一种新型萨芬芽孢杆菌脂肪酶的鉴定、生产和纯化

Krishna P. Patel, S. Parikh
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引用次数: 3

摘要

脂肪酶是由植物、动物和微生物等多种来源产生的细胞外酶。微生物是胞外脂肪酶的最佳来源,在各种工业过程中被广泛应用。在本研究中,从印度古吉拉特邦Aravalli地区炼油厂收集的受石油污染的土壤中分离出脂肪酶生产者。在三丁酸甘油酯琼脂等选择性培养基上进行筛选。采用16S rRNA法对分离物进行鉴定,定量测定脂肪酶活性。以橄榄油为底物,采用深层发酵法生产脂肪酶。脂肪酶活性最高的菌株为萨芬芽孢杆菌。采用(nh4) 2so4沉淀法和离子交换色谱法纯化了白螺旋藻脂肪酶,并用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法对其进行了表征。该酶的比活性为13.71 U/mg,产率为16.16%,脂肪酶分子量为55kDa。
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Identification, production, and purification of a novel lipase from Bacillus safensis
Lipase is an extracellular enzyme produced by various sources such as plants, animals, and microorganisms. Microorganisms are the best source of extracellular lipase and are used in various industrial processes for their potential application. In this research study, lipase producers were isolated in oil-contaminated soil collected from oil refineries of Aravalli district situated in Gujarat, India. It was screened on a selective medium like tributyrin agar. The isolate was identified using the 16S rRNA method and lipase enzyme activity was quantitatively assayed. Submerged fermentation was used for the production of lipase using olive oil as a substrate. Maximum lipase activity was found from Bacillus safensis . Lipase from B. safensis was purified using (NH 4 ) 2 SO 4 precipitation and ion exchange chromatography and characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis. B. safensis exhibited specific activity of 13.71 U/mg with a yield of 16.16% and the molecular weight of lipase is 55kDa was obtained.
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