Dr. Savas G. Anastassiadis

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Jun 2, 2005 - Thermo Electron Company (Boston, USA). ... Received two recognition bonus-awards for outstanding ... I received the Nomination Award as International Scientist of the Year ... International Biographical Centre in Cambridge (Nicholas S. Law, ..... FINAL LIST OF POSTERS - BIOCHEMICAL ENGINEERING ...
Résumé of Dr. Savas G. Anastassiadis

Pythia Institute of Biotechnology (Owner - Director) Tel. 0035974681575, Mobile 00359895761696

Research in Biotechnology Co. Vat#. 108851559 Avgi Sohos, 57002 Thessaloniki, Greece, 00359895761696 (BG) Assistant Professor at Democritus University of Thrace Phone/Fax: +30 (2395) 051470, Phone: +30-23950-51324 Mobile: +30-6973-801395, +30-6972227424, 6983-75287 www.pythia-biotechnology.com Vat# BG202174635 E-mail: [email protected], [email protected] http://www.researchgate.net/profile/Savas_Anastassiadis/

Education and Scientific Experience 10/2000 to 09/2014 (Lecturer, C1 Professor, at the present Assistant C2 Professor) - Research and Teaching at the Environmental Engineering Department of Democritus University of Thrace (Xanthi, Greece).  I acted as a consultant for the Greek Government during the EU presidency.  I started a personal company nam.ed Research in Biotechnology Vat#. 108851559 (Greece).  Grounder and Owner of Pythia Institute of Biotechnology (Large buildings of former Preliminary School, Avgi, Thessaloniki, Greece) – Research and Development activities and pilot scale production (Novel conditioner EcoPlant).  Grounder of a subsidiary of Pythia Institute of Biotechnology (Sandanski, Bulgaria, Vat# BG202174635) – Production company (in near future)  Research and Development cooperation and publishing with the Institute of Biochemistry and Physiology of Microorganisms of the Russian Academy of Sciences (Pushchino, Moscow Region, Russia).  Co-Instructor of Diploma and Ph.D. students as Visitor Professor at the Institute of Biochemistry and Physiology of Microorganisms of the Russian Academy of Sciences (Pushchino, Moscow Region, Russia). 01/2000 - 07/2000 Serving in the Greek Army (Poros and Navy Hospital of Salamina). 10/1998 - 10/1999 Postdoctoral Research associate at Tufts University, Biotechnol. Center (USA) 09/1996 – 12/1998 Senior researcher at Cargill and ThermoFibergen (ThermoElectron) (USA) 03/1994 - 08/1996 Postdoctoral Research Associate, Massachusetts Institute of Technology (MIT), Department of Chemical Engineering (USA). Academic advisor: Bayer Professor Dr. Gregory Stephanopoulos 01/1990 - 04/1994 Dr. Nat. Sc. (Ph.D. Thesis): Biotechnology Research Institute, Research Center Jülich (RCJ), Germany (former Nuclear Research Center Jülich, KFA, Germany) and University of Münster (Germany). Topic of the Ph.D. work: "Continuous Production of Citric acid and Gluconic acid with yeast and yeast-like Microorganisms" Academic Supervisors: Prof. Dr. Hans-Jürgen Rehm (University of Münster, Germany) Prof. Dr. Christian Wandrey (University of Bonn and KFA Jülich) Ph.D. thesis was financed by Haarmann & Reimer, a daughter company of the company Bayer, Leverkusen, Germany 1988 - 1989 Master Thesis (Diplomarbeit): "Microbial Events at the Soaking of Soybeans" Institute of Molecular Microbiology and Biotechnology (former Institute of Microbiology), University of Münster, Germany. Academic Supervisor: Prof. Dr. Hans-Jürgen Rehm 1982 - 1988 M.S. in Biology (Diploma), University of Münster, Germany Major: Microbiology Minor: Biochemistry and Molecular and General Biology 1981 - 1982 Münster Junior College, Münster, Germany 1

1973 - 1979 1967 - 1973

Sochos High School, Thessaloniki, Greece Preliminary School, Avgi and Thessaloniki, Greece

Research in Industry 09/1996 – 07/1997

Work on contract basis for an agency (on assignment, Lab Support Co.), including 4 months in a laboratory in the industry (CARGILL, Minneapolis, USA). The project focused on the continuous production of citric acid by yeast strains of Yarrowia lipolytica (ATCC strains) in chemostat. According to biotechnology books and bibliography, it would have been very difficult to develop an economic commercial continuous production of citric acid. The obtained results of up to 190 g/l of continuously produced citric acids surpass the production of current industrial batch and fed-batch fermentations by Aspergillus niger. Continuous fermentation ran for more than 40 days. 02/1997 - 12/1998 Work as scientific consultant and researcher at Thermo Fibergen Inc., a Thermo Electron Company (Boston, USA). I organized and set up the laboratory for fermentation and microbiological research and development. I tested various renewable carbon sources as substrate for microbial growth and production and I developed and optimized new fermentation processes for the continuous production of organic acids by new microorganisms, which have very important applications for environmental and other industrial purposes (biodegradable plastics) More than 110 g/l of Llactic acid have been produced continuously with a yield of about 90% at very low residence times. Volumetric productivities of up to 16 g/(l h) and 140 g/l of lactic acid have been achieved. Furthermore, I developed and optimized new fermentation processes for the continuous production of a novel ecological noncorrosive biodegradable deicer. The continuous process was running more than nine months without any problems. Compared with existing industrial processes, the new developed continuous processes have many advantages and are very promising for an industrial production. They base on free growing cells in chemostat and no biomass retention is necessary. Biomass retention can increase the volumetric productivity. Cheap carbon sources, such as molasses and glucose derived from paper sludge have been used successfully (patent application). Traditional fermentations for the production of lactic acid use homofermentative lactic acid bacteria, which require complex media and are sensitive to high osmotic pressures and stress conditions. Existing industrial fermentation processes for organic acids are generally batch or fed batch, because a stable continuous process is very complicated and difficult to develop. Furthermore, I did preliminary theoretical consulting work for the fermentation of recombinant proteins. During my studies at the University of Münster (Germany), I also had visited many special laboratory courses in molecular biology, immunology, Genetics, Protozoa, Biochemistry etc. Because of my longtime experiences in research and teaching, I can also start working successfully with other scientists in new research areas, such as Molecular biology, Genetics, and Fermentation of cell cultures and recombinant proteins. 2003-Today Research and Development and production at Pythia Institute of Biotechnology Citric acid production by yeasts from glycerol and other carbon sources Gluconic acid production by yeast-like fungi Microbial production of cellulose from atmospheric CO2 Production and distribution of a novel soil conditioner named EcoPlant L-Lysine fermentation Bioethanol production

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Special Recognition and achievements        

