Abstract: With the rapid advancement of synthetic biology and microbial metabolic engineering, the construction of microbial cell factories has achieved significant progress, facilitating the transition from traditional fossil resource-dependent industrial production to green biosynthesis based on renewable resources. Microbial cell factories enable the biosynthesis of chemicals, offering substantial environmental and economic benefits while providing critical technological support for sustainable production of bulk chemicals, high-value-added compounds, and biofuels. However, microbial metabolic engineering still faces numerous challenges, including the accumulation of intermediate metabolites, insufficient catalytic activity of key enzymes, and low yields and conversion rates of target products, all of which hinder its industrial application. To address these challenges, transcription factor-based biosensors have emerged as essential tools. Their ability to sense and transmit signals allows dynamic optimization of microbial metabolic fluxes, enhanced activity of critical enzymes, and high-throughput screening of superior strains, effectively mitigating these bottlenecks. This review systematically summarized the classification and characteristics of transcription factor-based biosensors, highlighting key strategies for optimizing signal sensing elements, signal transduction components, and output modules. Furthermore, it explored their practical applications in dynamic metabolic flux control, metabolic pathway optimization, and high-throughput screening. Finally, this review provided a perspective on future directions in this field, aiming to guide further advancements in biosensor development and applications.

Key words: transcriptional regulators, biosensors, dynamic regulation, high-throughput screening, synthetic biology