title: In planta biocatalysis screen of P450s identifies 8-methoxypsoralen as a substrate for the CYP82C subfamily, yielding original chemical structures.
abstract: An in vivo plant screen that allows for the analysis of exogenously applied substrates against transgenic Arabidopsis lines overexpressing individual cytochrome P450s has been developed. By deploying this screen with a subset of 91 P450s, we have identified an original substrate for members of the CYP82C subfamily. The therapeutic compound 8-methoxypsoralen was hydroxylated by plants overexpressing CYP82C2 or CYP82C4, forming 5-hydroxy-8-methoxypsoralen. Additionally, plants further modified this product to create a glycosylated compound, likely the compound 5-O-beta-D-glucopyranosyl-8-methoxypsoralen. The discovery of adducts of therapeutic compounds demonstrates the potential of this biocatalysis screening approach to create compounds that may be of pharmacological value. Additionally, this platform provides a means to expand the general knowledge base of P450 enzyme/substrate combinations and may provide valuable tools for a vast array of biocatalytic and bioremediation processes.
authors.affiliation: Ceres, Inc., 1535 Rancho Conejo Boulevard, Thousand Oaks, CA 91320, USA. tkruse@ceres-inc.com <tkruse@ceres-inc.com>
authors.forename: Tanya
authors.initials: T
authors.lastname: Kruse
authors.forename: Kwongling
authors.initials: K
authors.lastname: Ho
authors.forename: Hye-Dong
authors.initials: HD
authors.lastname: Yoo
authors.forename: Thomas
authors.initials: T
authors.lastname: Johnson
authors.forename: Matt
authors.initials: M
authors.lastname: Hippely
authors.forename: Joon-Hyun
authors.initials: JH
authors.lastname: Park
authors.forename: Richard
authors.initials: R
authors.lastname: Flavell
authors.forename: Steve
authors.initials: S
authors.lastname: Bobzin
mesh_terms: D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D002853:Chromatography, Liquid; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D006031:Glycosylation; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008730:Methoxsalen / Q000737:chemistry* / Q000378:metabolism*; D010944:Plants / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D036226:Seedlings / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D003577:Cytochrome P-450 Enzyme System; D008730:Methoxsalen