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Applied and Environmental Microbiology, December 2001, p. 5735-5739, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.12.5735-5739.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Engineering Cytochrome P450 BM-3 for Oxidation of Polycyclic Aromatic Hydrocarbons

Qing-Shan Li,1 Jun Ogawa,1 Rolf D. Schmid,2 and Sakayu Shimizu1,*

Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan,1 and Institut für Technische Biochemie, Universität Stuttgart, D-70459 Stuttgart, Germany2

Received 25 June 2001/Accepted 24 September 2001

Cytochrome P450 BM-3, a self-sufficient P450 enzyme from Bacillus megaterium that catalyzes the subterminal hydroxylation of long-chain fatty acids, has been engineered into a catalyst for the oxidation of polycyclic aromatic hydrocarbons. The activities of a triplet mutant (A74G/F87V/L188Q) towards naphthalene, fluorene, acenaphthene, acenaphthylene, and 9-methylanthracene were 160, 53, 109, 287, and 22/min, respectively. Compared with the activities of the wild type towards these polycyclic aromatic hydrocarbons, those of the mutant were improved by up to 4 orders of magnitude. The coupling efficiencies of the mutant towards naphthalene, fluorene, acenaphthene, acenaphthylene, and 9-methylanthracene were 11, 26, 5.4, 15, and 3.2%, respectively, which were also improved several to hundreds fold. The high activities of the mutant towards polycyclic aromatic hydrocarbons indicate the potential of engineering P450 BM-3 for the biodegradation of these compounds in the environment.

* Corresponding author: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan. Phone: 81-75-753-6115. Fax: 81-75-753-6128. E-mail: sim{at}kais.kyoto-u.ac.jp.

Applied and Environmental Microbiology, December 2001, p. 5735-5739, Vol. 67, No. 12
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.12.5735-5739.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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