA-regulated gene promoters Aishwarya Saxena, MSc Biotechnology Thesis Research Performed Under Dr Thamarai Schneiders, Department of Infection and Pathway Medicine, University of Edinburgh, Edinburgh, Scotland UK

Antibiotics are used to fight infectious diseases and are also utilised in industrial and agricultural processes. This widespread use of antibiotics together with the ability of microbial pathogens to quickly adapt to changing environments

escalates

the

problem

of

resistance

development.

Transcriptional regulators from the AraC/XylS family play key role in adaptation to environmental stresses. These regulators allow the finetuning of many bacteria cellular processes that increases their survival. Alteration in expression of these regulators that in turn modulate their regulon can develop superoxide resistance or multiple antibiotic resistance (mar) like phenotypes. This mar phenotype is conferred by reducing the permeability of the cells. MarA/SoxS/Rob are examples of global regulators that are found in the Enterobacteriaceae family. These regulators are highly conserved and share high homology amongst each other. RamA is another such regulator, first discovered in Klebsiella pneumoniae, where it plays a major role in regulating the mar phenotypes. MarA, SoxS, Rob and RamA regulate expression of same set of genes, but with different affinities. A recent study has shown that RamA-mediated regulation in K. pneumoniae controls genes involved in both antibiotic resistance and virulence. Page | 28   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Therefore this study was aimed at determining the binding site of the different genesin order to regulate transcription. In order to determine this sequence, the promoter sequencesof candidate genes were fused to GFP to

assess

expression

in

the

presence

and

absence

of

RamA.

Subsequently,fluorometric readings were taken to measure the levels of activation of various promoters from RamA-regulated genes. As previously shown, our resultsconfirms activation of different promoter sequences. Hypobaric hypoxia inducedskeletal muscle loss: A biochemical and molecular approach Akanksha Agrawal, Rajendra Prasad, Geetha Suryakumar, Richa Rathor Defence Institute of Physiology and Allied Sciences, DRDO, Lucknow Road, Timarpur, Delhi-110054 [email protected]

Background: High altitude exposure leads to a number of physiological, biochemical and molecular changes in skeletal muscle. The present study was planned to elucidate the various proteolytic pathways and the role of calcium in muscle protein degradation. Material and Methods: Twenty four male Sprague- Dawley rats (200220g) were divided into four groups (I, II, III, and IV) and were exposed to altitude of 7620 m for 0, 6, 12 and 24 hrs respectively for hypobaric hypoxia (HH) exposure. On completion of exposure rats were sacrificed and muscle was excised for analysis. Results and Conclusion: Numerous changes were observed at24 hrs HH exposure. At 24 hrs HH exposure, ROS was increased significantly and that was accompanied by decrease in GSH leveland total thiol content Page | 29   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

which attributed for antioxidant function in rats.Disturbances in oxidantantioxidant redox status leads to increase protein oxidation in terms ofincreased protein carbonyl content, advanced oxidized protein products (AOPP), POOH content, bityrosine content, tryptophan oxidation which was observed in HH exposed rats. Further, GRP78 (ER Chaperone), HSP70 (Cytoplasmic Chaperone) and HSP60 (Mitochondrial Chaperone) were also increased in HH exposed rats that depicts protein misfolding. Along with this calpain activity, proteosome activity and acid phosphatase activity was also increased in 24hr HH exposure, which further represents increase in proteolytic pathways like ubiquitin-proteosome, calpain and lysosomalautophagy for degradation of misfolded protein. One of the interesting finding of present study is elevated cytosolic calcium in HH exposure group which could be responsible for increase calpain activity and ultimately resultant into skeletal muscle loss at high altitude.

Energy use and economy analysis of bioethanol production from wheat straw in India Akash Kumar, Archana Mishra, Dr. SanjoyGhosh Department of Biotechnology, IIT Roorkee [email protected], kakash,[email protected]

Conventional fossil fuels such as coal, oil and natural gases are main sources of energy in the present era. However, foreseeable depletion of fossil fuels resulted in huge recognition of biofuels which are derived from recently dead biological material. Bioethanol has shown its potential as one of the most promising alternative for gasoline as transportation fuel among biofuels, though ethanol production from traditional feedstock like corn is limited. Therefore, alternate feedstock sources are needed to produce Page | 30   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

bioethanol. Among these, lignocellulosic biomasses are most promising as these are abundant, cheap and have high polysaccharide content. The cellulose and hemicellulose, which typically comprise two-third of the dry biomass, are polysaccharides that can be hydrolyzed to sugars and eventually to ethanol by fermentation. A suitable pretreatment step is required to increase the digestibility of total available fermentable sugars in the biomass at industrial scale. Some common examples of lignocellulosic biomasses are rice straw, wheat straw, bagasse and kans grass. As India is 2nd largest producer of wheat, wheat straw can be the most suitable lignocellulosic feedstock for Indian scenario. In addition, wheat straw has shown highest energy efficiency. This paper focuses on economy and energy analysis of bioethanol production from wheat straw in India. Energy consumption and generation of each step involving ethanol production from wheat straw will be discussed. It may help Government of India to achieve its goal of 20% blending of bioethanol in gasoline by promoting research based on wheat straw for bioethanol production. Keywords: Biofuel, bioethanol, lignocellulosic biomass, wheat straw

Adaptation to Salt Stress in moderately Salt-Tolerant Strain of the Yeast Zygosaccharomycesbisporous Akshya Sharma, Dr.Sukesh Chander Sharma Department of Biochemistry, Panjab University, Chandigarh, 160014 [email protected]

Many early studies reported that yeasts caused spoilage of various highsugar foods and high salted foods such as honey, raw sugar cane, jams .The dominant yeasts isolated from these foods belongs to the genus Zygosaccharomyces, the majorly of which are Z. rouxii, Z. bailii and Z. Page | 31   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

bisporus. The yeast Zygosaccharomycesbisporous is commercially known for its osmotolerance and moderately halotolerance. When exposed to NaCl, the cells experience both osmotic stress and ion toxicity. Specifically, this research work is focused on the evaluation of NaCl-induced stress responses of a food spoilage yeast Zygosaccharomycesbisporous, particularly with regard to yeast cell growth, viability and various biochemical mechanisms. Experiments were designed by pretreating cells with different sodium chloride concentrations (NaCl; 0.0M, 0.5M, 0.75M, 1.0M) growing in defined YEPD media and it was clearly observed that growth of cells was reduced with increased concentration of stress agent from 0.0M to 1.0M NaCl. Further, we assessed and compared the growth, percent viabilty, and measured the intracellular polyols (mainly glycerol), osmolytes (like trehalose, glutathione) and cations (Na+ or K+) accumulated during growth leading to ‘osmotic adjustment’ as a major element in accomplishing tolerance in the absence and presence of 1.0M NaCl concentration. It was observed that salt stress is accompanied by an increase in intracellular level of trehalose, glutathione, glycerol as compared to control cells. Moreover, elevated level of antioxidant enzymes like Superoxide Dismutase (SOD), Catalase (CAT) scavengeexcess of Reactive Oxygen Species (ROS) generated due to salt stress which down regulates the oxidative injury to cells. Thus, the study of the physiological and molecular mechanisms underlying yeast osmoregulation and long term osmoadaptation relate to the importance of these aspects in biotechnology and in agriculture but yet poorly understood. For instance, improving crop resistance to soil salinity stresses is a long-standing goal of agricultural biotechnology. Thus, the understanding of ion homeostasis mechanisms and the creation of salt tolerant yeasts and plants by metabolic engineering Page | 32   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

is a very important goal of salt stress research. The information hidden in yeast genes may be successfully transferred to other microorganisms, plants and other higher eukaryotes, resulting in the improvement of resistance capabilities. Thus, heterologous expression of genes associated with salt stress resistance remains one of the most important goals in salt stress research. Also, there is almost no substantial knowledge on basic areas of physiology, biochemistry and even genetics related with salt stress in these and other yeasts suggesting the study of more halotolerant yeasts, since they represent potentially good models to unveil some of the physiological and molecular mechanisms underlying yeast long-term extreme tolerance to salt, which cannot be approached in less tolerant yeasts such as S. cerevisiae. Keywords: Genetics, Agriculture, Biotechnology, Biochemistry, Stress tolerance, Physiology

Effects of Processing on the Proximate Composition and Energetic Values in Two Fish Species from Iran Southern Waters Ali Aberoumand Behbahan Khatam Alanbia University of Technology, Behbahan, Iran. [email protected]

It is imperative to conserve nutrients of seafood products and also is a major consumer concern related to seafood preparation. It is important to determine the retention of nutrients in fish cooked using several common domestic practices, namely boiling, baking, and frying. The aim of this study was impact different cooking methods (frying, boiling and baking) Page | 33   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

on

the

proximate

and

mineral

composition

of fish species

Scomberoides commersonnianus and Spyraena jello . The established AOAC methods were followed for composition bio chemical of fish. Protein and lipid content were founded higher in baked and fried in fish S.commersonnianus (74.29%) and (20.20%) and fish S. jello (88.12%) and (17.77%) respectively. Ash content in fish S.commersonnianus varies from 9.80% to 15.34%, and in fish S. jello from 5.83% to 7.68%. The highest protein, the lowest fat and energy contents were found in boiled in both fish species; therefore, boiling can be recommended as the best cooking method for healthy diet. Comparison of nutrients contents of boiled fillets of two fish species showed that fish S. jello found best. Keywords: Scom beroides lysan, Sphyraenidae, cooking methods, proximate composition

Microsatellite marker based identification of turmeric (Curcuma longa L.) cultivars Ambika Sahoo, Sudipta Jena, BasudebaKar, SanghamitraNayak Centre of Biotechnology, School of Pharmaceutical Sciences, Siksha O Anusandhan University, Bhubaneswar-751003, Odisha, India. [email protected]

Turmeric (Curcuma longa L.) of the family Zingiberaceae is one of the most economically important plant due to its use in food, medicine and cosmetic industries. Due to high morphological similarity and existence of many synonyms, the cultivar identification in turmeric is a major constrain affecting its commercialization. Microsatellite markers are presently being Page | 34   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

used as preferred molecular markers for cultivar identification in several plant species for their uniqueness of not being influenced by environmental factors. In this study, molecular identification work was carried out on eight elite cultivars using EST-SSR markers. Fifty EST-SSR primers were screened against eight cultivars of turmeric (Suroma, Roma, Lakadong, Megha, Alleppy Supreme, Kedaram, Pratibha and Suvarna) out of which 11 primers showed polymorphic banding pattern. The polymorphic information content (PIC) of these primers ranged from 0.13 to 0.48. However only 3 SSR loci (CSSR 14, CSSR 15 and CSSR 18) gave reproducible unique banding pattern clearly distinguishing the cultivars ‘Lakadong’ and ‘Suvarna’ from other cultivars tested. A total of 40 alleles were amplified from these three selected polymorphic SSR primers. Consistent matching of 3 SSR primers were validated with multiple number of registered cultivars of ‘Lakadong’ and ‘Suvarna’. Further, two identified cultivars (Lakadong and Suvarna) were also precisely differentiated based on phylogenetic tree when analyzed with other 94 genotypes of turmeric. The novel SSR markers can be used for identification and authentication of two high drug yielding turmeric cultivars ‘Lakadong’ and ‘Suvarna’.

Plant Biotechnology: Future Viewpoint Ambuj Kumar Bajpai Bhagwant Univesity, Ajmer (Raj.)-305 004 [email protected]

Plant biotechnology has made useful strides in thc past some years encompassing within its fold the spectacular developments in plant molecular biology and genetic engineering. Some of the most vexing problems to be faced in agricultural ecosystems could be solved with the Page | 35   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

introduction of transgenic crops endowed with feature for insect pest resistance, herbicide tolerance and resistance to viral diseases. Attention is now being focused on the development of transgenic plants having industrial,

economic,

pharmaceutical,

nutritional

and

environmental

importance. In the next millennium, crops will assist as factories for the synthesis of valuable metabolites and organic compounds. Agronomically important

characters,

such

as

drought

tolerance,

adeptness

in

photosynthesis, nutrient use and nitrogen fixation will be manipulated in the next century to embellish the genetic and physiological potential of the crops. Recent developments in the genome chaining of Arabidopsis, rice and maize will have far extending intimation for future agriculture. Structural and functlonal genomics of plant species will effectively rebellionise the complexion of agricultural biotechnology as well as human health care.It is requisite that the developing world adopts these fast-changing technologies soon and harness their unprecedented imminent for the benefit of the mankind.

Keywords:

Plant

biotechnology,

Molecular

engineering, plant genomics,

breeding,

plant

transgenic plants, gene-transfer

techniques,

Page | 36   

genetic

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Differential nucleosome occupancy in plant genomes; Are promoter regions significantly depleted of nucleosomes? Amish Kumar and Gitanjali Yadav* National Institute of Plant Genome Research, Aruna Asaf Ali Road, New Delhi-110067, India [email protected]

Wrapping of DNA around the nucleosomes core particle is the key to the chromatin packaging as well as for the regulation of gene expression. A comparative study of nucleosome occupancy patterns, at the genome wide level may help to understand chromatin structure and its functional control over the cell. Whole genome nucleosome occupancy maps are available for several organisms and provide sufficient knowledge base to accurately predict the site of occurrence of nucleosomes over a new genome sequence. For example, yeast genome shows differential organization of nucleosomes in the upstream regulatory region of genes, resulting in the existence of distinct nucleosome free regions (NFRs) just before the transcription start site (TSS). This study was designed to test whether such NFRs are present in plant genomes as well, and whether the DNA itself guides differential nucleosome occupancy. Thus in order to understand the role of DNA sequence in nucleosome occupancy, we hypothesized that “There are fewer nucleosomes in promoters compared to other regions of the plant genome”. For this, we analyzed two plant genomes namely, Zea mays (a monocot) and Solanum tuberosum L. (a dicot) to construct genome wide nucleosome prediction maps. Per base nucleosome positioning was compared over the promoter and non-promoter regions of both the genomes. Statistical tests reveal that nucleosomes are indeed

Page | 37   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

depleted in promoters as compared to random, or other genomic regions of plants.

Standardization of DNA isolation and PCR protocol for RAPD analysis of different wheat (Triticum aestivum L.) genotypes Shinde Amol S* Lokmangal College of Agricultural Biotechnology, Wadala, Solapur, Maharashtra

Molecular analysis of plant require pure and high yield of DNA samples. Generally wheat DNA having problems of contamination of polyphenols and secondary metabolites and contaminated DNA is not properly amplifies in PCR. To avoid this problem here we standardized DNA isolation method for different wheat genotypes by referring of CTAB and SDS methods and also optimized RAPD protocol. The DNA extraction method involves grinding samples with extraction buffer containing 120mM Tris HCL 300 mM Nacl and 25 mM EDTA. All steps carried out at room temperature and resulted DNA quantity ranged from 158ng/µl to 425ng/µl and the purity (ratio) was between 1.6-1.9 indicating minimal levels of contaminating metabolites. The technique is ideal for isolation of DNA from different wheat genotypes and the DNA isolated was used for Randomly Amplified Polymorphic DNA (RAPD) analysis. RAPD protocol was optimized based on the use of higher concentration of MgCl2 (2.5mM), dNTPs (0.4mM), lower concentrations of primer (10 pmole) and Taq polymerase (1 unit), 30ng of template DNA and an annealing temperature of 37°C, resulted optimal amplification. Reproducible amplifiable products were observed in all PCR reactions. Thus the results indicate that the optimized protocol for Page | 38   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

DNA isolation and PCR was amenable to wheat genotypes, which are suitable for further work on diversity analysis. Key words: Triticum aestivum L., RAPD, PCR.

Effect of nicotine and caffeine on α-synuclein aggregation Ankur Bhuva*, Ratnika Sethi, Ipsita Roy. Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), S.A.S. Nagar, Punjab [email protected]

Parkinson’s disease (PD) is an age related neurodegenerative disorder. Neuropathological features of PD are that it shows a progressive loss of dopaminergic neurons in the substantia nigra in the mid-brain. It is marked by presence of lewy bodies (LB) and lewy neurites. Lewy bodies are fibrillar intracytoplasmic inclusions containing amyloid like fibrils composed of small protein called α-synuclein. α-Synuclein is a major component of lewy bodies. Various environmental and genetic factors are associated with etiology of PD. Oxidative stress, mitochondrial dysfunction, neuroinflammation, protein aggregation and neuro-degeneration have important role in pathogenesis of PD. Depending on the environmental conditions natively unfolded monomer forms fibrils, soluble oligomers, or amorphous aggregates. α-Synuclein fibrillation kinetics is represented by sigmoidal curve and is divided into three phases viᵶ. lag phase, exponential phase and stationary phase which follow a nucleation dependent fibrillation. αSynuclein oligomers can bind to lipid membrane and disrupt it and also penetrate into the membrane and form pore like structures. Epidemiological studies have shown an inverse relation of progression of disease to cigarette smoking and coffee drinking. Non smoker has twice the risk of PD Page | 39   

 

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than smoker. Cigarette smoke contains many compounds like nicotine, anabasine, cotinine, hydroquinone, and nornicotine. Among all, nicotine is a possible candidate for several reasons. Nicotine stimulates striatal dopamine neurons. Nicotine promotes dopamine release by stimulating nicotinic acetylcholine receptors. This indicates neuroprotective effect of nicotine. Caffeine is the main component of coffee and has a neuroprotective effect due to its ability to antagonize adenosine A2A receptor. A previous study of individual effect of nicotine and caffeine on yeast model of PD has shown prominent effect on aggregation of αsynuclein. Our study deals with the combined effect of nicotine and caffeine on aggregation of α-synuclein in-vitro and in a yeast model of PD.

Biochemical and Proteome Response of Rice to Salt Stress Arajmand Frukh1, T O Siddiqi1, Altaf Ahmad2 1 2

Department of Botany, Faculty of Science, Jamia Hamdard, New Delhi, India

Department of Botany, Faculty of Life Science, Aligarh Muslim University, Aligarh, India [email protected]

Abiotic stresses like heat, cold, freezing, drought, salinity, flooding or oxidizing agents are serious threats to agriculture and the environment which have been exacerbated in the current century by global warming and industrialization. According to FAO statistics, more than 800 million hectares of land throughout the world are currently salt-affected. Continuing salinization of arable land is expected to have overwhelming global impact, resulting in a 30% loss of agricultural land over the next 25 years and up to 50% loss by 2050. Over the last few decades there has been tremendous progress in understanding the molecular, biochemical, and physiological basis of stress tolerance in plants. Abiotic stress leads to a series of

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

morphological, physiological, biochemical and molecular changes in plants that adversely affect growth and productivity. An important consequence of salt stress is the excessive generation of reactive oxygen species (ROS) such as superoxide anion, hydrogen peroxide (H2O2) and the hydroxyl radicals (OH-). In plant cells, ROS are generated in high amounts by both constitutive and inducible routes, but under normal situations, the redox balance of the cell is maintained via the constitutive action of a wide range of antioxidant mechanisms that have evolved to remove ROS. Enzymatic ROS scavenging mechanisms in plants include SOD (superoxide dismutase), catalase, APX (ascorbate peroxidase), GR (glutathione reductase). By using hydroponic rice seedling culture system, we investigated the enzymatic antioxidative defense response of ten rice genotypes to screen out tolerant variety under various levels of salt treatment. To improve our understanding of the complex mechanisms involved in the response of rice to salt stress, a proteomic approach was used to identify proteins in the leaves of two genotypes (sensitive and tolerant) differing in their response to salt stress. Results indicate complex changes in leaf proteome in response to salt stress.

Page | 41   

 

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KIXBASE: A KIX domain database and web server for prediction and analysis. Archana Yadav, Jitendra K. Thakur* and Gitanjali Yadav* National Institute of Plant Genome Reaseach, Aruna Asaf Ali Marg, New Delhi [email protected]

In today’s era of ever-increasing biological data, it is essential to maintain databases that not only integrate and cluster the data as a warehouse but as well help analyze it. Such resources are invaluable tools for researchers as they provide a useful platform to retrieve biological information like sequences, structures, structure classes, pathways etc., thus aiding in the research to a great extent. KIXBASE is a global repository as well as a prediction tool for KIX domains. These domains are present in coactivators and play a central role in the transcription process. KIXBASE has two main parts, a web server and a database. To date, there is no other web resource that provides information on KIX domains. The KIX prediction program detects KIX domains in any organism on the basis of profile hidden markov models developed through alignment of known KIX sequences along with additional filtering criteria for improving accuracy. KIXBASE incorporates the most widely used programs like PSIPRED-3.5 and CLUSTALW2 in the webserver for further annotation and quality assessment of the predictions. Users can upload batch entries for protein, genomic or EST sequences in FASTA format and carry out the detection of potential KIX domains, generate the secondary structure for domain of interest and examine conservation with other KIX domains. The backend prediction algorithm is also used for development of the KIX database, which contains 1891 KIX proteins representing 427 organisms spanning metazoans, fungi and plants, comprising the largest online collection of KIX Page | 42   

 

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sequences. The prediction algorithm is highly accurate, robust and very efficient in handling multiple input types for KIX prediction.

