Cytochrome P450 promiscuity leads to a bifurcating biosynthetic pathway for tanshinones

doi:10.1111/nph.13790

	Cytochrome P450 promiscuity leads to a bifurcating biosynthetic pathway for tanshinones
	Cui, Guanghong 1; 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
刊名	NEW PHYTOLOGIST
	2016-04-01
卷号	210 期号:2 页码:525-534
关键词	Cytochrome P450 (Cyp) Monooxygenases Diterpenoid Biosynthesis Enzymatic Promiscuity Metabolic Pathways Salvia Miltiorrhiza Bunge Synthetic Biology
ISSN号	0028-646X
DOI	10.1111/nph.13790
文献子类	Article
英文摘要	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.
WOS关键词	SALVIA-MILTIORRHIZA ; SACCHAROMYCES-CEREVISIAE ; ARABIDOPSIS-THALIANA ; YEAST EXPRESSION ; GENE-CLUSTER ; P450 ; MILTIRADIENE ; METABOLISM ; CULTURES ; CLONING
WOS研究方向	Plant Sciences
语种	英语
出版者	WILEY-BLACKWELL
WOS记录号	WOS:000373380700017
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/170646]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者	Huang, Luqi
作者单位	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
推荐引用方式 GB/T 7714	Cui, Guanghong,Guo, Juan,Ma, Xiaohui,et al. Cytochrome P450 promiscuity leads to a bifurcating biosynthetic pathway for tanshinones[J]. NEW PHYTOLOGIST,2016,210(2):525-534.
APA	Cui, Guanghong.,Guo, Juan.,Ma, Xiaohui.,Cai, Yuan.,Ma, Ying.,...&Huang, Luqi.(2016).Cytochrome P450 promiscuity leads to a bifurcating biosynthetic pathway for tanshinones.NEW PHYTOLOGIST,210(2),525-534.
MLA	Cui, Guanghong,et al."Cytochrome P450 promiscuity leads to a bifurcating biosynthetic pathway for tanshinones".NEW PHYTOLOGIST 210.2(2016):525-534.