Construction of cell factories for high production of ginsenoside Rh_2 in Saccharomyces cerevisiae.
10.19540/j.cnki.cjcmm.20210914.101
- Author:
Yu-Song SHI
1
;
Dong WANG
2
;
Rong-Sheng LI
2
;
Xue-Li ZHANG
2
;
Zhu-Bo DAI
2
Author Information
1. School of Biology and Biological Engineering, South China University of Technology Guangzhou 510006, China Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences Tianjin 300308, China Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences Tianjin 300308, China.
2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences Tianjin 300308, China Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences Tianjin 300308, China.
- Publication Type:Journal Article
- Keywords:
Saccharomyces cerevisiae;
ginsenoside Rh_2;
synthetic biology;
triterpenoid saponin
- MeSH:
Fermentation;
Ginsenosides;
Humans;
Panax/genetics*;
Panax notoginseng;
Saccharomyces cerevisiae/genetics*;
Uridine Diphosphate Glucose
- From:
China Journal of Chinese Materia Medica
2022;47(3):651-658
- CountryChina
- Language:Chinese
-
Abstract:
Ginsenoside Rh_2 is a rare active ingredient in precious Chinese medicinal materials such as Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Panacis Quinquefolii Radix. It has important pharmacological activities such as anti-cancer and improving human immunity. However, due to the extremely low content of ginsenoside Rh_2 in the source plants, the traditional way of obtaining it has limitations. This study intended to apply synthetic biological technology to develop a cell factory of Saccharomyces cerevisiae to produce Rh_2 by low-cost fermentation. First, we used the high protopanaxadiol(PPD)-yielding strain LPTA as the chassis strain, and inserted the Panax notoginseng enzyme gene Pn1-31, together with yeast UDP-glucose supply module genes[phosphoglucose mutase 1(PGM1), α-phosphoglucose mutase(PGM2), and uridine diphosphate glucose pyrophosphorylase(UGP1)], into the EGH1 locus of yeast chromosome. The engineered strain LPTA-RH2 produced 17.10 mg·g~(-1) ginsenoside Rh_2. This strain had low yield of Rh_2 while accumulated much precursor PPD, which severely restricted the application of this strain. In order to further improve the production of ginsenoside Rh_2, we strengthened the UDP glucose supply module and ginsenoside Rh_2 synthesis module by engineered strain LPTA-RH2-T. The shaking flask yield of ginsenoside Rh_2 was increased to 36.26 mg·g~(-1), which accounted for 3.63% of the dry weight of yeast cells. Compared with those of the original strain LPTA-RH2, the final production and the conversion efficiency of Rh_2 increased by 112.11% and 65.14%, respectively. This study provides an important basis for further obtaining the industrial-grade cell factory for the production of ginsenoside Rh_2.
