Combinatorial Alanine Substitution Enables Rapid Optimization of Cytochrome P450BM3 for Selective Hydroxylation of Large Substrates

Jared C. Lewis; Simone M. Mantovani; Yu Fu; Christopher D. Snow; Russell S. Komor; Chi-Huey Wong; Frances H. Arnold

doi:10.1002/cbic.201000565

Combinatorial Alanine Substitution Enables Rapid Optimization of Cytochrome P450BM3 for Selective Hydroxylation of Large Substrates

Jared C. Lewis, Simone M. Mantovani, Yu Fu, Christopher D. Snow, Russell S. Komor, Chi-Huey Wong, Frances H. Arnold

Research output: Contribution to journal › Article › peer-review

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Abstract

Made for each other: Combinatorial alanine substitution of active site residues in a thermostable cytochrome P450BM3 variant was used to generate an enzyme that is active with large substrates. Selective hydroxylation of methoxymethylated monosaccharides, alkaloids, and steroids was thus made possible (see Scheme). This approach could be useful for improving the activity of enzymes that show only limited activity with larger substrates. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Original language	English (US)
Pages (from-to)	2502-2505
Number of pages	4
Journal	ChemBioChem
Volume	11
Issue number	18
DOIs	https://doi.org/10.1002/cbic.201000565
State	Published - Nov 24 2010
Externally published	Yes

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title = "Combinatorial Alanine Substitution Enables Rapid Optimization of Cytochrome P450BM3 for Selective Hydroxylation of Large Substrates",

abstract = "Made for each other: Combinatorial alanine substitution of active site residues in a thermostable cytochrome P450BM3 variant was used to generate an enzyme that is active with large substrates. Selective hydroxylation of methoxymethylated monosaccharides, alkaloids, and steroids was thus made possible (see Scheme). This approach could be useful for improving the activity of enzymes that show only limited activity with larger substrates. {\textcopyright} 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.",

author = "Lewis, {Jared C.} and Mantovani, {Simone M.} and Yu Fu and Snow, {Christopher D.} and Komor, {Russell S.} and Chi-Huey Wong and Arnold, {Frances H.}",

note = "KAUST Repository Item: Exported on 2020-10-01 Acknowledged KAUST grant number(s): KUS-F1-028-03 Acknowledgements: J.C.L. is supported by a U.S. National Institutes of Health Pathways to Independence Award (1K99M087551-01A1). S.M.M. is supported by the Fundacao Coordenadoria de Aperfeicoamento de Pessoal de Nivel Superior (CAPES; 1756-09-5). This work was supported by the U.S. National Institutes of Health (2R01 M068664-05A1), the U.S. Department of Energy, Office of Basic Science, grant DE-FG02-06ER15762, and King Abdullah University of Science and Technology (KAUST), Award No. KUS-F1-028-03. This publication acknowledges KAUST support, but has no KAUST affiliated authors.",

year = "2010",

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doi = "10.1002/cbic.201000565",

language = "English (US)",

volume = "11",

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AU - Lewis, Jared C.

AU - Mantovani, Simone M.

AU - Fu, Yu

AU - Snow, Christopher D.

AU - Komor, Russell S.

AU - Wong, Chi-Huey

AU - Arnold, Frances H.

N1 - KAUST Repository Item: Exported on 2020-10-01 Acknowledged KAUST grant number(s): KUS-F1-028-03 Acknowledgements: J.C.L. is supported by a U.S. National Institutes of Health Pathways to Independence Award (1K99M087551-01A1). S.M.M. is supported by the Fundacao Coordenadoria de Aperfeicoamento de Pessoal de Nivel Superior (CAPES; 1756-09-5). This work was supported by the U.S. National Institutes of Health (2R01 M068664-05A1), the U.S. Department of Energy, Office of Basic Science, grant DE-FG02-06ER15762, and King Abdullah University of Science and Technology (KAUST), Award No. KUS-F1-028-03. This publication acknowledges KAUST support, but has no KAUST affiliated authors.

PY - 2010/11/24

Y1 - 2010/11/24

N2 - Made for each other: Combinatorial alanine substitution of active site residues in a thermostable cytochrome P450BM3 variant was used to generate an enzyme that is active with large substrates. Selective hydroxylation of methoxymethylated monosaccharides, alkaloids, and steroids was thus made possible (see Scheme). This approach could be useful for improving the activity of enzymes that show only limited activity with larger substrates. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

AB - Made for each other: Combinatorial alanine substitution of active site residues in a thermostable cytochrome P450BM3 variant was used to generate an enzyme that is active with large substrates. Selective hydroxylation of methoxymethylated monosaccharides, alkaloids, and steroids was thus made possible (see Scheme). This approach could be useful for improving the activity of enzymes that show only limited activity with larger substrates. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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JO - ChemBioChem

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IS - 18

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Combinatorial Alanine Substitution Enables Rapid Optimization of Cytochrome P450BM3 for Selective Hydroxylation of Large Substrates

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