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Received two recognition bonus-awards for outstanding achievements and innovative activities (patenting) from Nuclear Research Center Jülich (KFA) in Germany (6 patents, Ph.D. thesis). Financing of my Ph.D. thesis at the RCJ (KFA) by Haarmann and Reimer, a Bayer co. (1990-1994). Postdoctoral Research Associate at MIT and Tufts University (USA). Inclusion of biographical profile in 2006-2007 Edition of Who’s Who in Science and Engineering because of the reference value of outstanding achievements (chronicle of lives and career of the world’s most accomplished scientists and engineers). Cooperation and affiliation with the Institute of Biochemistry and Physiology of Microorganisms and with the Institute of theoretical and experimental biophysics of the Russian Academy of Sciences (Publishing, Research and Development) (2005 - present) Consulting for the Greek Government in the area of biotechnology (Future developments and possibilities) during the Greek European Presidency (2002-2003). I was elected and received a grant (44000 Euro) for innovative technologies (patents) from the Greek General Secretary of Research and Technology (G.G.E.T.), which was accomplished successfully (2003-2006). Invitation from the Russian Academy of Sciences for participating with POSTER PRESENTATION at the AT THE FOURTH CONGRESS OF RUSSIAN Y.A. OVCHINNIKOV SOCIETY OF BIOTECHNOLOGISTS, Pushchino Scientific Centre, Russia (December 6-7, 2006). Member of the RUSSIAN Y.A. OVCHINNIKOV SOCIETY OF BIOTECHNOLOGISTS Positive comments from leading International Journals and scientists Reviewer of significant research and review articles at leading International Journals Member of the Editorial Advisory Board in the Journal Recent Patents on Biotechnology (Bentham Science Publisher, Journals with impact factor 4.7-5.762). Monography titled “L-lysine fermentation”. Recent Patents on Biotechnology, Vol. 1, issue 1, p. 11-24 (Review article). Request from Bentham Science Publisher (a leading international publisher of high quality scientific journals and books; Impact Factor 4.8-5.762). Anastassiadis Savas and Morgunov Igor G. (2007). Gluconic acid fermentation. Recent Patents on Biotechnology, Vol. 1, issue 2. Bentham Science Publisher (Review article) On January 12, 2007, I received the Nomination Award as International Scientist of the Year 2007 (confirmation of nomination on 2nd November 2007). It is nominated to few illustrious individuals whose achievements and leadership stand out in the International Scientific Community as decreed by the Research and Advisory Board sitting at the International Biographical Centre in Cambridge, England (Outstanding Scientists of the 21st Century and Living Science). On January 12, 2007, I received the Outstanding Scientists of the 21st Century Award from International Biographical Centre in Cambridge (IBC), England (will include Outstanding Scientists of the 21st Century Diploma and Medal). Chosen as candidate and seriously considered for inclusion in the special 25th Silver Edition of Who’s Who in the World 2007, scheduled to be published in November 2007 (selected). Compilation for inclusion in the upcoming 10th Anniversary Edition of Who's Who in Science and Engineering 2008-2009, scheduled to be published in December 2007. On February 22, 2007, I attended the International Conference “EU-Russia Co-operation: Priorities for Science & Technology 2007-2013” after invitation from Directorate of EU (president of biotechnology) and the STATE UNIVERSITY – HIGHER SCHOOL OF ECONOMICS, MOSCOW, RUSSIA. Co-Instructor of Diploma thesis and Ph.D. students as Visitor Professor at the Institute of Biochemistry and Physiology of Microorganisms of the Russian Academy of Sciences On March 2, 2007, I received the Nomination as LEADING SCIENTISTS of the WORLD 2007 from Awards Board of the IBC in Cambridge, England (will include Outstanding Scientists of the 21st Century Diploma and Medal). (Ratification on September 28th 2007) Named as a Member of the International Biographical Centre, Cambridge, England, IBC (Nicholas S. Law, Director General) Personal presentations on TV and Radio stations and articles have been written about my scientific career at several News Papers. 3

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On March 30, 2007, I received the selection to be included in the PRESTIGIOUS PUBLICATION OF “2000 OUTSTANDING INTELLECTUALS OF THE 21st CENTURY” of the International Biographical Centre in Cambridge (Nicholas S. Law, Director General) On 27th April 2007, I received the IBC’s 21st Century Award for Achievement for outstanding contribution from the International Biographical Centre in Cambridge, England On 25th May 2007, I received the IBC’s Archimedes Award for reaching such goals and being recognized for endeavours both nationally and internationally from the International Biographical Centre in Cambridge, England On 8th June 2007, I received the invitation of IBC to be automatically included in the Cambridge Blue Book of Foremost International Scientists from the International Biographical Centre in Cambridge, England On July 20, 2007, I received the Honor placket of Recognition for the contribution to the Science from the Pontian Association of Chorygi, Kilkis (Greece) On July 21, 2007, I received (at the 2nd International Conference of Biotechnology 2007 of Pythia Institut of Biotechnology) a certificate (placket) of official membership of the RUSSIAN Y.A. OVCHINNIKOV SOCIETY OF BIOTECHNOLOGISTS from the President, Dr. R. G. Vasilov On July 21, 2007, I received (at the 2nd International Conference of Biotechnology 2007 of Pythia Institute of Biotechnology) the Honor certificate (placket) of congratulations from the General Director of NTM HOLDING Co. (Russia) Mr. German E. Karachevkiy (Future joined business and cooperation; Biodiesel production from plant oils and Joined production of biotechnological bulk and fine chemicals) On July 31, 2007, I received an invitation to serve as Guest Editor and contribute a special topic issue to the exciting journal CURRENT PHARMACEUTICAL BIOTECHNOLOGY, in view of my eminent contributions in the field of pharmaceutical biotechnology, being as an authority in the field responsible for soliciting top class reviews from eminent scientist/researches whom I will invite. The reviews will care various important aspects of the related topic. On 7th December 2007, I received the Nomination of “Top 100 SCIENTISTS 2008” as a noted and eminent professional in the field of science, from the International Biographical Centre (IBC) in Cambridge, England and ratification of nomination. On 11th January 2008, Inclusion in the new and revolutionary Biographical Directory: Outstanding People of the 21st Century, from the IBC in Cambridge, England. On 11th January 2008, I have received an invitation for the selection of 2000 outstanding intellectuals of the 21st century. Member of the Society of Biochemistry and Molecular biology of Russia since 2008 (Moscow society branch). On 8th February 2008, I received the decision for inclusion in 2000 OUTSTANDING SCIENTISTS 2008/2009 (Publication in mid 2009), from the IBC in Cambridge, England. Consideration for inclusion in the upcoming 2009 Edition of Who's Who in the World®, which is scheduled for publication in November 2008. February 29th, Inclusion in the Dictionary of International Biography 2008 (IBC, England) (November 28, 2008) On 21st March 2008, I received the granting of the IBC’s 21st Century Award for Achievement On 25th April 2008, I received the Ratification of nomination as Leading Scientists of the World 2008 (IBC in Cambridge, England) On 25th April 2008, I received the nomination for the international accolade “International Professional of the Year 2008” On July 11 2008, I received the IBC award FOREMOST PROFESSIONALS of the WORLD 2008 On July 12, I received the award FOREMOST SCIENTISTS of the WORLD 2008 (IBC) On July 25 2008, I received the forwarding as one of the FOREMOST SCIENTISTS of the World 2008 On August 8, 2008, I received the invitation for inclusion in the 2000 OUTSTANDING INTELLECTUALS OF THE 21ST CENTURY On August 22 2008, I received the selection for the inclusion in the IBC 2000 Outstanding Scientists 2008/2009 4

