Dashuai Li1, Qiang Zhang1, Zhijiang Zhou1, Fanglong Zhao1, Wenyu Lu2,3,4. 1. School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People's Republic of China. 2. School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People's Republic of China. wenyulu@tju.edu.cn. 3. Key Laboratory of System Bioengineering (Tianjin University), Ministry of Education, Tianjin, 300072, People's Republic of China. wenyulu@tju.edu.cn. 4. SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin, 300072, People's Republic of China. wenyulu@tju.edu.cn.
Abstract
OBJECTIVES: To achieve heterologous biosynthesis of dammarenediol-II, which is the precursor of dammarane-type tetracyclic ginsenosides, by reconstituting the 2,3-oxidosqualene-derived triterpenoid biosynthetic pathway in Escherichia coli. RESULTS: By the strategy of synthetic biology, dammarenediol-II biosynthetic pathway was reconstituted in E. coli by co-expression of squalene synthase (SS), squalene epoxidase (SE), NADPH-cytochrome P450 reductase (CPR) from Saccharomyces cerevisiae, and SE from Methylococcus capsulatus (McSE), NADPH-cytochrome P450 reductase (CPR) from Arabidopsis thaliana. Sequences of transmembrane domains were truncated if necessary in each of the genes. Different sources of SE/CPR combinations were tested, during which two CPRs were detected to be new reductase partners of McSE. When the gene encoding dammarenediol-II synthase was co-expressed with the 2,3-oxidosqualene expression modules, dammarenediol-II was detected and the production was 8.63 mg l(-1) in E. coli under the shake-flask conditions. CONCLUSIONS: Two E. coli chassis for production of dammarenediol-II were established which could be potentially applied in other triterpenoid production in E. coli when different oxidosqualene cyclases (OSCs) introduced into the system.
OBJECTIVES: To achieve heterologous biosynthesis of dammarenediol-II, which is the precursor of dammarane-type tetracyclicginsenosides, by reconstituting the 2,3-oxidosqualene-derived triterpenoid biosynthetic pathway in Escherichia coli. RESULTS: By the strategy of synthetic biology, dammarenediol-II biosynthetic pathway was reconstituted in E. coli by co-expression of squalene synthase (SS), squalene epoxidase (SE), NADPH-cytochrome P450 reductase (CPR) from Saccharomyces cerevisiae, and SE from Methylococcus capsulatus (McSE), NADPH-cytochrome P450 reductase (CPR) from Arabidopsis thaliana. Sequences of transmembrane domains were truncated if necessary in each of the genes. Different sources of SE/CPR combinations were tested, during which two CPRs were detected to be new reductase partners of McSE. When the gene encoding dammarenediol-II synthase was co-expressed with the 2,3-oxidosqualene expression modules, dammarenediol-II was detected and the production was 8.63 mg l(-1) in E. coli under the shake-flask conditions. CONCLUSIONS: Two E. coli chassis for production of dammarenediol-II were established which could be potentially applied in other triterpenoid production in E. coli when different oxidosqualene cyclases (OSCs) introduced into the system.
Authors: Anita Loeschcke; Dennis Dienst; Vera Wewer; Jennifer Hage-Hülsmann; Maximilian Dietsch; Sarah Kranz-Finger; Vanessa Hüren; Sabine Metzger; Vlada B Urlacher; Tamara Gigolashvili; Stanislav Kopriva; Ilka M Axmann; Thomas Drepper; Karl-Erich Jaeger Journal: PLoS One Date: 2017-12-27 Impact factor: 3.240