2016-Microb Cell Fact_生物学_自然科学_专业资料.pdfVIP

Yang et al. Microb Cell Fact Microbial Cell Factories DOI 10.1186/s12934-016-0409-7 RESEARCH Open Access Combinatorial engineering of hybrid mevalonate pathways in Escherichia coli for protoilludene production 1† 1† 1,2 1* Liyang Yang , Chonglong Wang , Jia Zhou and Seon‑Won Kim Abstract Background: Protoilludene is a valuable sesquiterpene and serves as a precursor for several medicinal compounds and antimicrobial chemicals. It can be synthesized by heterologous expression of protoilludene synthase in Escheri- chia coli with overexpression of mevalonate (MVA) or methylerythritol‑phosphate (MEP) pathway, and farnesyl diphos‑ phate (FPP) synthase. Here, we present E. coli as a cell factory for protoilludene production. Results: Protoilludene was successfully produced in E. coli by overexpression of a hybrid exogenous MVA pathway, endogenous FPP synthase (IspA), and protoilludene synthase (OMP7) of Omphalotus olearius. For improving protoil‑ ludene production, the MVA pathway was engineered to increase synthesis of building blocks isopentenyl diphos‑ phate (IPP) and dimethylallyl diphosphate (DMAPP) by sequential order permutation of the lower MVA portion (MvL), the alteration of promoters and copy numbers for the upper MVA portion (MvU), and the coordination of both por‑ tions, resulting in an eicient entire MVA pathway. To reduce the accumulation of mevalonate observed in the culture broth due to lower eiciency of the MvL than the MvU, the MvL was further engineered by homolog substitution with the corresponding genes from Staphylococcus aureus. Finally, the highest protoilludene production of 1199 mg/L was obtained from recombinant