In planta biocatalysis screen of P450s identifies 8-methoxypsoralen as a substrate for the CYP82C subfamily, yielding original chemical structures.
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.
Kruse
Tanya
T
Ceres, Inc., 1535 Rancho Conejo Boulevard, Thousand Oaks, CA 91320, USA. tkruse@ceres-inc.com <tkruse@ceres-inc.com>
Ho
Kwongling
K
Yoo
Hye-Dong
HD
Johnson
Thomas
T
Hippely
Matt
M
Park
Joon-Hyun
JH
Flavell
Richard
R
Bobzin
Steve
S
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
D003577:Cytochrome P-450 Enzyme System; D008730:Methoxsalen