Discovery of a non-stereoselective cytochrome P450 catalyzing either 8 alpha- or 8 beta-hydroxylation of germacrene A acid from the Chinese medicinal plant, Inula hupehensis

doi:10.1111/tpj.13760

CORC > 武汉植物园 > 中国科学院武汉植物园

	Discovery of a non-stereoselective cytochrome P450 catalyzing either 8 alpha- or 8 beta-hydroxylation of germacrene A acid from the Chinese medicinal plant, Inula hupehensis
	Gou, Junbo 1,2; Hao, Fuhua 3; Huang, Chongyang 3; Kwon, Moonhyuk 4; Chen, Fangfang 2; Li, Changfu 2; Liu, Chaoyang 3; Ro, Dae-Kyun 4; Tang, Huiru 3,5; Zhang, Yansheng 2
刊名	PLANT JOURNAL
	2018
卷号	93 期号:1 页码:92-106
关键词	C12 8-fused STLs cytochrome P450 (CYP) germacrene A acid C8-hydroxylase Inula hupehensis inunolide sesquiterpene lactones (STLs)
ISSN号	0960-7412
DOI	10.1111/tpj.13760
英文摘要	Sesquiterpene lactones (STLs) are C15 terpenoid natural products with alpha-methylene gamma-lactone moiety. A large proportion of STLs in Asteraceae species is derived from the central precursor germacrene A acid (GAA). Formation of the lactone rings depends on the regio-(C6 or C8) and stereoselective (alpha- or beta-)hydroxylations of GAA, producing STLs with four distinct stereo-configurations (12,6 alpha-, 12,6-, 12, 8 alpha-, and 12,8 beta-olide derivatives of GAA) in nature. Curiously, two configurations of STLs (C12,8 alpha and C12,8 beta) are simultaneously present in the Chinese medicinal plant, Inula hupehensis. However, how these related yet distinct STL stereo-isomers are co-synthesized in I. hupehensis remains unknown. Here, we describe the functional identification of the I. hupehensis cytochrome P450 (CYP71BL6) that can catalyze the hydroxylation of GAA in either 8 alpha-or 8 beta-configuration, resulting in the synthesis of both 8 alpha- and 8 beta-hydroxyl GAAs. Of these two products, only 8 alpha-hydroxyl GAA spontaneously lactonizes to the C12,8 alpha-STL while the 8 beta-hydroxyl GAA remains stable without lactonization. Chemical structures of the C12,8 alpha-STL, named inunolide, and 8 beta-hydroxyl GAA were fully elucidated by nuclear magnetic resonance analysis and mass spectrometry. The CYP71BL6 displays 63-66% amino acid identity to the previously reported CYP71BL1/2 catalyzing GAA 6aor 8 beta-hydroxylation, indicating CYP71BL6 shares the same evolutionary lineage with other stereoselective cytochrome P450s, but catalyzes hydroxylation in a non-stereoselective manner. We observed that the CYP71BL6 transcript abundance correlates closely to the accumulation of C12,8-STLs in I. hupehensis. The identification of CYP71BL6 provides an insight into the biosynthesis of STLs in Asteraceae.
资助项目	National Natural Science Foundation of China[31370339] ; National Natural Science Foundation of China[81230090] ; National Natural Science Foundation of China[81520108030] ; Ministry of Science and Technology of China[2011YQ120035] ; Natural Sciences and Engineering Research Council of Canada (NSERC) ; Next-Generation Bio-Green 21 Program (SSAC) from Rural Development Administration, Republic of Korea[PJ01105501] ; Basic Science Research Programme through the National Research Foundation of Korea (NRF) - Ministry of Education[2017R1A6A3A03003409]
WOS研究方向	Plant Sciences
语种	英语
出版者	WILEY
WOS记录号	WOS:000418228000008
内容类型	期刊论文
源URL	[http://202.127.146.157/handle/2RYDP1HH/3366]
专题	中国科学院武汉植物园
通讯作者	Tang, Huiru; Zhang, Yansheng
作者单位	1.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 2.Chinese Acad Sci, Wuhan Bot Garden, CAS Key Lab Plant Germplasm Enhancement & Special, Wuhan 430074, Hubei, Peoples R China 3.Univ Chinese Acad Sci, Wuhan Inst Phys & Math, Natl Ctr Magnet Resonance Wuhan,Key Lab Magnet Re, State Key Lab Magnet Resonance & Atom & Mol Phys, Wuhan 430071, Hubei, Peoples R China 4.Univ Calgary, Dept Biol Sci, 2500 Univ Dr NW, Calgary, AB T2N 1N4, Canada 5.Fudan Univ, Collaborat Innovat Ctr Genet & Dev, State Key Lab Genet Engn, Shanghai 200438, Peoples R China
推荐引用方式 GB/T 7714	Gou, Junbo,Hao, Fuhua,Huang, Chongyang,et al. Discovery of a non-stereoselective cytochrome P450 catalyzing either 8 alpha- or 8 beta-hydroxylation of germacrene A acid from the Chinese medicinal plant, Inula hupehensis[J]. PLANT JOURNAL,2018,93(1):92-106.
APA	Gou, Junbo.,Hao, Fuhua.,Huang, Chongyang.,Kwon, Moonhyuk.,Chen, Fangfang.,...&Zhang, Yansheng.(2018).Discovery of a non-stereoselective cytochrome P450 catalyzing either 8 alpha- or 8 beta-hydroxylation of germacrene A acid from the Chinese medicinal plant, Inula hupehensis.PLANT JOURNAL,93(1),92-106.
MLA	Gou, Junbo,et al."Discovery of a non-stereoselective cytochrome P450 catalyzing either 8 alpha- or 8 beta-hydroxylation of germacrene A acid from the Chinese medicinal plant, Inula hupehensis".PLANT JOURNAL 93.1(2018):92-106.