We have a continuing interest in applying the current knowledge of cytochrome P450cam substrate recognition to engineer the enzyme for the biotransformation of unnatural substrates with the long-term aim of applications in the synthesis of fine chemicals and bioremediation of environmental contaminants. Comparisons of the structure of target substrates with that of camphor, the natural substrate, led to the design of active-site mutants with greatly enhanced activity for the oxidation of chlorinated benzenes and selectivity of (+)-α-pinene oxidation. The crystal structures of the F87W/Y96F/V247L mutant with 1,3,5-trichlorobenzene or (+)-α-pinene bound have revealed the enzyme–substrate contacts and provided insights into the activity and selectivity patterns. The structures have also provided a novel basis for further engineering of P450cam for increased activity in the oxidation of the highly inert pentachlorobenzene and hexachlorobenzene, and increased selectivity of (+)-α-pinene oxidation.
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Conference Article|
June 01 2003
Engineering substrate recognition in catalysis by cytochrome P450cam
S.G. Bell;
S.G. Bell
*Department of Chemistry, Inorganic Chemistry Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QR, U.K.
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X. Chen;
X. Chen
†Laboratory of Structural Biology, School of Life Sciences and Engineering, Tsinghua University, Beijing 100084, People's Republic of China
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F. Xu;
F. Xu
†Laboratory of Structural Biology, School of Life Sciences and Engineering, Tsinghua University, Beijing 100084, People's Republic of China
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Z. Rao;
Z. Rao
†Laboratory of Structural Biology, School of Life Sciences and Engineering, Tsinghua University, Beijing 100084, People's Republic of China
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L.-L. Wong
L.-L. Wong
1
*Department of Chemistry, Inorganic Chemistry Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QR, U.K.
1To whom correspondence should be addressed (e-mail luet.wong@chem.ox.ac.uk).
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Publisher: Portland Press Ltd
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2003 Biochemical Society
2003
Biochem Soc Trans (2003) 31 (3): 558–562.
Citation
S.G. Bell, X. Chen, F. Xu, Z. Rao, L.-L. Wong; Engineering substrate recognition in catalysis by cytochrome P450cam. Biochem Soc Trans 1 June 2003; 31 (3): 558–562. doi: https://doi.org/10.1042/bst0310558
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