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A diverse family of thermostable cytochrome P450s created by recombination of stabilizing fragments

Li, Yougen and Drummond, D. Allan and Sawayama, Andrew M. and Snow, Christopher D. and Bloom, Jesse D. and Arnold, Frances H. (2007) A diverse family of thermostable cytochrome P450s created by recombination of stabilizing fragments. Nature Biotechnology, 25 (9). pp. 1051-1056. ISSN 1087-0156. https://resolver.caltech.edu/CaltechAUTHORS:20150317-105811561

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Abstract

Thermostable enzymes combine catalytic specificity with the toughness required to withstand industrial reaction conditions. Stabilized enzymes also provide robust starting points for evolutionary improvement of other protein properties3. We recently created a library of at least 2,300 new active chimeras of the biotechnologically important cytochrome P450 enzymes. Here we show that a chimera's thermostability can be predicted from the additive contributions of its sequence fragments. Based on these predictions, we constructed a family of 44 novel thermostable P450s with half-lives of inactivation at 57 °C up to 108 times that of the most stable parent. Although they differ by as many as 99 amino acids from any known P450, the stable sequences are catalytically active. Among the novel functions they exhibit is the ability to produce drug metabolites. This chimeric P450 family provides a unique ensemble for biotechnological applications and for studying sequence-stability-function relationships.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1038/nbt1333DOIArticle
http://dx.doi.org/10.1038/nbt1207-1488dDOIErratum
http://rdcu.be/cldcPublisherFree ReadCube access -- Article
http://rdcu.be/cldePublisherFree ReadCube access - Erratum
ORCID:
AuthorORCID
Snow, Christopher D.0000-0002-7690-3519
Bloom, Jesse D.0000-0003-1267-3408
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 2007 Nature Publishing Group. Received 30 May; accepted 6 August; published online 26 August; corrected after print 12 December 2007. The authors thank Christopher R. Otey and Marco Landwehr for their assistance. This work was supported in part by National Institutes of Health Grant R01 GM068664-01, Army Research Office Contract DAAD19-03-0004, Eli Lilly & Co. and a Howard Hughes Medical Institute predoctoral fellowship (to J.D.B.). Author Contributions: Y.L., A.M.S. and F.H.A. designed the research; Y.L. and A.M.S. performed the research; Y.L., D.A.D., A.M.S., C.D.S. and J.D.B. contributed to the analytical tools; Y.L., D.A.D., A.M.S., C.D.S., J.D.B. and F.H.A. analyzed data; Y.L., D.A.D., A.M.S., C.D.S. and F.H.A. wrote the paper.
Funders:
Funding AgencyGrant Number
NIHR01 GM068664-01
Army Research Office (ARO)DAAD19-03-0004
Eli Lilly and Co.UNSPECIFIED
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Issue or Number:9
Record Number:CaltechAUTHORS:20150317-105811561
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20150317-105811561
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:55853
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:17 Mar 2015 18:20
Last Modified:17 Dec 2019 00:38

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