Functional characterization of CYP81C16 involved in the tanshinone biosynthetic pathway in Salvia miltiorrhiza  

作　　者：REN Li LUO Linglong HU Zhimin MA Ying WANG Jian CHENG Yatian JIN Baolong CHEN Tong TANG Jinfu CUI Guanghong GUO Juan HUANG Luqi 

机构地区：[1]State Key Laboratory of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100000,China

出　　处：《Chinese Journal of Natural Medicines》2023年第12期938-949,共12页中国天然药物（英文版）

基　　金：This work was supported by the National Key R&D Program of China(Nos.2020YFA0908000,2018YFA0900600);the National Natural Science Foundation of China(Nos.82003904,81822046);the Fundamental Research Funds for the Central public welfare research institutes(No.ZZ13-YQ-083);a Scientific and Technological Innovation Project of China Academy of Chinese Medical Sciences(No.CI2021A04110);Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(No.ZYYCXTD-D-202005);a key project at central government level(the ability to establish sustainable use of valuable Chinese medicine resources;2060302,China).

摘　　要：Danshen,the dried roots and rhizomes of Salvia miltiorrhiza Bunge(S.miltiorrhiza),is widely used in the treatment of cardiovascular and cerebrovascular diseases.Tanshinones,the bioactive compounds from Danshen,exhibit a wide spectrum of pharmacological properties,suggesting their potential for future therapeutic applications.Tanshinone biosynthesis is a complex process involving at least six P450 enzymes that have been identified and characterized,most of which belong to the CYP76 and CYP71 families.In this study,CYP81C16,a member of the CYP71 clan,was identified in S.miltiorrhiza.An in vitro assay revealed that it could catalyze the hydroxylation of four para-quinone-type tanshinones,namely neocryptotanshinone,deoxyneocryptotanshinone,and danshenxinkuns A and B.SmCYP81C16 emerged as a potential broad-spectrum oxidase targeting the C-18 position of para-quinone-type tanshinones with an impressive relative conversion rate exceeding 90%.Kinetic evaluations and in vivo assays underscored its highest affinity towards neocryptotanshinone among the tested substrates.The overexpression of SmCYP81C16 promoted the accumulation of(iso)tanshinone in hairy root lines.The characterization of SmCYP81C16 in this study accentuates its potential as a pivotal tool in the biotechnological production of tanshinones,either through microbial or plant metabolic engineering.

关 键 词：Salvia miltiorrhiza TANSHINONE Biosynthetic pathway Cytochrome P450 HYDROXYLATION Neocryptotanshinone 

分 类 号：R284[医药卫生—中药学]