Urlacher, Vlada B.Makhsumkhanov, AkhmadjanSchmid, Rolf D.2006-06-012016-03-312006-06-012016-03-312005263638626http://nbn-resolving.de/urn:nbn:de:bsz:93-opus-26603http://elib.uni-stuttgart.de/handle/11682/837http://dx.doi.org/10.18419/opus-820Wild type cytochrome P450 monooxygenase from Bacillus megaterium (P450 BM-3) has low activity for the hydroxylation of beta-ionone (>1 min-1). Substitution of phenylalanine by valine at position 87 increased the beta-ionone hydroxylation activity up to 100-fold (115 min-1). For further activity improvement methods of site-directed and random mutagenesis were applied. The R47L Y51F F87V mutant, designed by site-directed mutagenesis and the A74E F87V P386S mutant, obtained after two rounds of error-prone PCR, exhibit an increase in activity up to 300-fold compared to the wild type enzyme. All mutants converted -ionone regioselectively to 4-hydroxy-beta-ionone.eninfo:eu-repo/semantics/openAccessCytochrom P-450 , Monooxygenasen , Biokatalyse , Biokonversion , Biotransformation , Hydroxylierung , Mutagenese , Genbibliothek540P450 BM-3 , beta-ionone , regioselectivity , mutagenesis , gene librarypreprint2015-12-10