title: Heterologous biosynthesis of triterpenoid dammarenediol-II in engineered Escherichia coli.
abstract: OBJECTIVE
  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.
authors.affiliation: School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People's Republic of China.
authors.forename: Dashuai
authors.initials: D
authors.lastname: Li
authors.affiliation: School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People's Republic of China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People's Republic of China.
authors.forename: Zhijiang
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People's Republic of China.
authors.forename: Fanglong
authors.initials: F
authors.lastname: Zhao
authors.affiliation: School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People's Republic of China. wenyulu@tju.edu.cn.|Key Laboratory of System Bioengineering (Tianjin University), Ministry of Education, Tianjin, 300072, People's Republic of China. wenyulu@tju.edu.cn.|SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin, 300072, People's Republic of China. wenyulu@tju.edu.cn.
authors.forename: Wenyu
authors.initials: W
authors.lastname: Lu
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D053898:Biosynthetic Pathways; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D013186:Farnesyl-Diphosphate Farnesyltransferase / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D009251:NADPH-Ferrihemoprotein Reductase / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D012503:Saponins / Q000096:biosynthesis*; D050603:Squalene Monooxygenase / Q000235:genetics / Q000378:metabolism; D014315:Triterpenes
chemical_list: D001426:Bacterial Proteins; D005656:Fungal Proteins; D010940:Plant Proteins; D012503:Saponins; D014315:Triterpenes; C106188:dammarenediol; D050603:Squalene Monooxygenase; D009251:NADPH-Ferrihemoprotein Reductase; D013186:Farnesyl-Diphosphate Farnesyltransferase
keywords: 2; 3-oxidosqualene; Dammarenediol-II; Escherichia coli; NADPH-cytochrome P450 reductase; Squalene; Triterpenoid