Active Site Substitution A82W Improves the Regioselectivity of Steroid Hydroxylation by Cytochrome P450 BM3 Mutants As Rationalized by Spin Relaxation Nuclear Magnetic Resonance Studies

V. Rea; A.J. Kolkman; E.R. Vottero; E.J. Stronks; K.A. Ampt; M. Honing; N.P.E. Vermeulen; S.S. Wijmenga; J.N.M. Commandeur

doi:10.1021/bi201433h

Active Site Substitution A82W Improves the Regioselectivity of Steroid Hydroxylation by Cytochrome P450 BM3 Mutants As Rationalized by Spin Relaxation Nuclear Magnetic Resonance Studies

V. Rea, A.J. Kolkman, E.R. Vottero, E.J. Stronks, K.A. Ampt, M. Honing, N.P.E. Vermeulen, S.S. Wijmenga, J.N.M. Commandeur

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

Cytochrome P450 BM3 from Bacillus megaterium is a monooxygenase with great potential for biotechnological applications. In this paper, we present engineered drug-metabolizing P450 BM3 mutants as a novel tool for regioselective hydroxylation of steroids at position 16β. In particular, we show that by replacing alanine at position 82 with a tryptophan in P450 BM3 mutants M01 and M11, the selectivity toward 16β-hydroxylation for both testosterone and norethisterone was strongly increased. The A82W mutation led to a ≤42-fold increase in V

Original language	English
Pages (from-to)	750-760
Journal	Biochemistry
Volume	51
Issue number	3
DOIs	https://doi.org/10.1021/bi201433h
Publication status	Published - 2012

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Rea, V., Kolkman, A. J., Vottero, E. R., Stronks, E. J., Ampt, K. A., Honing, M., Vermeulen, N. P. E., Wijmenga, S. S., & Commandeur, J. N. M. (2012). Active Site Substitution A82W Improves the Regioselectivity of Steroid Hydroxylation by Cytochrome P450 BM3 Mutants As Rationalized by Spin Relaxation Nuclear Magnetic Resonance Studies. Biochemistry, 51(3), 750-760. https://doi.org/10.1021/bi201433h

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title = "Active Site Substitution A82W Improves the Regioselectivity of Steroid Hydroxylation by Cytochrome P450 BM3 Mutants As Rationalized by Spin Relaxation Nuclear Magnetic Resonance Studies",

abstract = "Cytochrome P450 BM3 from Bacillus megaterium is a monooxygenase with great potential for biotechnological applications. In this paper, we present engineered drug-metabolizing P450 BM3 mutants as a novel tool for regioselective hydroxylation of steroids at position 16β. In particular, we show that by replacing alanine at position 82 with a tryptophan in P450 BM3 mutants M01 and M11, the selectivity toward 16β-hydroxylation for both testosterone and norethisterone was strongly increased. The A82W mutation led to a ≤42-fold increase in V",

author = "V. Rea and A.J. Kolkman and E.R. Vottero and E.J. Stronks and K.A. Ampt and M. Honing and N.P.E. Vermeulen and S.S. Wijmenga and J.N.M. Commandeur",

year = "2012",

doi = "10.1021/bi201433h",

language = "English",

volume = "51",

pages = "750--760",

journal = "Biochemistry",

issn = "0006-2960",

publisher = "American Chemical Society",

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Rea, V, Kolkman, AJ, Vottero, ER, Stronks, EJ, Ampt, KA, Honing, M, Vermeulen, NPE, Wijmenga, SS & Commandeur, JNM 2012, 'Active Site Substitution A82W Improves the Regioselectivity of Steroid Hydroxylation by Cytochrome P450 BM3 Mutants As Rationalized by Spin Relaxation Nuclear Magnetic Resonance Studies', Biochemistry, vol. 51, no. 3, pp. 750-760. https://doi.org/10.1021/bi201433h

TY - JOUR

T1 - Active Site Substitution A82W Improves the Regioselectivity of Steroid Hydroxylation by Cytochrome P450 BM3 Mutants As Rationalized by Spin Relaxation Nuclear Magnetic Resonance Studies

AU - Rea, V.

AU - Kolkman, A.J.

AU - Vottero, E.R.

AU - Stronks, E.J.

AU - Ampt, K.A.

AU - Honing, M.

AU - Vermeulen, N.P.E.

AU - Wijmenga, S.S.

AU - Commandeur, J.N.M.

PY - 2012

Y1 - 2012

N2 - Cytochrome P450 BM3 from Bacillus megaterium is a monooxygenase with great potential for biotechnological applications. In this paper, we present engineered drug-metabolizing P450 BM3 mutants as a novel tool for regioselective hydroxylation of steroids at position 16β. In particular, we show that by replacing alanine at position 82 with a tryptophan in P450 BM3 mutants M01 and M11, the selectivity toward 16β-hydroxylation for both testosterone and norethisterone was strongly increased. The A82W mutation led to a ≤42-fold increase in V

AB - Cytochrome P450 BM3 from Bacillus megaterium is a monooxygenase with great potential for biotechnological applications. In this paper, we present engineered drug-metabolizing P450 BM3 mutants as a novel tool for regioselective hydroxylation of steroids at position 16β. In particular, we show that by replacing alanine at position 82 with a tryptophan in P450 BM3 mutants M01 and M11, the selectivity toward 16β-hydroxylation for both testosterone and norethisterone was strongly increased. The A82W mutation led to a ≤42-fold increase in V

U2 - 10.1021/bi201433h

DO - 10.1021/bi201433h

M3 - Article

SN - 0006-2960

VL - 51

SP - 750

EP - 760

JO - Biochemistry

JF - Biochemistry

IS - 3

ER -

Active Site Substitution A82W Improves the Regioselectivity of Steroid Hydroxylation by Cytochrome P450 BM3 Mutants As Rationalized by Spin Relaxation Nuclear Magnetic Resonance Studies

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