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November 7, 2008: International Einstein Award for Scientific Achievement (IBC) November 28, 2008, 2011: 2000 Outstanding Intellectuals of the 21st Century – 2009-2010 and 2011 December 31, 2010, Inclusion in the Dictionary of International Biography 2011 (IBC, England) Nomination for “Top 100 Educators 2011” (IBC, England) The international Plato Award for Educational Achievement (7th October 2011), (IBC, England) Honor award of Recognition for the contribution to the Science from the Cultural Association of Avgi (Birth place), Thessaloniki (Greece)

References – Recommendations 1. Univ. Professor Dr. Hans-Jürgen Rehm Institute of Molecular Microbiology and Biotechnology Corrensstrasse 3 – 48149 Münster – Germany Tel. +49 (251) 83 – 39 820 Fax: +49 (251) 83 – 38 388 E-mail: [email protected] http://mibi.uni-muenster.de 2. Professor Dr. Christian Wandrey Director of Institute of Biotechnology 2 Research Center Jülich (Forschungszentrum Jülich GmbH; former Nuclear Research Center, KFA) Leo-Brandt-Strasse - D-52425 Jülich, Germany Tel. 011-49-2461-613118 Fax. 011-49-2461-613870 E-mail: [email protected] 3. Bayer Professor Dr. Gregory Stephanopoulos Department of Chemical Engineering, Massachusetts Institute of Technology (MIT) Room 56-469, Cambridge, MA 02139, USA Tel.: +1-617-253-4583 Fax: +1-617-253-3122, E-mail: [email protected] 4. Dr. Igor G. Morgunov Head of the laboratory of Aerobic Metabolism of Microorganisms, Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, 142290, Pushchino, Moscow Region, Russia). Tel. (095) 9257448 Fax. (095)9563370, Mobile: (007) -916-77-35-53-1, E-mail: [email protected]

Academic Teaching   

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Pythia Institute of Biotechnology (Owner) - Biotechnology and Microbiology - Research and Teaching (2003-present) Co-Instructor of Diploma thesis and Ph.D. Students as Visitor Professor at the Institute of Biochemistry and Physiology of Microorganisms of the Russian Academy of Sciences (2006-2007) (Dr. Igor G. Morgunov) Environmental Engineering Department of Democritus University of Thrace, (Xanthi, Greece) Teaching Environmental Microbiology, Ecology (class and laboratory), Biology (class and laboratory) and Biotechnology (for Ph.D. students) as Lecturer and Assistant Professor; From October 2000 (C1) to present (C2 Epicurean professor). Civil Engineering Department of Democritus University of Thrace, (Xanthi, Greece) – Teaching Ecology for Engineers (from Academic year 2010-2011) and exams until today Environmental Engineering Department of Democritus University of Thrace, (Xanthi, Greece) Instruction at the present of more than 20 diploma theses (since 2005) Tufts University (Boston, USA), Instructed students in laboratory (Biotechnology Center); 1999 Massachusetts Institute of Technology (MIT, Boston, USA), Department of Chemical Engineering; 3/1994 – 8/1996 - Supervised students in undergraduate research programs and for Bachelor Thesis and helped Ph.D. students in laboratory (Bayer Prof. Dr. Gregory Stephanopoulos)



Research Center Jülich, RCJ (KFA, former Nuclear Research Center Jülich), Institute for Biotechnology (Germany); 1990 – 1994 - Supervised Bachelor and Master Students



University of Bonn (Germany); 1990 – 1994 - Teaching Assistant for laboratory courses in biotechnology and technical chemistry (Prof. Dr. Christian Wandrey)



University of Münster (Germany); 1985 – 1989 - Helped to organize several microbiology courses in the Institute of Microbiology (Prof. Dr. Hans-Jürgen Rehm)

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Special Skills 

Extensive international experiences in Research and Development and teaching in Fermentation process technology and Industrial and Classical Microbiology



Biology, classical and industrial microbiology, biotechnology, chemical engineering, ecology



Selective isolation and screening of microorganisms for industrial applications and innovations



Classical mutagenesis, analytical methods



Fermentation technology, development, design and optimization of biotechnological processes



Development of continuous fermentation processes (chemostat) with and without biomass retention



Utilization of wild type and genetically engineered microorganisms in batch and continuous cultures



Cultivation and storage of microorganisms (bacteria, yeasts, fungi)



Extensive training in sterile and analytical techniques (enzymatic, GC, HPLC etc.)



Extensive fermentation experience with biomass retention in various types and scales of bioreactors



Fermentation media and operating parameter optimization in continuous and batch cultures



Additional qualification in Reaction Technique, Biotechnology and Chemical Engineering



Elucidation of the regulatory mechanisms of microbial biosynthesis, physiology and pathways



Technology transfer, cooperation with industrial partners and patenting



Academic instruction for undergraduate courses and Bachelor and Master Theses



Extensively experienced in laboratory set up, research organization and design, in starting new projects and in leading a research group



Teaching of microbiology, biology, ecology and biotechnology



Start up of a Research biotechnology company and of Pythia Institute of Biotechnology (Greece)



Starting industrial production of a novel superior fertilizer at Pythia Institute of Biotechnology (Greece) International cooperation – Research and Development – Teaching (Institute of Biochemistry and Physiology of Microorganisms of the Russian Academy of Sciences)

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Initiation of private biotechnology business (production of novel biological fertilizer)



Music compositions

Languages: English, German and Greek Hobbies and other interests   

Playing piano and keyboards, Music compositions (copyrighted music in USA and Greece), Text writer. Member of Associations (e.g. RUSSIAN Y.A. OVCHINNIKOV SOCIETY OF BIOTECHNOLOGISTS and Member of the Society of Biochemistry and Molecular biology of Russia since 2008, Moscow society branch etc.) Honorary member and scientific advisor of the Association of Biofarmers of Drama (Greece)

I. Patents, Conference presentations, Scientific works and Publications 1. ANASTASSIADIS S., AIVASIDIS A. and WANDREY CH., 1992; "Verfahren zur fermentativen Gewinnung von Gluconsäure und dafür geeignete Mikroorganismen", ("Process for the production of gluconic acid by fermentation and the microorganisms suitable for the process"). German Patent Application No. P 42 17 674.3-41 (09/09/1992) 2. ANASTASSIADIS S., AIVASIDIS A. and WANDREY CH., 1999. "Process for the production of gluconic acid with a strain of Aureobasidium pullulans (DE BARY) Arnaud. US Patent No. 5,962,286 (Oct. 5, 1999) 3. ANASTASSIADIS S., AIVASIDIS A. and WANDREY CH., 1993; "Fermentationsverfahren zur kontinuierlichen Citronensäuregewinnung". (“Process for the continuous production of citric acid by fermentation”). German patent No. P 43 07 517.7 (03/18/1993) 6

