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Title: Optimization Of Medium Components And Process Parameters For Enhanced Production Of Lactase By A Bacterium Isolated From Dairy Effluent
Researcher: T.C.Venkateswarulu
Guide(s): Vidya Prabhakar Kodali, Bharath Kumar Ravuru
University: Vignans Foundation for Science Technology and Research
Completed Date: 2017
Abstract: Lactose intolerance is characterized by indigestion of milk sugar called as lactose. newlineIndigestion of milk and milk based products in lactose intolerants can be treated with newlinesupplementing lactase enzyme. Bacteria that produce lactase have been used to treat the newlinelactose intolerants. Commercially, this enzyme production from bacterial sources was not newlinereported for higher production. In the present study, dairy effluent was screened for newlinelactase producing bacteria by biochemical methods and found 46 lactase positive isolates. newlineThe highest lactase yielding isolate was named as VUVD001 and this isolate was newlinecharacterized by morphological, biochemical and molecular methods. The VUVD001 newlineisolate was stable at extreme pH and temperature conditions. The o-nitrophenyl-and#946;-Dgalactopyranoside (ONPG) disc analysis was performed for the confirmation of lactase newlineproducing activity. The produced lactase was partially purified followed by zymogram newlineanalysis for confirmation of extracellular activity. The isolate VUVD001 was identified newlineas Bacillus subtilis by 16S rRNA gene sequencing and this strain was able to survive in a newlinetemperature range of 20-55 oC, pH range of 5-8 and a salt concentration upto 8% of NaCl. newlineIn addition, the production of lactase was improved by designing new medium through newlineoptimization of nutrient components by one-factor-at-a-time (OFAT) and statistical newlinemethods. Lactose and yeast extract were selected as preferable carbon and nitrogen newlinesources for lactase production. Further, addition of tryptophan and MgSO4 showed newlineenhanced lactase production. Statistical analysis proved to be a powerful tool in exploring newlineoptimum fermentation conditions. The individual and interactive role of lactose, yeast newlineextract, magnesium sulfate and tryptophan concentration on lactase production was newlineexamined by central composite design. The submerged fermentation with B. subtilis newlineVUVD001 strain produced an enhanced lactase activity of 63.54 U/ml in optimized medium.
Pagination: 124
Appears in Departments:Department of Biotechnology

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