Effect of Environmental Pollutant Bisphenol-A (BPA) on Human Health Archita Chowdhury,# Sandip Pal* Post-Graduate Department of Zoology, Asutosh College (University of Calcutta), West Bengal, India #

Bisphenol-A

[email protected]; *[email protected]

(BPA),

a

carbon-based

synthetic

compound,

exhibits

hormone-like properties and is present ubiquitously in the environment and in human tissues due to its widespread use and biological accumulation, respectively. It is present in polycarbonate plastic and epoxy resins, which can be used in impact-resistant safety equipment and baby bottles, as protective coatings inside metal food containers. BPA is an environmental estrogen (Xenoestrogen) which can mimic estrogen to interact with estrogen receptors α and β leading to changes in cell proliferation, apoptosis or migration and thereby, contributing to cancer development and progression. At the genomic level, BPA is involved in multiple oncogenic signaling pathways, such as the STAT3, MAPK and PI3K/AKT pathways and also interact with other steroid receptors (such as androgen receptor) and plays a role in prostate cancer development. It has a vital role in cancers of breast, ovary and prostate. BPA induces apoptosis by activation of Caspase-3. It also regulates the generation of Reactive Oxygen Species (ROS) by increasing intracellular calcium. ERK and JNK are also involved in BPA induced apoptosis. Now-a-days, BPA becomes highly carcinogenic for human. So, people can significantly lower the levels Page | 43   

 

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of BPA by strictly avoiding many packaged foods and beverages and also by changing the preparation and storage of food and by using BPA-free plastics.

Targeting Wnt pathway with Recombinant sFRP1 loaded Composite Nanoparticles for Cancer Therapy Archita Ghoshal 1 , Upashi Goswami 2, and Siddhartha Sankar Ghosh 1,2 1

Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati-39, Assam, India

2

Centre for Nanotechnology, Indian Institute of Technology Guwahati, Guwahati-39, Assam, India [email protected]

In this study, we report the anti-tumor activity of recombinant human Secreted Frizzled-related protein1 (sFRP1). In non-cancerous cells, sFRP1 blocks the Wnt pathway. However, due to epigenetic silencing of sFRP1 in a multitude of cancer types, Wnt cascade is aberrantly upregulated. Herein, we cloned the human sFRP1 gene bacterial expression vector pGEX-4T2 in order to express GST tagged sFRP1 in Escherichia coli BL21(DE3). Expression conditions were optimized to obtain maximum yield. To solubilize the recombinant protein from inclusion bodies, the process was modulated using detergents. Thereafter, we purified the GST-sFRP1 with glutathione agarose affinity chromatography. Using homology modeling, we determined that the structure of binding domain of sFRP1 does not change significantly, when tagged with GST. Docking analyses revealed that binding of sFRP1 to corresponding Wnt ligand remains potentially unaffected upon GST tagging. After using bioinformatics tools to analyze potential binding, we proceeded to confirm the same experimentally. The Page | 44   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

purified and characterized GST-sFRP1 was used for treatment of cervical cancer cells (HeLa), where significant anti-proliferative effects were observed, signifying its therapeutic potential. The implication of canonical Wnt signaling was ascertained by examining the expression levels of certain genes and protein lying downstream of Wnt binding. Further, we proceeded to enhance the therapeutic efficacy of the recombinant protein by coating it over gold-nanocluster embedded chitosan-alginate composite. Sustained release of the protein from the nanoparticles ensured augmented efficacy, whereas the remarkable luminescence properties of the gold nanoclusters were exploited for imaging purposes. Further, a module of co-therapy was engendered, whereby the protein-nanoparticle composite was coupled with conventional chemotherapeutic drug cisplatin, to achieve heightened anti-cancer response. In summary, this recombinant sFRP1 based nanosystem has the potential to pave the way for a new dawn in cancer therapy. References:  Archita Ghoshal, Upashi Goswami, Amaresh Kumar Sahoo, Arun Chattopadhyay, and Siddhartha Sankar Ghosh (2015). Targeting Wnt Canonical Signaling by Recombinant sFRP1 Bound Luminescent AuNanocluster Embedded Nanoparticles in Cancer Theranostics, ACS Biomaterials Science and Engineering, Accepted on Nov 2, 2015.  Archita Ghoshal, Siddhartha Sankar Ghosh (2014). Expression, Purification, and Therapeutic Implications of Recombinant sFRP1, Applied Biochemistry and Biotechnology, Vol 175(4), 2087-2103.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Study of prospective model to treat pharmaceutical components in Hospital sewage Argho Bakshi1,2 ,Deblina Mitra2 1. Centre for Biotechnology, Anna University, Chennai 600085, Tamil Nadu 2. School of Biosciences and Technology, Vellore 632014, Tamil Nadu

Generally 10-40 % of the dosage intake of drugs by patients undergoes bio-transformation in the body and the remaining 60 - 90 % is flushed out of the human body along with urine. Hence In hospitals and other large scale health care facilities the sewage matter constitutes a large amount of multivariate drugs which are part of the dosage of the indwelling patients along with a variety of disease causing microbes. The waste treatment plant in hospitals operates by segregating the wastes according to particle sizes along with other physical and chemical processes. Hence in the liquid slurry its a mix of microbes and drugs which results in formation of multiple drug resistant microbes and also part of the drugs gets mixed with urban drainage posing a great threat to the water quality difficult to treated by the municipality treatment plants in regards the current infrastructural buildup in the Indian subcontinent. Focusing mainly on the unused drugs, an innovative way is to treat the waste before it becomes more waste. Since drugs are mainly found in urine, if the urine is treated with a filter to screen out the drugs and harmful chemicals before release to the aquatic ecosystem, it removes chances of cross contamination as well as prevents growth of multiple drug resistant microbes. The filter shall comprise of gels along with activated charcoal and chelating and encapsulating agents in a composite matrix which traps the drugs and passes the remaining liquid. The filters can be periodically changed and its filtration matrix can be flushed with detergents, salts and a consortium of various physical and

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

chemical means to render the drugs harmless. Our study deals with the various aspects related to the designing of the filtration unit. Genetic engineering approach in favour of increased resistant starch formation in Indica rice cultivar Arindam Chakraborty* and Dr Asitava Basu Advanced laboratory of plant genetic engineering, Indian Institute of Technology, Kharagpur Kharagpur

Diabetes Type II is spreading like a pandemic especially in eastern and southern Asian countries. The Indian epidemiology of the disease is a grave concern with the highest number of detected diabetic patients. Food habit like taking rice as staple food is attributed as one of the main reasons of such a scenario due to its high glycemic index. One of the strategies to overcome this problem may be increase production of resistant starch in rice by genetic engineering approach. In fact, resistant starch is produced from cooked rice upon retrogradation. However, the amount of resistant starch formation in Indica rice varieties is very low (~ 3%) which is only 30% of recommended daily requirement for its beneficial effects.

The

major determinant of resistant starch production in rice on retrogradation is the proportionate presence of amylose with respect to amylopectin. Increase in the ratio of these constituent molecules was demonstrated to favour the production of resistant starch in rice on retrogradation. Thus, the present study aimed at to alter the ratio of constituent molecule to achieve the targeted resistant starch production in rice through RNA interference (RNAi) of rice branching enzyme. Agrobacterium mediated transformation of RNAi cassette of rice branching enzyme 3 under endosperm specific Page | 47   

 

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promoter resulted generation of three independent transgenic rice lines evidenced by Southern hybridization. The endogenous enzyme activity of the enzyme was found to decrease in a range of 77%-63% compared to control plant lines with a concomitant increase of amylose fraction in T0 seeds by 8% to 12.76 % of total starch as evidenced by biochemical and powder X-RAY diffraction analysis of rice in T0 seeds. This was found to affect the maltose release as well as hydrolysis index with a reduction by ~ 43% and ~ 30 % respectively when compared to control lines as evidenced by in vitro digestion.

Wnt mediated signaling pathways and their therapeutic use for the treatment of Parkinson’s disease by generation of midbrain dopaminergic neurons Aroshi Mitra, Sohini Dutt, Pramiti Mukhopadhyay, Department of Biotechnology, St. Xavier’s College, Kolkata (Autonomous), 30, Mother Teresa Sarani, Kolkata, West Bengal [email protected]

Wnt (naming and discovery involved wingless gene and Int1 gene) are a conserved family of secreted lipid-modified signaling glycoproteins. They play a key role in regulating multiple stages in the development of neurons and differentiation of stem cells. The signaling pathways primarily involved in the developing ventral midbrain are the Wnt/βcatenin pathway (triggered by Wnt1, Wnt2 and Wnt3a) and the Wnt/Rac1 pathway in dopaminergic cell (activated by Wnt5a). Shh (Sonic Hedgehog) and Wnt1 are the two key morphogens for midbrain dopaminergic (mDA) neuron development. The expression of Shh (which provides ventral identity to the neural tube) is necessary for midbrain dopaminergic neuron development in the midbrainPage | 48   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

hindbrain region (MHR). Wnt1 and Wnt5a work in tandem with both synergistic and antagonistic actions in the development of mDA neurons. Development of MHR and generation of mDA neurons by the pathway involving the expression of homeoprotein engrailed1 (En1) requires Wnt1, which thus facilitates specification, proliferation and neurogenesis of mDA progenitor. Wnt5a promotes morphogenesis, but inhibits progenitor proliferation and enhances the differentiation of mDA neuroblasts. Degeneration of mDA neurons in Parkinson’s disease (PD) has led to speculations, within the field of regenerative medicine, of using Wnt and their signaling pathway as major target for stem cell replacement and drug development strategies. Development of small molecules which modulate different branches of Wnt signaling (e.g. R-Spondins (activator) or Frizzledrelated protein (inhibitor)) can potentially regulate stem cell behavior and cause reprogramming to form mDA. Sequential Wnt treatment leads to an increase in the number of stem cells that differentiate into Tyrosine hydroxylase (TH+) cells and acquire expression of transcription factors critical for mDA neurons development (Foxa2, Lmx1a, Pitx3 and Nurr1). Wnt signaling pathways in the midbrain and in stem cells for generation of mDA neurons may thus have clinical implication in the treatment of PD.

Page | 49   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Nanotechnology: A Boon To Cervical Cancer Arpita Wagle, Tejal Patil, Aishwarya Iyer SIES Graduate School of Technology Sri Chandrasekaendra Saraswathy Vidyapuram Nerul, Navi Mumbai [email protected] ; [email protected]; [email protected]

Cancer is the leading cause of death worldwide. Cervical cancer is a disease in which malignant (cancer) cells form in the tissues of the cervix. Human papilloma virus (HPV) infection is the major risk factor for cervical cancer. Although preventive vaccines for cervical cancer are successful, treatment of cervical cancer is far less satisfactory because of multidrug resistance and side effects. Nanomaterials and nanotechnologies could provide potential solutions. Early detection of cervical cancer enables tumours to be efficiently removed by surgical procedures, leading to increased survival rate. The current method of detecting cervical cancer by Pap smear can only achieve 50% sensitivity, whereas nanotechnology has been used to detect HPVs with greatly improved sensitivity. In cervical cancer treatment, nanotechnology has been used for the delivery of anticancer drugs to increase treatment efficacy and decrease side effects. Nanodelivery of HPV preventive and therapeutic vaccines has also been investigated to increase vaccine efficacy. Overall, these developments suggest that nanoparticles-based vaccine may become the most effective way to prevent and treat cervical cancer, assisted or combined with some other

nanotechnology-based

therapy.

The

recent

application

of

Nanotechnology to the diagnosis and treatment of cervical cancer as well as the development of HPV vaccines can be a boon to humans. Keywords: Cancer, cervix, human papilloma virus, nanotechnology, nanomaterials. Page | 50   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Computational insight of K1444N substitution in NF1 gene associated neurofibromatosis type I disease: A molecular modeling and dynamics Ashish Agrahari, George Priya Doss. C* Medical Biotechnology Division, School of Biosciences and Technology, VIT University, Vellore 632014, Tamil Nadu, India E-mail: [email protected]; [email protected]

Neurofibromatosis type 1 (NF1) is one of the most common and complex autosomal dominant disorders, with a global occurrence of at least 1 in 4,000. NF1 gene encodes neurofibromin protein, which seems to be a negative

regulator

of

the RAS

signal

transduction

pathway.

Neurofibromatosis type I known as von Recklinghausen disease is a consequence of mutation in the NF1 gene. In this work, we utilized a set of different in silico prediction methods to screen 99 nsSNPs related with NF1 gene. In this way, we distinguished the most deleterious and pathogenic nsSNPs associated with Neurofibromatosis Type 1.To get more insight into the three dimensional structure of neurofibromin protein after mutation, homology structure of the native type as well as a K1444N mutant, were modeled using Modeler 9.13 software. Molecular dynamics simulation of 10 ns was commenced to reveal the impact of the mutation on its structure and function. The obtained results describe that the protein with the K1444N mutation had variable structural conformation and dynamic behavior than native neurofibromin protein. We believe that our computational study might be useful in rationalizing the deleterious mutations in Neurofibromatosis type1 and the advancement of novel pharmacological strategy. Page | 51   

 

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Keywords: Neurofibromatosis type 1, K1444N, homology modeling, Molecular dynamics

Faster and efficient algorithm for detecting overlapping and nested palindrome sequences in DNA sequences Akshita Bhandari1, AshutoshGupta2 1. Department of CSE NIIT University Alwar, India 301705 2. Department of Biotechnology NIIT University Alwar, India 301705 [email protected]; [email protected]

Inverted repeats (IR) or Palindromic sequences, (PR) are sequences which make complementary pairing with themselves. They are found in genomes of both eukaryotes and prokaryotes genomes. Palindromic sequences (PS) in DNA sequences involve important cellular or biological processes such as DNA–protein binding, DNA replication and DNA transposition and several other processes. Development of bioinformatics tools that are capable of detecting PS accurately and efficiently can enable genome-wide studies of PS patterns in both prokaryotes and eukaryotes. Different from conventional string-comparison approaches, we have proposed a linear time algorithm that uses bitwise operators for comparison of nucleotide bases. Bitwise operators are extremely fast

and most powerful in

operations. Proposed algorithm is then implemented as a PERL based program for PS detection. In comparison with other existing tools, our program shows a very high accuracy in detecting PR in very less time. Page | 52   

 

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Moreover, proposed approach proves to be very efficient for finding PS in larger genome data sets like chromosomes of 1 of Arabidopsis thaliana. Proposed algorithm took 1.27 minutes in execution in comparison to conventional palindromes finders like EMBOSS palindrome tool (Stable version 6.3.1) which took about 35.53 minutes for the same. Proposed approach accurately detected all the Inverted Repeats whereas EMBOSS missed various overlapping and nested palindromes. Other convention tools like BioPHP crashed during execution on large genomic data. The proposed algorithm further shows the frequency of each palindrome in the sequence. This can help in generating highly specific patterns that can be used to discriminate DNA from different species, by clustering together sequences from the same species.

Study of haematological parameters in catfish Pangasianodon hypophthalmus fed with probiotic supplemented diets Ashwini A. Vhatkar Research Scholar, Department of Zoology The Institute of Science 15, Madame Cama Road, Mumbai [email protected]

Bhavita Chavan Associate Professor, Department of Zoology The Institute of Science 15, Madame Cama road, Mumbai K. Pani Prasad Principal Scientist, AEHM Division Central Institute of Fisheries Education Panch Marg, Off Yari Road Versova, Andheri (W), Mumbai Page | 53   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

The present study was undertaken to assess the influence of dietary supplementation of probiotic bacteria Bacillus subtilis and Saccharomyces cerevisiae on hematological parameters of the catfish Pangasianodon hypophthalmus. The fishes were fed for 60 days with two different diets supplemented

with

bacterial

suspension

of

Bacillus

subtilis

and

Saccharomyces cerevisiae respectively at x108 CFUg-1. The control was maintained without the probiotic. Blood samples were collected at the intervals of 0, 15, 30, 45 and 60 days. The haematological parameters such as total erythrocyte count (RBC), total leucocyte count (WBC), haematocrit (Hct), haemoglobin (Hb) and haematological indices (MCV, MCH and MCHC) were examined. Haematological parameters showed elevated levels in fishes fed with diet supplemented with probiotic Bacillus subtilis. The results indicate that Bacillus subtilis proves to be a better probiotic tool than the Saccaromyces cerevisiae which may be preferred for aquaculture practices. Key words: Probiotic, Bacillus subtilis, Saccharomyces cerevisiae, Pangasianodon hypophthalmus, Haematological parameters

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Assessment of variability ofcoronarin D content in Hedychium coronarium Koen. from different locations of Eastern India through validated HPTLC method. Asit Ray, Biswabhusan Dash, Noohi Nasim, Sanghamitra Nayak. Centre of Biotechnology, Siksha O Anusandhan University, Bhubaneswar, Odisha, India [email protected]

Hedychium coronarium Koen, commonly known as white ginger lily is an important medicinal herb and extensively used in Ayurvedic system of traditional Indian medicineas a remedy for antirheumatic, carminative, febrifuge, stomachic, tonic and for lancinating pain.H. coronarium is a rich source of sesquiterpene and diterpenes with a wide range of biological and pharmacological activities. The pharmacologically bioactive compound of H.coronarium is coronarin D, a labdane diterpene which shows chemopreventive activity against cancer. It inhibits constitutive and inducible nuclear factor kβ pathway leading to potentiation of apoptosis and suppression

of

osteoclastogenesis.In

the

present

study, Hedychium coronarium collected from various locations of Eastern India was evaluated on the basis of their coronarin D content. Coronarin D content in the rhizomes was estimated by HPTLC densitometry. The extraction was checked using different solvents: n-hexane, acetone and methanol.Among all the solvents, acetone was found to be a good extraction solvent that allowed higher amount of coronarin D.Significant variation in content of coronarin D was marked among germplasms of H.coronarium ranging from 3.96 ± 0.04 to 42.67 ± 0.05 mg/g w/w in acetone extract of H.coronarium rhizomes. The highest content of coronarin D was present in Simlipal Biosphere Reserve (Orissa) accession whereas Page | 55   

lowest

content

was

found

in

Sargachi

(West

 

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Bengal)

3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

accession.The method will be useful for rapid screening purpose of H.coronarium for elite selection, genotype assessment and for crop improvement programme.

Toxicological Studies of the Aqueous Leaves Extracts of Combretum micranthum on Rats Auwalu Muttaka1, Lawan Jamilu Abdullahi2, Muhammed Sani Sule*3 1

Department Biotechnology, School of School of Biotechnology and Biosciences, Lovely Professional University, Phagwara, India-144411

2

Department of Pharmaceutical Chemistry, School of Pharmaceutical Sciences, Lovely Professional University, Phagwara, India-144411 3

Department of Biochemistry, College of Health Sciences, Bayero University, Kano, Nigeria [email protected], [email protected], [email protected]

In order to assess the toxicity of aqueous leaves extract of C. micranthum against liver disease, extracts of 500 mg/kg and 1500 mg/kg were administered orally to rats in two groups respectively, and a third group was used as control. Administration of the aqueous extracts for the determination of acute toxicity did not produce any mortality in the rats for dosages of up to 5000 mg/kg. The rats treated with a daily dose of 500 mg/kg for seven days had serum liver enzyme activities significantly higher than those in control rats whereas those treated with a daily dose of 1500 mg/kg for seven days had serum liver enzymes activities even higher than those treated with 500 mg/kg dose rats. Thus, both 500 and 1500 mg/kg doses for seven days induce liver toxicity. Key words: C. micranthum, toxicity, liver enzymes, mortality

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Metal Stabilization of Denovo Designed Collagen Mimetic Peptides for Biomedical Applications Avanish Singh Parmar1, Vikas Nanda2 1

Department of Physics, Indian Institute of Technology (Banaras Hindu University), Varanasi, India

2

Center for Advanced Biotechnology and Medicine, Department of Biochemistry and Molecular BiologyRobert Wood Johnson Medical School, Rutgers University, Piscataway, New Jersey 08854, USA [email protected]

We explore the design of metal binding sites to modulate triple-helix stability of collagen and collagen-mimetic peptides.

Globular proteins

commonly utilize metals to connect tertiary structural elements that are well separated in sequence, constraining structure and enhancing stability. It is more challenging to engineer structural metals into fibrous protein scaffolds, which lack the extensive tertiary contacts seen in globular proteins.

