Optimization of UDP-glucose supply module and production of ginsenoside F1 in Saccharomyces cerevisiae.
10.19540/j.cnki.cjcmm.20190829.101
- Author:
Jin-He WANG
1
;
Dong WANG
2
;
Wei-Xian LI
2
;
Ying HUANG
2
;
Zhu-Bo DAI
2
;
Xue-Li ZHANG
2
Author Information
1. College of Biotechnology,Tianjin University of Science & Technology Tianjin 300457,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:
Sacchromyces cerevisiae;
UDP-glucose;
ginsenoside F1;
synthetic biology
- MeSH:
Ginsenosides/metabolism*;
Glucose;
Panax;
Saccharomyces cerevisiae/metabolism*;
Uridine Diphosphate Glucose
- From:
China Journal of Chinese Materia Medica
2019;44(21):4596-4604
- CountryChina
- Language:Chinese
-
Abstract:
Ginsenoside F1 is a rare ginsenoside in medicinal plants such as Panax ginseng,P. notogingseng and P. quinquefolius. It has strong pharmacological activities of anti-tumor,anti-oxidation and anti-aging. In order to directly produce ginsenoside F1 by using inexpensive raw materials such as glucose,we integrated the codon-optimized P.ginseng dammarenediol-Ⅱ synthase(Syn Pg DDS),P.ginseng protopanaxadiol synthase(Syn Pg PPDS),P. ginseng protopanaxatriol synthase(Syn Pg PPTS) genes and Arabidopsis thaliana cytochrome P450 reductase(At CPR1) gene into triterpene chassis strain BY-T3. The transformant BY-PPT can produce protopanaxatriol. Then we integrated the Sacchromyces cerevisiae phosphoglucomutase 1(PGM1),phosphoglucomutase 2(PGM2) and UDP-glucose pyrophosphorylase 1(UGP1) genes into chassis strain BY-PPT. The UDP-glucose supply module increased UDP-glucose production by 8. 65 times and eventually reached to 44. 30 mg·L-1 while confirmed in the transformant BY-PPT-GM. Next,we integrated the UDPglucosyltransferase Pg3-29 gene which can catalyze protopanaxatriol to produce ginsenoside F1 into chassis strain BY-PPT-GM. The transformant BY-F1 produced a small amount of ginsenoside F1 which was measured as 0. 5 mg·L-1. After the fermentation process was optimized,the titer of ginsenoside F1 could be increased by 900 times to 450. 5 mg·L-1. The high-efficiency UDP-glucose supply module in this study can provide reference for the construction of cell factories for production of saponin,and provide an important basis for further obtaining high-yield ginsenoside yeast cells.
