Laboratory evolution of P450 alkane monooxygenases

CATL 22

Rudi Fasan, rfasan@caltech.edu and Frances H. Arnold, frances@cheme.caltech.edu. Division of Chemistry and Chemical Engineering, California Institute of Technology, 1200 E. California Blvd., Mail Code 210-41, Pasadena, CA 91125
Engineering multi-domain, multi-cofactor cytochrome P450 catalytic systems to function efficiently on nonnative substrates is difficult, yet crucial for future applications of these versatile oxygenases. We have applied laboratory evolutionary strategies towards the engineering of cytochrome P450-BM3 for the controlled oxidation of alkanes, a major chemistry goal with important industrial applications. Here we report on the development of engineered enzyme variants that show excellent catalytic properties in the hydroxylation of alkane substrates as challenging as propane and ethane. While yielding useful biocatalysts for chemical synthesis, these studies also provide fascinating insights into the mechanisms underlying evolution of natural enzymes.