Development of a nonauxotrophic L-homoserine hyperproducer in Escherichia coli by systems metabolic engineering

L-Homoserine is a valuable amino acid as a platform chemical in the synthesis of various important compounds. Development of microbial strains for high-level L-homoserine production is an attractive research direction in recent years. Herein, we converted a wild-type Escherichia coli to a non-auxotrophic and plasmid-free hyper-producer of L-homoserine using systematically metabolic engineer strategies. First, an initial strain was obtained through regulating L-homoserine degradation pathway and enhancing synthetic flow. To facilitate L-homoserine production, flux-control genes were tuned by optimizing the copy numbers in chromosome, and transport system was modified to promote L-homoserine efflux. Subsequently, a strategy of cofactors synergistic utilization was proposed and successfully applied to achieve L-homoserine hyperproduction. The final engineered strain could efficiently produce 85.29 g/L L-homoserine, which was the highest production level ever reported from a plasmid-free, antibiotic-free, inducer-free and nonauxotrophic strain. These strategies used here can be consid-ered for developing microbial cell factory of other L-aspartate derivatives.

Automated summary

In this study, a wild-type Escherichia coli was converted to a high-producer of L-homoserine using metabolic engineer strategies. By regulating the degradation pathway, optimizing copy numbers, and modifying the transport system, a final engineered strain was able to efficiently produce a high level of L-homoserine.