TY - JOUR
T1 - Alkene epoxidation catalyzed by cytochrome P450 BM-3 139-3
AU - Farinas, Edgardo T.
AU - Alcalde, Miguel
AU - Arnold, Frances
N1 - Funding Information:
The authors thank Dr Nathan Dalleska for assistance with the GC/MS and Dr Bernhard Hauer for the chiral analysis of styrene oxide. This research is funded by the National Science Foundation.
PY - 2004/1/12
Y1 - 2004/1/12
N2 - We recently reported conversion of cytochrome P450 BM-3, a medium-chain (C12-C18) fatty acid monooxygenase, into a highly efficient alkane hydroxylase by directed evolution [Nat. Biotechnol. 2002, 20, 1135]. P450 BM-3 mutant 139-3 exhibited high activity towards a variety of fatty acid and alkane substrates, including C3-C8 alkanes. We report here that mutant 139-3 is also active on benzene, styrene, cyclohexene, 1-hexene, and propylene. Benzene is converted to phenol, while styrene is converted to styrene oxide. Propylene oxidation generates only propylene oxide, but cyclohexene oxidation produces a mixture of cyclohexene oxide (85%) and 2-cyclohexene-1-ol (15%), and 1-hexene is converted to the allylic hydroxylation product, 1-hexene-3-ol. Initial rates of NADPH oxidation for 139-3 in the presence of the substrates greatly (17- to >100-fold) surpass the wild-type in all cases. However, NADPH consumption is only partially coupled to product formation (14-79%). This cytochrome P450 epoxidation catalyst is a suitable starting point for further evolution to improve coupling and activity.
AB - We recently reported conversion of cytochrome P450 BM-3, a medium-chain (C12-C18) fatty acid monooxygenase, into a highly efficient alkane hydroxylase by directed evolution [Nat. Biotechnol. 2002, 20, 1135]. P450 BM-3 mutant 139-3 exhibited high activity towards a variety of fatty acid and alkane substrates, including C3-C8 alkanes. We report here that mutant 139-3 is also active on benzene, styrene, cyclohexene, 1-hexene, and propylene. Benzene is converted to phenol, while styrene is converted to styrene oxide. Propylene oxidation generates only propylene oxide, but cyclohexene oxidation produces a mixture of cyclohexene oxide (85%) and 2-cyclohexene-1-ol (15%), and 1-hexene is converted to the allylic hydroxylation product, 1-hexene-3-ol. Initial rates of NADPH oxidation for 139-3 in the presence of the substrates greatly (17- to >100-fold) surpass the wild-type in all cases. However, NADPH consumption is only partially coupled to product formation (14-79%). This cytochrome P450 epoxidation catalyst is a suitable starting point for further evolution to improve coupling and activity.
KW - Biocatatysis
KW - Cytochrome P450
KW - Directed evolution
KW - Epoxidation
KW - Monooxygenase
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U2 - 10.1016/j.tet.2003.10.099
DO - 10.1016/j.tet.2003.10.099
M3 - Article
AN - SCOPUS:0346964466
SN - 0040-4020
VL - 60
SP - 525
EP - 528
JO - Tetrahedron
JF - Tetrahedron
IS - 3
ER -