Regulation of β-mercuryl alcohol metabolic flow in Saccharomyces cerevisiae cells.
10.19540/j.cnki.cjcmm.20200506.112
- Author:
Er-Kun CHAO
1
;
Guang-Tao QIAN
1
;
Meng-Chu SUN
2
;
Xin-Yao SU
2
;
Zhi-Hui ZHU
1
;
Wei SHENG
3
;
Cai-Xia WANG
2
;
Jian-Ping XUE
3
Author Information
1. College of Life Sciences, Huaibei Normal University Huaibei 235000, China Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
2. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
3. College of Life Sciences, Huaibei Normal University Huaibei 235000, China.
- Publication Type:Journal Article
- Keywords:
CRISPR/CAS9;
Saccharomyces cerevisiae;
synthetic biology;
β-amyrin
- MeSH:
Ethanol;
Fermentation;
Metabolic Engineering;
Saccharomyces cerevisiae;
genetics
- From:
China Journal of Chinese Materia Medica
2020;45(16):3819-3825
- CountryChina
- Language:Chinese
-
Abstract:
In this study, citrate synthase gene(CIT2), and malate synthase gene(MLS1) were successfully knocked out in β-amyrin-producing yeast cells by using CRISPR/CAS9. The promoter of phosphoglucose isomerase gene(PGI1) was replaced by that of cytochrome c oxidase subunit Ⅶa(Cox9)to weaken its expression, aiming to channel more carbon flux into the NADPH-producing pathway. The fermentation results showed that CIT2 deletion had no effect on the β-amyrin production. Compared with the control strain, the production of β-amyrin was increased by 1.85 times after deleting MLS1, reaching into 3.3 mg·L~(-1). By replacing the promoter of PGI1, the β-amyrin yield was 3.75 times higher than that of the control strain, reaching up to 6.7 mg·L~(-1). This study successfully knocked out the CITT2 and MLS1 genes and weakened the PGI1 gene by using CRISPR/CAS9, which directly influenced the production of β-amyrin and provided some reference for the the metabolic engineering of triterpernoid producing strain.
