Cytochrome P450 promiscuity leads to a bifurcating biosynthetic pathway for tanshinones

被引：210

作者：

Guo, Juan ^{[1
]}

Ma, Xiaohui ^{[1
,2
]}

Cai, Yuan ^{[1
,3
]}

Ma, Ying ^{[1
]}

Zhan, Zhilai ^{[1
]}

Zhou, Yongjin J. ^{[3
]}

Liu, Wujun ^{[3
]}

Guan, Mengxin ^{[4
]}

Yang, Jian ^{[1
]}

Cui, Guanghong ^{[1
]}

Kang, Liping ^{[1
]}

Yang, Lei ^{[5
]}

Shen, Ye ^{[1
]}

Tang, Jinfu ^{[1
]}

Lin, Huixin ^{[1
]}

Ma, Xiaojing ^{[1
]}

Jin, Baolong ^{[1
]}

Liu, Zhenming ^{[4
]}

Peters, Reuben J. ^{[6
]}

Zhao, Zongbao K. ^{[3
]}

Huang, Luqi ^{[1
]}

机构：

[1] China Acad Chinese Med Sci, Natl Resource Ctr Chinese Mat Med, State Key Lab Breeding Base Daodi Herbs, Beijing 100700, Peoples R China

[2] Chengdu Univ Tradit Chinese Med, Coll Pharm, Chengdu 611137, Peoples R China

[3] Chinese Acad Sci, Dalian Inst Chem Phys, Div Biotechnol, Dalian 116023, Peoples R China

[4] Peking Univ, Sch Pharmaceut Sci, State Key Lab Nat & Biomimet Drugs, Beijing 100191, Peoples R China

[5] Shanghai Chenshan Bot Garden, Plant Sci Res Ctr, Shanghai Key Lab Plant Funct Genom & Resources, Shanghai 201602, Peoples R China

[6] Iowa State Univ, Roy J Carver Dept Biochem Biophys & Mol Biol, Ames, IA 50011 USA

来源：

NEW PHYTOLOGIST | 2016年 / 210卷 / 02期

基金：

美国国家卫生研究院; 中国国家自然科学基金;

关键词：

cytochrome P450 (CYP) monooxygenases; diterpenoid biosynthesis; enzymatic promiscuity; metabolic pathways; Salvia miltiorrhiza Bunge; synthetic biology; SALVIA-MILTIORRHIZA; YEAST EXPRESSION; P450; MILTIRADIENE; CLONING; SMCPS;

D O I：

10.1111/nph.13790

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

Cytochromes P450 (CYPs) play a key role in generating the structural diversity of terpenoids, the largest group of plant natural products. However, functional characterization of CYPs has been challenging because of the expansive families found in plant genomes, diverse reactivity and inaccessibility of their substrates and products. Here we present the characterization of two CYPs, CYP76AH3 and CYP76AK1, which act sequentially to form a bifurcating pathway for the biosynthesis of tanshinones, the oxygenated diterpenoids from the Chinese medicinal plant Danshen (Salvia miltiorrhiza). These CYPs had similar transcription profiles to that of the known gene responsible for tanshinone production in elicited Danshen hairy roots. Biochemical and RNA interference studies demonstrated that both CYPs are promiscuous. CYP76AH3 oxidizes ferruginol at two different carbon centers, and CYP76AK1 hydroxylates C-20 of two of the resulting intermediates. Together, these convert ferruginol into 11,20-dihydroxy ferruginol and 11,20-dihydroxy sugiol en route to tanshinones. Moreover, we demonstrated the utility of these CYPs by engineering yeast for heterologous production of six oxygenated diterpenoids, which in turn enabled structural characterization of three novel compounds produced by CYP-mediated oxidation. Our results highlight the incorporation of multiple CYPs into diterpenoid metabolic engineering, and a continuing trend of CYP promiscuity generating complex networks in terpenoid biosynthesis.

引用

页码：525 / 534

页数：10

共 42 条

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