Role of residue 87 in substrate selectivity and regioselectivity of drug-metabolizing cytochrome P450 CYP102A1 M11

E.R. Vottero; V. Rea; J. Lastdrager; M. Honing; N.P.E. Vermeulen; J.N.M. Commandeur

doi:10.1007/s00775-011-0789-4

Role of residue 87 in substrate selectivity and regioselectivity of drug-metabolizing cytochrome P450 CYP102A1 M11

E.R. Vottero, V. Rea, J. Lastdrager, M. Honing, N.P.E. Vermeulen, J.N.M. Commandeur

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

Abstract

CYP102A1, originating from Bacillus megaterium, is a highly active enzyme which has attracted much attention because of its potential applicability as a biocatalyst for oxidative reactions. Previously we developed drug-metabolizing mutant CYP102A1 M11 by a combination of site-directed and random mutagenesis. CYP102A1 M11 contains eight mutations, when compared with wild-type CYP102A1, and is able to produce human-relevant metabolites of several pharmaceuticals. In this study, active-site residue 87 of drug-metabolizing mutant CYP102A1 M11 was mutated to all possible natural amino acids to investigate its role in substrate selectivity and regioselectivity. With alkoxyresorufins as substrates, large differences in substrate selectivities and coupling efficiencies were found, dependent on the nature of residue 87. For all combinations of alkoxyresorufins and mutants, extremely fast rates of NADPH oxidation were observed (up to 6,000 min

Original language	English
Pages (from-to)	899-912
Number of pages	13
Journal	Journal of Biological Inorganic Chemistry - JBIC
Volume	16
Issue number	6
DOIs	https://doi.org/10.1007/s00775-011-0789-4
Publication status	Published - 2011

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Vottero, E. R., Rea, V., Lastdrager, J., Honing, M., Vermeulen, N. P. E., & Commandeur, J. N. M. (2011). Role of residue 87 in substrate selectivity and regioselectivity of drug-metabolizing cytochrome P450 CYP102A1 M11. Journal of Biological Inorganic Chemistry - JBIC, 16(6), 899-912. https://doi.org/10.1007/s00775-011-0789-4

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title = "Role of residue 87 in substrate selectivity and regioselectivity of drug-metabolizing cytochrome P450 CYP102A1 M11",

abstract = "CYP102A1, originating from Bacillus megaterium, is a highly active enzyme which has attracted much attention because of its potential applicability as a biocatalyst for oxidative reactions. Previously we developed drug-metabolizing mutant CYP102A1 M11 by a combination of site-directed and random mutagenesis. CYP102A1 M11 contains eight mutations, when compared with wild-type CYP102A1, and is able to produce human-relevant metabolites of several pharmaceuticals. In this study, active-site residue 87 of drug-metabolizing mutant CYP102A1 M11 was mutated to all possible natural amino acids to investigate its role in substrate selectivity and regioselectivity. With alkoxyresorufins as substrates, large differences in substrate selectivities and coupling efficiencies were found, dependent on the nature of residue 87. For all combinations of alkoxyresorufins and mutants, extremely fast rates of NADPH oxidation were observed (up to 6,000 min",

author = "E.R. Vottero and V. Rea and J. Lastdrager and M. Honing and N.P.E. Vermeulen and J.N.M. Commandeur",

year = "2011",

doi = "10.1007/s00775-011-0789-4",

language = "English",

volume = "16",

pages = "899--912",

journal = "Journal of Biological Inorganic Chemistry - JBIC",

issn = "0949-8257",

publisher = "Springer Verlag",

number = "6",

}

Role of residue 87 in substrate selectivity and regioselectivity of drug-metabolizing cytochrome P450 CYP102A1 M11. / Vottero, E.R.; Rea, V.; Lastdrager, J.; Honing, M.; Vermeulen, N.P.E.; Commandeur, J.N.M.

In: Journal of Biological Inorganic Chemistry - JBIC, Vol. 16, No. 6, 2011, p. 899-912.

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

TY - JOUR

T1 - Role of residue 87 in substrate selectivity and regioselectivity of drug-metabolizing cytochrome P450 CYP102A1 M11

AU - Vottero, E.R.

AU - Rea, V.

AU - Lastdrager, J.

AU - Honing, M.

AU - Vermeulen, N.P.E.

AU - Commandeur, J.N.M.

PY - 2011

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AB - CYP102A1, originating from Bacillus megaterium, is a highly active enzyme which has attracted much attention because of its potential applicability as a biocatalyst for oxidative reactions. Previously we developed drug-metabolizing mutant CYP102A1 M11 by a combination of site-directed and random mutagenesis. CYP102A1 M11 contains eight mutations, when compared with wild-type CYP102A1, and is able to produce human-relevant metabolites of several pharmaceuticals. In this study, active-site residue 87 of drug-metabolizing mutant CYP102A1 M11 was mutated to all possible natural amino acids to investigate its role in substrate selectivity and regioselectivity. With alkoxyresorufins as substrates, large differences in substrate selectivities and coupling efficiencies were found, dependent on the nature of residue 87. For all combinations of alkoxyresorufins and mutants, extremely fast rates of NADPH oxidation were observed (up to 6,000 min

U2 - 10.1007/s00775-011-0789-4

DO - 10.1007/s00775-011-0789-4

M3 - Article

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JO - Journal of Biological Inorganic Chemistry - JBIC

JF - Journal of Biological Inorganic Chemistry - JBIC

SN - 0949-8257

IS - 6

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