title: Production of bioactive ginsenosides Rh2 and Rg3 by metabolically engineered yeasts.
abstract: Ginsenosides Rh2 and Rg3 represent promising candidates for cancer prevention and therapy and have low toxicity. However, the concentrations of Rh2 and Rg3 are extremely low in the bioactive constituents (triterpene saponins) of ginseng. Despite the available heterologous biosynthesis of their aglycone (protopanaxadiol, PPD) in yeast, production of Rh2 and Rg3 by a synthetic biology approach was hindered by the absence of bioparts to glucosylate the C3 hydroxyl of PPD. In this study, two UDP-glycosyltransferases (UGTs) were cloned and identified from Panax ginseng. UGTPg45 selectively transfers a glucose moiety to the C3 hydroxyl of PPD and its ginsenosides. UGTPg29 selectively transfers a glucose moiety to the C3 glucose of Rh2 to form a 1-2-glycosidic bond. Based on the two UGTs and a yeast chassis to produce PPD, yeast cell factories were built to produce Rh2 and/or Rg3 from glucose. The turnover number (kcat) of UGTPg29 was more than 2500-fold that of UGTPg45, which might explain the higher Rg3 yield than that of Rh2 in the yeast cell factories. Building yeast cell factories to produce Rh2 or Rg3 from simple sugars by microbial fermentation provides an alternative approach to replace the traditional method of extracting ginsenosides from Panax plants.
authors.affiliation: CAS-Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, China.
authors.forename: Pingping
authors.initials: P
authors.lastname: Wang
authors.affiliation: CAS-Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, China.
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Wei
authors.affiliation: CAS-Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, China; State Key Laboratory of Genetic Engineering, Department of Microbiology, School of Life Sciences and Institute of Biomedical Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Fan
authors.affiliation: State Key Laboratory of Drug Research, Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 201203, China.
authors.forename: Qunfang
authors.initials: Q
authors.lastname: Liu
authors.affiliation: CAS-Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.affiliation: CAS-Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, China.
authors.forename: Chengshuai
authors.initials: C
authors.lastname: Yang
authors.affiliation: CAS-Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.affiliation: CAS-Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, China; State Key Laboratory of Genetic Engineering, Department of Microbiology, School of Life Sciences and Institute of Biomedical Sciences, Fudan University, Shanghai 200433, China.
authors.forename: Guoping
authors.initials: G
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Drug Research, Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 201203, China. Electronic address: jmyue@mail.shcnc.ac.cn.
authors.forename: Jianmin
authors.initials: J
authors.lastname: Yue
authors.affiliation: CAS-Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, China. Electronic address: yanxing@sibs.ac.cn.
authors.forename: Xing
authors.initials: X
authors.lastname: Yan
authors.affiliation: CAS-Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, China. Electronic address: zhouzhihua@sippe.ac.cn.
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Zhou
mesh_terms: D036145:Ginsenosides / Q000096:biosynthesis / Q000235:genetics; D005964:Glucosyltransferases / Q000096:biosynthesis / Q000235:genetics; D060847:Metabolic Engineering; D005894:Panax / Q000201:enzymology / Q000235:genetics*; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism
chemical_list: D036145:Ginsenosides; D010940:Plant Proteins; C097367:ginsenoside Rg3; C055305:ginsenoside Rh2; D005964:Glucosyltransferases
keywords: Ginsenoside Rg3; Ginsenoside Rh2; Panax plants; Protopanaxadiol producing chassis; UDP-glycosyltransferase