4. ANASTASSIADIS S., 1994. "Zymotiki methodos gia tin sinechi paragogi tou kitrikou oxeos”; (“Process for the continuous production of citric acid by fermentation”). Greek Patent No. 940100098 (24.02.1994) 5. ANASTASSIADIS S., AIVASIDIS A. and WANDREY CH., 1994. "Fermentationsverfahren zur kontinuierlichen Citronensäuregewinnung" (“Process for the continuous production of citric acid by fermentation”). Austrian Patent No. 473/94 (03/07/1994) 6. ANASTASSIADIS S., AIVASIDIS A. and WANDREY CH., 2001. "Process for the continuous production of citric acid by fermentation". US Patent No. 6,303,351 (October 16, 2001) 7. ANASTASSIADIS S., 1998. “Continuous fermentation of lactic acid” Final Report for US Patent application (at Thermo Fibergen Inc., a ThermoElectron Co., Boston, USA) 8. ANASTASSIADIS Savas (2007). Production and composition of an ecological biological plant antifreezing, growth stimulating and plant protecting fertilizer of microbial origin (Growth stimulating, protective, antifreezing fertilizer for trees, flowers and other plants). Greek Patent application (2007). Pythia Institute of Biotechnology (Avgi-Sohos, Thessaloniki, Greece)

II. International Conferences 1. ANASTASSIADIS S., AIVASIDIS A. AND WANDREY CH., 1992. "Continuous Gluconic Acid Fermentation with Yeast-Like Molds". 10th Dechema Meeting of Biotechnologists, June 1 to 3 1992, Karlsruhe , F.R.G (Poster). Biotechnology Conferences 5-VCH Verlagsgesellschaft 1992, 537-540 2. ANASTASSIADIS S., AIVASIDIS A., WANDREY CH. AND H.-J. REHM, 1992. "Continuous production of citric acid with yeasts and gluconic acid with yeast-like molds". VAAM-Tagung`93, Leipzig, Germany (Conference paper, presentation) 3. SAVAS ANASTASSIADIS, ALEXANDER AIVASIDIS (Department of Environmental Engineering, Democritus University of Thrace, GREECE) and CHRISTIAN WANDREY (Institute of Biotechnology of Research Center Jülich, GERMANY). Continuous citric acid fermentation by Candida oleophila. FINAL LIST OF POSTERS - BIOCHEMICAL ENGINEERING CONFERENCE - POSTER SESSION I - Monday, June 11, 2001 (USA) 4. Dr. Savas Anastassiadis (July 23, 2006) - Organization of the 1nd International Initiation ceremony and small Conference of Biotechnology of Pythia Institute of Biotechnology (Avgi Sohos, Thessaloniki, Greece) and oral presentation entitled: Future perspectives of Biotechnology 5. Dr. Savas Anastassiadis (December 6-7, 2006) – “Perspectives for microbial production of gluconic acid and L-lysine” POSTER PRESENTATION AT THE FOURTH CONGRESS OF RUSSIAN Y.A. OVCHINNIKOV SOCIETY OF BIOTECHNOLOGISTS (Invitation by the Russian Academy of Sciences) Pushchino Scientific Centre, Russia 6. On February 22, 2007, I attended the International Conference "EU-Russia Co-operation: Priorities for Science & Technology 2007-2013 after invitation from Directorate of EU (president of biotechnology) and the STATE UNIVERSITY – HIGHER SCHOOL OF ECONOMICS, MOSCOW, RUSSIA. 7. Dr. Savas Anastassiadis (July 19-21, 2007) – I organized and carried out the 2nd International Conference of Biotechnology and Alternative Energies of Pythia Institute of Biotechnology (Avgi Sohos, Thessaloniki, Greece). Oral presentation entitled: Continuous production of organic acids (e.g. Citric acid, Gluconic acid, L-Lactic acid, Succinic acid etc.)” and novel plant protecting agent (Presence of Science, Politics, Industry and Arts (Exhibition)): Chairman: Dr. Igor G. Morgunov (RAS) 8. Dr. Savas Anastassiadis (July 19-21, 2007) - Poster presentation entitled: Production of a novel antifreezing, growth and production promoting and disease protecting fertilizer for plants and trees (Pythia Institute of Biotechnology). 2nd International Conference of Biotechnology and Alternative Energies of Pythia Institute of Biotechnology (Avgi Sohos, Thessaloniki, Greece)

III. Literature Reviews and Final Research projects and Reports in industry and academy 1. ANASTASIADIS S., 1989, “Tempe-Fermentation” (Soybean fermentation), Literature Review work in German, which was translated in English by the German Government (University of Münster, Germany), page 1-50 Academic supervisor: Prof. Dr. Hans-Jürgen Rehm 2. ANASTASSIADIS Savas and Stephanopoulos Gregory (1994-1996) Final Research Report for continuous L-lysine production by coryneform bacteria (MIT, USA) (100 g/l of L-lysine in chemostat), (Chemical Engineering Department, MIT, USA) Academic advisor: Prof. Dr. Gregory Stephanopoulos Autonomous Senior Scientist 7

3. ANASTASSIADIS Savas and Stephanopoulos Gregory (1994-1996) Final Research Report for the optimization of the effect of betaine on L-lysine fermentation by coryneform bacteria as a function of glucose concentration 4. ANASTASSIADIS Savas and Stephanopoulos Gregory (1994-1996) Final Research Report on the continuous production of citric acid by yeasts (230-250 g/l citric acid in continuous repeated batch in 48-54 h and 120 g/l in repeated batch cultures) Autonomous Senior Scientist 5. ANASTASSIADIS Savas (1997) Final Research Report on the continuous production of citric acid by yeasts (200 g/l citric acid in chemostat) (CARGILL, Minneapolis, USA). The project focused on the continuous production of citric acid by yeast strains of Yarrowia lipolytica (ATCC strains) in chemostat. Autonomous Senior Scientist 6. ANASTASSIADIS S., 1998, “Continuous fermentation of lactic acid” Final Research Report for US Patent application (at Thermo Fibergen Inc., a ThermoElectron Co., Boston, USA) (150 g/l Llactic acid) Autonomous Senior Scientist 7. ANASTASSIADIS S., 1998, “Continuous production of a biodegradable non-corrosive deicer composition” Final Research Report for US Patent application (at Thermo Fibergen Inc., a ThermoElectron Co., Boston, USA) (150 g/l L-lactic acid) Autonomous Senior Scientist 8. ANASTASSIADIS SAVAS, 2000-2001. Finalization of a project regarding to the acceleration of wine fermentation and writing of a final report in German Language at the Environmental Engineering of Democritus University of Thrace (Greece). The wine fermentation is completed within 24-30 hours. Autonomous Senior Scientist Academic advisor: Prof. Dr. Alexander Aivasidis

IV. International Books 1.

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Savas Anastassiadis (Pythia Institute of Biotechnology), Igor G. Morgunov and Tatiana Finogenova (Russian Academy of Sciences) (2006-2007). Industrial Microbiology and Biotechnology. Review book (Under preparation on Request from an international publisher) RESEARCH SIGNPOST accepted Anastassiadis Savas, Kamzolova Svetlana V., Morgunov Igor G. and Rehm Hans-Jürgen (2007). Comparative study of the effect of iron on Citrate-producing yeast growing on different substrates. Book Chapter at “Communicating Current Research and Educational Topics and

