Engineering Escherichia coli for l-Threonine Hyperproduction Based on Multidimensional Optimization Strategies

Exploring effective remodeling strategies to further improve the productivity of high-yield strains is the goal of biomanufacturing. However, the lack of insight into host-specific metabolic networks prevents timely identification of useful engineering targets. Here, multidimensional engineering strategies were implemented to optimize the global metabolic network for improving l-threonine production. First, the metabolic bottleneck for l-threonine synthesis was eliminated by synergistic utilization of NADH and an enhanced ATP supply. Carbon fluxes were redistributed into the TCA cycle by rationally regulating the GltA activity. Subsequently, the stress global response regulator UspA was identified to enhance l-threonine production by a transcriptomic analysis. Then, l-threonine productivity was improved by enhancing the host’s stress resistance and releasing the inhibitory reaction of glucose utilization. Eventually, the l-threonine yield of THRH16 reached 170.3 g/L and 3.78 g/L/h in a 5 L bioreactor, which is the highest production index reported. This study provides rational guidance for increasing the productivity of other chemicals.