Characterization of Panax ginseng UDP-Glycosyltransferases Catalyzing Protopanaxatriol and Biosyntheses of Bioactive Ginsenosides F1 and Rh1 in Metabolically Engineered Yeasts

doi:10.1016/j.molp.2015.05.010

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	Characterization of Panax ginseng UDP-Glycosyltransferases Catalyzing Protopanaxatriol and Biosyntheses of Bioactive Ginsenosides F1 and Rh1 in Metabolically Engineered Yeasts
	Wei, Wei 2; Wang, Pingping 2; Wei, Yongjun 2; Liu, Qunfang1 ; Yang, Chengshuai 2; Zhao, Guoping 2; Yue, Jianmin1 ; Yan, Xing 2; Zhou, Zhihua 2
刊名	MOLECULAR PLANT
	2015-09-07
卷号	8 期号:9 页码:1412-1424
关键词	UDP-glycosyltransferase triterpenoids protopanaxatriol ginsenoside F1 ginsenoside Rh1 Panax ginseng
ISSN号	1674-2052
DOI	10.1016/j.molp.2015.05.010
文献子类	Article
英文摘要	Ginsenosides, the main pharmacologically active natural compounds in ginseng (Panax ginseng), are mostly the glycosylated products of protopanaxadiol (PPD) and protopanaxatriol (PPT). No uridine diphosphate glycosyltransferase (UGT), which catalyzes PPT to produce PPT-type ginsenosides, has yet been reported. Here, we show that UGTPg1, which has been demonstrated to regio-specifically glycosylate the C20-OH of PPD, also specifically glycosylates the C20-OH of PPT to produce bioactive ginsenoside F1. We report the characterization of four novel UGT genes isolated from P. ginseng, sharing high deduced amino acid identity (>84%) with UGTPg1. We demonstrate that UGTPg100 specifically glycosylates the C6-OH of PPT to produce bioactive ginsenoside Rh1, and UGTPg101 catalyzes PPT to produce F1, followed by the generation of ginsenoside Rg1 from F1. However, UGTPg102 and UGTPg103 were found to have no detectable activity on PPT. Through structural modeling and site-directed mutagenesis, we identified several key amino acids of these UGTs that may play important roles in determining their activities and substrate regio-specificities. Moreover, we constructed yeast recombinants to biosynthesize F1 and Rh1 by introducing the genetically engineered PPT-producing pathway and UGTPg1 or UGTPg100. Our study reveals the possible biosynthetic pathways of PPT-type ginsenosides in Panax plants, and provides a sound manufacturing approach for bioactive PPT-type ginsenosides in yeast via synthetic biology strategies.
资助项目	National Basic Research Program of China (973 program)[2012CB721103] ; National Basic Research Program of China (973 program)[2015CB755703] ; Science and Technology Commission of Shanghai Municipality[14JC1407000]
WOS关键词	MEDICAGO-TRUNCATULA ; CRYSTAL-STRUCTURES ; ANTHOCYANIN BIOSYNTHESIS ; ARABIDOPSIS-THALIANA ; CLITORIA-TERNATEA ; DAMMARENEDIOL-II ; GLUCOSYLTRANSFERASE ; TRANSCRIPTOME ; SYNTHASE ; REVEALS
WOS研究方向	Biochemistry & Molecular Biology ; Plant Sciences
语种	英语
出版者	CELL PRESS
WOS记录号	WOS:000360980300010
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/276401]
专题	天然药物化学研究室中科院受体结构与功能重点实验室新药研究国家重点实验室
通讯作者	Zhou, Zhihua
作者单位	1.Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Mat Med, State Key Lab Drug Res, Shanghai 201203, Peoples R China 2.Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Plant Physiol & Ecol, CAS Key Lab Synthet Biol, Shanghai 200032, Peoples R China;
推荐引用方式 GB/T 7714	Wei, Wei,Wang, Pingping,Wei, Yongjun,et al. Characterization of Panax ginseng UDP-Glycosyltransferases Catalyzing Protopanaxatriol and Biosyntheses of Bioactive Ginsenosides F1 and Rh1 in Metabolically Engineered Yeasts[J]. MOLECULAR PLANT,2015,8(9):1412-1424.
APA	Wei, Wei.,Wang, Pingping.,Wei, Yongjun.,Liu, Qunfang.,Yang, Chengshuai.,...&Zhou, Zhihua.(2015).Characterization of Panax ginseng UDP-Glycosyltransferases Catalyzing Protopanaxatriol and Biosyntheses of Bioactive Ginsenosides F1 and Rh1 in Metabolically Engineered Yeasts.MOLECULAR PLANT,8(9),1412-1424.
MLA	Wei, Wei,et al."Characterization of Panax ginseng UDP-Glycosyltransferases Catalyzing Protopanaxatriol and Biosyntheses of Bioactive Ginsenosides F1 and Rh1 in Metabolically Engineered Yeasts".MOLECULAR PLANT 8.9(2015):1412-1424.