In the collagen triple helix, the structural adjacency of the

carboxy-termini of the three chains makes this region an attractive target for introducing metal binding sites. We engineered His3 sites based on structural modeling constraints into a series of designed homotrimeric and heterotrimeric peptides, assessing the capacity of metal binding to improve stability and in the case of heterotrimers, affect specificity of assembly. Notable enhancements in stability for both homo and heteromeric systems were observed upon addition of zinc(II) and several other metal ions only when all three histidine ligands were present. Metal binding affinities were consistent with the expected Irving- Williams series for imidazole. Unlike other metals tested, copper(II) also bound to peptides lacking histidine ligands. Acetylation of the peptide N-termini prevented copper binding, indicating proline backbone amide metal-coordination at this site. Copper Page | 57   

 

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similarly stabilized animal extracted Type I collagen in a metal specific fashion, highlighting the potential importance of metal homeostasis within the extracellular matrix. Based on the observed broad spectrum effects of copper on many collagen-like systems, CMPs could be used as models to understand how other metals impact collagen structure in both industrial and biological contexts. It can have major implication in hepatolenticular degeneration (Wilson’s disease) in which copper accumulates in tissues and these peptides might be used to reduce the concentration of copper.

Influence of manipulated rumen fermentation using buffers on ruminal environment in crossbred calves Awadhesh Kishore1 and Prakash Chandra Department of Animal Husbandry and Dairying Raja Balwant Singh College, Bichpuri, Agra, INDIA [email protected]

Present investigation is an effort to study the effect of manipulation of rumen fermentation using buffers in crossbred calves on rumen environment i,e, pH, content of different VFA and acetate to propionate (A:P) ratio and total nitrogen (TN), ammonia nitrogen, (AN), urea nitrogen (UN), and non urea and ammonia nitrogen (NAUN). Twelve crossbred calves for the purposes were selected (age 131-221 d; live weight LW 57.593.9 kg). All of them received grass mixture as green fodder and wheat straw as dry fodder. Concentrate mixture contained barley grain, and mustard cake. On the basis of different phenotypic traits the animals were divided into three groups. One animal from each group was randomly allotted to one of the four treatments viz. T1, T2, T3 and 4. Buffer in the form                                                              Page | 58   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

of sodium bicarbonate (S) and magnesium oxide (M) in combination @ 0.00 & 0.00, 0.50 & 0.25, 1.00 & 0.50, 1.50 & 0.75 of assumed dry matter intake to be 4% of LW or 0.0 & 0.0, 0.2 & 0.1, 0.4 & 0.2 and 0.6 & 0.3% of LW were given in T1, T2, T3 and 4, respectively. The ruminal study trial was started on day 30 of the experiment for 5 consecutive days. The rumen liquor samples were collected after 0, 2, 4, 6, 8 and 10 hours (h) of feeding buffer. The samples were analysed to TVFA, pH, acetic acid (AA), butyric acid (BA), iso-butyric acid (IBA), iso-valeric acid (IVA), propionic acid (PA), valeric acid (VA), A:P ratio, TN, AN, UN, and NAUN. The data recorded were subjected to statistical analysis using suitable models. The results of the study indicated that pH was fell down, TVFA, BA, VA & A:P ratio increased and AA, PA & AN decreased in rumen liquor due to buffer supplementation. Pattern of pH showed decreasing up to 10h, TVFA up to 6h, IBA, AN and UN up to 8h in rumen liquor after feeding buffer. The overall conclusion can be made on the basis of the observations that the addition of buffer addition in calf nutrition was responsible to manipulate ruminal environment in such a way so that so that it became much helpful to produce surplus fat due to appropriate pH and A:P ratio in the rumen ecosystem. Keywords: Buffer, Calf, Magnesium oxide. Nutrition, Rumen, Sodium bicarbonate.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Cytotoxic and Inflammatory responses induced by Outer Membrane Vesicles (OMVs) associated proteases of Vibrio cholerae Ayan Mondal1, Rima Tapader1, Sun Nyunt Wai2 & Amit Pal1 1

Divisions of Pathophysiology, National Institute of Cholera and Enteric Diseases, Kolkata, India

2

Department of Molecular Biology, Umea Centre of Microbial Research (UCMR), Umea University, Umea, Sweden. [email protected]

Microbial extracellular proteases have been shown to play a role in pathogenesis. The major protease secreted by Vibrio cholerae is hemagglutinin protease (HAP). In an earlier study, we showed that purified HAP from V. cholerae non-O1, non-O139 strain showed haemorrhagic response in rabbit ilealum and increased intestinal short circuit current in Ussing’s chamber. Besides HAP, the other major well characterized metalloprotease in V. cholerae is Vibrio cholerae protease (PrtV). In an earlier study we reported the presence of a novel 59 kDa serine protease VesC in V. cholerae O1 strain and showed its role in pathogenesis in rabbit ileal loop assay. V. cholerae uses different secretion systems to transport important virulence factors to the cell envelope and the extracellular milieu. Outer membrane vesicles (OMVs) may also serve as a mechanism for delivering active virulence factors into host cells. Toxins delivery mediated by OMVs is recognized as a potent virulence mechanism for many bacterial pathogens. Our present study demonstrated that both HAP and VesC are secreted in association with OMVs in an active form. These proteases associated OMVs showed cytotoxic and inflammatory response in human cultured intestinal epithelial cells and induced fluid accumulation in mice ileal loop (MIL). VesC associated OMVs induces haemorrhagic fluid response in MIL assay and an increased IL-8 response in T84 cells. Page | 60   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Haemorrhagic fluid was abolished and IL-8 expression was reduced in VesC deleted strain. In conclusion, our study shows that V. cholerae OMVs secrete biologically active proteases in an active form and may play a role in pathogenesis.

Relative Structural Analysis of LpxC enzyme: A promising drug target Ayon Pal1, Subhasis Mukhopadhyay2 andAsim K Bothra3 1

Department of Botany, RaiganjUniversity, Raiganj-733134, Uttar Dinajpur, West Bengal, India;

2

Department of Biophysics, Molecular Biology and Bioinformatics, University of Calcutta, 92 APC Road, Kolkata-700009, West Bengal, INDIA; 3

Cheminformatics Bioinformatics Laboratory, Department of Chemistry, Raiganj University, Raiganj-733134, Uttar Dinajpur, West Bengal, India [email protected]

Multidrug defiant organisms result in severe mortality and stand as a big challenge worldwide. The gram negative bacilli (GNB) in particular are a massive cause of concern and no new antimicrobial directed against these resistantorganisms have emerged in recent times. Many GNB are linked with serious nosocomial infections and have surfaced as superbugs with high levels of resistance due to novel mechanisms of resistance like extended

spectrum

beta-lactamases,

carbapenemasesetc.

The

LpxCenzyme is a vital component of the Lipid A biosynthetic pathway existing universally in all the gram-negative bacteria which catalyzes the first committed step of the Lipid A biosynthesis providing structural and physiological integrity to the gram negative eubacteria. The LpxC enzyme has been found to display a unique two domain ‘β-α-α-β sandwich’fold. The LpxC enzyme being a zinc metalloamidase is different from other metalloenzymes for lacking the typical zinc binding motifs. Page | 61   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

A thorough structural analysis of the LpxC enzyme was performed to elucidate the relative structural properties of this enzyme. Homology modelling of the LpxC enzyme fromsixteen highly pathogenic organisms was carried outusing the PDB entries 3P3G and 3UHM as template along with model validation, RMSD analysisand analysis of enzyme surface clefts.The RMSD values were found to range between 0.2Å to 2.0Å. Certain vital structural variations within the enzyme was observeddue to the occurrence of different types of secondary structural elements(SSE) and different residues interacting with the zinc atom. Analysisof clefton the enzyme surface demonstrated the presence of specific residues within the substrate bindingcleft. Preference for different SSE within the major cleft was also noticed in the different LpxC enzyme tertiary structures. The LpxC enzymes, though universally present in all gram-negative bacteria show some degree of sequence and structural difference that has prevented the development of a near universal inhibitor targeted towards it. Detailed analysis of the substrate-binding site and inhibitor binding interactions by docking studies will help to explore the different features of this promising antibacterial target.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Design Of Novel Allosteric Inhibitors Against Human Fructose-1,6Bisphosphatase B. Saravana Gowtham & M.S. Shahul Hameed Department of Biotechnology, Bannari Amman Institute of Technology, Sathyamangalam - 638401, Erode District, Tamil Nadu, India. [email protected]

Fructose-1,

6-

bisphosphatase,

a

key

rate

limiting

enzyme

in

gluconeogenesis has over the years emerged as an important target for anti-diabetic drugs. Allosteric inhibition of Fructose-1, 6- bisphosphatase at cyclic AMP binding site has led to the development of many novel antidiabetic drugs. However it has been observed that targeting AMP binding sites leads to cross reactive inhibition of many other AMP dependent enzymes in cell. A more prudent approach to this problem would be to identify, validate and target novel allosteric sites in Fructose-1, 6bisphosphatase. Identifying novel allosteric sites through computational methods has become more reliable with the advent of powerful MD based simulation methods. In this study we have predicted two novel allosteric sites in Fructose-1, 6- bisphosphatase using such computational methods. The mechanism by which allosteric inhibition occur at these sites have been investigated. The shortest communication paths between the allosteric sites and the active sites were then identified using Monte Carlo simulation based methods. Virtual screening for ligands against these novel allosteric sites resulted in identification of several compounds with high binding affinity. The ligands with promising binding affinity, as determined from

free

energy

calculations,

were

then

subjected

to

further

pharmacophore modelling to design drugs with better pharmacokinetic properties.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Key Words: Fructose-1, 6- bisphosphatase, Anti-diabetic drug, Allosteric site, Virtual screening, Pharmacophore modelling.

SOCS1 mediated inhibition of STAT1 phosphorylation in Mycobacterium tuberculosis infected macrophages: an in-silico analysis Bala Tripura Sundari A Madduri, Sumedha Mukherjee, Arunima Sharma And Somdeb Bose Dasgupta School of Biomedical Engineering, Indian Institute of Technology (BHU), Varanasi, India [email protected]

Tuberculosis, caused by Mycobacterium tuberculosis still affects one-third of world population each year more so due to extreme and total drug resistant parasites that have set therapeutics at limbo. Generally macrophage activation through inflammatory cytokines IFN-

and TNF-

causes shuttling of engulfed pathogens to lysosomes for elimination. But, mycobacteria induced over-expression of Suppressor of cytokine signaling (SOCS) proteins, by a yet unknown mechanism hinder downstream IFNsignaling and thereby prevent phagosome-lysosome fusion. The plausible mechanisms could be SOCS1 mediated disruption of the IFNassembly,

inhibited

STAT1

or

JAK

phosphorylation,

receptor

proteasomal

degradation of phosphorylated STAT1, hindered nuclear translocation of phosphorylated STAT1, or all combined. Here we ascertain the possibility of SOCS protein in inhibiting STAT1 phosphorylation by virtue of Kinase Inhibitory Region (KIR) present at N-terminus of the protein; identified from NCBI protein database. Crystal structure of Mus musculus SOCS3 served as template in generating homology model of SOCS1-KIR motif using SWISS-MODEL. The residues on STAT1, phosphorylated during IFNPage | 64   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

signaling were highlighted in Raswin and also mutated in-silico to their phosphomimetic counterparts. Using AutoDock, SOCS1-KIR motif was docked onto the control and phosphomimetic residue models of STAT1. Applying energy minimization algorithms, we found that KIR establishes the most stably-docked structure with control STAT1 than its phosphomimetic counterpart. This indicated that KIR prevents phosphorylation of STAT1 rather than dephosphorylating of already phosphorylated STAT1. Further on in vitro experiments with recombinant proteins would confirm our insilico data. This would allow an insight into the SOCS1 mediated dampening of IFN-g signaling during mycobacterial infection.

Estimation Of The Levels Of Lipid Peroxidation & Total Antioxidant Capacity In Hyperthyroid, Hypothyroid & Euthyroid Subjects – A Cross Sectional Study Basant Joshi Department of Biochemistry,GMC, Haldwani, Uttarakhand

Introduction: Thyroid hormones (TH), body’s prime regulator of metabolism, has a pro-oxidant effect and plays a crucial role in the regulation of mitochondrial oxidative metabolism with a consequent lipid peroxidative (LPO) response. Free radicals may cause lipid per oxidation & damage macromolecules & cellular structure of the organism. Antioxidants are reducing agents & limit oxidative damage to biological structures by passivating free radicals. Any imbalance due to an excess oxidant formation or lowered antioxidants leads to oxidative stress damaging lipids, proteins & nucleic acids which leads to inflammation followed by tissue injury and cell death.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Objectives: Objectives of the present study were to estimate the levels of serum T3, T4, TSH and To Estimate the levels of serum Malondialdehyde (MDA) and Total Antioxidant Capacity. Methodology: Subjects were approached, explained & consent form was taken along with 3ml of Fasting blood. Serum T3, T4 & TSH by chemiluminescence assay, LPO by Okawa H et al (modified) and TAC by D Koracevic et al were estimated in 40 eu-thyroids, 20 hyperthyroids and 30 hypothyroids subjects of age group of 25-50 years. Statistical Analysis: Values were expressed as Mean±SD. Data were analysed by one way ANOVA. The level of significance was considered at P12µg/dl, T3 > 1.50 ng/ml & TSH < 0.49 µIU/ml; those of low level had T4 < 4.50 µg/dl, T3 < 0.75 ng/ml & TSH as > 4.67 µIU/ml & subjects as normal had T4, T3 & TSH as 4.50 to 12.0 µg/dl, 0.75 to 1.50 ng/dl & 0.49 to 4.67 µIU/ml respectively. Conclusion: We suggest that hyperthyroidism is more prone for lipid peroxidation that need more supplementation of antioxidants because of low antioxidant level to improve the level of lipid peroxidation

as compared to

hypothyroidism. The measurement of the total anti-radical potential is also very useful for the determination of the sum of pro- and antioxidants in healthy individuals who are exposed to a pathogenic environment.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Novel Exploitation of Dried distillers grains with soluble (DDGS) for Human Consumption Bedotroyee Chowdhury & Dr.Lakshmishri Roy*1 Dept.of Food Technology.Techno India, Salt Lake, Sector-, Kolkata, INDIA [email protected]

Now days there is worldwide interest in developing a sustainable economy where bio based chemicals are the lead actors. For bio refinery application the byproducts streams from distilleries and bio ethanol plant such as dried distiller’s grains with soluble (ddgs) are presently being explored. Recently with the ever increasing demand of variety food products there is significant expansion and extent of work being done in the food industry. DDGS is abundantly available coproduce of ethanol production that is inexpensive. Grains offer a variety of nutrients. Wheat flour is used in most of breads because of its nutrients component and availability but different problems are associated with this flour such as allergies and loss of nutrient components due to milling and refining. Distillers dried grains with soluble is coproduct of bio fuel ethanol and potable ethanol production using cereal as the biomass for fermentation. The DDGS utilized in the present investigation is from rice grain based ethanol industry.Characterization studies revealed it contains about 86 - 93 % dry matter, 26 - 34 % crude proteins and 3-13 % fat. Hence an attempt was made to use DDGS for bread production. Cereal based ddgs is high in fibre but low in gluten forming protein, so in this study the bread is tested with various ddgs substitution level with flour (5gm, 10gm, 10gm, ......30gm in 10gm flour). DDGS samples were subjected to a series of treatments for bleaching of the

sample.

The

samples

were

subsequently

utilized

for

bread

production.Determination of the optimised proportion of ddgs with flour in bread making process was also conducted .The products obtained were Page | 67   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

evaluated for textural and sensory parameters.It was found that the product with DDGS supplementation was at par with the conventionally produced bread. Evaluation of IR Drying technique for production of ginger powder from Blanched And Un-blanched Ginger Slab Trishita Bose, Dr.Debabrata Bera1* & Dr.Lakshmishri Roy*1 1

Department of Food and Biochemical Engineering, Jadavpur University

Dept.of Food Technology.Techno India, Salt Lake, Sector-5, Kolkata, INDIA [email protected]

Drying characteristics of ginger slices by infra red moisture analyzer at different temperatures were studied. Ginger slabs were water blanched at 800 C for three minutes. An effective and efficient IR moisture analyzer is used to study the drying characteristic of blanched and unblanched ginger slab at three temperature levels and observed the moisture removal rate within every 1-5 minutes interval and then 5-15 minutes interval. The size of sliced ginger is an average of 2 mm each. Experiments were carried out of three different drying temperatures of (60°, 90° and 105°). The moisture removal increased with increase in temperature. Thin layer drying models such as Newton Equation, Handerson&Pabis and Page Equation was fitted from the experimental data for unblanched and Newton Equation for blanched sliced ginger and validated on the basis of determination of coefficient (R2). Drying took place in the falling rate period and Wang and Singh and Logarithmic models adequately described the drying behaviour of both blanched and unblanched ginger slabs. The drying rate of unblanched samples was significantly (P< 0.05) faster than the blanched ginger slabs.Temperature dependence of the effective diffusivity was adequately described by the Arrhenius equation.

Page | 68   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Color of ginger powder changed after drying for unblanched sliced ginger while it almost remained the same for blanched sliced ginger. Lightness of ginger rhizomes decreased with an increase in drying temperature for all samples except sliced and blanched samples.According to the calculating value of Angle of repose the flowability of the of the unblanched ginger powder obtained was free flowing while for blanched ginger powder it was very free flowing. The acceptable value of diffusivity, activation energy and Hausner Ratio was also obtained for both the unblanched and blanched ginger.

The Role of Pak1 In UV B Induced Premalignant Skin Lesions Beesetti PS Swarna Latha, Mavuluri Jayadev, Rayala Suresh Kumar Indian Institute of Technology Madras [email protected] 21

P

activated kinase 1 (Pak1), a major mitogen-responsive serine/threonine

signaling kinase is a well-known regulator of cytoskeletal remodeling that contributes to tumor formation. Alterations in Pak1 expression have been documented in many type of cancers and its role in DNA damage response (DDR) was well reported. DNA damage response to UV involves the activation of Ataxia Telangiectasia mutated and Rad3 related (ATR) a serine/threonine kinase and its functional Dysregulation leads to increased tumourogenesis. Yet, the knowledge of ATR transcriptional regulation was feebly known. In our current study Pak1 overexpression and knockdown clones were generated in keratinocytes using viral transduction and observed a significant change in ATR expression with Pak1 confirmed by real-time quantitative RT-PCR and Western blot analysis. In addition Active Pak1 mutant is more efficient in ATR regulation indicating that ATR as the Page | 69   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

probable transcriptional target of Pak1. Further our studies with ATR promoter using luciferase reporter assays and by Deletion analysis we identified that 652bp fragment is the core promoter sequence being regulated by Pak1.In conclusion, our results specify for the first time, that Pak1 is able to regulate the ATR expression, considering ATR to be one of the possible target of Pak1 in DNA damage response. Keywords: P21 activated kinase 1 (Pak1), Ataxia Telangiectasia mutated and Rad3 related (ATR), keratinocytes.

Microbial fuel cell: A promising technology for Wastewater Treatment and generation of Electricity Bimala Dhakal, Jarina Joshi Central Department of Biotechnology, Tribhuvan University Nepal

Microbial Fuel cell (MFC) can be a great demand for waste water treatment in future. Alternatively, the increasing demand of energy can be fulfilled by this technique in the future if the performance of MFC is improved. In this research, a MFC was constructed by using graphite felt immobilized with neutral red as anode and a platinum coated platinum wire as cathode. Anolyte used was municipal wastewater and catholyte was phosphate buffer pH 7. The mixed culture of organism dominantly present in the wastewater was used in MFC.The COD of wastewater was reduced by 69.96% when MFC was run for days. Similarly the level of reducing sugar lowered by 86.66% after several days. MFC showed a open circuit voltage of 513mv on the fifth day and remained almost constant after that day which indicates that MFC can generate significant power along with wastewater treatment.The maximum power generated was found to be Page | 70   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

24.45w/m3when 1% H2O2 was supplied as a source of oxygen in the cathode compartment. The result was found to be effective when cellulose acetate was used as membrane compared with the Nafion membrane. However the fuel cell showed a constant voltage rate when cheese whey was provided as the substrate and Saccharomyces cerevisiae was the organism indicating there was no utilization of the feed by organism. The result indicates microbial fuel cell technology to be a new approach for wastewater treatment when mixed culture is used, as it produces sustainable clean energy by minimizing COD level. Keywords: Microbial Fuel cell, Wastewater treatment, Electricity generation, Mixed microbial culture.

