Glycolysis is the primary metabolic pathway in all living organisms. Maintaining the balance of glycolysis flux and biosynthetic pathways is the crucial matter involved in the microbial cell factory. Few regulation systems can address the issue of metabolic flux imbalance in glycolysis. Here, we designed and constructed a bifunctional glycolysis flux biosensor that can dynamically regulate glycolysis flux for overproduction of desired biochemicals. A series of positive-and negative-response biosensors were created and modified for varied thresholds and dynamic ranges. These engineered glycolysis flux biosensors were verified to be able to characterize in vivo fructose-1,6-diphosphate concentration. Subsequently, the biosensors were applied for fine-tuning glycolysis flux to effectively balance the biosynthesis of two chemicals: mevalonate and N-acetylglucosamine. A glycolysis flux-dynamically controlled Escherichia coli strain achieved a 111.3 g/L mevalonate titer in a 1L fermenter.

糖酵解是所有生物体的主要代谢途径。维持糖酵解通量和生物合成途径的平衡是微生物细胞工厂所涉及的关键问题。很少有调节系统可以解决糖酵解中代谢通量不平衡的问题。在这里，我们设计并构建了一种双功能糖酵解通量生物传感器，可以动态调节糖酵解通量以过量生产所需的生化物质。针对不同的阈值和动态范围创建和修改了一系列正响应和负响应生物传感器。这些工程化的糖酵解通量生物传感器经验证能够在体内表征1,6-二磷酸果糖浓度。随后，生物传感器用于微调糖酵解通量，以有效平衡两种化学物质的生物合成：甲羟戊酸和 N-乙酰氨基葡萄糖。糖酵解通量动态控制的大肠杆菌菌株在 1L 发酵罐中实现了 111.3 g/L 的甲羟戊酸滴度。