期刊
BEILSTEIN JOURNAL OF ORGANIC CHEMISTRY
卷 18, 期 -, 页码 881-888出版社
BEILSTEIN-INSTITUT
DOI: 10.3762/bjoc.18.89
关键词
artificial pathway; ent-kaurene; Escherichia coli; overproduction; terpentetriene
资金
- National Science Foundation of China [82073746]
- Thousand Youth Talents Program of China
- Jiangsu Specially Appointed Professor Program
- Jiangsu Double Innovation Plan
This study successfully constructed two truncated pathways to efficiently synthesize clerodane and ent-kaurane skeletons using synthetic biology. This method may be a promising approach for preparing other privileged diterpene scaffolds.
The clerodane and ent-kaurane diterpenoids are two typical categories of diterpenoid natural products with complicated polycyclic carbon skeletons and significant pharmacological activities. Despite exciting advances in organic chemistry, access to these skeletons is still highly challenging. Using synthetic biology to engineer microbes provides an innovative alternative to bypass synthetic challenges. In this study, we constructed two truncated artificial pathways to efficiently produce terpentetriene and ent-kaurene, two representative clerodane and ent-kaurane diterpenes, in Escherichia coli. Both pathways depend on the exogenous addition of isoprenoid alcohol to reinforce the supply of IPP and DMAPP via two sequential phosphorylation reactions. Optimization of these constructs provided terpentetriene and ent-kaurene titers of 66 ?? 4 mg/L and 113 ?? 7 mg/L, respectively, in shake-flask fermentation. The truncated pathways to overproduce clerodane and ent-kaurane skeletons outlined here may provide an attractive route to prepare other privileged diterpene scaffolds.
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