Fingerprint analysis of Zingiber zerumbet(L) Smith by HPTLC coupled with Chemometric analysis Biswabhusan Dash, Asit Ray, SupravaSahoo, SanghamitraNayak Centre of Biotechnology, School of Pharmaceutical Sciences, Siksha O Anusandhan University, Bhubaneswar-751003, Odisha, India. [email protected]

A

simple,

reliable

HPTLC

method

was

developed

for

chemical

fingerprinting of Zingiber zerumbet and for quantitative estimation of bioactive constituent zerumbone. 36 batches of Z.zerumbet were collected from five ecoregions of Eastern India. Zerumbone content varied within the range 52.4 to 214.6 mg/g w/w in methanolic extract of Z.zerumbet rhizomes. Zerumbone content was in the following order: Lower Gangetic plains moist deciduous forests> Brahmaputra valley evergreen forest> Odisha semi-evergreen forests> Sundarbans freshwater swamp forests> Eastern Highlands moist deciduous forest. Relative Standard Deviation

Page | 71   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

(RSD) of the Relative Peak Areas (RPA) and Relative Retention Times (RRT) of 8 characteristic peaks (one of them identified as zerumbone) in repeatability, precision, and stability test were 60 diseases of which 16 are major ones. Turcicum Page | 120   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

leaf blight (TLB) caused by E. turcicum, is considered a serious disease where climatic conditions are cool with high relative humidity. Severe losses in grain yield ranging from 25 to 90 per cent have been reported due to this disease in India. Seventy two hybrids developed by line x tester design along with 18 parents and three checks were screened against TLB under artificial epiphytotics. Disease score and percent disease indexes were recorded at the time of tasseling. The disease scoring was done on 1-5 scale. Nineteen hybrids possessing a disease score of 0, 33 hybrids with a score of 1 and 20 with a score of 2 were categorised as resistant. At the time of tasseling, the range of per cent disease indexes was between 0 per cent and 28.6 per cent. Twenty days after tasseling (DAT), one hybrid (PDM-8 PDM-32) was highly resistant (score 1) and 15 hybrids were resistant (score 2). Twenty nine hybrids had a disease score of 3 indicating moderate resistance. Twenty four crosses were susceptible (score of 4) while three crosses were highly susceptible (score 5), After 20 DAT, the per cent disease index ranged from 22.86 to 68.57 per cent. Thirty four hybrids showed less than 45 per cent disease index falling under moderately tolerant category and twenty six hybrids showed more than 50 per cent disease index falling under susceptible category. After 20DAT, parents PDM-8, PDM-60 and PDM-254 had disease index of 50% indicating their susceptibility to TLB.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

Effects of Antioxidants on Acute pancreatitis in caerulein induced Rat model Jeyaparvathi Somasundaram Department of Biotechnology, Lady Doak College, Madurai. Tamil Nadu, India. [email protected]

Acute pancreatitis is an inflammatory disease of the pancreas and a major cause of morbidity worldwide. The clinical course of acute pancreatitis ranges from mild abdominal discomfort to life threatening severe disease. The most common causes of acute pancreatitis are heavy alcohol consumption and gallstone disease, together accounting for the majority of all cases. In India, alcohol is the predominant cause of acute pancreatitis. The mechanism by which alcohol causes acute pancreatitis remains unclear, and despite being a clear risk factor for acute pancreatitis. The present studies sought the level of digestive enzyes associated with the onset or severity of caerulin induced acute pancreatitis in humans. In the present study, we hypothesized that the administration of an antioxidant complex could decrease the level of digestive enzymes in an experimental rat model. The aim of this study was to assess the possible pro tective effect of antioxidant against cerulein-induced acute pancreatic inflammation (AP) and oxidative injury. Key words: Pancreatitis, Acute, alcohol, antioxidant

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Adipocytokines: The molecular link between Type 2 Diabetes and Obesity Joyita Banerjee, Neetu Mishra, Yogita Dhas Symbiosis School of Biomedical Sciences, Symbiosis International University, Pune412115 [email protected]

Type 2 Diabetes mellitus (T2DM), a non- insulin dependent metabolic disorder, has become one of the most global public health concerns. The International Federation of Diabetes estimated that India will become a home for 109 million of diabetic people by 2035 from 65.1 million in 2013 in the age between 20-79 years. Rapid urbanization and sedentary life style has increased the prevalence of obesity, which has become a global threat and plays significant role in developing diabetes mellitus. Since last decade, extensive researches have been carried out to find the correlation between T2DM and obesity by different factors such as insulin resistance, abdominal fat, free fatty acids, triglycerides, proinflammatory cytokines, genetic and environmental factors. There is still a need to explore the underlying molecular mechanisms of various adipocytokines in linking T2DM and obesity. The objective of this study is to find out the possible roles of adipocytokines as well as certain non-adipokines in interlinking T2DM and obesity by analyzing the recent researches in this field. Some adipocytokines such as, adiponectin, leptin, peroxisome proliferatoractivated receptors, apelin, etc. found to have anti-hyperglycemic role while tumor necrosis factor-alpha, interleukin-6, resistin, retinol binding protein-4, dipeptidyl peptidase-4, plasminogen activator inhibitor-1, visfatin, free fatty acid, angiotensin

and toll-like receptors found to have pro-hyperglycemic

role. Adiponectin among all the above factors has been found to play crucial and definite role in linking T2DM and obesity. Adipocytokines could Page | 123   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

become novel targets for therapeutic interventions to treat T2DM and development of obesity. Keywords: Type 2 Diabetes mellitus, obesity, adipocytokines, adiponectin

Recurrent Spontaneous Abortions: PBMC and serum cytokine profile in a cohort of Indian women Jyothi V.Mallia1, Anurupa Maitra2, Parag Tamhankar3, Anushree D. Patil4 and Nafisa Balasinor5 1

Assistant professor, SICES College of Arts, Science and Commerce, Ambernath (W) , 2,Dept. of Molecular endocrinology, 3Genetic Research center , 4Dept. of Clinical

Research, 5Dept. of Neuroendocrinology National Institute for Research in Reproductive Health, Parel, Mumbai -12 [email protected]

Cytokines are regulatory peptides or glycoproteins that can be produced by virtually every nucleated cell type in the body and have pleiotropic regulatory effects on hematopoietic, endocrine, neural and many other cell types. Cytokines have been classified on the basis of their biological responses into pro – or anti-inflammatory cytokines, depending on their effects on immunocytes. Cytokines acts in networks or cascades. Cytokines act at all stages of pregnancy from implantation to parturition. Recurrent spontaneous abortion (RSA) is the spontaneous loss of two or more consecutive pregnancies before 20 weeks of gestation. Our study investigates whether cytokines from PBMC and serum differ between non pregnant Recurrent Spontaneous Abortions (RSA) women and nonpregnant fertile women. Further, the role of 3 stimulants (PHA-M, Con-A and Anti Cd3) in cytokine production have been also assessed. PBMC was separated from 25 RSA and 25control women and incubated along with 3

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

stimulants. Serum was also collected. Cytokine expression was analyzed by ELISA and compared.

ELISA of 12 cytokines TGFβ production was

found to be higher in both PBMC and serum in non pregnant RSA women, followed by TNFα. Anticd3 was found to be effective stimulants compared to PHA – M and Con-A for increased cytokine production in PBMC.

Extracellular Biosynthesis of Metal Nanoparticles using Holarrhena antidysenterica and Aegle marmelos and their Antimicrobial activity. Suhas Pednekar, R.S. Chaughule and Kalpana Dabhade* Department of Biotechnology, Ramnarain Ruia College, Matunga, Mumbai-19. (Affiliated to University of Mumbai) [email protected]

Biosynthesis of nanoparticles is under exploration due to wide bio-medical application and research interest. Use of Holarrhena antidysenterica and Aegle marmelos leaf broth in the extracellular synthesis of pure metallic Gold and Silver nanoparticles is reported at room temperature. On treatment of aqueous solution of silver nitrate and chloroauric acid with Holarrhena antidysenterica and Aegle marmelos leaf extract, the synthesis of Gold and Silver nanoparticles was carried out. The biosynthesized gold and silver nanoparticles were confirmed by UV-spectroscopy and TEM analysis. The silver and gold nanoparticles were found to be spherical in shape. The results showed that leaf extracts of Holarrhena antidysenterica and Aegle marmelos are good bioreductants for the synthesis of Gold and Silver nanoparticles and active against clinically isolated human pathogens Staphylococcus aureus and E.coli.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Exploration of phytocompounds for the treatment of drug resistant breast cancer: experimental & computational approach KanikaVerma and K. Ramanathan* Industrial Biotechnology Division School of Bio Sciences and Technology, VIT University, Vellore- 632014, Tamil Nadu, India. [email protected]

Paclitaxel is the most effective chemotherapeutic agent used for the treatment of breast cancer. However, in clinical trials resistance to paclitaxel is one of themajor obstacles for cancer chemotherapy. Most importantly, resistance due to β-tubulin mutations(R306C) has been intensely debated in recent years. With the rise of paclitaxel resistant mutation in β-tubulin there is a need to add a novel inhibitor from natural source, as they have less possibility of getting resistance furthermore less side effects. Keeping this in mind we have utilized experimental and computational techniques to isolate the potent inhibitor for target protein βtubulin. Initially, phytocompounds from Chenopodium album plant were extracted and the structures were identified with the help of GC-MS technique.Subsequently, the Bioavailability, toxicity, bioactivityand docking analysis

were

examined

MOLINSPIRATION,

OSIRIS,

by

using PASS

the

programs

and

YASARA

such

as

algorithm

respectively.The results from our computational analysis certainly indicates that phytocompoundErgosta-7, 22-dien-3-ol could be a novel inhibitor for breast cancer treatment and provide direction for future research.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Micropropogation and Phytochemical Analysis of Asparagus racemosus (Shatavari) K.E. Thakur1, R.R. Kamble1, R.S. Choudhary1,* N.S.Chavan1 Department of Plant Biotechnology, K. K. Wagh College of Agricultural Biotechnology, Nashik, Maharashtra, 422003. [email protected]

Asparagus racemosuscommonly known as Shatavari in Marathi and Hindi. It is monocot medicinal plant & vegetative propogation in nature. Shatavari is well known drug in Ayurveda.It increases the intelligency &promote learning or memory. Micropropagation and phytochemical analysis of Asparagus racemosus was investigated in the present study. In vitro propogation ofAsparagus racemosus; shoot apex and nodal segment used. Shatavari explants were kept in MS Medium with sucrose (3%), Agar (0.8%) and pH is adujusted at 5.8 andmaintained at temperature 25±2 0C, light intensity is 1500 to 2000 lux and humidity 50 to 60 % with regular fumigation processes. For callus formation the MS-medium with growth regulator 2,4-D (0.1mg/L), BAP (0.2mg/L), IAA (0.2mg/L) and NAA (0.1mg/L) gave vigorous callus. For shoot formation using MS-medium with growth regulator BAP (0.1-1.5mg/L) in combination with IAA (0.15 mg/L).The highest percentage of shoot proliferation observed in media code AS-5 with BAP (1.5mg/L) and IAA (0.15mg/L). Root formation using MS-medium with growth regulator IAA (0.1mg/L to 2.5mg/L) and Kinetine (0.1mg/L). The highest root formation observed in media code AS- 10 with IAA (3.0mg/L) and Kinetine (0.1mg/L).For phytochemical analysisthe methanolicleaves extract used and it shows the presence of phytosteroids, saponins,carbohydrates,

flavanoids,

phenolic

compound,

steroids,protein,oil and fatty acid. Keywords: BAP, IAA, Kinetine, Phytochemical analysis, Shatavari. Page | 127   

 

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tannins,

 

3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

Phenotypic characterization and Metabolite profiling of Black cherry and Black pear tomatoes Mr.KunalDhokne University of Hyderabad, Prof.CR Rao Road Gachibowli, Hyderabad, Telangana, 500046. [email protected]

Tomato (Solanum lycopersiconL.) is an important vegetable crop with numerous uses with a high nutritional value such as dietary carotenoids serve as precursor for vitamin A. The consumption of tomatoes and tomato product has been associated with a reduction in risk of contracting some type of cancer and other chronic diseases. Metabolite profiling is necessary to understand their role at different level of fruit development stages and interaction among them.In our study, we have selected Black Cherry and Black Pear heirloom tomatoes and Arkavikas as control. To investigate about these heirloom lines we have framed our objective in two parts (i) phenotypic characterization of these heirloom lines and (ii) metabolite profiling of the lines to understand role of metabolite in fruit ripening and different developmental stages of fruit.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Tissue culture of Ageratum houstonianum (Asteraceae) M. Mohammadi, F. Chalabian, S. Arbabian Department of Biology, Faculty of Biological Sciences, Tehran North Branch, Islamic Azad University, Tehran, Iran. [email protected]

Various

explants

(apical

meristem,

epicotyl,

hypocotyl

and

root)

ofAgeratumhoustonianumplantlets (grown in MS medium), were cultured on MS media supplemented with various combinations and concentrations (0.4–4mgL−1) of IAA and Kin. Generation of calli and regeneration of root, stem and leaf from the calli were used for assessment. Among the explants, the apical meristems were responding for regeneration of plantlets, and also were the bestexplants for callus induction. MS medium supplemented with 1 mg L−1 Kin /0.4 mg L−1 IAA, 1 mg L−1 Kin /1 mg L−1 IAA, 2 mg L−1 Kin / 2 mg L−1 IAA and 0.4 mg L−1 Kin / 0.4 mg L−1 IAAconcentrations were observed to be responsive to regenerate plantlets.However, 4 mg L−1Kin and 2 mg L−1IAAwas recorded as the best medium. In this research, hypocotylexplants did not respond for callus induction, but regenerated root and stem only in MS medium supplemented with 0.4mg L−1 Kin / 1 mg L−1IAAconcentrations, whichgave best response for stem induction in leaf and root explants. Keywords: Ageratum houstonianum, Apical meristem, Tissue culture.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Cellulase activity, structure prediction & comparative analysis from various fungal isolates Madhavi M. Motankar1, Amisha Shirodker Pednekar2, Nidhi Tallur3 1

Department of Biotechnology and

2&3

Department of Botany, Parvatibai Chowgule

College Autonomous Gogol, Margao Goa, India [email protected] & [email protected]

Cellulases have wide industrial applications and chiefly produced by microbial sources during their growth on cellulosic materials like twigs, leaves etc. To determine the cellulase activity qualitatively, eukaryotic, achlorophyllous, ubiquitous fungi from litter and soil were collected from various areas of Salcete, Goa. Aspergillus niger, Aspergillus sp., Cheilymenia fimicola, Fusarium sp., Ganoderma applanatum, Ganoderma sp., Marasmius haematocephalus, Mucor sp., Mycena acicula, Penicilium sp., Rhizopus sp., Schizophyllum commune and Trichoderma sp. were isolated and identified. Screening of cellulase producing fungi was confirmed by zone of clearance. Amongst the isolated fungi from soil, the maximum zone of clearance of 3.5cm was observed in Rhizopus sp., followed by Aspergillus niger with 2.9cm and the maximum zone clearance of 1.8cm was observed in Trichoderma sp. followed by Schizophyllum commune with 1.7 cm from the fungi collected from litter. Cellulase showed maximum activity at 300C at pH 6. Cellulase structure prediction was carried out seeing for maximum amount of amino acid sequences and multiple sequence alignment comparison using Clustal-W and T-coffee was done showing closeness of the various fungal isolates as sources of cellulase. Further research should consider increasing quantity of Cellulase and finding it’s use in bio-pharmaceutics.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Potential of bioactive peptides of protein hydrolysate from the edible oyster Lamellidens marginalis for nutraceutical therapy Madhushrita Das1, Ankhi Haldar2, Roshni Bhattacharya2, Tanmoy Kumar Dey2, Pubali Dhar2 and Jana Chakrabarti3 1.Department of Chemical Technology, University of Calcutta, 92, A.P.C Road, Kolkata, West Bengal, (e-mail: [email protected]); Phone No. 9903149530 2. Laboratory of Food Science and Technology, Food and Nutrition Division, University of Calcutta, 20B Judges Court Road, Alipore, Kolkata, West Bengal, India, PIN- 700027. 3. Department of Biotechnology (Division of Biological Science), Presidency University (Erstwhile Presidency College), 86/1, College Street, Kolkata-73, West Bengal.

Higher incidences of Protein Energy Malnutrition due to lower protein intake are quite prevalent among the children in our Country. But increase in population over the last decade has led to greater pressure on the existing animal protein sources. So the need of the hour is to search for efficient utilization of unconventional low cost animal protein resources. Moreover, current research focuses on dietary proteins as good sources of bioactive peptides with broad spectrum of pharmacological activities. Molluscs, as a group are regarded as under-exploited source of health-benefit molecules and have great prospect. The freshwater mussel Lamellidens marginalis is well accepted as food all over India. Furthermore, ethno-medicinal uses of the flesh of Lamellidens among the rural people to treat hypertension have been documented. Here, an attempt has been made to evaluate the potential of Lamellidens as functional food. Mussels were collected from freshwater ponds. Shells were removed and fleshes were preserved at 20˚C for analyses. Tissue homogenate was prepared for proximate studies. Mussel Protein hydrolysates consists of short peptides were prepared using food grade protease-Alcalase 2.4L to investigate functional Page | 131   

 

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properties.

3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India Ferric Reducing Anti-oxidant Power & DPPH Anti-oxidant

assays were performed. Anti-hypertensive property was evaluated by measuring Angiotensin Converting Enzyme (ACE) inhibition assay. Proximate analysis indicates that mussel meat contains moderate amount of protein, carbohydrate but less amount of fat. Moisture content is quite high but ash content is very low. Phospholipids content is significantly high. Substantial amount of eicosapentaenoic acid & docosahexaenoic acid are present. Functionality analyses of protein hydrolysate show increase in fat absorption,

emulsification,

foaming

capacity

and

protein

solubility.

Progressive anti-oxidant and anti-hypertensive properties have also been documented. Present investigation has demonstrated that bioactive peptides derived from Lamellidens marginalis provide opportunity for utilizing them in various food formulations and pharmaceuticals and thus can be used for neutraceutical therapy. Keywords: Functional food, Functional properties, Lamellidens marginalis, Protein hydrolysates Acknowledgment:

This work is funded by The University Grants

Commission (UGC), New Delhi, India

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

“A Novel Biological Route to Synthesis of Silver Nanoparticles by Using Green Technology as New Generation of Plant Virus Inhibitors” Magar Nakul Divakar1 and Bhamare Deepak Prashant2 1

K.K.Wagh College of Agricultural Biotechnology, Nashik 2

K.k.waghCollege of agricultural biotechnology

A cost effective and environment friendly technique for green synthesis of silver nanoparticles using spore crystal mixture of Bt (Bacillus thuringensis) and also by using plant extract (Neem, Banana), their application in plant virus inhibition was standardized. silver nanoparticles were successfully synthesized from 1mM AgNO3 via a green synthesis process, and characterized using UV–VIS absorption spectroscopy.Spray application of 50 ppm aqueous solution of silver nanoparticles on cluster bean leaves inoculated with sunhemp rosette virus (SHRV) showed complete suppression of the disease, suggesting that silver nanoparticles are effective antiviral agent. Toxicity studies of silver nanoparticles on plant pathogens open the door for a new range of plant antimicrobial agents.

Antisense Technology Mahnoor Patel M.Sc. Biotechnology, Department of Biotechnology, Veer Narmad South Gujarat University, Udhna Magdalla Road, Surat, Gujarat, India [email protected]

Diseases are often connected on the expression of some disease causing gene which is important to produce that “protein”. If the expression of this gene can be disputed then the disease can be cured. Antisense technology is a method of disputing the production of protein. It may be used to treat Page | 133   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

some therapeutics for diseases in whose pathology is the production of protein plays a major role. Antisense technology is important tool in the inhibition of that particular gene expression. The principle behind it is that the antisense nucleic acid sequence base pairs with its complementary sense RNA strand is inserted and prevents the RNA from translated into protein. The complementary nucleic acid sequence is complementary synthetic oligonucleotide, often oligodeoxy ribonucleotide, or longer antisense RNA sequence. Key words: Antisense Technology, Protein inhibition, RNAi therapeutics

Edible Vaccine: The Future of Oral Vaccination Mahnoor Patel, Sana Madhani, Somya Jajodia M.Sc. Biotechnology, Department of Biotechnology, Veer Narmad South Gujarat University, Udhna Magdalla Road, Surat, Gujarat, India [email protected]

Edible vaccine is cost effective, easy in administer, easy in storage and safe vaccine delivery system. Edible vaccines are prepared by molecular pharming using the science of genetic engineering. Selected genes which are pathogenic can be isolated from the source and then introduced into the plants. That plant is now called transgenic plant and it is able to produce the encoded protein. That protein is pathogenic which is used to give to the people as edible vaccine. That protein is used for the therapeutic purposes. It is less costly rather than another vaccine. Resistance to genetically modified food may affect the future of edible vaccine. Key words: Edible, Molecular pharming, Vaccine Page | 134   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India Nanorobot: The Future of Medical Science Mahnoor Patel, ZarinIftekhar, ShivangiJariwala, NeetuBohra M.Sc. Biotechnology, Department of Biotechnology, Veer Narmad South Gujarat University, Udhna Magdalla Road, Surat, Gujarat, India [email protected]

Nanorobotics is the branch of nanotechnology. Nanorobotics is important in creating machines or robots which is close to the microscopic scale of nanometer

(10-9 m). A nanorobot is a tiny machine and it is designed

specifically to perform a specific or particular task repeatedly and with great precision at nanoscale dimensions. The diameter of nanorobot is about 0.5 to 3 microns and it is constructed out of parts which have dimension range of 1 to 100 nm. Nanorobotics has strong potential to revolutionized healthcare, to treat disease in future. Nanotechnology changes the shape of the industry, broadening the product development and interrelationship between

pharmaceuticals,

biotechnology

and

healthcare

Nanorobots are used for various purposes in medical science. Key words: Nanorobot, Nanotechnology

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industry.

