Extraction, partial purification and characterization of kiwifruit enzyme

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Dec 30, 2017 - same family of thiol proteases as papain and bromelain. The enzyme was estimated at ..... cathepsins and stem bromelain. Journal of Molecular.
The Pharma Innovation Journal 2018; 7(1): 312-315

ISSN (E): 2277- 7695 ISSN (P): 2349-8242 NAAS Rating: 5.03 TPI 2018; 7(1): 312-315 © 2018 TPI

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Received: 29-11-2017 Accepted: 30-12-2017 Swati Sharma Department of Food Science and Technology, YSP University of Horticulture and Forestry, Nauni, Solan (HP), India Devina Vaidya Department of Food Science and Technology, YSP University of Horticulture and Forestry, Nauni, Solan (HP), India Neerja Rana Department of Basic Sciences, YSP University of Horticulture and Forestry, Nauni, Solan (HP), India

Correspondence Swati Sharma Department of Food Science and Technology, YSP University of Horticulture and Forestry, Nauni, Solan (HP), India

Extraction, partial purification and characterization of kiwifruit enzyme Swati Sharma, Devina Vaidya and Neerja Rana Abstract Kiwifruit enzyme is a plant cysteine protease enzyme abundantly present in kiwifruit. Protease enzymes are multifunctional class of enzymes and henceforth can be used in different sectors of food industry such as for milk coagulation, for meat tenderization etc. So, present study was conducted for extraction, partial purification and characterization of kiwifruit enzyme from kiwifruit. Kiwifruit enzyme is belonging to same family of thiol proteases as papain and bromelain. The enzyme was estimated at various stages of fruit maturity followed by partial purification and characterization. The supernatant was precipitated with different concentration of ammonium sulphate i.e., from 10-90 %. Characterization of protease enzyme was done by using response surface methodology (RSM). Results showed that maximum enzyme activity was observed at immature stage of fruit and with 60% of ammonium sulphate concentration. The enzyme had an optimum temperature of 45°C, functioned best at pH 8.0. This enzyme can be exploited commercially in food industry. Keywords: kiwifruit enzyme, protease, extraction, partial purification, characterization

Introduction Kiwifruit (Actinidia chinensis) is native to Southern China (Ferguson, 1984) [5] and was originally called “Yang Tao” in china and “Chinese Gooseberry” in rest of world. Its cultivation were spread from China in the early 20th century, when Isabel Fraser introduced seed to New Zealand. In India, the plant was first grown in Bangalore, where it did not fruit, but the plants introduced in Shimla started bearing in 1969 (Dadlani et al., 1971) [4]. Kiwifruit is very popular in human diet due to its pleasant taste and high content of vitamin C, minerals (potassium, phosphorus, iron) and low calorific value. Kiwifruits are good sources of folate, potassium and contain large amounts of vitamin E in the seeds. Moreover, kiwifruit is known to contain a highly active protease enzyme, actinidin (Kaur et al., 2010) [9]. The proteases are mainly obtained from microbial sources for industrial purposes. Though microbial proteases are the predominant source of industrial enzymes due to their broad biochemical diversity, rapid growth of microorganisms and limited space required for cell cultivation, it involves advanced technology in the field of biotechnology and microbiology (Rao et al., 1998) [13]. The most familiar proteases of animal origin are pancreatic trypsin, chymotrypsin, pepsin and renins. In the view of limited availability of livestock, high price of rennet, religious concern (e.g. Islam and Judaism), diet (vegetarian) or ban on recombinant calf rennet. In addition to these, proteolytic plant enzymes are superior to microbial derived enzymes as well as to animal derived enzymes mainly because of safety problems such as pathogenicity or other disadvantageous effects (Chen et al., 2006) [3]. Henceforth the plant sources would be a possible alternative of microbial and animal proteases. Kiwifruit enzyme is a type of cysteine protease enzyme found in kiwifruits. This enzyme is part of papain-like peptidase C1 family (Baker et al., 1980) [1] and consists of 220 amino acids with an apparent molecular weight about 23.0 kDa (Kamphuis et al., 1985) [8]. It has wide substrate specificity, hydrolyzes most strongly the amide and ester bonds at the carboxyl side of a lysine residue and it is active at pH range 4-10 (Hussain et al., 2003; Morimoto et al., 2006) [7, 11]. Henceforth kiwifruit enzyme as a proteolytic enzyme can be used in food industry for milk coagulation (cheese preparation in dairy industry), for reducing cereal proteins in bakery industry and for meat tenderization. So the present study was conducted for extraction, partial purification and characterization of kiwifruit enzyme.

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  Materials and methods Procurement of Kiwifruits Kiwifruits (Actinidia chinensis) were procured from Kiwifruit Orchad, Department of Fruit Science, Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni-Solan Himachal Pradesh (INDIA). Kiwifruits were procured at three stages of fruit maturity i.e. one week before attaining the commercial harvest stage (TSS