Improvement of alkaline lipase from Proteus vulgaris
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- 作者：Yaowei Fang ; Yingjian Lu ; Fengxia Lv ; Xiaomei Bie ; Haizhen Zhao ; Yu Wang ; Zhaoxin Lu
- 关键词：Proteus vulgaris lipase ; Directed evolution ; DNA shuffling ; Error-prone PCR
- 刊名：Enzyme and Microbial Technology
- 出版年：2009
- 出版时间：5 February 2009
- 年：2009
- 卷：44
- 期：2
- 页码：84-88
- 全文大小：286 K
文摘
To expand the functionality of lipase from Proteus vulgaris (PVL) we have used error-prone PCR and DNA shuffling methods to create PVL mutants with improved lipase activity. One desirable mutant with three amino acids substitutions was obtained. The mutated lipase was purified and characterized. The activity of the mutant lipase EF3.3 was 3.5 times higher than that of the wild-type (WT-PVL). The mutational effect is interpreted according to a simulated three-dimensional structure for the mutant lipase. Amino acid substitution at position 102 was determined to be critical for lipase activity, while the residue at positions 197 and 229 had only marginal effect.