3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Effects of Cadmium Stress on Biochemical Traits of Lallemantia royleana Majid Rostami Assistant Professor of Crop Physiology, Department of Agronomy, Malayer University [email protected]

Lallemantia royleana (Nazboo, Black psyllium or Balangu) is an important medicinal plant of Iran which is belonged to Lamiaceae family. The plant is very interesting for traditional medicine in different area of world especially Asian countries and extracted mucilage from the seed of this plant has many applications in food science and also modern pharmacology. In order to study the effects of cadmium stress on biochemical traits of Lallemantia an experiment was conducted in experimental greenhouse of Malayer University based on completely randomized design (CRD) with four replications. Different concentration of cadmium (0, 2.5, 5, 10 mg.l-1) was made using CdCl2 and used for irrigation of lallemantia pots. Based on results effects of different levels of cadmium on antioxidant enzymes of Lallemantia were significant but the observed trend wasn’t similar. By increasing the concentration of cadmium amounts of superoxide dismutase (SOD), ascorbate peroxidase (APOX) and peroxidases (POX) activity increased linearly but the effects of cadmium on catalase (CAT) activity was different. By increasing the cadmium concentration up to 5 mg.l-1 the activity of CAT increased but in higher concentration (i.e. 10 mg.l-1) CAT activity decreased significantly. The trend of change in activity of antioxidant enzymes was similar in root and leaf of Lallemantia but in all of the measurement the observed values for root were higher than leaves. Significant increase of antioxidant enzymes mean that oxidative stress started in the plants because of cadmium toxicity.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Keyword: Enzymes, heavy metals, medicinal plant, oxidative stress

Identification of novel two component lanthipeptides in threeStreptomyces species by microbial genome database mining Mangal Singh*, Dipti Sareen Department of Biochemistry, Panjab University, Chandigarh

*email: [email protected], Frequent use of antibiotics leads to the generation of new antibiotic resistant strains. This poses a need for further identification of novel antibiotics, trapped in a cycle of discovery and resistance emergence. Lantibiotics are a class of antimicrobial peptides that can circumvent this problemowing to their advanced mode of action, which take decades for a pathogen mechanism to evade.One such example is of nisin, which is being used in food preservation for more than 50 years without development of any significant resistance. Nisin consist of a single posttranslationally processed peptide while two-component lantibiotics contain twofunctionally distinct peptides thatact in synergy to provide bactericidal activity. Because of the synergistic action,they display similar or higher activity than nisin. Both activity-based and in silicoapproaches have been adopted for the discovery of novel lantibiotics. In this work, a strategy was employed to identify novel cryptic lantibiotic clusters by identification of lantibiotic transporter (LanT) analogues, followed by a detailed in silicoanalysis of the nearby genetic region for the potential lantibiotic biosynthetic genes. This strategy resulted in identification ofnovel twocomponent lanthipeptide clusters in three Actinomycete representatives, Streptomyces hygroscopicus ATCC 53653, S. bingchenggensis BCW-1

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

and S.viridochromogenes DSM 40736. To date, two-component lantibiotics have only been isolated and confined to Firmicutes.

Atomic Force Microscopy as a tool for Chronic Obstructive Pulmonary Disease Research Manoj Madheswaran Swansea University, United Kingdom [email protected]

Chronic obstructive pulmonary disease (COPD) is the third leading cause of death in world. It causes irreversible damage to lungs and affects the quality of life. Currently there is no cure for COPD and currents treatments are concerned about controlling the symptoms. Treatment for COPD is still challenge and increases death rate gradually. It is necessary to find new effective drugs to clear airway blockages and improve the life style of patients. The gel forming mucins are the major polymeric component of mucus. These mucins are glycoprotein contains sugar molecules and forms highly diverse in structure and more conserved. By understanding the glycobiology of mucin proteins can lead to development of suitable drugs. The major aim of the study was to develop and begin optimization of Atomic Force Microscopy (AFM), as a nanotechnology, to visualize the potential global structural changes to mucins during an inflammatory response. The first objective was to use a cultured cell lines to the model to model the inflammatory response in lung post infection and obtain secreted mucins. For this A549 human lung cell carcinoma epithelial cell line was incubated with bacterial virulence factor lipopolysaccharide (LPS) and the cytokine TNF- α. The second objective was to isolate secreted mucins, post inflammatory response from cell culture media. This mucin extraction would Page | 138   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

be dominated by secreted mucins MUC5AC and MUC5B. With the mucins molecules prepared the third and major objective was to develop a protocol to AFM to visualize the mucin molecules for differential structures during the inflammatory response. Much of the experimental time spent during the research was devoted to AFM optimization. Keywords: Chronic obstructive pulmonary disease, Mucus, glycoprotein, nanotechnology, Atomic force microscopy.

Nanopore based forth generation DNA sequencing Maurya Yamini Omprakash MSC biotechnology V.N.S.G.U.Surat [email protected]

Nanopore-based sequencers, as the fourth-generation DNA sequencing technology, havethe potential to quickly and reliably sequence the entire human genome.Nanopores are being hailed as a potential next-generation DNA sequencer that could provide cheap, high throughput DNA analysis. The techniques being investigated for use in DNA sequencing and mapping applications. A crucial impasse to the success of nanopores as a reliable DNA analysis tool is the fast and stochastic nature of DNA translocation. The single-molecule techniques used by this technology allow us to further study the interaction between DNA and protein, as well as between protein and protein. Nanopore analysis opens a new door to molecular biology investigation at the single-molecule scale. The significant advantages of nanopores include label-free, ultra-long reads (104–106bases), high throughput, and low material requirement. At present, the rapid

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

development of semiconductor technology makes it possible to fabricate nanopore-based DNA sequencing microchips of high speed. A UV-Spectrophotometric assay for determination of artemisinin Mayengbam Nganthoi* and Keithellakpam Sanatombi Department of Biotechnology, Manipur University, Imphal, 795003. [email protected]

Abstract: A simple UV- Spectrophotometric method has been developed for determination of artemisinin content in four Artemesia cultivars. The developed method was validated according to ICH guidelines. The developed method was validated in terms of accuracy, precision, linearity, limit of detection, limit of quantification and reproducibility. The proposed method can be used for determination of artemisinin from petroleum ether, n-hexane and toluene extracts of leave, stem and flower tissues of Artemisia plants. Key words: UV- Spectrophotometry; validation; Artemisia; Artemisinin.

Evaluation of rice genotype under Different Forms of Nitrogen in Rice (Oryza sativa) Wallalwar M. R.1*, Rashmi Upadhyay1, Verulkar S.B. 2 1 Ph.D. Scholar, Department of Plant Molecular Biology and Biotechnology,Indira Gandhi KrishiVishwavidyalaya, Raipur,Chhattisgarh. 2 Professor and Head, Department of Plant Molecular Biology and Biotechnology,Indira Gandhi KrishiVishwavidyalaya, Raipur,Chhattisgarh [email protected]

The main driver for crop improvement over the years has been yield and yield improvement has accelerated primarily not only due to introduction of scientific approach but also through use of fertilizers. Among fertilizers, Page | 140   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

nitrogen (N) is one of the most limiting nutrients in natural and agricultural ecosystems. The majority of plant-useable N is consumed as nitrate (aerated soils) and as ammonium (poorly aerated/submerged soil). Increasing frequency of drought and change in water availability drastically changes available predominant form of N as nitrate. Rice genotypes show significant variability for N uptake (external efficiency) and N utilization (internal efficiency).Influx of nutrient such as nitrate and ammonium into roots is governed by highandlow-affinity transport system (uptake, compartmentation,translocation and remobilization) and regulated by N form and concentration.Integration of physiological, molecular and agronomical approach in evaluation of cultivars under different nutrient and water regimes for development of varieties that use N more efficiently, 122 lines of F14 generation of recombinant inbred lines(RILs) from the cross of DanteswariXDagaddeshi

along

with

few

parents

were

evaluated

andrevealed that some of the genotype shows strong correlation with particular type of N fertilizer. Further 32 lines with parents were selected on basis of their nitrogen use efficiency(NUE) and yield, were further assessed under nitrate, ammoniaand control condition, showed that some genotype respond well to particular form of nitrogen with variation in phenotypic data (as compared to control) with higher yield and NUE. Fundamental molecular mechanismsofNUEinvolve number of gene in nitrate and ammonia transport system. The expression analysis of selectedgenes involve in nitrate and ammoniautilization among 10 selected line,5 lines showed positive correlation with phenotypic data.The identification of genotype with great nitrate use efficiency,certainly have key role in N efficiency particularly under aerobic and water stress condition.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Embryoscope: Advanced Embryo Monitoring BMI Kajol Patel, Shreya Parekh, Mahnoor Patel, Vibhuti Surati M.Sc. Biotechnology, Department of Biotechnology, Veer Narmad South Gujarat University, Udhna Magdalla Road, Surat, Gujarat, India [email protected]

Embryoscope is a machine which is used to monitor embryo from the time of conception. It allows fertility specialist to select the most viable embryo during in vitro fertilization. This instrument reduces the risk of taking embryo from the incubator for observation. It can monitor 12 embryos at once and take photos of each embryo every 5 to 20 min during the entire incubation period. With the help of this instrument fertility expert can also monitor the abnormality in embryo. The embryoscope is an incubator with an integrated camera. The detection of abnormality can be done without harming the embryo. Key words: Embryoscope, Embryo monitor

Chemical Synthesis of the Repeating Unit of Type Ia Group B Streptococcus Capsular Polysaccharide For Carbohydrate Based Antibacterial Vaccine Development Prolay K. Mondal, Guochao Liao, Mohabul A. Mondal†, and Zhongwu Guo* *Department of Chemistry, Wayne State University, Detroit, MI 48202, USA, † Department of Chemistry, University of Gour Banga, Malda, WB 732103, India [email protected], [email protected]

The structure of the capsular polysaccharide (CPS) of serotype Ia Group B Streptococcus (GBS) has been characterized for years, but its repeating unit, which is a challenging pentasaccharide with a branch and a difficult αPage | 142   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

sialic acid linkage, has not been synthesized yet. In this report, an effective synthesis was developed for the serotype Ia GBS CPS repeating unit, which had a reactive functionality linked to its main-chain reducing end to enable further elaboration, such as coupling with carrier proteins. The target molecule was accomplished by a convergent [2 + 3] glycosylation strategy employing a sialodisaccharide as donor and a branched trisaccharide as acceptor. The strategy was designed to suit the synthesis of oligomers of the repeating unit.

Gene expression of morphinan alkaloids in Papaverorientale L. Mohammad Reza Naghavi* and Seyed Mohammad Hashemi Department of Agronomy and Plant Breeding, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran [email protected]

In the present study, the expression level of various genes (COR, SalAT, SalR, T6ODM, CODM and Salsyn) in themorphinan pathway for response to methyl jasmonate and salicylic acid at different time points (6, 12, 24, 48 h) was examined. Afterwards,in vitro production of thebaine, morphine and codeine was investigated. A. rhizogenes strains ATCC15834 on the hypocotyl explant were able to produce transformed roots with the highest frequency (%26).Quantitative reverse-transcription PCR analysis showed that the

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

expression of SalR, Salsyn, SalAT, T6ODM and CODMwere up-regulated significantly with MJ treatment at 48 h. Moreover, expression levels ofSalsyn, T6ODM and CODM increased considerably in SA at 48 h treatment and SalRwas down-regulated at 24 h.

Furthermore, High-performance liquid

chromatography analysis showed that thebaine and morphine in MJ elicitation at 48 h and codeine at 24 h was significantly increased by 2.63-fold (3.08 mg/g), 6.18-fold (5.38 mg/g) and 3.67-fold (2.57 mg/g) respectively. Also, the production of morphineafter SA treatment for 48 h was increased by 4.22-fold (2.87 mg/g) and codeine after 24 h wasincreased by 2.59-fold (1.61 mg/g) in comparison with control treatment.Our findings suggest that Orientale poppy hairy roots may produce morphine, codeine and thebaine using methyl jasmonate and salicylic acid as an abiotic elicitor and can be as a good source for production of referred commercial alkaloids. Keywords:Morphinan alkaloids, Elicitation, Gene expression, Hairy root culture, Papaverorientale L.,

Immunological detection of Pythium aphanidermatum, a fungal pathogen found in Rhizome rot disease in Turmeric Monalisa Ray1, Swagatika Dash1, Abtar Mishra1, Sanghamitra Nayak1, Shikha Singh1* 1

Centre of Biotechnology, Siksha O Anusandhan University, Khandagiri, Bhubaneswar751003, Orissa, India [email protected]; [email protected]

Pythium aphanidermatum is a fungal pathogen responsible for rhizome rot, a devastating disease affecting the productivity of turmeric around the world. In the present study, we have discussed detection of the disease at an early stage of infection by serological methods. In these methods, polyclonal

antibody

(pAb)

raised

against

the

polypeptide

fraction

recognized the fungal proteins with high sensitivity (approx. 50 ng). In

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

enzyme-linked immunosorbent assay (ELISA), the antiserum dilution up to 1:5000 detected fungal antigens at a concentration of 7 mg/mL and at 1:2000 antiserum dilutions, antigens at a concentration as low as 2 mg/mL could be detected. The immunodominant P. aphanidermatum proteins were identified by Western blot at different bands ranging from 15-70 kDa. The antibody was specific since they cross-reacted with the fungal proteins isolated from all the infected samples collected from different agroclimatic zones of India. Importantly, as shown by ELISA tests and immunoblot analysis, the pre-bleed serum did not cross-react with the fungal proteins. Using these serological approaches, the presence of P. aphanidermatum was detected in rhizomes of turmeric infected with rhizome rot, even at a very low level of infection. Hence, the developed polyclonal antibody raised in rabbit against polypeptides secreted by P. aphanidermatum is a useful tool to specifically detect the presence of the fungal pathogen at an early stage. These serological approaches may allow us to propose development of a rapid, reliable, and easy to perform diagnostic kit for early detection of rhizome rot in turmeric. Keywords: Pythium aphanidermatum, Rhizome rot, ELISA, Western blot, Polyclonal antibody.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Overview of Recent Advancements on Biobutanol Production Monika Mate*, Swapnali De, Vishakha Bais, V.P. Bhange Department of Biotechnology, Priyadarshini Institute of Engineering & Technology, Nagpur- 19. [email protected]

The increasing cost of petroleum products, its non sustainability, political crisis at source locations, environmental hazard, is now focusing interest on renewable, sustainable and environmental safe fuels as alternatives. The greatest area that brings about the rise in price of fossil fuels in the transportation sector that use gasoline and diesel. Biofuels in particular are receiving climbing attention due to the increase in industrialization, population, global demand for energy driven by fast depletion of conventional fuels. For best evaluation of use of biofuel bio based butanol is superior to ethanol with regard to having higher energy content and less corrosive. Biobutanol or biobased butanol is also called biogasoline is a second generation alcoholic fuel. Also the advantage is that it utilizes lignocellulosic waste as feedstock which is mainly the non edible plant material which includes keen crops of wood and grass and agro forest residues. The production and consumption of biobutanol is expected to reduce the consumption of oil and natural gases by automobile industry and also reduces emission of greenhouse gases that are harmful to environment. It suits internal combustion engine more than bioethanol and can be used as a fuel and also used as a direct replacement for gasoline and also acts as a rich fuel extender. This poster focuses on recent advancements that are coupled with the newer approaches for biobutanol production that renews the hopes on having a cleaner and energy-efficient commercial process.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India Serratio Peptidase

Summary: Many of 110 strains of Serratia, isolated from soil, water, milk and dairy equipment, were biochemically closely related to the coliaerogenes bacteria. Acid and gas was formed from glucose in 14 days at 30° by 53% and from lactose and MacConkey's broth by about 40%. All except one strain gave——++ IMViC reactions. Selected 3 Strains Strain No. 1- Serratia marcescens – NCIM- 2600 Strain No.2- Customer No.5029 E-15-( ATCC /IMTECH )- MTCC -7298 Date:14/09/2010 (Procured on) AAATC 2716 R ST005.From: MTCC C , Invoice No.MTCC/10/09/5029 Strain No. 3-Isolated from Dairy Equipment – Chandigarh (UT) India No.2 Strain from MTCC selected to Developed for commercial scale Production , submitted project proposal to DSIR – ON 2ND May 2011. Principally Proposal was accepted on innovative and technical aspects but unfortunately turn down due to financial company budget so We did not stop the work , transferred all findings to AETL- Mumbai. Developed in house by Strain improvement Techniques and standardization of Process followed by scaling up An inverse relationship was observed between depth of pigmentation and carbohydrate fermentation. Complete loss of pigment in mutant strains was not uncommon, and was associated with loss of proteolytic properties and increase of saccharolytic activity. The majority of the strains had psychrophilic characteristics: 75% grew at 3–5°. Most strains showed moderate growth at 37°, but only 7 formed red pigment at that temperature. strains resembled Serratia marcescens in Page | 147   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

morphology, containing minute coccoid rods smaller than those of coliaerogenes bacteria. IDENTIFICATION 

Gram stain negative



Facultative anaerobes



Motile



Rod shaped



0.5-0.8 microns x 1.0-5.0 microns



Produces

characteristic

red

pigment, Prodiginines ( Prodigiosin) 

Highly mucoid colonies



Oxidase negative



Reduce nitrate



Indole negative



Gives dark pink pigmentation



Vogues-Proskauer positive



Simmon's citrate positive



Produces DNAse, Gelatinase, Lipase, Lecithinase, Chitinase and Esterase enzymes



Utilizes malonate



Serratia is often confused with the organism Enterobacter

Identified and Propagated Seed prepared in adaptation Media Fermentation on Casein/Dextrose Media: at 30 0 C, Ph: 7.0. DO: 40% for 45 Hrs. Shown 8000 pu/ml activity stable by set Assay procedure and conditions Page | 148   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

Caseiene quality required as substrate: total protein content not less than 85%. The medicinal use of Serratiopeptidase is very well known and very well documented. Recent Japanese patents even suggested that oral Serratio peptidase may help to cure or prevent such viral diseases as AIDS and hepatitis B and C. But perhaps it’s most spectacular application is in reversing cardiovascular disease. In fact, serrapeptase appears so effective in unblocking carotid arteries that one researcher- Dr.Hans Nieper, the late, eminent internist from Hannover, Germany-called it a "miracle"enzyme.

