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Title: A method to rationally increase protein stability based on the charge-charge interaction, with application to lipase LipK107
Authors: Zhang, L.
Tang, X.
Cui, D.
Yao, Z.
Gao, B.
Jiang, S.
Yin, B.
Yuan, Y.A. 
Wei, D.
Keywords: Designed protein stabilization
Electrostatic stabilization
Lipase stabilization
Surface charge-charge interaction
Issue Date: 2014
Citation: Zhang, L., Tang, X., Cui, D., Yao, Z., Gao, B., Jiang, S., Yin, B., Yuan, Y.A., Wei, D. (2014). A method to rationally increase protein stability based on the charge-charge interaction, with application to lipase LipK107. Protein Science 23 (1) : 110-116. ScholarBank@NUS Repository.
Abstract: We report a suite of enzyme redesign protocol based on the surface charge-charge interaction calculation, which is potentially applied to improve the stability of an enzyme without compromising its catalytic activity. Together with the experimental validation, we have released a suite of enzyme redesign algorithm Enzyme Thermal Stability System, written based on our model, for open access to meet the needs in wet labs. Lipk107, a lipase of a versatile industrial use, was chosen to test our software. Our calculation determined that four residues, D113, D149, D213, and D253, located on the surface of LipK107 were critical to the stability of the enzyme. The model was validated with mutagenesis at these four residues followed by stability and activity tests. LipK107 mutants D113A and D149K were more resistant to thermal inactivation with ~10°C higher halfinactivation temperature than wild-type LipK107. Moreover, mutant D149K exhibited significant retention in residual activity under constant heat, showing a 14-fold increase in the halfinactivation time at 50°C. Activity tests showed that these mutants retained the equal or higher specific activity, among which noteworthy was the mutant D253A with as much as 20% higher activity. We suggest that our protocol could be used as a general guideline to redesign protein enzymes with increased stabilities and enhanced activities. © 2013 The Protein Society.
Source Title: Protein Science
ISSN: 1469896X
DOI: 10.1002/pro.2388
Appears in Collections:Staff Publications

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