Trends in Applied Microbiology, FORMATEX Microbiology series No. 1, Vol. 1 (Book)” Industrial and Food Microbiology, p. 308-314, A. Mendez-Vilas (Ed.). ISBN-13: 978-13:978-84-611-9422-3

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S.V. Kamzolova, S.G. Anastassiadis2, A.R. Fatyhkova, N.P. Golovchenko, and I.G. Morgunov (2010). Strain and process development for citric acid. production from glycerol-containing waste of biodiesel manufacture. FORMATEX Microbiology Book Series No. 2, Vol. 2 (Book), "Current Research, Technology and Education Topics in Applied Microbiology and Microbial Biotechnology" MICROBIOLOGY BOOK SERIES - Number 2. A. Mendez-Villas (Ed.), http://www.formatex.info/microbiology2/1020-1028.pdf Anastassiadis Savas (Pythia Institute of Biotechnology) (2007). Industrial Microbiology. Book Chapter at Clinical Neuropharmacology (Κλινική Νευροφαρμακολογία - Κεφάλαιο: Βιομηχανική Μικροβιολογία) (Author: Evangelos Dimitriadis, Aristotelis University of Thessaloniki, Greece), 1st edition, Publisher Siokis Anastassiadis Savas (Pythia Institute of Biotechnology) (2007). Biotechnology. Book Chapter at Clinical Neuropharmacology (Κλινική Νευροφαρμακολογία - Κεφάλαιο: Βιοτεχνολογία) (Author: Evangelos Dimitriadis, Aristotelis University of Thessaloniki, Greece), 1st edition, Publisher Siokis On July 31, 2007, I received an invitation to serve as Guest Editor and contribute a special topic issue to the exciting journal CURRENT PHARMACEUTICAL BIOTECHNOLOGY, in view of my eminent contributions in the field of pharmaceutical biotechnology, being as an authority in the field responsible for soliciting top class reviews from eminent scientist/researches whom I will invite. The reviews will care various important aspects of the related topic. Invitation to edit a an encyclopedia (5 books) from Bentham Sciences Publisher

V. International publications in Journals with reviewers and analytical publications 1.

ANASTASSIADIS S., AIVASIDIS A. and WANDREY CH. (2002). Citric acid production by Candida oleophila under intracellular nitrogen limitation. Applied Microbiology and Biotechnology 60 (1-2): 81-87 (DOI 10.1007/s00253-002-1098-1, Band 60 (1-2), October 21, 2002 Impact factor: 3.69 8

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ANASTASSIADIS S., AIVASIDIS A. and WANDREY CH. (2003). Continuous gluconic acid production by isolated yeast-like mould strains of Aureobasidium pullulans. Applied Microbiology and Biotechnology 61(2): 110-117 (s00253-002-1180-8) Impact factor: 3.69 Anastassiadis, S., Wandrey, C. and Rehm, H.-J. 2005 (July). Continuous citric acid fermentation by Candida oleophila under nitrogen limitation at constant C/N ratio. World Journal of Microbiology and Biotechnology, Volume 21, Number 5, p. 695-705, ISSN: 0959-3993 (Paper) 1573-0972 (Online), DOI: 10.1007/s11274-004-3850-4 Impact fector: 1.26 Anastassiadis, S., Aivasidis, A., Wandrey, C. and Rehm, H.-J. 2005 (June 2). Process optimization for continuous gluconic acid fermentation by isolated yeast-like strains of Aureobasidium pullulans. Biotechnology & Bioengineering, Volume 91, Issue 4, Pages 494-501; PMID: 15937884, Published Online: 2 Jun 2005 Impact factor: 3.65 Anastassiadis, S. and Rehm, H.-J. 2005. Continuous citric acid secretion by a high specific pH dependent active transport system in yeast Candida oleophila ATCC 20177. Electronic Journal of Biotechnology. Vol. 8 No. 2, Issue of August 15, 2005 Impact factor: 0.83 Anastassiadis, S. and Rehm, H.-J. 2005. Continuous citric acid secretion by a high specific pH dependent active transport system in yeast Candida oleophila ATCC 20177. Electronic Journal of Biotechnology. Vol. 8 No. 2, Issue of August 15, 2005 BIP Article Anastassiadis S. Candida citric acid fermentation. Industrial Bioprocessing, Volume 27, Issue 9, September 2005, Pages 6-7 Anastassiadis, S. and Rehm, H.-J. 2006. Citric acid production from glucose by yeast Candida oleophila ATCC 20177 under batch, continuous and repeated batch cultivation. Electronic Journal of Biotechnology, Vol. 9 No. 1, Issue of January 15, 2006 Impact factor: 0.83 Anastassiadis, S. and Rehm, H.-J. 2006. Citric acid production from glucose by yeast Candida oleophila ATCC 20177 under batch, continuous and repeated batch cultivation. Electronic Journal of Biotechnology, Vol. 9 No. 1, Issue of January 15, 2006 BIP Article Anastassiadis, S. and Rehm, H.-J. 2006. Oxygen and temperature effect on continuous citric acid secretion in Candida oleophila. Electronic Journal of Biotechnology, Vol. 9 No. 4, Issue of July 15, 2006 Impact factor: 0.83 Anastassiadis, S. and Rehm, H.-J. 2006. Oxygen and temperature effect on continuous citric acid secretion in Candida oleophila. Electronic Journal of Biotechnology, Vol. 9 No. 4, Issue of July 15, 2006 BIP Article Anastassiadis, S. and Rehm, H.-J. 2006. Continuous gluconic acid production by Aureobasidium pullulans with and without biomass retention. Electronic Journal of Biotechnology, Vol. 9 No. 5, Issue of October 15, 2006 Impact factor: 0.83 Anastassiadis, S. and Rehm, H.-J. 2006. Continuous gluconic acid production by Aureobasidium pullulans with and without biomass retention. Electronic Journal of Biotechnology, Vol. 9 No. 5, Issue of October 15, 2006 BIP- Article Anastassiadis, S. and Rehm, H.-J. 2006. Continuous gluconic acid production in cascading of fermenters. Applied Microbiology and Biotechnology (accepted on 14 May 2006, DOI 10.1007/s00253-006-0499-y). Vol. 73(3), p. 541-8. (Epub 2006 Aug 12) Impact factor: 3.69 Anastassiadis Savas (2007). L-lysine fermentation. Recent Patents on Biotechnology, Vol. 1, issue 1, p. 11-24 (Review article). Request from Bentham Science (a leading international publisher of high quality scientific journals and books; Impact Factor 4.8-5.762). (ISSN: 1872-2083 - Volume 1, Issue 1, 2007, p. 11-24)

16. Anastassiadis Savas and Morgunov Igor G. (2007). Gluconic acid production. Recent Patents on Biotechnology, Vol. 1, Issue 2. Bentham Science Impact Factor 4.8-5.762 17. Anastassiadis Savas, Kamzolova Svetlana V., Morgunov Igor G. and Rehm Hans-Jürgen (2007). Comparative study of the effect of iron on Citrate-producing yeast growing on different substrates. Book Chapter at “Communicating Current Research and Educational Topics and

