文章基本信息

标题：Probing the promiscuity of ent-kaurene oxidases via combinatorial biosynthesis
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作者：Sibongile Mafu ; Meirong Jia ; Jiachen Zi 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2016
卷号：113
期号：9
页码：2526-2531
DOI：10.1073/pnas.1512096113
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：The substrate specificity of enzymes from natural products’ metabolism is a topic of considerable interest, with potential biotechnological use implicit in the discovery of promiscuous enzymes. However, such studies are often limited by the availability of substrates and authentic standards for identification of the resulting products. Here, a modular metabolic engineering system is used in a combinatorial biosynthetic approach toward alleviating this restriction. In particular, for studies of the multiply reactive cytochrome P450, ent-kaurene oxidase (KO), which is involved in production of the diterpenoid plant hormone gibberellin. Many, but not all, plants make a variety of related diterpenes, whose structural similarity to ent-kaurene makes them potential substrates for KO. Use of combinatorial biosynthesis enabled analysis of more than 20 such potential substrates, as well as structural characterization of 12 resulting unknown products, providing some insight into the underlying structure–function relationships. These results highlight the utility of this approach for investigating the substrate specificity of enzymes from complex natural products’ biosynthesis.
关键词：labdane-related diterpenoids ; cytochrome P450 monooxygenases ; natural products ; enzyme specificity ; gibberellins