Proteolytic enzymes (also known as proteinases or peptidases) are ubiquitous in nature, being found in animals, plants, bacteria, and fungi. Human beings produce such well known peptidases as trypsin and chymotrypsin to help digest our food, but we also generate countless others to control virtually every regulatory mechanism in our bodies. For example, various peptidases are involved in initiating blood clotting (thrombogenesis) and also in dissolving clots (fibrinolysis); in evoking an immune response and quelling it; and in both promoting and halting inflammation. The mechanism in each case is the ability of the enzyme to cut or cleave a protein target into two or more pieces, usually at very specific cleavage sites. The same mechanism makes it possible for peptidases to inactivate HIV, the AIDS-associated virus, by pruning the viral proteins necessary for infectivity. We all had some kind of function in the childhood with formation of pus around it, so have you wondered what exactly is pus and why it is caused ? Pus is basically whitish-yellow, yellowbrown or yellow exudates that are produced at the time of pyogenic

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

bacterial infection. Normally a sore doesn’t stop weeping continuously for three days gets worse and form pus. Formation of pus Pus is formed from the living and dead cells traveling into the inter-cellular spaces all around the concerned cells. Purulent, pyrogenic or suppurative element results in pus. Pus boasts of a protein rich thin fluid identified as dead cells and liquor puris, which are major part of innate immune reaction of the body. How to treat or get relief from pus Once you discover pus coming out from the wound, a few things are required to do, which includes: * One of the best and effective things to keep the pus area clean is to gently wash the wound with mild water and soap. This would result in removing the bacteria completely along with the accumulated pus. * Bandage is another thing to get relief from the pus, since it absorbs a relatively high amount of pus from unhygienic area and protects the wound from additional injury or dirt. Bandage will also help in holding the moisture, which is imperative in the process of healing. It is suggested to change the bandages at least thrice a day. One can find scores of specialized bandages in the market, but a conventional adhesive bandage with a tape also works pretty well. * Before wrapping the wound, it is suggested to rub a topical anti-biotic ointment or solution like Polysporin, Neosporin or the one comprising the Page | 150   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

bactitracin ingredient. This antibiotic helps to seal the moisture and keeps the additional germs out. A person may need to eat oral drugs if the infection gets worse. * Another measure to treat the wound filled with pus is to wrap it by a moist cloth as many times as you can throughout the day. This will gently make pus coming down to the surface, therefore, softening the skin in order to drain and rupture the skin without infuriating the virus or causing severe pain. * If the outside layer of the wound is discharging pus, it can result in forming an abscess. It is suggested not to rip the scab off. You should frequently soak it in lukewarm water and wrap the scab up with a bandage in order to keep it damp. This way it would be much easy to drain the pus that will lead to heal the scab faster Patent Position : Planned to patent the process when achieve the optimum. Technology Trends

:

It was always in lime light as hot cake Molecule utility wise since long ago so Biotechnologists and Microbiologists were constantly on R&D Globally, Japan ( Nagase) and Chialdezang S. Korea were active and Developed process in 1972 but still on work to find economical means downstream for getting therapeutically quality product without much recovery loss and feasible yield. In India no one trying only depending on outcome of other countries if possible so it’s positively needed in India someone (R&D Lab) to take up quickly the venture to fulfill the demand. Technology Forecasting: Page | 151   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Technology will be surely demonstrated and translated to commercial scale production with time bound schedule to produce quality with desired specifications in economical mode and viable yield (3.5 gr. /L) better than available technologies yet anywhere for innovating therapeutic applications. So with other formulations to enhance the range of usage. Standards / Specifications: Strain:

Serratia marcescens (E-15)

Yield:

- 3.5- 5.0 gr/ lit (finished product/L fermented

Broth) Fermentation cycle:

35 hrs

Fermented Broth activity:

25-28 spu /mg (8000 -9000pu/ml)

Finished product’s Purity:

88 %

Specifications of Enzyme Physical state: granules

Free flowing powder /granules/Enter coated

pH:

7.5- 8.5

Temp:

(optima) 30-45o C

Fermentation yield:

3.5-4.5gr/L

LOD:

31-3.5%

B.D.:

0.3-.45

Ash content:

1.5 – 2.5%

Tenacity:

1.6

Odor:

pleasant

Color:

off white to white crystalline free flowing Powder

Peptidase Activity:

not less than 2500 spu/mg

Stability:

24 months, consistent

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Sustainability:

positive in gut and intestine

Knowledge Gaps : 1. Clinical Testing of finished product through recognized institution. 2. Stability of product storage as well as formulation. 3. Oral / gut/ Intestinal sustainability Critical Elements : 1. Contamination free and controlled broth foaming Fermentation Cycle 2. Clarification of Filtrate 3. Selection of Ultra filter Cartridge Mol. Wt cut off pore size to get Pure Enzyme 4. Stability of activity as Bulk or formulated product 5. Sustainability of finished product 6. Scaling up to commercial scale 7. Strain maintenance and improvement 8. Yield consistency 9. Impurity removal ( associated other Proteins) 10.

Odor and pyroxene removal

11.

Supportive and smooth Technology transfer

Enabling Technologies: The medical use of enzymes as anti-inflammatory agents goes back many years. The proteolytic enzymes in common use today derive from bacteria (serra peptidase grown from Serratia marcescens cultures), plants (bromelain from pineapple stem and papain from papaya) and animal sources (trypsin and chymotrypsin from hogs or cattle). They're all generally useful, but for many applications serrapeptase appears to be the Page | 153   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

most useful of them al l. In one study serrapeptase was compared to trypsin, chymotrypsin and pronase (another microbial peptidase) in a rat model of scalding, which is known to induce abnormal activation of fibrinolysis. Serrapeptase was far more effective than any other enzyme in repressing fibrinolysis in this model, in agreement with its documented clinical efficacy as an antiInflammatory agent. As it naturally secretes a secondary metabolite in form of Red color Pigment Identified as “Prodigiosin” Prodigiosins are a family of bright red colored bacterial pigment and derive their name from the miraculous (prodigious) events associated with their occurrence. They indeed seem to be living up to their name as a host of activities

such

as

anti-microbial,

anti-malarial,

anti-cancer

and

immunosuppressive have been associated with them. Out of these, immunosuppressive and anti-cancer activity has received more importance as it has a clinical promise. Prodigiosins, isolated mostly from Gram negative bacteria are characterized by a common pyrryldipyrrylmethene structure with varying side chains. The review discusses the mechanisms involved in the anti-cancer activity of this class of compounds. In vitro, prodigiosins have been shown to primarily target the cancer cells independently of the status while little or no effect has been observed on normal cells. In addition, prodigiosins are effective in cancer cells with multidrug resistance phenotype and defects in the apoptotic pathways. These make prodigiosins attractive candidates for further development. Though the molecular targets of prodigiosins have not been clearly defined, they have been found to target different signaling pathways possibly through induction of DNA double strand breaks and/ or neutralization of pH Page | 154   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

gradients leading to changes in cell cycle proteins and apoptosis. The review will discuss the recent findings related to the mechanism involved in the anti-cancer activity of this class of molecules. On further study of pigment secreted revealed that it consititute as bipyrrole structure having Mol.Wt. 323.432 g/mol. And Mol formula: C20 H25 N3O.

Prodigiosin is Secondary metabolite produced naturally beside secretion of primary Peptidase Enzyme. Many industries globally though few in India Manufacturing commercially viable therapeutic Enzyme “Peptidase by Fermentation process using Microbial Strain : Serratia Marsesance coco bacili.so facing problem to remove or get rid of the red colour from the Broth filtrate and finished product as this colour hinders the purified product Serratio Peptidase for pharma formulation. During quick growth phase of Strain ( MTCC-8708) lot of dark pink color pigment is generated, this pigment is not desired for producing Enzyme finished product for Pharma formulations. So We had to remove it by repeated Salting out/Dialysis or Activated carbon adsoption.The removal of pigments process decreases’ Recovery efficiency due to Enzyme activity loss.and the pigment was also lost as it is washed out. A novel thought evolved to extract the pigment and collect separately which will provide colorless Peptidase of better yield (80% of secreted in Broth). And beside it Pigment is obtained Separately which can be as production of Prodigiosin by purification which is Page | 155   

known for effective usage as Anti  

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

biofouling /inhibitor Agent. so a automatic concept for taking up as R&D Project at our Lab. initiated successfully. To make a innovative and commercially viable and successful project, We collaborated with two prestigious Academic Institutions NIT-Agartala and NIT- Durgapur having expertise in Bio processing Biocolours and Engineering.Dr. Biswanath Bhunia and Dr. Tamal mandal both are continuously working on applied and Industrial Biotechnology, We AETL – has proved its potential as pioneer Enzymes manufacturing and R&D unit. Business wise also promising. The task of maximum production of Pigment followed by Peptidase production. Extraction and separating color from Enzyme filtrate completely is our first objective. Then collection of Pigment and purify Prodigiosin is 2nd objective. So to exploit inhibitory activity of Prodigiosin against gram+ Bacteria make Bulk powder of maximum activity to provide clinical formulation as antibacterial, anti-fungal, anti neoplastic, anti-proliferative, anti oxidant and anti malarial parasites Exercises done successfully on line towards achievement of said project: 1) Cultivation of Serratia Strain on Ground nut / Peas as substrate 2) Maximum Pink Pigment accumulated 3) Test protocol followed for Prodigiosin production found positive 4) Separation of Pigment Lab scale process 5) Qualitative and quantitative analysis being followed 6) The molecule has great business potential so producer of Serratio peptidase can explore this for double profitable business.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

A Novel In Silico Drug Designing Approach to identify herbal active principles to combat Rabies Viral adherence and pathogenesis 1

Muralidharan. V, 2Deecaraman.M, 3Kumarasamy.P, 4Sureshkannan.S, 5Sujatha Mohanakrishnan.

1

Phd research scholar, Department of Biotechnology, AMET University, Kanathur603112, India

2

Dean, Department of Industrial Biotechnology, Dr. MGR Educational & Research Institute, Maduravoyal, Chennai-600095, India. 3

Professor & Head, Department of Bioinformatics & ARIS cell, Madras veterinary college, Chennai 600007, India.

4

Associate Professor & Head, Veterinary Public health & Epidemiology, Veterinary college & Research Institute, Tirunelveli-627358, India.

5

Assistant Librarian, Department of Bioinformatics & ARIS Cell, Madras Veterinary College-60007 [email protected]

Rabies is a fatal central nervous system (CNS) disease responsible for approximately 60,000 annual deaths worldwide, making it the tenth most common lethal infectious disease. It is known to be present on all continents except Antartica, and

infects domestic and wild animals

including humans. Rabies still poses a significant health problem in many countries in Asia and Africa, where 95% of human deaths occur even though safe, effective vaccines for both human and veterinary use exist. The causative agent is neurotropic virus consisting of non segmented, negative-stranded RNA contained within a bullet shaped envelope of the rhabdoviridae family and the genus Lyssavirus. Neuroinvassiveness and neurotropism are the main features that define the pathogenesis of rabies. Although Rabies Virus (RV) pathogenicity is a multigenic trait involving Page | 157   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

several elements of the RV, Genome, the Rabies Virus (RV) Glycoprotein (G) plays a major role in the RV pathogenesis. The main objective of this research work is to find a natural compound which inhibits the attachment to the target cells by interaction with cell membrane components. It is now possible through computer based bioinformatics procedures, to identify and structurally modify a natural compound to design a drug with desired properties and to assess its therapeutic effects theoretically. Discovery studio 2.5 provides a set of protocols for predicting and analyzing the intreraction between protein and ligands. Molecular Docking experiments were carried out among six selective herbal active principles viz. Scutellarin, Lobeline, Cichoric Acid, Diterpene, Lactone, Andrographolide and Alangin. The Molecular interaction analysis based on dock score and Potential means energy value, Diterpenoid was identified to have highest dock score 71.08 and -25.332 pmf value and this compound could be used in the process of drug development to control Rabies virus adherence and pathogenesis. Keywords:

Rabies,

Glycoprotein,

pathogenesis

neurotropism,

Neuroinvassiveness, Genome, Acetylcholine.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Biosynthesis, characterization of gold nanoparticles using M. piperita leaf extract and their anticancer activity Nabeel Ahmad1 and Prof. Rajiv Dutta2* 1. School of Biotechnology; IFTM University, Lodhipur Rajput Moradabad U.P. India 2. Institute of Bioscience and Bioengineering, ShriRamswroop Memorial University, Barabanki (UP), India. [email protected]; [email protected]

The present study involves the novel approach for the synthesis of nanomaterial holds infinite possibilities as nanotechnology has a major impact on medicine and its application. Leaf extract of Mentha piperita is a very good bioreductant for gold chloride (HAuCl4·4H2O) which leads to the synthesis of gold nanoparticles. Leaf extract of Mentha piperita was mixed with gold chloride (HAuCl4·4H2O) and UV-vis spectroscopy and FTIR were used to detect the synthesis of nanoparticles. TEM and SEM were used for size, shape and structural morphology of synthesized nanoparticles. Synthesized gold nanoparticles were in range of less than 100nm and were spherical in hexagonal in shape. The gold nanoparticles also showed potent cytotoxic effect against A549 lung cancer cell lines with an IC 50 value of 59 μg/ml/24 h by the MTT assay. Keywords: Nanotechnology, Gold chloride, Biosynthesis, Characterization, Cytotoxicity,etc.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India Ebola Vaccines: - “VSV-Zebov” Nayak Jitendra Jayprakash Department of Biotechnology, V.N.S.G. University, Surat-395007 [email protected]

New Ebola vaccine is 100% successful in clinical trials. This could be the beginning of the end for Ebola. A new single dose of vaccine shows the effectiveness against Ebola after just one week. The results of a fast tracked clinical trial involving the people, offer the most promising evidence so far that we may be able to protect against the haemorrhagic fever. Known as VSV-ZEBOV, the vaccine is now going through further trials in younger patients, and if all goes to plan could become the first licensed vaccine against Ebola. We believe that the world is on the verge of an efficacious Ebola vaccine. The trial, which was supported by the drug firms, is now continuing in children between the ages of 13 and 17. In the future, the researchers also want to see if they can offer protection to 6-12 year olds. Keywords: - EBOLA, Vaccine-(VSV-ZEBOV), trials, drugs.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Cloning Of Ciprofloxacin Resistance and Amylase Producing Property in Dh5-Α Strains of Escherichia Coli Neeraj K. Gaur School of Biotechnology and Bioinformatics – D.Y. Patil University, Belapur Navi Mumbai [email protected]

One of the major causes for obesity in humans is incomplete digestion of starch which increases the absorption of Disaccharides. The disaccharides cannot undergo catabolism to release energy, so if they are absorbed from the intestines, the molecules will directly take part in fatty acid synthesis. The aim of this study was to counter starch indigestion by transforming gut microbes with amylase producing gene. It also aims to develop antibiotic resistance property in them ensuring the survival of transformed organism during any treatment of broad spectrum antibiotics. Microorganisms with the desired properties were screened from various soil samples; their plasmids DNA were isolated and respective genes were PCR amplified using sequence specific primers. The PCR products were restriction digested by EcoRI enzyme and incorporated into pUC18 vectors. The recombined vectors were used to transform host DH5-α strains of Escherichia Coli. Both the antibiotic resistance and amylase producing property were observed in the transformed host organisms. One set of host cells were transformed using both the recombinant vectors, they were grown on Nutrient agar supplemented with 10 ppm ciprofloxacin and enriched with 2% starch. The double transformed E. coli cells were able to form large number of colonies on the antibiotic supplemented media and displayed large clear zones for iodine test which indicates starch digestion because of the amylase produced by the organisms.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Further research and experimentation is required in the form of in-vivo studies on mouse models using various host micro-organisms, this will facilitate and encourage future research on transformation of gut microorganisms.

In-Silico Approach to Control Pathogenesis of Kyasanur Forest Disease Neha. N. Patil, Shivalingsarj Desai, Sharanappa A, Deepak Yaraguppi Department of Biotechnology, B.V.B. College of Engineering & Technology, Hubli580031 [email protected]

Drug is an organic small molecule that activates or inhibits the function of a biomolecule such

as

a protein,

which

in

turn

results

in

a

therapeutic benefit to the human beings. Drugs are essential for the prevention and treatment of disease. Drug development is demanding. But the process of drug design, development and commercialization is a tedious, time-consuming and cost-intensive process. To fulfil these challenges, several multidisciplinary approaches are required for the process of drug development; collectively these approaches would form the basis of In Silico approach in drug design. Kayasanur Forest disease is a Monkey fever and is a tick bone disease. It is caused by a flavivirus. The mode of infection to humans is through tick bite and Haemaphysalis spinigera is the major vector. Statistical survey has revealed that in the year 2003 to 2012 there were 3263 suspected cases, with 823 confirmed cases and 28 deaths. Therefore there arises a serious necessity to find a drug molecule to cure this disease. At the first, we identified the proteins involved in the disease for which we obtained FASTA sequence using Page | 162   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

NCBI. Sequence alignment of the proteins was carried out by using BLAST (Basic Local Alignment Search Tool), This tool compares the query sequence with the sequences in database. For each comparison, it gives the E- value, identity and score, later from the identified sequences we identify the protein by structural and functional characteristics using different bioinformatics tools, later drugs will be developed for the identified protein to control the pathogenesis of Kyasanur Forest Disease (KFD).

Fabrication of Glycated Hemoglobin Biosensor Based on Au Nanoparticles Embedded Nitrogen Doped Graphene Nanosheet Nidhi Chauhan1* and Utkarsh Jain2 1

Amity Institute of Nanotechnology, Amity University, Noida - 201303, Uttar Pradesh, India

2

Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore, India [email protected]

The glucose level measurement in the diabetic patients without short term variations is significant. The glycated hemoglobin (HbA1c) level in blood provides this long-term glucose level analysist. In the present work, we have successfully established the method for fabrication of enzyme sensor based on nitrogen-doped graphene–AuNPs modified FTO electrode. Nitrogen-doped graphene–AuNPs modified FTO electrode was immobilized with fructosyl amino-acid oxidase (FAO) enzyme. The biosensor showed optimum response, when operated at +0.2 V. The biosensor exhibited excellent sensitivity and low detection limit, fast response time and wide linear range (from 0.4 to 2000 μM). The enzyme electrode was used 100 times over 4 months, when stored at 4 oC. Page | 163   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Key words— Biosensor, glycated hemoglobin, graphene

Basic and Applied Research in Biology and Medicine: Delivery of antiinflammatory nutraceuticals by nanoparticles Nidhi Shukla*, Harshada Shenoy*, Riddhi Gadre* & Shraddha Dhamale* SIES Graduate School of Technology Sri Chandrasekaendra Saraswathy Vidyapuram, Plot1-C D & E, Sector V, Nerul, Navi Mumbai

Nanotechnology is the engineering of functional systems at the molecular scale. Nanotechnology is able to create many new materials and devices with a vast range of applications, such as in medicine, electronics, biomaterials energy production, and consumer products. Most chronic illnesses, including cancer, diabetes, and cardiovascular and pulmonary diseases are mediated through chronic inflammation. Thus, suppressing chronic inflammation has the potential to delay, prevent, and even treat various chronic diseases, including cancer. Various nutraceuticals from fruits, vegetables, vitamins, spices, legumes, and traditional Chinese and Ayurvedic medicine have been shown to safely suppress proinflammatory pathways; however, their low bioavailability in vivo limits their use in preventing and treating cancer. We describe here the potential of nanotechnology to fill this gap. Several nutraceuticals, including green tea polyphenols, coenzyme Q, quercetin, thymoquinone and others, have been packaged

as

nanoparticles

and

proven

to

be

useful

in

“nano-

chemoprevention” and “nano-chemotherapy.” Keyword:

Nanotechnology, Chronic Illness, Chronic Inflammation,

Nutraceuticals, Nanoparticles.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Computational study of blocking nucleotide binding domain of P-glycoprotein by flavonoids from Oroxylum indicum Neeraj Gaur, Nirav Shah* and Harshvardhan Rao School of Biotechnology and Bioinformatics, D. Y Patil University, Plot No-50, Sec-15, CBD Belapur, Navi Mumbai [email protected]

Malignant neoplasm involved abnormal cell growth and by inchmeal proliferated

over

other

parts

of

the

body.

Repeated

usage

of

chemotherapeutic drugs in cancer cannot overlay its complete cytotoxic effect. Cancereous cells have intrinsic ability to efflux these drugs by over expression of P-glycoprotein, an ATP-binding cassette transporter. Binding of flavanoids (phytochemicals) at the nucleotide-binding domain of Pglycoprotein is known to abbreviate the amount of drug effluxesOroxylum indicum, a medicinal plant is known for its anti cancerous properties. In the present study, we focused on screening flavonoid compounds from the plant that would interact at the nucleotide binding domain of P-glycoprotein to block the transport. Homology modelling of human P-glycoprotein has been performed based on crystal structure of 4M2S (Chain A) and the protein was energy minimized and equilibrated using GROMACS software. A molecular docking was performed between modelled P-glycoprotein and the flavonoids using iGemDock program. Many of the flavonoids showed high binding affinity to the nucleotide binding domain of P-glycoprotein. The interaction profiles of 24 flavonoid compounds showed Baicalein 6glucuronide as a promising molecular recognition point at the nucleotide binding domain with a binding energy of -102.4 kcal/mol. Moreover, Asn94, Ser829, Thr834 were hydrogen bonded with the above polyphenol and formed a stabilised molecular complex. Thus, the present work highlights the specificity of plant polyphenol compounds towards blocking the Page | 165   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

nucleotide binding domain by using computational methods. This will further assist the biologists to study the phytochemicals of Oroxylum indicum as a promising lead to regulate the efflux mechanism in cancerous cells

The effect of Anulom vilom on self confidence of an individual. Varsha V Andhare and Nisha Velayudhan The Institute of Science 15, Madam Cama Road, Fort, Mumbai 400032 [email protected]

Ancient Indian breathing technique of Pranayam can have beneficial effects on the health of an individual. Practicing anulom vilom helps an individual to fill the lungs with appropriate amount of air i.e. oxygen. A study was performed with a total of 30 post graduation students who were segregated into two groups –control and test. The test groups would practice anulom vilom regularly for 15 mins once in the morning and once in the evening for a period of 15 days whereas the control group continued a normal routine and strictly did not practice any exercise. The study basically concentrated on finding out the confidence improvement of these students. It was observed that the self confidence of the individual practicing anulom vilom improves whereas the one in the control group did not show any significant change. Hence, it can be concluded that practicing anulom vilom is a very useful aspect for any individual for improving ones personality especially for students. Keywords: Anulom vilom, confidence, personality, etc.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India Full Blown Human Body on a Tiny Chip Nitu Bohra M.Sc. Biotechnology (SEM – 8) Veer Narmad South Gujarat University Surat [email protected]

Recently, an exciting new multidisciplinary scientific field has emerged around a concept which has come to be coined “Organ on a chip”.This chip is tiny representation of human organs,such as lungs,hearts and intestine,made from cells embedded on plastic about the size of a computer memory stick. Channels lined by living vascular cells then mimic the body’s circulatory system. Human bodies on chips would have applications for beyond drug development,toxins effect on neural system and recreation of body system. The chip is about the size of a microscope slide and in contains one or more open or closed chamber in which cells can be seeded. The material the “chip” is made up of different kinds of material like silicone-based rubber(so called PDMS). The surface of the chip can be coated with extracellular matrix proteins, these are proteins that normally reside in the intercellular space of tissues and form structure which help create the tissue architecture. The Organ-on-chip approach is particularly relevant for human- specific diseases of which no animal models are known. Organ like the brain and immune system are very typically human. For this reason representative cell line or animal model system are lacking for many human serological and psychiatric diseases, as well as for diseases in which the immune system plays a critical role,ranging widely from auto-immune disease all the way to cancer. Just a few eg. Amytrophic lateral sclerosis, multiple sclerosis, depression, Page | 167   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

schizophrenia and psychosis, Alzheimer, diabetes, rheumatic diseases and diseases like SLE, skin diseases, fibrotic diseases, and all types of cancer. However, in view of the very human nature of our immune system which is involved in the evolution and progression of basically every disease, development of drugs for nearly all diseases may benefit from a the Organon-Chip approach. This technology proving to be long term technology.

