Characterization of a Self‐Sufficient Cytochrome P450 Monooxygenase from Deinococcus apachensis for Enantioselective Benzylic Hydroxylation

A self‐sufficient cytochrome P450 monooxygenase from Deinococcus apachensis (P450DA) was identified and successfully overexpressed in Escherichia coli BL21(DE3). P450DA would be a member of the CYP102D subfamily and assigned as CYP102D2 according to the phylogenetic tree and sequence alignment. Puri...

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Published inChembiochem : a European journal of chemical biology Vol. 21; no. 13; pp. 1820 - 1825
Main Authors Xie, Lingzhi, Chen, Ke, Cui, Haibo, Wan, Nanwei, Cui, Baodong, Han, Wenyong, Chen, Yongzheng
Format Journal Article
LanguageEnglish
Published Germany Wiley Subscription Services, Inc 01.07.2020
Subjects
Alcohols
characterization
chiral benzylic alcohols
Cytochrome
Cytochrome P450
Cytochrome P450 monooxygenase
Cytochromes P450
Deinococcus
E coli
Enantiomers
enantioselectivity
Hydroxylation
NADH
NADP
Nicotinamide adenine dinucleotide
Nucleotide sequence
Phylogeny
Substrates
enantioselectivity
cytochrome P450 monooxygenase
chiral benzylic alcohols
hydroxylation
characterization
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Summary:A self‐sufficient cytochrome P450 monooxygenase from Deinococcus apachensis (P450DA) was identified and successfully overexpressed in Escherichia coli BL21(DE3). P450DA would be a member of the CYP102D subfamily and assigned as CYP102D2 according to the phylogenetic tree and sequence alignment. Purification and characterization of the recombinant P450DA indicated both NADH and NADPH could be used by P450DA as a reducing cofactor. The recombinant E. coli (P450DA) strain was functionally active, showing excellent enantioselectivity for benzylic hydroxylation of methyl 2‐phenylacetate. Further substrate scope studies revealed that P450DA is able to catalyze benzylic hydroxylation of a variety of compounds, affording the corresponding chiral benzylic alcohols in 86–99 % ee and 130–1020 total turnover numbers. A self‐sufficient cytochrome P450 monooxygenase from Deinococcus apachensis (P450DA) was identified and successfully overexpressed in Escherichia coli BL21(DE3). Further substrate scope studies revealed that P450DA is able to catalyze benzylic hydroxylation of a variety of compounds, affording the corresponding chiral benzylic alcohols in 86–99 % ee and 130–1020 total turnover numbers.
Bibliography:These authors contributed equally to this work.
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ISSN:1439-4227
1439-7633
DOI:10.1002/cbic.201900691