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

Abstract

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.

Keywords

Thermosensitive biosensor / Dynamic metabolic regulation / Corynebacterium glutamicum / Recombinant protein production / Synthetic biology

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Funding

National Natural Science Foundation of China

Key Research and Development Project of Henan Province

RIGHTS & PERMISSIONS

Jiangnan University