Metagenomic analysis of microbial diversity and prevalence of pathogens in fish from Mumbai Onkar Naik1, R. Shashidhar2, Devashish Rath3, J. R. Bandekar2 & Archana Rath1 1

Department of Biotechnology, University of Mumbai, Vidyanagari, Santacruz (East), Mumbai-400098, INDIA. 2

Food Technology Division, Bhabha Atomic Research Centre, Trombay, Mumbai400085, INDIA.

3

Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai400085, INDIA. [email protected]; [email protected]

Mumbai being a coastal city with a large population, the demand for sea food is very high. It is also a major exporter of fish all over the world. So it is imperative to study the microbiological quality of fish we consume. Vast majority of microbes are non-pathogens, yet they contribute to the evolution of antibiotic resistance in pathogens by serving as an accessible reservoir of resistance genes. Hence prevalence study of pathogenic and nonpathogenic microbes in fish will give further insights for future antibiotic resistant studies. Most microbes cannot be cultured under normal lab conditions, hence non-culturable methods like metagenomics is used to analyse the total microbial population in a niche. Using metagenomic analysis and taxonomic profile the overall distribution of microbial Page | 168   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

population in marine fish sampled in Mumbai was studied. The fish microflora was dominated by Proteobacteria with 92.07% of total OTUs representation. Firmicutes with only 2.97% of total OTUs was followed by Bacteroidetes (2.64% OTUs). Significant microbial diversity was observed with

detection

of

28

families

including

many

rare

groups

like

Helicobacteraceae, Xanthomonadaceae, Planococcaceae, Aminiphilaceae etc. having low but uniform distribution. Within the identified species, high abundance of fish pathogens like Flavobacterium spp., Psychrobacter spp., Aeromonas spp., Arcobacter spp. and significant presence of groups with most known potential human pathogenic species such as Vibrio, Shewanella, Photobacterium, Enterobacteriaceae was found. Prevalence of 42.90% pathogenic families is indicative of fish being a major carrier of pathogenic micro-organisms. Many of the pathogens belonging to these families can be transmitted to human beings through food. Presence of pathogenic micro-organisms like Photobacterium damselae, Shewanella algae, Vibrio metschnikovii, Acinetobacter johnsonii suggests a high possibility of lateral gene transfer of antibiotic resistant determinants from non-pathogenic to pathogenic microbes. Relatively high abundance of Acinetobacter indicates significant impact of anthropogenic activities in this ecosystem. Key words:- Acinetobacter, anthropogenic effects, lateral gene transfer, metagenomic analysis, micro-organisms, Mumbai, seafood, V3 region

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Use Of Mustard Oil Cake For Protease Production By Bacillus Subtilis And Its Application Pallavi S. Badhe, Ravindra V. Adivarekar Department of Fibres and Textile Processing Technology Institute of Chemical Technology, N.P. Marg, Matunga, Mumbai, 400019 [email protected]

A Gram positive, motile, mesophilic Bacillus subtilis strain capable of producing extracellular protease was isolated from abattoir waste material, collected from Wadala local market, India. The efficiency of mustard oil cake as a substrate for protease production by Bacillus subtilis strain in submerged fermentation (SmF) has been studied. The mustard oil cake was selected, as it is rich in protein, cheap, abundantly available being an agroindustrial waste and can be constructively used as one of the main constituent in medium formulation. Different physical and chemical parameters such as pH, temperature, and substrate concentration and incubation time were optimized. The protease activity was found to be maximum when pH of medium containing 4% of mustard oil cake was 11, at 40ºC, after 70hrs of submerged cultivation. Further, the growth curve studies indicated that the production of extracellular protease was initiated in the early lag phase and was maximum in the late exponential phase. The application of this protease enzyme was studied for removal of blood stain and chocolate stain. Stain removing tendency of protease under study was higher as compared to commercial and standard detergents.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

A review on biofilm formations and its impact Lathika Gopalan, Michelle Sawant, Panchami Pradeep Kumar SIES Graduate School of Technology Sri Chandrasekaendra Saraswathy Vidyapuram, Plot1-C D & E, Sector V, Nerul, Navi Mumbai-400706. [email protected], [email protected], [email protected]

Biofilms are communities of micro organisms attached to surfaces which can be found in medical industrial and natural settings. The ability of many bacteria to adhere to surfaces and to form biofilms has major implications in a variety of fields including the food industry, which creates a persistant source of contamination. The formation of biofilm is determined not only by the nature of attachment surface but also by the characteristics of bacterial cell and by environmental factors biofilm microbes are typically surrounded by an extracellular matrix that provides structure and protection to the community. Microbes growing in a biofilm have a characteristic architecture generally comprised of macro colonies (containing thousands of cells) surrounded by fluid-filled channels. Biofilm grown microbes are also notorious for their resistance to a range of antimicrobial agents including clinically relevant antibiotics. Biofilms play an important role by acting as a protective barrier for the microorganisms, it acts as a barrier to protect the soil and ground water from contaminationand has many other advantages. On a contrasting note it also has many disadvantages like these include biofouling, biocorrossion, equipment damage, product contamination and threat to public health as well.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Microbiological control programmes at industrial level should aim at reducing both the detrimental effects of micro organisms on the process and the environmental impact associated to the use of biocides. Keywords: biofilms, biofouling, biocorrosion, microbiological control.

An Innovative Attempt to Restrain Environment Stress Using Locally Developed Ecofriendly (Botanical / Green) Pesticides Dr.Pankaj Audichya1, Phool Chand Kumawat2 and Mahesh BG3 Reliance Foundation, Research & Development Farm, Mouda (Nagpur), MaharashtraIndia pankaj.audichya@reliancefoundation

57353 MT chemical pesticides were consumed in India during the year 2014-15. According to Cornell environment scientist David Pimente, only 0.1 % of applied chemical pesticides reach the target pests, leaving the bulk (99.9 %) of the pesticides to impact the environment. Maximum chemical pesticides are not easily degradable, they persist in air, soil, leach to ground and surface water, leads biodiversity loss, gene erosion, infertility of lands, population change, thus disrupt ecosystem. Awareness on harmful effects of synthetic chemical pesticides on environment has encouraged researchers of Reliance Foundation to look for better alternatives to synthetic chemical pesticides. An ecofriendly and least cost extraction and purification technology was invented at Research & Demonstration Farm, Mouda. During the year 2014-15 & 2015-16 rigorous studies were conducted using 6 own developed purified botanical / natural pesticide formulations. Among the treatments Mouda CG-Plus reduced the highest disease and pest(70-90%) followed by Mouda Bio-Shooter (7087%), Mouda Bio-10(70-80%),Mouda Bio-CEB(63-74%), Mouda BioPage | 172   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Chrym( 50-71%), Mouda Bio-5plus(50-63%). These botanicals were developed by using different parts of plants. In the present study 3.2 kg a.i. chemical pesticide in cotton, 4 kg a.i. in chilli, 4.64 kg a.i. in brinjal, 4 kg a.i. in cauliflower and 0.6 kg a.i. in mustard was impeded to penetrate in the wide range of environment. These botanical pesticides performed similar to commercial

standard.

No

adverse

effect

on

beneficial

insects

( Coccinellidae, honeybees) was observed. It was concluded that botanical pesticides affect only target pest, break down into harmless compounds and decomposed quickly, their use maintains the biological diversity of predators as a result, and they provide residue free food, water and safe environment to live.

Lipid production from algal culture under different growth conditions & its quality analysis Paramjeet S.1,, Manasa P.2, K. Narasimhulu 3 National Institute of Technology, Warangal, Telangana [email protected]

Microalgae cells have the potential to rapidly accumulate lipids that contain fatty acids important for high value fatty acids . There is currently no standard extraction method for the determination of the fatty acid content of microalgae. This has caused a few problems in microlagal biofuel research due to the bias derived from different extraction methods. Therefore, this study aims to evaluate three different methods for lipid extraction. Significant differences were observed among all the three methods in regards to extract yield and fatty acid composition. The Modified Bligh & Dyer extraction technique stood out best for effective extraction of microalgal lipids for long chain saturated fatty acids. Page | 173   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Different

strains

consumed

different

nitrate

sources

at

different

concentrations. As a result of which the nitrogen stress induction is not uniform for all the strains. Chlorella vulgaris yielded better biomass when NaNO3 was used as nitrogen source compared to other strains and nitrogen sources i.e. KNO3 and Urea in BG11 media. Chlorella vulgaris also produced highest lipid content while Scenedesmus obliquus being least productive. Chlorella vulgaris showed the best combination with NaNO3 as a nitrogen sources with BG11 broth media for biomass and lipid productivity. The study also indicated that optimal biomass yield is obtained at pH 7. However, the maximum lipid output obtained at pH 8 .This clearly indicates that higher pH leading to stress in the culture & inducing lipid production. Keywords: Microalgae oil, Fatty acid, Extract yield, Lipid profile

Structural analysis of autolysin enzymes of bile soluble and bile insoluble Streptococcus pneumoniae reveals altered number of choline-binding sites. Dr.Peela Sree Ramchandra Murthy, Dr. S. Sujatha Department of Microbiology,Jawaharlal Institute of Postgraduate Medical Education & Research, Pondicherry [email protected]; [email protected] 

Bile solubility assay was considered as the gold standard for the identification of Streptococcus pneumoniae. The complete structure of the enzyme autolysin responsible for the phenotype is recently determined. It was proposed the dimerization of the sub-units and the complete binding of choline (of teichoic acid in the cell walls) to all the ten choline-binding Page | 174   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

regions are two independent steps necessary for the enzymatic activity of the protein. Reports suggest that bile insoluble pneumococci are frequently isolated from the clinical samples. The complete structure of the enzyme in bile insoluble pneumococci is not determined to validate the roles of each domain. Aims and objectives: The study was done to correlate the structural changes in autolysin enzymes of bile soluble and insoluble pneumococci. Materials and methods: Two sequences were extracted from NCBI database-Streptococcus pneumoniae R6 strain (bile soluble; accession number AE007317.1) andstrain 1508/92 (accession number AJ419973.1; reported to be bile insoluble). Amino acid sequences were translated for the sequences and aligned using BLAST. The protein structures were predicted using SWISSMODEL programme (swissmodel.expasy.org). Results and discussion: Homo-dimerization of the proteins were identified in both the organisms. However, there are 6 choline binding regions (CBRs) in chain A and 4 choline binding regions (CBRs) in chain B of R6 strain, whereas only 3 each choline binding sites were observed in chains A and B in strain 1508/92. E275N mutation in strain 1508/92 caused loss of two CBRs, one in each chain. G289 and T290 deletions were highly conserved in bile insoluble pneumococci, but are suggested to be not responsible for enzyme inactivity. This shows that the reduced number of choline binding sites can be the possible explanation for the bile insoluble phenotype. Page | 175   

 

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Conclusion: The study identified that dimerization can still occur in bile insoluble pneumococci, while the number of choline binding sites can determine the bile solubility phenotype.

Study of Antioxidant activity of Bacterial Carotenoids Department of Microbiology, Smt CHM College Ulhasnagar- 3 Dr. Padma Deshmukh, Pranali Shete [email protected]

Synthetic pigments have many disadvantages over naturally occurring pigments like toxicity and accumulation. Hence the search for novel natural pigments is the prevailing inclination of the industry. Carotenoids, which comprise of 600 different pigments, rangingfrom yellow to red in color has multifaceted applications. Major sources of carotenoids are plants, bacteria and fungi. Humans, who cannot produce them, require carotenoids not only as precursor of vitamin A, but also for other health benefits like anti cancer, anti oxidant activity. Plant based carotenoids are subject to seasonal variation and also limited in range; plant based diet provides only 40 different carotenoids, hence major repertoire of carotenoids is still to be explored. Bacteria can be a potential source of carotenoids in terms of variance, stability and constant output by using bioprocess technology. In present study pigmented bacteria have been isolated from different fresh water and marine water samples. Around 30 isolates producing pigment in the range of yellow to orange were used for further studies. Extraction and purification of the pigment has been done using solvent solvent extraction. Page | 176   

 

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Screening of potential isolates was done using absorption maxima of purified pigments. Extracts showing absorption maxima in the range of 430nm to 490nm, which matches with the absorption range of carotenoids, were selected for further studies. Confirmation of presence of carotenoids is done using various chemical tests like hexane test, acidified pyridine test etc. All chemical tests gave positive result for the purified extracts. Purified extracts were used to study antioxidant activity using DPPH. Extracts were found to have above 60% antioxidant activity. Key words : Carotenoids, DPPH

Identification and Molecular Cloning of the Putative TerpeneSynthase Gene from Stackebrandtia nassauensis DSM 44728 Pranav Bhaskar*, Dipti Sareen Department of Biochemistry, Panjab University, Chandigarh 160014, Chandigarh, India [email protected]

The comprehensive study of microbial secondary metabolites extends wonderful possibilities for the discovery of novel natural products which often present promising biological activities. Actinomycetes are already remarked for the production of an enormous variety of natural products which constitute an affluent source of bioactive compounds of potential commercial and pharmaceutical value, consisting biosynthetic pathways of various metabolites being used in human and veterinary medicine, agriculture and manufacturing. Terpenoids constitute the largest and the most varied group of natural products but only few terpenesynthases have been characterized from actinomycetes till date. However, recent extensive bacterial genome sequencing and bioinformatics analysis have rendered more than a hundred distinct predicted terpene synthase genes. Using Page | 177   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

BLASTp tool, and pentalenene synthase from Streptomyces sp. UC5319 strain as a query, we have found theterpene synthase homolog gene inStackebrandtia nassauensis DSM 44728 genome, snas_1127. The putative gene product of S. nassauensis, SNAS_1127, consists of two sets of universally conserved Mg2+ binding motifs, the aspartate-rich motif and the downstream NSE/DTE triad,

235

domain and the corresponding459DDLYP and

92

DDHFLE

NDLFSYQREin N-terminal

602

NDLFSYQKEmotifs in C-

terminal domain.Like all other terpene synthases, the conserved motifs in each domain are separated by 142 amino acids. SNAS_1127 was found to contain 59% and 62% amino acid sequence identity with germacradienol/ geosmin

synthase

Streptomyces

from

griseusNBRC

Streptomyces 13350,

avermitilisMA-4680

respectively;

indicating

and that

SNAS_1127 is a putative germacradienol/ geosmin synthase.To establish the function of this putative gene, it was cloned in pET28a(+) vector and was expressed in E. coli BL21(DE3) as N-terminal-His6-tagged protein. The expressionanalysis of the expressed protein would enable us to assign the role that it plays in terpene biosysnthesis.

Exploring of potentialbacteriophages against multi-drug resistant clinical pathogens a*

Prasanth.M, aRamesh.N,bBulentBozdogan,aGothandam.K.M.

a

Antibiotic Resistance Laboratory and HTS Lab, School of Bio-Sciences and technology, VIT University, Vellore, Tamil Nadu, India.

b

Medical Microbiology Department, Adnan Menderes University, Aydin, 09100, Turkey. [email protected]

Multiple drug resistant Gram negative clinical bacteria are a very serious concern worldwide because of its increasing prevalence in hospital settings. Recent attention has focused towards phage therapy to overcome Page | 178   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

this threat. Aim of our study was to isolate and characterize the highest lytic capability phage as a model to evaluate the potential on phage therapy. Total of 120 clinical bacterial isolates were collected from clinical centres, screened and identified by 16s rRNA identification. Antibiogram, minimal inhibitory concentration (MIC), and transconjugation experiments were performed. Prevalence of resistance genes (CTX-M, IMP, VIM, OXA, KPC, and NDM) among the clinical isolates was identified and sequenced. For isolation of bacteriophages, water samples were collected from Ganges river, Kaveri river, and water treatment plants (from Chennai, Karur and Bangalore). Potential phages were isolated by agar overlay method, parallel streaking and spot test, and lytic phages were characterized. All the 120 clinical isolates received were found to be multi-drug resistant by disk diffusion and identified as 62 Escherichia coli, 16 Klebsiella pneumoniae, 14 Pseudomonas aeruginosa, 9 Acinetobacter baumannii, 6 Enterobacter hormaechei, 6 Enterobacter cloacae, 4 Proteus mirabilis, 3 Proteus rettgeri. MIC

results

for

meropenem,

showed

MIC50=0.12

µg/mland

MIC90=16µg/ml. PCR results showed that 22 of 120 carried blaNDM, 5 oxa181, and 2 imp4. Lytic phages were isolated against MDRE. coli, K. pneumoniae, P. aeruginosa, A. baumannii and E. cloacae. Phages that showed highest lytic capability were characterized. Detailed analysis was done to examine the MOI (10, 1, 0.1, and 0.001) of phage particles required to decrease the bacterial load, frequency of appearance of phageresistant mutants, latent period and burst size. Phage genome size, protein profiling, and microscopic identification are to be done. Our results showed the potential of lytic phages to eliminate drug resistant pathogens and potential candidates for phage cocktails.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Correlation studies in Sunflower (Helianthus annuus L.) A. D. Khilare, K. R. Kamble and P. B. Bhusari College of Agriculture, Latur VasantraoNaikMarathwadaKrushiVidyapeeth, Parbhani [email protected]

The experimental material consisted of 112 sunflower exotic germplasm accessions and three checks (SS-2038, LSF-8(1), and Morden) obtained from the Germplasm Maintenance Unit (GMU) of Oilseeds Research station, Latur. The details of the exotic germplasm accessions and check varieties sown during kharif 2014. In order to find out the association or correlations between the characters considered in the present study. Phenotypic and genotypic correlation coefficients were worked out for the quantitative characters. Positive and significant phenotypic and genotypic correlations were observed between yield and yield contributing component viz., plant height (G 0.418, P 0.414), head diameter (G 0.57, P 0.573), hull content (G 0.183, P 0.160), 100 seed weight (G 0.272, P 0.259) and seed filling percentage (G 0.228, P 0.242), while negative and non-significant correlation with days to 50 per cent flowering (G -0.020 and P -0.022), volume weight (G -0.102 and P -0.102) and days to maturity G(-0.003 and P -0.003) at both genotypic and phenotypic levels. However, positive but non-significant correlation was noted only with oil content (G 0.121, P 0.120) at both genotypic and phenotypic levelsSeed yield per plant was highly positively correlated with 100 seed weight, head diameter, plant height and seed filling percentage. The genetic improvement in yield can be achieved by applying strong selection to above characters which was genetically correlated. Among the yield components days to 50 per cent flowering, days to maturity and plant height were positively and significantly

Page | 180   

 

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correlated between each other indicating that simultaneous selection for these characters might be important in seed yield.

Novel recombinant epoxide hydrolases for the production of enantiopure drug intermediates Priya Sainia*, Shadil Ibrahim Wania and Dipti Sareena a

Department of Biochemistry, Panjab University, Sector 14, Chandigarh 160 014, India e-mail:[email protected]

Epoxide hydrolases (EH; 3.3.2.x) are ubiquitous enzymes catalyzing the ring opening of epoxides to their equivalent vicinal diols,being the green alternative epoxides

to and

asymmetric

chemical

methods.Enantiomerically

diols

produced

are

thus

key

chiral

pure

synthons

for

pharmaceutical drugs. EHs are co-factor independent enzymes as well as have extensive substrate range. Microbial EHs offer unlimited supply of biocatalysts so they are being explored for novel EHs. Inthe present study, two novel EHs have been identified from the actinomycetes; Streptomyces griseus NBRC 13350 and Stackebrandtia nassauensis DSM 44728 by genome database mining. The two EH genes (sgehand sneh) were clonedin pET28a(+) and overexpressed inE. coli BL21(DE3). Whereas, (His)6-SGEH was found in soluble fraction, (His)6SNEH

formed

soluble

aggregates.

