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Combinatorial Alanine Substitution Enables Rapid Optimization of Cytochrome P450 BM3 for Selective Hydroxylation of Large Substrates
Author(s) -
Lewis Jared C.,
Mantovani Simone M.,
Fu Yu,
Snow Christopher D.,
Komor Russell S.,
Wong ChiHuey,
Arnold Frances H.
Publication year - 2010
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201000565
Subject(s) - hydroxylation , alanine , chemistry , active site , enzyme , cytochrome p450 , stereochemistry , cytochrome , monosaccharide , substitution (logic) , combinatorial chemistry , protein engineering , biochemistry , amino acid , computer science , programming language
Made for each other: Combinatorial alanine substitution of active site residues in a thermostable cytochrome P450 BM3 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.