Dynamic regulation is a promising strategy for fine-tuning metabolic fluxes in microbial cell factories. However, few of these synthetic regulatory systems have been developed for central carbon metabolites. Here we created a set of programmable and bifunctional pyruvate-responsive genetic circuits for dynamic dual control (activation and inhibition) of central metabolism in Bacillus subtilis. We used these genetic circuits to design a feedback loop control system that relies on the intracellular concentration of pyruvate to fine-tune the target metabolic modules, leading to the glucaric acid titer increasing from 207 to 527 mg l⁻¹. The designed logic gate-based circuits were enabled by the characterization of a new antisense transcription mechanism in B. subtilis. In addition, a further increase to 802 mg l⁻¹ was achieved by blocking the formation of by-products. Here, the constructed pyruvate-responsive genetic circuits are presented as effective tools for the dynamic control of central metabolism of microbial cell factories.

动态调节是微调微生物细胞工厂代谢通量的一种很有前景的策略。然而，这些合成调节系统中很少有针对中心碳代谢物开发的。在这里，我们创建了一组可编程和双功能丙酮酸响应基因电路，用于枯草芽孢杆菌中枢代谢的动态双重控制（激活和抑制）。我们使用这些遗传回路设计了一个反馈回路控制系统，该系统依赖于丙酮酸的细胞内浓度来微调目标代谢模块，从而使葡萄糖二酸滴度从 207 增加到 527 mg l ^-1。设计的基于逻辑门的电路是通过对枯草芽孢杆菌中一种新的反义转录机制的表征来实现的. 此外，通过阻止副产物的形成，进一步增加至 802 mg l ^{-1 。}在这里，构建的丙酮酸响应遗传回路被提出作为动态控制微生物细胞工厂中枢代谢的有效工具。