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Engineering Nonaqueous Solvent-Compatible Enzymes

Arnold, Frances H. and Chen, Keqin and Economou, Chara and Chen, Wayne and Martinez, Pascal and Yoon, Kyung Pyo and Van Dam, Mariana (1993) Engineering Nonaqueous Solvent-Compatible Enzymes. In: Biocatalyst Design for Stability and Specificity. ACS Symposium Series. No.516. American Chemical Society , Washington, DC, pp. 109-113. ISBN 9780841225183.

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Given current powerful tools for protein molecular engineering and a continually improving understanding of protein folding and stability, it is possible to construct vastly improved industrial enzyme catalysts. Enzymes are better suited to industrial syntheses if they can be made stable and active in high concentrations of polar organic solvents. Sitedirected mutagenesis has been used to test two "design rules" for stabilizing enzymes in organic media. We have shown that surface charge substitution and the introduction of metal-chelating sites provide simple and generally-applicable mechanisms for enzyme stabilization. A variant of α-lytic protease containing two surface charge substitutions is 27 times more stable than wild-type enzyme in 84% DMF. Random mutagenesis and rapid screening techniques have been used to isolate enzyme variants with enhanced catalytic activity in polar organic solvents. Our first experiments resulted in a variant of subtilisin E that is 38 times more active in 85% DMF than the wild-type enzyme. Subsequent rounds of random mutagenesis and screening have yielded a variant 256 times more active in 60% DMF.

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Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 1993 American Chemical Society. Received March 31, 1992. Published in print 11 December 1993. This research is supported by the Catalysis and Biocatalysis Program of the Advanced Industrial Concepts Division of the U. S. Department of Energy and by the Office of Naval Research. F.H.A. gratefully acknowledges an NSF PYI award and a fellowship from the David and Lucile Packard Foundation.
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Department of Energy (DOE)UNSPECIFIED
Office of Naval Research (ONR)UNSPECIFIED
David and Lucile Packard FoundationUNSPECIFIED
Series Name:ACS Symposium Series
Issue or Number:516
Record Number:CaltechAUTHORS:20180405-140056909
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Official Citation:Engineering Nonaqueous Solvent-Compatible Enzymes Frances H. Arnold, Keqin Chen, Chara Economou, Wayne Chen, Pascal Martinez, Kyung Pyo Yoon, and Mariana Van Dam Biocatalyst Design for Stability and Specificity. December 11, 1993, 109-113 DOI:10.1021/bk-1993-0516.ch008
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:85654
Deposited By: Ruth Sustaita
Deposited On:05 Apr 2018 22:05
Last Modified:03 Oct 2019 19:34

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