ChemBioChem
10.1002/cbic.201900691
COMMUNICATION
2019-2579), Program for Outstanding Youth of Zunyi Medical
University (17zy-001), National First-Rate Construction
Discipline of Guizhou Province (Pharmacy) (YLXKJS-YX-04)
and the Fifth Batch of Talent Base in Guizhou Province (2016).
Table 1. Substrate scope of the P450DA-catalyzed benzylic hydroxylation.[a]
Keywords: cytochrome P450 monooxygenase •
characterization • benzylic hydroxylation • enantioselective
hydroxylation • chiral benzylic alcohols
Yield
2
ee
2 (%)
Entry
1
Substrate
Product
TTN[c]
130
(%)[b]
[b]
[
1]
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10
99(S)
92(R)
86(R)
95(R)
1
a
2a
2b
2c
2d
01018.
[
[
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2
3
4
72
77
62
940
1010
810
1b
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1c
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1d
[
[
5]
6]
F. Hannemann, A. Bichet, K. M. Ewen, R. Bernhardt, BBA-Gen.
Subjects 2007, 1770, 330-344.
5
6
78
58
96(R)
96(R)
1020
760
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1
e
2e
2f
1
f
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[
a] Reactions were carried out in 5 mL PB (50 mM, pH = 7.5) containing 10 g
cdw/L the resting cells of E. coli (P450DA) and 2 mM substrate 1, at 20 C for
4 h. [b] Determined over the formed products 2 by chiral HPLC at 24 h. [c]
TTNs were calculated from the analytical yields of 2. The P450DA
o
2
[8]
[9]
R. S. Matson, R. S. Hare, A. J. Fulco, BBA-Lipid. Lipid. Met. 1977, 487,
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-
1
concentration expressed in E. coli cell was 0.153 μmol·g cdw .
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3
In summary, we cloned the gene, expressed and
characterized a self-sufficient P450 monooxygenase from D.
apachensis. Sequence analysis suggested the P450DA was a
member of CYP102D subfamily. Our results also revealed the
P450DA could utilize both NADH and NADPH as a reducing
cofactor, as well as a preference of NADH over NADPH. In
addition, P450DA catalyzed benzylic hydroxylation of various
[
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a
promising biocatalyst for the synthesis of chiral benzylic alcohols
through direct C-H hydroxylation.
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Acknowledgements
Chem. 2012, 14, 440-446; f) A. Li, S. Wu, J. P. Adams, R. Snajdrova, Z.
Li, Chem. Commun. 2014, 50, 8771-8774; g) A. Eichler, Ł. Gricman, S.
Herter, P. P. Kelly, N. J. Turner, J. Pleiss, S. L. Flitsch, ChemBioChem
We are grateful for financial support from the National Natural
Science Foundation of China (No. 21961048 and 21562054),
Guizhou Science and Technology Department (QKHRC-2016-
2
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Yanagisawa, H. Sugimoto, Y. Shiro, Y. Watanabe, Catal. Sci. Technol.
017, 7, 3332-3338; i) A. Rentmeister, F. H. Arnold, R. Fasan, Nat.
Chem. Biol. 2008, 5, 26.
2
4029, QKHPTRC-2016-5801, QKHJC-2019-1467 and QKHZC-
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