Multi-module engineering to guide the development of an efficient L-threonine-producing cell factory

The rapid development of high-productivity strains is fundamental for bio-manufacture in industry. Here, Multi-module metabolic engineering was implemented to reprogram Escherichia coli, enabling it to rapidly transitioning from zero-producer to hyperproducer of L-threonine. Firstly, the synthesis pathway of L-threonine was rationally divided into five modules, and the rapid production of L-threonine was achieved by optimizing the expression of genes in each module. Subsequently, the capture and fixation of CO₂ was enhanced to improve L-threonine yield. Dynamically balancing cell growth and yield by quorum-sensing system resulted in the accumulation of L-threonine up to 34.24 g/L. Ultimately, the THR36-L19 strain accumulated 120.1 g/L L-threonine with 0.425 g/g glucose in a 5 L bioreactor. This is the highest yield for de novo producing L-threonine reported to date and without the use of exogenous inducers and antibiotics in the fermentation process. It also provided an effective technological guidence for the zero-to-overproduction of other chemicals.