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Engineered Alkane-Hydroxylating Cytochrome P450BM3 Exhibiting Nativelike Catalytic Properties

Authors

  • Rudi Fasan Dr.,

    1. Department of Chemistry and Chemical Engineering, California Institute of Technology, 1200 California Blvd. MC 210-41, Pasadena, CA 91125, USA, Fax: (+1) 626-568-8743
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  • Mike M. Chen,

    1. Department of Chemistry and Chemical Engineering, California Institute of Technology, 1200 California Blvd. MC 210-41, Pasadena, CA 91125, USA, Fax: (+1) 626-568-8743
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  • Nathan C. Crook,

    1. Department of Chemistry and Chemical Engineering, California Institute of Technology, 1200 California Blvd. MC 210-41, Pasadena, CA 91125, USA, Fax: (+1) 626-568-8743
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  • Frances H. Arnold Prof. Dr.

    1. Department of Chemistry and Chemical Engineering, California Institute of Technology, 1200 California Blvd. MC 210-41, Pasadena, CA 91125, USA, Fax: (+1) 626-568-8743
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  • This work was supported by a Swiss National Science Foundation fellowship to R.F., a U.S. NSF Fellowship to M.M.C. and by the U.S. Army Research Office, ARO Contract DAAD19-03-D-0004. We thank Dr. Christopher Snow for providing the homology model of P450BM3 reductase and Dr. Matthew W. Peters, Dr. Peter Meihnold, and Dr. Marco Landwehr for helpful discussions regarding the biotransformations and for providing access to DasGip fermenter.

Abstract

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Divide, evolve, and conquer: A domain-based strategy (see scheme) was used to engineer high catalytic and coupling efficiency for propane hydroxylation in a multidomain cytochrome P450 enzyme. The engineered enzymes exhibit high total activities in whole-cell bioconversions of propane to propanol under mild conditions, using air as oxidant.

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