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<p>Dynamic metabolic regulation is crucial for optimizing microbial cell factories. To address the limitations of chemical inducers, this study developed a temperature-responsive synthetic biology toolkit for <i>Corynebacterium glutamicum</i>. A high-performance, heat-inducible biosensor was engineered by optimizing the CI<sup>857</sup> repressor and its cognate promoter, yielding a variant (CI<sup>857</sup>-M3/H1) with a 107-fold dynamic range and minimal background leakage. Additionally, a cold-inducible RNA thermometer was implemented using the <i>Escherichia coli csapA</i> 5’UTR. These components were integrated into a dual-functional genetic circuit enabling bidirectional metabolic control. Finally, the optimized heat-inducible sensor was applied to the production of three secretory proteins with distinct characteristics (AmyE, XylA, and VHH), and the scale-up cultivation of AmyE was successfully achieved in 1 L shake-flasks. This work provides an efficient, inducer-free strategy for precise metabolic regulation, offering a scalable and cost-effective tool for advanced biomanufacturing.</p>

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Engineering a temperature-programmable biosensor toolkit for recombinant protein production in Corynebacterium glutamicum

  • 期刊论文

  • 发表日期：

    2026年5月6日

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  • Haofei Xu,
  • Yanbo Li,
  • Yiran Gan,
  • Songzhou Liu,
  • Bin Lin,
  • Shijun Cen,
  • Yankun Yang,
  • Chunli Liu,
  • Xiuxia Liu,
  • Zhonghu Bai

摘要

Dynamic metabolic regulation is crucial for optimizing microbial cell factories. To address the limitations of chemical inducers, this study developed a temperature-responsive synthetic biology toolkit for Corynebacterium glutamicum. A high-performance, heat-inducible biosensor was engineered by optimizing the CI857 repressor and its cognate promoter, yielding a variant (CI857-M3/H1) with a 107-fold dynamic range and minimal background leakage. Additionally, a cold-inducible RNA thermometer was implemented using the Escherichia coli csapA 5’UTR. These components were integrated into a dual-functional genetic circuit enabling bidirectional metabolic control. Finally, the optimized heat-inducible sensor was applied to the production of three secretory proteins with distinct characteristics (AmyE, XylA, and VHH), and the scale-up cultivation of AmyE was successfully achieved in 1 L shake-flasks. This work provides an efficient, inducer-free strategy for precise metabolic regulation, offering a scalable and cost-effective tool for advanced biomanufacturing.