Trends in Applied Microbiology, FORMATEX Microbiology series No. 1, Vol. 1 (Book)” Industrial and Food Microbiology, p. 308-314, A. Mendez-Vilas (Ed.). ISBN-13: 978-13:978-84-611-9422-3

18. Anastassiadis S. (1993). Institute of Biotechnology 2, Research Center Jülich (KFA) in Germany. "(Application No. 7; "Determination of organic acids, especially citric and isocitric acid, in fermentation solutions and fruit juices", Published in: Chromatography methods 1993, (Macherey-Nagel GmbH + Co. KG Düren, Postfach 101352, D-52348 Düren, Germany). 19. Trends in Food Science & Technology 12 (2001) 349–350. New Patents 20. Anastassiadis S. 2007. L-Lysine Fermentation - Patent Survey Review Article. Feedinfo News Service Scientific Reviews. November 2007. Available from URL: http://www.feedinfo.com, Request from Feedinfo News Service 21. Anastassiadis Savas, Kamzolova S.V., Morgunov Igor G. and Finogenova Tatiana (2008). Citric acid production patent review. Recent Patents on Biotechnology 2008, Vol. 2(2), 107-123 9

22.

23.

24.

25.

26.

(Review article in the patent area of Biotechnology; Request from Recent Patents on Biotechnology, Bentham Science, Impact Factor 4.8-5.762), DOI: 10.2174/187220808784619757, http://www.eurekaselect.com/92714/article Savas Anastassiadis, Svetlana V. Kamzolova, Igor G. Morgunov, Hans-Jurgen Rehm (2008). Iron effect on gluconic acid production by Aureobasidium pullulans. The Open Biotechnology Journal, Volume 2 (ISSN: 1874-0707). pp.195-201 (7) S.V. Kamzolova, S.G. Anastassiadis2, A.R. Fatyhkova, N.P. Golovchenko, and I.G. Morgunov (2010). Strain and process development for citric acid. production from glycerol-containing waste of biodiesel manufacture Microscopy Book Series titled "Current Research, Technology and Education Topics in Applied Microbiology and Microbial Biotechnology" A. Mendez-Villas (Ed.), http://www.formatex.info/microbiology2/1020-1028.pdf Svetlana V. Kamzolova1*, Alina R. Fatykhova1, Emiliya G. Dedyukhina1, Savas G. Anastassiadis2, Nikolay P. Golovchenko1 and Igor G. Morgunov1. Citric Acid Production by Yeast Grown on Glycerol-Containing Waste from Biodiesel Industry. Food Technol. Biotechnol. 49(1) 65–74 (2011) Impact factor: 0.98 Kamzolova Svetlana V., Anastassiadis Savas, Morgunov Igor G and Finogenova T. (2008) Citric acid production (2007) Mikrobiologia (Russian), (submitted) E.A. Dimitriadis, S. Anastassiadis, Α. Katranias, D. Mixmizos. SOURCES OF

INTERINDIVIDUAL VARIATION IN RED BLOOD CELL CHOLINESTERASE ACTIVITY (Laboratory of Neuropathology, A’ Neurologic Clinic, AHEPA, A. U. Th) (under submission) 27. Savas Anastassiadis (2008). Succinic acid production patent review. Recent Patents on Biotechnology. Accepted (under preparation) 28. 6 Review articles under preparation (2014)

VI. Publications under preparation and under submission 1. Anastassiadis Savas and Kamzolova Svetlana V., Citric acid fermentation by yeasts. Review (under preparation) 2. Loupa G. and Anastasiadis S. 2006. INDOOR / outdoor PARTICULATE matter and d bioaerosol levels IN NATURALLY VENTILATED OFFICES (under submission) 3. Anastassiadis, S. and Rehm, H.-J. 2005. Production of gluconic acid under batch, fed batch and continuous cultivation (under preparation) 4. Anastassiadis, S. and Rehm, H.-J. 2005. The phenomena of oscillations in citric acid production by yeast Candida oleophila ATCC 20177. Ovchinnikov bulletin of biotechnology and physical and chemical biology (English and Russian) (under submission) 5. Savas Anastassiadis1*, Svetlana V. Kamzolova2 and Igor G. Morgunov2 (2008). Manganese effect on continuous gluconic acid production by yeast-like mold Aureobasidium pullulans (under preparation and submission, Applied Microbiology and Biotechnology) 6. Anastassiadis Savas, Svetlana V. Kamzolova and Morgunov G. Igor (2007). Succinic acid production by various microorganisms, e.g. bacteria and yeasts (under preparation) (Pythia Institute of Biotechnology, Democritus University of Thrace and Institute of Biochemistry and Physiology of Microorganisms of the Russian Academy of Sciences) 7. ANASTASSIADIS et al. (2007) Several Publications under preparation (Continuous fermentation of citric acid, gluconic acid, L-lysine, L-lactic acid, succinic acid etc.) 8. Kamzolova Svetlana V., Morgunov Igor G., Anastassiadis Savas, and Finogenova Tatiana (2007). Biochemical aspects for citric acid production in yeasts (Review article) 9. Anastassiadis Savas, Svetlana V. Kamzolova and Morgunov G. Igor (2007). Pharmaceutical applications of organic acids (Review article) Under preparation

VIII. Analytical chromatography applications (HPLC methods) 1. ANASTASSIADIS S. (1993). Institute of Biotechnology 2, Research Center Jülich (KFA) in Germany. "(Application No. 7; "Determination of organic acids, especially citric and isocitric acid, in fermentation solutions and fruit juices", Published in: Chromatography methods 1993, (Macherey-Nagel GmbH + Co. KG Düren, Postfach 101352, D-52348 Düren, Germany). 2. ANASTASSIADIS S. (1994), Ph.D. thesis, Institute of Biotechnology 2, Research Center Jülich (KFA) in Germany New HPLC application for selective sugar analysis and separation of chemically related organic acids 3. ANASTASSIADIS S. (1996-7). ThermoFibergen Inc. (ThermoElectron Co.), Boston, USA. New HPLC application for selective separation of organic acids, in especially lactic acid, acetic acid, succinic acid etc. and sugar analysis (separation of glucose and gluconic acid) (Senior scientist) 10