Improperly

folded

SNEH

was

transformed to completely active soluble form by fusing it with trigger factor (TF) chaperone. Both (His)6-SGEH and TF-(His)6-SNEH were purified by IMAC using Ni-NTA. HPLC analysis of SGEH showed 80% enantiomeric excess of the product diol with 2,6-dichlorophenylglycidyl ether and phenylglycidyl

ether

epoxides.

TF-SNEH

catalysed

hydrolysis

of

benzylglycidyl ether and α-methylstyrene oxide to 86% and 73% Page | 181   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

enantiomeric excess of the product diol, respectively. Enantioselectivity of these EHs will be further enriched by further optimizations.

Phytoremediation R Mira Nishvanthi Bannari Amman Institute of Technology, Sathyamangalam, Alathukombai (pt), Erode, Tamil Nadu [email protected]

Land, surface waters, and ground water worldwide, are increasingly affected by contaminations from industrial, research experiments, military, and agricultural activities either due to ignorance, lack of vision, carelessness, or high cost of waste disposal and treatment. The rapid build-up of toxic pollutants (metals, radionuclide, and organic contaminants in soil, surface water, and ground water) not only affects natural resources, but also causes major strains on ecosystems. Phytoremediation is a method to solve environmental contamination. This green technology that involved “tolerant plants” has been utilized to clean up soil and ground water

from

heavy

metals

and

other

toxic

organic

compounds.

Phytoremediation involves growing plants in a contaminated matrix to remove environmental contaminants by facilitating sequestration and/or degradation (detoxification) of the pollutants. Phytoremediation is a cost effective, environmental friendly, aesthetically pleasing approach most suitable for developing countries.Abundant amount of metals along with their ores are present in the soil. The presence of large quantity of metals in soil will lead to adverse environmental effect. There is a need for the removal of these metals in soil by using eco-friendly techniques.Plants can also clean up contaminants like pesticides, explosives, and oils present in

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

soil. The plants store the contaminants in the roots, stems, or leaves. And convert them to less harmful chemicals within the plant shoot or root zone. At last they are made to vapors, which are released into the air.

Unraveling Gene/ Genome Evolution in Eukaryotes using NGS technology: A Study in Drosophila Species Radhika Khanna and SujataMohanty Department of Biotechnology, Jaypee Institute of Information Technology, A-10, Sector 62, Noida, Uttar Pradesh [email protected]

The genotype environmental interaction can result in alteration in gene or genome structure of an organism, which can well be understood by doing comparative study among sympatric and allopathic species populations. By applying Next Generation sequencing approach, we can comprehensively explore the structural variation arising in various protein-coded nuclear genes. Drosophila has been well established eukaryotic model organism used for evolutionary studies.During the present study, evolutionary relation with respect to five enzyme encoded genes such as Gpdh , Amy,Gld, Marf and sina were studied between four sympatric species of Drosophila (D.bipectinata,

D.takahashii,

D.biarmipes,

and

D.nasuta

collected

fromUnchehara,MadhyaPradesh) and four allopatric species(D.bipectinata, D.takahashii,D.melanogaster,D.ananassae). Zaprionus indianus collected from Ludhiana, Punjab was used as an outgroup species during the analysis work. By using NGS technology we generated sequences of above mentioned genes in four sympatric species of India and for allopatric comparison the sequences were retrieved from NCBI.The results were analyzed

Page | 183   

using

advanced

computational

sequence

analysis

 

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tools.

 

3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

Phylogenetic trees were constructed using Neighbor-Joining and Maximum Likelihood methods. The outcome of our study reflects that the environmental factors play a key role in bringing molecular changes; however the effect varies differentially on functional genes. Therefore the conserved genes or the markers may not be reliable always for the phylogenetics and molecular taxonomy studies for species population without exploring their eco-geographic distribution. Plant Tissue Culture: A Broad Field Dr.Rahul Vijay, Dr.Ankit Agrawal Madhya Pradesh Council of Science and Technology, Bhopal

Plant Tissue Culture is a very important technique for broad field. Nowadays this techniques have take best role in science.Modern plant tissue culture is performed under aseptic conditions under HEPA filtered air provided by a cabinet. Different techniques in plant tissue culture may offer certain advantages over traditional methods of propagation. Mainly Plant tissue culture techniques is used maintain or grow plant cells, tissue or organs under sterile conditions on a nutrient culture medium of known composition. Although some growers and nurseries have their own labs for propagating plants by the technique of tissue culture, a number of independent laboratories provide custom propagation services. Plant tissue culture is used widely in the plant sciences, forestry, and in horticulture. The commercial production of plants used as potting, landscape, and florist subjects, which uses meristem and shoot culture to produce large numbers of identical individuals. To conserve rare or endangered plant species. Keywords: Plant Tissue Culture, propagation, horticulture etc. Page | 184   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Molecular basis of male sterility and fertility restoration in rice Rajib Das, Jayalekshmy V. G2, and Siddhesh. R Nadkarni3,Chandan Roy4 Ph.D Scholar1,3,Associate Professor 2,Department of Plant Breeding and Genetics, College of Agriculture, Vellayani, Kerala Agricultural University, Assistant Professor4, Bihar Agriculture University, Sabour. [email protected]

Most of higher plant species are hermaphroditic and male-sterility is often considered as an accident of development. In fact among the multiple possible causes of male-sterility, the most frequently met in nature is cytoplasmic male-sterility (CMS) in a plant is unable to produce functional pollen. Recent molecular studies have shown that this trait is determined by additional genes created in plant mitochondrial genomes due to their high recombinogenic activity. The products of these genes interfere with the formation of male gametophytes leads to sterility. It occurs in many plant species and is often associated with chimeric mitochondrial open reading frames. In a number of cases, transcripts originating from these altered open reading frames are translated into unique proteins that appear to interfere with mitochondrial function and pollen development. Recently, a series of genes and proteins related to CMS, photoperiodsensitive male sterility, self-incompatibility, and other types of microspores deterioration have been characterized through genetics or proteomics. Especially the latter, offers us a powerful and high throughput approach to discern the novel proteins involving in male-sterile pathways which may help us to breed artificial male-sterile system. This represents an alternative tool to meet the critical challenge of further development of hybrid rice.

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Nuclear restorer (Rf) genes function to suppress the deleterious effects of CMS-associated mitochondrial abnormalities by diverse mechanisms. There are now several well-characterized CMS systems based on the restoration-maintenance relationship. It can be categorized into three types, namely, Wild Abortive (WA) type, Honglian (HL) type and Boro-Taichung (BT). Both HL and BT CMS belong to the gametophytic CMS systems, they share similar heredity models of Rf alleles. Their inheritance of the fertility restoration is controlled by one restorer gene whose effect in the male sterile cytoplasm is gametophytic. WA-CMS is reported to be restored with one to three genes with epistatic interactions. Key Words: Male Sterility, Genetics, Proteomics, Hybrid rice and CMS System.

Growth Kinetics of Chlorella Vulgaris and Botryococcus Braunii Based on Presence of Sodium Bicarbonate in Medium as Parameter Ojasvi Bhatnagara, Rajini Kanthb, Archana Rao Pc* a,b,c Department of Chemical engineering, BVRIT, Narsapur *[email protected]

Growth studies were conducted on microalgae species Chlorella Vulgaris and BotryococcusBraunii in a batch mode. In the present study the effect of sodium bicarbonate salt NaHCO3 in the culture medium on growth rate of the two selected microalgae species was investigated. For this purpose the growth kinetics of the aforementioned two species under varying concentrations of sodium bicarbonate salt in the medium (0ppm, 15ppm, and 70ppm) was investigated. Growth was conducted in batch mode for 7 days with temperature, pH, and light intensity maintained constant. Specific growth rate, doubling time and dry weight of biomass obtained were Page | 186   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

calculated. Maximum specific growth rate showed by Chlorella vulgaris was observed at 70ppm bicarbonate, while BotryococcusBraunii showed best specific growth rate at 15ppm bicarbonate. Keywords: microalgae, bicarbonate, growth kinetics, batch culture

Transgenic Plastids in Basic Research and Plant Biotechnology Bammidi Ramesh Sri Sivani Degree College. Srikakulam. Andrapradesh, INDIA. 532001 [email protected].

Facile methods of genetic transformation are of outstanding importance for both basic and applied research. For many years, transgenic technologies for plants were restricted to manipulations of the nuclear genome. More recently, a second genome of the plant cell has become amenable to genetic engineering: the prokaryotically organized circular genome of the chloroplast. The possibility to directly manipulate chloroplast genomeencoded information has paved the way to detailed in vivo studies of virtually all aspects of plastid gene expression. Moreover, plastid transformation technologies have been intensely used in functional genomics by performing gene knockouts and site-directed mutageneses of plastid genes. These studies have contributed greatly to our understanding of the physiology and biochemistry of biogenergetic processes inside the plastid compartment. Plastid transformation technologies have also stirred considerable excitement among plant biotechnologists, since transgene expression from the plastid genome offers a number of most attractive advantages, including high-level foreign protein expression and transgene containment due to lack of pollen transmission. This review describes the Page | 187   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

generation of plants with transgenic plastids, summarizes our current understanding of the transformation process and highlights selected applications of transplastomic technologies in basic and applied research. Keywords; chloroplast, plastid transformation, reverse genetics, functional genomics, plant biotechnology. Decoding chromatin dynamics during B cell fate commitment Ravi Boya, Anurupa Devi Yadavalli, SameenaNikhat, Jagan M.R.Pongubala Department of Animal Biology, School of Life Sciences, University of Hyderabad, Hyderabad [email protected]

Organization of the genome in 3D nuclear-space is known to play a crucial role in regulation of gene expression. However, the chromatin architecture that impinges on the B cell-fate choice of multi-potent progenitors remains unclear. By employing in situ Hi-C, we demonstrate that distinct sets of genomic loci undergo a developmental switch between permissive and repressive compartments during B-cell fate commitment. Correspondingly, we show that topologically associating domains (TADs) represent coregulated functional subunits of chromatin and display considerable structural alterations as a result of changes in the cis-regulatory interaction landscape. The extensive rewiring of cis-regulatory interactions is closely associated with differential gene expression programs. Further, our results highlight the regulatory role of Ebf1 and its collaborative factors in chromatin reorganization and transcription regulation. Together, our studies reveal that alterations in promoter and cis-regulatory interactions underlie changes in higher order chromatin architecture, which in turn determines cell-identity and cell-type specific gene expression patterns. Page | 188   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Role of glutathione in plant defense signaling network: A multistep crosstalk with salicylic acid and ethylene Riddhi Datta1, 2 and Sharmila Chattopadhyay1* 1

Plant Biology Laboratory, Drug Development/Diagnostics & Biotechnology Division,

CSIR- Indian Institute of Chemical Biology, 4, Raja S.C. Mullick Road, Kolkata 700 032, India 2

Present address: Department of Botany, Government General Degree College, New Town, Rajarhat, Kolkata. [email protected]

Plants are continuously threatened by a range of biotic and abiotic stress factors. Consequently, they have evolved highly sophisticated defense strategies. The role of glutathione (GSH) in plant defense is an established fact. However, its association with other established signaling molecules in the defense signaling network remains to be evaluated. In this study, we developed chloroplast-targeted gamma-glutamylcysteine synthetase (γECS) over-expressed transgenic tobacco (NtGp line) and constructed a forward subtracted cDNA (SSH) library using NtGp line as a tester. Proteomic profiling of NtGp line was also performed by employing twodimensional gel electrophoresis (2-DE) coupled to MALDI-TOF-TOF MS/MS analysis. NtGp11 was further tested for tolerance against drought stress.

Interestingly,

in

addition

to

SA-related

transcripts,

1-

aminocyclopropane-1-carboxylate oxidase (ACC oxidase), a key enzyme of ethylene (ET) biosynthesis, was identified in the SSH library. Proteomic profiling of NtGp line also corroborated with the transcriptomic profile and several

defense-related

accumulation.

proteins

Fascinatingly,

were

induction

identified of

with

increased

1-aminocyclopropane-1-

carboxylate synthase (ACC synthase), which catalyzes the rate-limiting step of ET biosynthesis, was also noted. Protein gel blot analysis confirmed Page | 189   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

the enhanced accumulation of ACC oxidase in NtGp line. NtGp11 was next tested for stress tolerance. It exhibited tolerance to drought stress by increased germination rate, water retention, water recovery, chlorophyll, and proline content compared to wild-type plants. Our results demonstrated that GSH plays an important role in combating drought stress in plants by inducing stress related genes and proteins. Together, our data revealed that GSH is involved in multiple steps crosstalk through ET as well as SA to combat environmental stress.

Engineering Applied to the Biosciences and Biotechnology: Applications of Nanotechnology in Pollution Control Riddhi Gadre*, Nidhi Shukla*, Harshada Shenoy* & Shraddha Dhamale* SIES GRADUATE SCHOOL OF TECHNOLOGY Sri Chandrasekaendra Saraswathy Vidyapuram, Plot1-C D & E, Sector V, Nerul, Navi Mumbai

Nanotechnology is the engineering of functional systems at the molecular scale. Nanotechnology is able to create many new materials and devices with a vast range of applications, such as in medicine, electronics, biomaterials energy production, and consumer products. Nanomaterials represent a promising newtechnology for rapid environmental cleanup and pollution control. Research into and development of innovative engineered nanomaterials such as nanoscale zerovalent iron (nZVI) for site remediation has increased dramatically in the past decade. Because of the small particle size, nanomaterials achieve treatment rates that are significantly greater than micronscale treatment media. Another expected advantage of nanomaterials is that they are small enough to be mobile in aquifer systems and can be injected directly into the subsurface to treat Page | 190   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

contaminated source zones. Keyword: Nanotechnology, Nanomaterials, Zerovalent ion, Micronscale treatment.

Enhanced antimicrobial activity of transition metal and curcumin complexes S.A.Valia1, A.V.Gole2, S.D.Kamat3 and D.V.Kamat3 1 and 3 Department of Microbiology and 2 Department of Chemistry Mithibai College, Vile Parle west Mumbai [email protected]

The active principle of turmeric called curcumin has been of great interest to scientists due to its several beneficial characteristics including antiinflammatory and anticancer properties. However, the major stumbling block for curcumin has remained its solubility. References indicate that biological activity of curcumin increases with metal complex formation. Present investigation involved preparation of complexes of curcumin with iron as a metal. Iron (III) and curcumin in the molar ratio of 1:1, 1:2 and 1:3 were prepared using methanol as the solvent. A distinct colour change of the solution from yellow to brown was observed indicating formation of coordinate bond between the metal and curcumin. Complex formation was confirmed using UV and IR spectroscopy. The solubility characteristic of the complex was found to be very different from plain curcumin. The complex was found to be fairly soluble as compared to plain curcumin which was completely insoluble in water. Antimicrobial property of the complex was studied. All three complexes prepared in the ratio of 1:1, 1:2 and 1:3 were dissolved in methanol with a final concentration of 10mcg/mL. Page | 191   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

These solutions were aseptically incorporated in the wells in previously seeded Muller and Hinton agar plate. S aureus, B subtilis and E coli were used as test cultures.The complex in the ratio of 1:1 gave the largest zone of inhibition for all three test cultures. This indicates that the complex formed using 1:1 ratio of iron (III) and Curcumin is biologically more active than plain curcumin. Keywords: Curcumin, metal complex, Antimicrobial activity, Spectroscopy

Isolation and Characterization of Pigment Producing Halotolerant Bacteriaand Comparative Analysis of Enzyme Involved In Biosynthesis of Pigments S. Prathiba, Dr. G. Jayaraman School of Bio Sciences and Technology, VIT, University, Vellore -632014.

Sea water contains diverse microbial population. Microbial diversity arises due to the evolutionary changes in the microbes in response to stressful conditions like hyper salinity, low oxygen concentration, high or low temperatures, low nutrient availability, and presence of toxic compounds. High salinity led to the evolution of halobacteria. As an adaptive strategy, these organisms produce several metabolites (Enzymes, pigments, siderophores etc.) for their survival. Also, in industries, there is raising imposition for the use of metabolites which can exhibit activity in a wide range of salt concentration. The microbial pigments can be exploited, as they offer significant advantages like use in therapeutics/diagnostics, easy accessibility, GRAS (Generally Recognized As Safe) and it can be produced in short time. These microbial pigments have the potential to be used in pharmaceutical industries, food, cosmetics and textile industries. In the present study, soil samples were collected from Marakkanamsaltern, Page | 192   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

Tamil Nadu. A total of 6 pigment producing strains were isolated. Taxonomic characterization revealed that two of these strains were Bacillus sp. and four of them were Halobacillus sp. To start with,the pigments from Planoccoccus maritimusVITP21 were purified using preparative TLC and were partially characterized byconventional spectroscopic methods. The results revealed that the pigment could possibly be a carotenoid. As phytoene synthase is one of the important enzyme in the biosynthesis of this class of pigments, a comparative analysis of the aminoacid sequence and three dimensional structure of phytoene synthase was performed.

Changes in metabolites of floral scent volatiles emission and expressions of candidate genes in Polianthes tuberosa under daynight influence Saborni Maiti, Adinpunya Mitra Natural Product Biotechnology Group, Agricultural and Food Engineering Department, Indian Institute of Technology Kharagpur, Kharagpur - 721 302, West Bengal

Floral scent emission, in addition to colour, shape, surface morphology, and nectar guides leaves important cues to pollinators to assure reproduction. Common garden flower tuberose, Polianthes tuberosa L., is a tuberous monocot of tropical countries. It is popular for its long stick-like inflorescence bearing white flowers of intense fragrance which is frequently used as indoor cut-flower. To detect the chemical composition of emitted scent volatiles of tuberose, floral head-space technique was used for volatiles trapping for subsequent GC-MS analysis. Three major group of volatiles were identified namely, phenylpropanoids, terpenoids and fatty acid esters. Emission pattern was found clearly rhythmic and nocturnal under Page | 193   

normal

day-night

influence.

Two

candidate

genes

 

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of

3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

phenylpropanoids pathway and terpenoid-MEP pathway were isolated using degenerate primers and subsequent enzyme assays were carried out. Semi-quantitative expression study revealed concerted expression of these two key genes along with volatile emission.

Refractance Window Drying -capturing the essence of nature Sadhana Sharma Department of Food Engineering National Institute of Food Technology Entrepreneurship and Management (NIFTEM) Sonepat, Haryana, India [email protected]

Conventional drying methods, although effective are not highly efficient in retaining the quality and sensory attributes of the food product. These drying techniques are harsh, in terms of the severe heat exposure and long drying time required to dry the food. Some new techniques such as FreezeDrying and Spray-Drying are however comparatively milder, but they are commercially quite expensive. For encountering these limitations, a new method of drying, known as Refractance Window Drying (RW) came into existence.

RW is a low temperature drying system, which operates at

atmospheric condition. Thus this drying method is unique as there is minimum exposure of the food to damaging heat. As a result, the inherent sensory qualities which impart indispensable characteristics of a fresh food such as colour and taste are not lost or altered in any way. The secret behind this technology is the concept of Self-Limiting Infrared Window, which offers a “window” for the passage of infrared radiations generated as a result of the water being heated in the food. In the present study, various drying techniques were compared in terms of their effect on retaining nutrition and sensory qualities of tomatoes for the production of tomato Page | 194   

 

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3rd International Conference on Biotechnology and Bioinformatics (ICBB-2016), Pune, India

 

powder. It was found that RW system proved to be an efficient and suitable unit in terms of time, energy and cost for dehydrating moist material such as tomato paste. However much work is needed before a full scale commercial plant can be successfully actualized. Keywords: Convective Drying, Refractance Window Drying, Spray Drying, Freeze-Drying, tomato

Evaluation of anti-inflammatory and Insulin sensitising potential of Flaxseed oil in Diet induced obese model. Samina Bashir, Yadhu Sharma and Farah Khan* Department of Biochemistry, Hamdard University, New Delhi, INDIA [email protected], [email protected]

In this study, we investigated the effects of the Flax seed oil (linum usitatissimum) (FXO), on high fat diet–induced obesity and insulin resistance and evaluated its immunomodulatory effects after oral gavage in various concentrations (4, 8 and 16mg/kg body wt.) for 4 weeks. FXO was characterized using Gas chromatography – Mass spectrometry. Mice fed a high-fat diet (HFD, 60% energy as fat) and supplemented with dietary FXO (4 mg/kg body weight) reduced adiposity index and serum glucose levels (p < 0.05) and triglycerides (8 and 16 mg/kg body weight) compared to HFD obese model. All the groups showed an improvement in insulin sensitisation in comparison to HFD mice. A co-culture of 3T3-L1 adipocytes and RAW 264.7 macrophages was carried out to mimic the obese adipose tissue environment and a dose dependant shift in cytokine profile towards antiinflammatory (IL-4) state was observed with a decrease in (p < 0.05) proinflammatory ones (TNF-α). For immunomodulatory studies the groups showed a dose dependant increase (p