Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulans BTS-3

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• 作者：Kumar ; Satyendra ; Kikon ; Khyodano ; Upadhyay ; Ashutosh ; Kanwar ; Shamsher S. ; Gupta ; Reena
• 关键词：Bacillus coagulans ; Lipase ; Production ; Purification and characterization
• 刊名：Protein Expression and Purification
• 出版年：2005
• 出版时间：May, 2005
• 年：2005
• 卷：41
• 期：1
• 页码：38-44
• 全文大小：534 K

文摘

A thermophilic isolate Bacillus coagulans BTS-3 produced an extracellular alkaline lipase, the production of which was substantially enhanced when the type of carbon source, nitrogen source, and the initial pH of culture medium were consecutively optimized. Lipase activity 1.16 U/ml of culture medium was obtained in 48 h at 55 °C and pH 8.5 with refined mustard oil as carbon source and a combination of peptone and yeast extract (1:1) as nitrogen sources. The enzyme was purified 40-fold to homogeneity by ammonium sulfate precipitation and DEAE–Sepharose column chromatography. Its molecular weight was 31 kDa on SDS–PAGE. The enzyme showed maximum activity at 55 °C and pH 8.5, and was stable between pH 8.0 and 10.5 and at temperatures upto 70 °C. The enzyme was found to be inhibited by Al³⁺, Co²⁺, Mn²⁺, and Zn²⁺ ions while K⁺, Fe³⁺, Hg²⁺, and Mg²⁺ ions enhanced the enzyme activity; Na⁺ ions have no effect on enzyme activity. The purified lipase showed a variable specificity/hydrolytic activity towards various 4-nitrophenyl esters.