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.
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Acknowledgments
The authors acknowledge the financial support provided by the National Basic Research Program of China (“973” Program: 2012CB721105) and the Ministry of Science and Technology of China (“863” Program: 2012AA02A701).
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Supplementary Table 1—Plasmids and strains used in this study.
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Dashuai Li and Qiang Zhang have contributed equally to this work.
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Li, D., Zhang, Q., Zhou, Z. et al. Heterologous biosynthesis of triterpenoid dammarenediol-II in engineered Escherichia coli . Biotechnol Lett 38, 603–609 (2016). https://doi.org/10.1007/s10529-015-2032-9
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DOI: https://doi.org/10.1007/s10529-015-2032-9