Study of heterologous efficient synthesis of β-amyrin and high-density fermentation.
10.19540/j.cnki.cjcmm.20190129.011
- Author:
Meng-Chu SUN
1
;
Er-Kun CHAO
1
;
Xin-Yao SU
2
;
Min ZHU
3
;
Yong SU
1
;
Guang-Tao QIAN
1
;
Shi-Lin CHEN
2
;
Cai-Xia WANG
2
;
Jian-Ping XUE
4
Author Information
1. College of Life Sciences, Huaibei Normal University Huaibei 235000, China Institute of Chinese Medicine,China Academy of Chinese Medical Sciences Beijing 100700, China.
2. Institute of Chinese Medicine,China Academy of Chinese Medical Sciences Beijing 100700, China.
3. Institute of International Traditional Chinese Medicine Translational Medicine, Guangzhou University of Chinese Medicine Guangzhou 510000, China.
4. College of Life Sciences, Huaibei Normal University Huaibei 235000, China.
- Publication Type:Journal Article
- Keywords:
Saccharomyces cerevisiae;
oleanane-type triterpene saponins;
β-amyrin
- MeSH:
Fermentation;
Glycyrrhiza uralensis;
enzymology;
genetics;
Industrial Microbiology;
Intramolecular Transferases;
genetics;
Metabolic Engineering;
Oleanolic Acid;
analogs & derivatives;
biosynthesis;
Saccharomyces cerevisiae;
metabolism
- From:
China Journal of Chinese Materia Medica
2019;44(7):1341-1349
- CountryChina
- Language:Chinese
-
Abstract:
In this study, the synthetic pathway of β-amyrin was constructed in the pre-constructed Saccharomyces cerevisiae chassis strain Y0 by introducing β-amyrin synthase from Glycyrrhiza uralensis, resulting strain Y1-C20-6, which successfully produced β-amyrin up to 5.97 mg·L~(-1). Then, the mevalonate pyrophosphate decarboxylase gene(ERG19), mevalonate kinase gene(ERG12), 3-hydroxy-3-methylglutaryl-CoA synthase gene(ERG13), phosphomevalonate kinase gene(ERG8) and IPP isomerase gene(IDI1)were overexpressed to promoted the metabolic fluxto the direction of β-amyrin synthesis for further improving β-amyrin production, resulting the strain Y2-C2-4 which produced β-amyrin of 10.3 mg·L~(-1)under the shake flask fermentation condition. This is 100% higher than that of strain Y1-C20-6, illustrating the positive effect of the metabolic engineering strategy applied in this study. The titer of β-amyrin was further improved up to 157.4 mg·L~(-1) in the fed-batch fermentation, which was almost 26 fold of that produced by strain Y1-C20-6. This study not only laid the foundation for the biosynthesis of β-amyrin but also provided a favorable chassis strain for elucidation of cytochrome oxidases and glycosyltransferases of β-amyrin-based triterpenoids.
