Improvement of alkaline lipase from Proteus vulgaris T6 by directed evolution

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...

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Bibliographic Details
Published in:Enzyme and microbial technology 2009-02, Vol.44 (2), p.84-88
Main Authors: Fang, Yaowei, Lu, Yingjian, Lv, Fengxia, Bie, Xiaomei, Zhao, Haizhen, Wang, Yu, Lu, Zhaoxin
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Directed evolution
DNA shuffling
Error-prone PCR
Fundamental and applied biological sciences. Psychology
Methods. Procedures. Technologies
Protein engineering
Proteus vulgaris
Proteus vulgaris lipase
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Summary: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.
ISSN:0141-0229
1879-0909
DOI:10.1016/j.enzmictec.2008.10.018