Kruse Tanya, Ho Kwongling, Yoo Hye-Dong, Johnson Thomas, Hippely Matt, Park Joon-Hyun, Flavell Richard, Bobzin Steve
Ceres, Inc., 1535 Rancho Conejo Boulevard, Thousand Oaks, CA 91320, USA.
Chem Biol. 2008 Feb;15(2):149-56. doi: 10.1016/j.chembiol.2008.01.008.
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.
已开发出一种体内植物筛选方法，可用于分析外源施加的底物对过表达单个细胞色素P450的转基因拟南芥品系的作用。通过对91种P450的一个子集进行这种筛选，我们鉴定出了CYP82C亚家族成员的一种原始底物。治疗性化合物8-甲氧基补骨脂素被过表达CYP82C2或CYP82C4的植物羟基化，形成5-羟基-8-甲氧基补骨脂素。此外，植物进一步修饰该产物以产生一种糖基化化合物，可能是化合物5-O-β-D-吡喃葡萄糖基-8-甲氧基补骨脂素。治疗性化合物加合物的发现证明了这种生物催化筛选方法在创造可能具有药理价值的化合物方面的潜力。此外，该平台提供了一种扩展P450酶/底物组合一般知识库的方法，并可能为大量生物催化和生物修复过程提供有价值的工具。
