Manipulation of the precursor supply for high-level production of longifolene by metabolically engineered Escherichia coli

Authors

Yujin Cao, Qingdao Institute of Bioenergy and Bioprocess TechnologyFollow
Rubing Zhang, Qingdao Institute of Bioenergy and Bioprocess Technology
Wei Liu, Qingdao Institute of Bioenergy and Bioprocess Technology
Guang Zhao, Qingdao Institute of Bioenergy and Bioprocess Technology
Wei Niu, University of Nebraska - LincolnFollow
Jiantao Guo, University of Nebraska-LincolnFollow
Mo Xian, Qingdao Institute of Bioenergy and Bioprocess TechnologyFollow
Huizhou Liu, Qingdao Institute of Bioenergy and Bioprocess TechnologyFollow

ORCID IDs

Yujin Cao http://orcid.org/0000-0001-5561-2731

Date of this Version

1-2019

Document Type

Article

Citation

Scientific Reports (2019) 9:95

DOI:10.1038/s41598-018-36495-w

Comments

This article is licensed under a Creative Commons Attribution 4.0 International License,

Abstract

Longifolene is a naturally occurring tricyclic sesquiterpene widely used in many different fields. Up to now, this valuable terpene was mainly manufactured from the high-boiling fraction of certain pine resins. Microbial production can be a promising alternative to the extraction from natural plant sources. Here, we present the metabolic engineering strategy to assemble biosynthetic pathway for longifolene production in Escherichia coli. E. coli was rendered to produce longifolene by heterologously expressing a codon optimized longifolene synthase from Picea abies. Augmentation of the metabolic flux to farnesyl pyrophosphate (FPP) by different FPP synthases conferred a 1.8-fold increase in longifolene production. An additional enhancement of longifolene production (up to 2.64 mg/L) was achieved by introducing an exogenous mevalonate pathway. Under fed-batch conditions, the best-performing strain was able to produce 382 mg/L of longifolene in a 5 L bioreactor. These results demonstrated the feasibility of producing longifolene by microbial fermentation and could serve as the basis for the construction of more robust strains in the future.