title: Whole-cell microtiter plate screening assay for terminal hydroxylation of fatty acids by P450s.
abstract: A readily available galactose oxidase (GOase) variant was used to develop a whole cell screening assay. This endpoint detection system was applied in a proof-of-concept approach by screening a focussed mutant library. This led to the discovery of the thus far most active P450 Marinobacter aquaeolei mutant catalysing the terminal hydroxylation of fatty acids.
authors.affiliation: Institute of Technical Biochemistry, Universitaet Stuttgart, Allmandring 31, 70569 Stuttgart, Germany. bernhard.hauer@itb.uni-stuttgart.de.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Weissenborn
authors.forename: Sandra
authors.initials: S
authors.lastname: Notonier
authors.forename: Sarah-Luise
authors.initials: SL
authors.lastname: Lang
authors.forename: Konrad B
authors.initials: KB
authors.lastname: Otte
authors.forename: Susanne
authors.initials: S
authors.lastname: Herter
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Turner
authors.forename: Sabine L
authors.initials: SL
authors.lastname: Flitsch
authors.forename: Bernhard
authors.initials: B
authors.lastname: Hauer
mesh_terms: D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism*; D005692:Galactose Oxidase / Q000737:chemistry / Q000378:metabolism; D006900:Hydroxylation; D007850:Lauric Acids / Q000737:chemistry; D053520:Marinobacter / Q000201:enzymology; D016296:Mutagenesis; D009249:NADP / Q000737:chemistry / Q000378:metabolism; D046888:Tissue Array Analysis
chemical_list: D005227:Fatty Acids; D007850:Lauric Acids; C030358:lauric acid; D009249:NADP; D003577:Cytochrome P-450 Enzyme System; D005692:Galactose Oxidase