Research and Teaching Interests I. Classical and Industrial Microbiology and Biodiversity The appearance of carbohydrate-based microbial consumer products is a recent phenomenon. The vast majority of these products, including commodity, specialty and fine chemicals (e.g. carbonic acids such as citric, gluconic, lactic and itaconic acid, ethanol, pharmaceuticals, vitamins, recombinant proteins, enzymes, precursors for polymers, biodegradable plastics etc.) clearly demonstrate the tremendous economic potential for future biotechnological applications. The contribution of microorganisms to biotechnology is essential. The full potential of microorganisms to industrial applications is still not quantified. The isolation and screening of wild microorganisms and mutant strains, including bacteria, yeasts and fungi can be performed to assess their potential. Biotechnology will have a critical impact in the world’s economy and in solving future world problems encompassing environment, agriculture, health and energy. In order to quantify the full potential of microbial organisms, I am interested in:  The isolation, characterization and use of microbial wild and mutant strains of industrial importance  The production of important bulk, fine and specialty chemicals economically II. Fermentation process development and optimization Important factors in improving competitiveness of biological vs. chemical systems are yield and productivity enhancement. Sophisticated optimization of environmental process variables can have a tremendous improvement on fermentation processes. This was clearly illustrated in my work on the continuous production of citric and gluconic acid, where improvements of 10- to 20-fold were achieved and in my work on the continuous fermentation of L-lysine, succinic acid, L-lactic acid and novel fertilizer. In addition, the developed continuous processes have many advantages over the traditional batch and fed-batch operations of the last 100 years. As a result, I am interested in:  Analyzing and optimizing existing fermentation processes and introducing new processes  Studying the specific use of various carbon sources, including cheap agricultural wastes to produce specialty products such as carbonic acids, amino acids, food additives, fine chemicals and pharmaceuticals  Quantifying the use of various feeding strategies in fed-batch fermentations to simulate continuous chemostat operation III. Elucidation of microbial physiology and regulation The understanding of microbial physiology and regulation plays an important role in redirecting intermediary cellular metabolism to desirable products. Enzyme regulation and product secretion can be influenced by environmental factors such as trace elements, growth factors as well as by physical parameters. For example, iron-limiting yeast cultures overproduced citric acid, while iron sufficient cultures primarily formed biomass, as a result of aconitase regulation. Due to the complexity of living systems (a typical bacterial cell includes more than 1000 enzymes) it is essential to identify critical aspects of metabolite overproduction. This requires an insightful and systematic approach. As a result, I am interested in studying the:  Clarification of interactions between enzyme reactions, pathways and product secretion  Use of auxiliary carbon sources to increase the productivity  Ways to achieve maximum microbial production capacity by manipulating the environmental parameters and physiology  Combination of continuous and batch fermentation to use optimal conditions from both types  Selective isolation of high-producing mutant strains Furthermore, I am interested in starting biotechnology business for the production of biotechnological products and technology transfer. IV. Teaching (Class and laboratory) Biology, Microbiology (Classical, Environmental and Industrial Microbiology), Ecology and Microbial Ecology and Biotechnology (Environmental microbiology, Fermentation technology and Process development, Isolation and screening of microorganisms etc.)

Current Research and Development (Academy and Pythia Institute of Biotechnology Research Center, Cooperation with the Institute of Microbiology of the RAS) 11

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Research and Teaching at the Environmental Engineering Department of Democritus University of Thrace (Xanthi, Greece) Instruction of several diploma theses (currently about 20 diploma thesis students) Completed a Greek-German research project regarding the acceleration of most fermentation using Genetic Algorithms (Computer program). Glucose was completely consumed and wine fermentation accomplished within 24-30 hours, compared with more than 6 days that is necessary for wild most. Fermentation technology (Process development and optimization) and Waste minimization during the industrial production of biotechnological products Citric acid production from various agro-industrial waste sources, Gluconic acid from glucose and glucose containing sources, Succinic acid production from various agro-industrial waste sources, Lactic acid production L-Lysine fermentation Product separation and purification (Classical calcium hydroxide/acid separation, Ion exchange chromatography) Use of agro-waste byproducts such as molasses, glycerol etc. Isolation of new important microorganisms from natural ecosystems Strain development by classical mutagenesis, selection and screening of industrial microorganisms

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Atmospheric indoor and outdoor microbial pollution (methodology for rapid determination) Anticancer activities (Understanding and searching for cancer treatment; Involvement of microorganisms; Changing metabolism etc.) Note: My Research Center, Pythia Institute of Biotechnology is well equipped fulfilling international standards and it possesses new industrially important technologies. It has cooperation with the Institute of Biochemistry and Physiology of Microorganisms of the Russian Academy of Sciences. It plans to organize special summer courses for Russian and EU students on classical and industrial microbiology, fermentation technology and process development. Research Associate at Tufts University (Boston, USA) Writing proposals for grants and research on the microbial production of wild and novel biopolymers by wild and mutant microbial strains (emulsan, pullulan, cellulose etc), as well as on genetics. Several interesting mutant microbial strains were developed. Postdoctoral Research at MIT (Boston, USA) The primary scope of this work was to study and understand the formation of amino acids in Corynebacteria. New processes were developed and optimized for the continuous production of Llysine. Regulatory and auxotrophic mutant strains of Corynebacterium glutamicum and Brevibacterium lactofermentum (ATCC strains) were used. I. Studied the effects of fermentation parameters, such as pH, dissolved oxygen, agitation, temperature and media composition on process yield and productivity. 65 g/l of L-lysine have been achieved and maintained for long period of at residence times of 25 to 27 hrs. This concentration corresponds to 2.6 g/(l h) volumetric productivity or 0.1 g/(g h) specific productivity. About 40 g/l of L-lysine have also been achieved at a residence time of 14 hours, which corresponds to a volumetric productivity of 2.9 g/l h. Process yields approached 45% and were maintained at over 30% g/g at steady state and 100% conversion. These results compare very favorably with industrial fed-batch processes employing high production strains. Continuous production has been maintained for more than 70 days. II. Isolated high lysine production strains from chemostat experiments, and characterized them in shake flask and chemostat cultures along with ATCC strains. Faster growing strains have been isolated that can reduce operation time in shake flasks by a factor of 4. III. Studied effects of osmoprotectans, such as glycine-betaine on growth and lysine production in batch cultures of Corynebacteria (12 l fermenter) at various initial glucose concentrations. Optimum betaine 12

concentrations were found to reduce significantly lag phases, especially at high glucose concentrations and also to improve lysine production. IV. Isolated yeast-like molds of Aureobasidium pullulans from wild flowers (Massachusetts) which produce gluconic acid and were studied in batch and chemostat experiments. More than 200 g/l of gluconic acid was obtained in batch experiments. V. Preliminary development of fermentation process for the continuous production of trehalose (Disaccharide, 200 Dollar/Kg) by Corynebacteria. Experimental results indicated a feasible large-scale process. Trehalose can be used in order to keep dried food fresh after rehydration and in pharmaceuticals. VI. Developed various new processes for the semicontinuous production of citric acid using Candida strains. Citric acid concentrations of 240 g/l have been obtained in fed-batch and repeated fed-batch operation (12 l fermenter) within 140 hrs using different feeding strategies. The process yield was more than 70% at 100% conversion of glucose. This operation was maintained for a period of more than 2 months. Maximum volumetric productivities between 5 to 6 g/(l h) and specific productivities between 0.1 to 0.15 g/(g h) were achieved. Numerous repeated batch fermentations were performed with different Candida strains from ATCC in 1 l fermenters. 110 g/l of citric acid were produced repeatedly batch-wise in 42-hrs total fermentation time.

