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Laboratory evolution of cytochrome P450 BM-3 monooxygenase for organic cosolvents

Wong, Tuck Seng and Arnold, Frances H. and Schwaneberg, Ulrich (2004) Laboratory evolution of cytochrome P450 BM-3 monooxygenase for organic cosolvents. Biotechnology and Bioengineering, 85 (3). pp. 351-358. ISSN 0006-3592. doi:10.1002/bit.10896. https://resolver.caltech.edu/CaltechAUTHORS:20180824-074645775

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Abstract

Cytochrome P450 BM‐3 (EC 1.14.14.1) catalyzes the hydroxylation and/or epoxidation of a broad range of substrates, including alkanes, alkenes, alcohols, fatty acids, amides, polyaromatic hydrocarbons, and heterocycles. For many of these notoriously water‐insoluble compounds, P450 BM‐3's K_m values are in the millimolar range. Polar organic cosolvents are therefore added to increase substrate solubility and achieve high catalytic efficiency. Using P450 BM‐3 as a catalyst for these important transformations requires that we improve its ability to tolerate the cosolvents. By directed evolution, we improved the activity of P450 BM‐3 in the presence of dimethylsulfoxide (DMSO) and tetrahydrofuran (THF), achieving increases in specific activity up to 10‐fold in 2% (v/v) THF and 6‐fold in 25% (v/v) DMSO. The engineered P450 BM‐3's are also significantly more resistant to acetone, acetonitrile, dimethylformamide, and ethanol as cosolvents in the reaction.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1002/bit.10896DOIArticle
ORCID:
AuthorORCID
Arnold, Frances H.0000-0002-4027-364X
Schwaneberg, Ulrich0000-0003-4026-701X
Additional Information:© 2004 Wiley Periodicals, Inc. Received 17 March 2003; accepted 29 September 2003. Published online 31 December 2003. Contract grant sponsors: Caltech Summer Undergraduate Research Fellowship program (to T.S.W.); Maxygen Corporation.
Funders:
Funding AgencyGrant Number
Caltech Summer Undergraduate Research Fellowship (SURF)UNSPECIFIED
Maxygen CorporationUNSPECIFIED
Subject Keywords:P450 BM-3; CYP102; Bacillus megaterium; organic solvent resistance; random mutagenesis; directed evolution
Issue or Number:3
DOI:10.1002/bit.10896
Record Number:CaltechAUTHORS:20180824-074645775
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20180824-074645775
Official Citation:Seng Wong, T. , Arnold, F. H. and Schwaneberg, U. (2004), Laboratory evolution of cytochrome P450 BM‐3 monooxygenase for organic cosolvents. Biotechnol. Bioeng., 85: 351-358. doi:10.1002/bit.10896
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:89120
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:24 Aug 2018 14:54
Last Modified:16 Nov 2021 00:32

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