Ph.D. Thesis The primary task of my Ph.D. Thesis was to isolate and characterize microbial systems suitable for the continuous production of carboxylic acids of low molecular weight. I. Isolated about 500 carbonic acid producing strains from natural ecosystems and compared them with existing citric acid producing yeast strains. The isolated yeast-like molds of Aureobasidium pullulans were found to be ideal for gluconic acid production. II. Οptimized both the growth media composition (iron, manganese, copper, zinc, magnesium, phosphate etc.) for the above strains, as well as the process conditions (pH, temperature, dissolved oxygen, pressure and CO2). Competition effects between growth and production were determined in continuous cultures. Process optimization resulted in 1000% improvement for citric and gluconic acid production in chemostat. Various types of bioreactors such as stirred and fluidized bed fermenters, single and series vessels with and without biomass retention by microfiltration and immobilization were evaluated for their applicability in these processes. Furthermore, electrodialysis was tested for the separation of citric acid. This work resulted in two new continuous processes with high process yields and productivity, using free growing wild type cells. These new yeast-based continuous processes are highly advantageous compared to the traditional fungi-based processes of the last 100 years. Due to the industrial importance of these processes, numerous patent applications have been submitted (see enclosure). III. Achieved 150 g/l of citric acid in continuous culture at residence times of 80 to 100 hrs with Candida oleophila (Yarrowia lipolytica according to new classifications). The maximum process yield was 75% g/g. Maximum volumetric productivities of 2 g/l h or biomass-specific productivities of 0.1 g/g h have been determined. In repeated batch fermentations 170 g/l of citric acid have been obtained. IV. Overproduced 370 g/l of gluconic acid with isolated strains of Aureobasidium pullulans in continuous processes at residence times of 20 to 25 hrs with molar process yields of more than 95% (100% g/g). Maximum volumetric productivities of 20 g/l h or biomass-specific productivities of 10 g/g h were achieved. Even higher gluconic acid concentrations (500 g/l) have been attained in fed-batch mode. V. Illustrated for the first time that citrate secretion in yeast occurs by an energy consuming transport system, which is specific for citrate and not isocitrate. Developed a working hypothesis about the overproduction of citric acid in yeast that is linked with the proposed transport system and the yeast’s physiology. VI. Developed new HPLC methodologies to separate and quantify citric and isocitric acid or glucose and gluconic acid.

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Master Thesis (Diplomarbeit) Completed many curriculum courses in classical and applied Microbiology, Biology and special courses in Biochemistry, Molecular Biology, Genetics and Immunology. Composed a literature review on the solid-state fermentation of soybeans (Tempe) for a GermanIndonesian scientific cooperation. The objective of my Master Thesis was to study the microbial events that take place during the soaking of soybeans. Briefly, these include an initial acidification step followed by a solid-state fermentation using soybeans (Tempe or Tempeh). Tempe is a broadly used vitamin-rich meatanalogous food in Indonesia. I. Isolated hundreds of bacteria strains from various kinds of soybeans (Indonesia, China, USA and Brazil) and from soaking water (from Indonesia). These strains mostly included lactic acid bacteria, and were lyophilized for long-term studies. II. Screened the isolated strains for the acidification of soybeans during the soaking process. Some isolated strains acidified the soaking water to pH 4.5 within 4 to 5 hrs, compared with the traditional Indonesian soaking process that requires 24 hrs. III. Analyzed spontaneous fermentation byproducts resulting from the soaking of soybeans such as sugars, proteins and vitamins and spontaneous acidification. IV. Screened the isolated strains for their ability to produce various essential vitamins e.g. B1, B6, B12 in order to increase the nutritional value of Tempe. Vitamins were analyzed using auxotrophic mutant strains.

Current Research activities at the Environmental Engineering Department of Democritus University of Thrace  

Microbiological pollution of Indoor and Outdoor atmospheric environments Process development, process optimization and Minimization of waste materials during the industrial production of biotechnological bulk products, e.g. Citric acid (250 g/l), Gluconic acid (430 g/l in chemostat, 504 g/l in fed batch), Lactic acid (150 g/l), L-lysine (90 g/l), Pullulan, Biodiesel, Bioethanol, Hydrogen etc.  Biotechnological production of bulk chemicals using byproducts of industry, e.g. glycerol, plant and animal oils etc.  Biotechnological production of bulk chemical using renewable carbon sources such as plant and animal oils etc. Current Research activities at the Biotechnology Research Center (Avgi/Sohos-Thessaloniki)  Continuous production of citric acid by yeasts (200-250 g/l)  Continuous citric acid production of gluconic cid by yeast-like molds (430 g/l)  Continuous production of lactic acid (150 g/l)  Continuous production of succinic acid by bacteria (more than 40 g/l)  Continuous production of an ecological non-corrosive biodegradable deicer (140 g/l)  Continuous production of pullulan by yeast-like molds Aureobasidium pullulans  Continuous production of L-lysine by Corynebacterium strains  I developed at Pythia Institute of Biotechnology a new process for the production of a novel microbial fermentation product mixture (biodegradable, ecological, nontoxic etc.), which protects the plants and trees from freezing and enhances their growth and production. It also functions against plant pathogenic fungi, bacteria and insects; I started the commercial production of the new product.

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Reviewer of research articles at leading international journals 1. Biotechnology & Bioengineering 2. Chemical Papers Prof. Milan Polakovic, Editor-in-Chief, Chemical Papers E-mail: [email protected] 3. International Journal of Food Properties (IJFP) Shyam S. Sablani, Ph.D., Associate Professor of Food & Bioprocess Engineering Sultan Qaboos University, P.O. Box-34, Al-Khod 123, Muscat, Oman Tel: +968 2414 1289; Fax: +968 244 13 418 E-mail: [email protected] Email: [email protected] 4. Applied Biochemistry and Biotechnology E-mail: [email protected] 5. Current Microbiology (Springer) Erko Stackebrandt, Current Microbiology Editorial Office, E-mail: [email protected] Dr. Tom Edrington, Associate Editor, Current Microbiology, E-mail: [email protected] Dr. Ashvini Chauhan, Associate Editor, Current Microbiology, E-Mail: [email protected] 6. Recent Patents on Biotechnology (BIOT) Bentham Science Publishers Ltd. (Impact Factor 4.8-5.762) Ms. Tayyaba Zaidi, Manager Publications, E-mail: [email protected] Dr. Khurshid Zaman, Editor Fax: (+92-21)-4960510, Email: [email protected], URL: www.bentham.org/biot 7. Journal Open Biotechnology Bentham Science Publishers Ltd. (Impact Factor 4.8-5.762) Kazim Ali Baig Manager Publications ambreen Shakeel , [email protected], [email protected], [email protected] 8. Current Pharmaceutical Biotechnology (Bentham Science Publishers Ltd.) On July 31, 2007, I received an invitation to serve as Guest Editor and contribute a special topic issue to the exciting journal CURRENT PHARMACEUTICAL BIOTECHNOLOGY, in view of my eminent contributions in the field of pharmaceutical biotechnology, being as an authority in the field responsible for soliciting top class reviews from eminent scientist/researches whom I will invite. The reviews will care various important aspects of the related topic. Dr. Alain P. Rolland, Vice President Editor-in-Chief, Vice President, Product Development Vical, Inc., 10390 Pacific Center Court San Diego, CA 92121, 92121, United States, E-mail: [email protected] 9. 10.

Revista Latinoamericana de MICROBIOLOGÍA Brenda Valderrama, Ph .D., Editor FORMATEX (Spain) Journal of Industrial Microbiology & Biotechnology

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