Corynebacterium glutamicum, a microorganism
classified as generally recognized as safe for use in the industrial
production of food raw materials and additives, has encountered challenges
in achieving widespread adoption and popularization as microbial cell
factories. These obstacles arise from the intricate nature of manipulating
metabolic flux through conventional methods, such as gene knockout
and enzyme overexpression. To address this challenge, we developed
a CRISPR/dCpf1-based bifunctional regulation system to bidirectionally
regulate the expression of multiple genes in C. glutamicum. Specifically, through fusing various transcription factors to the
C-terminus of dCpf1, the resulting dCpf1-SoxS exhibited both CRISPR
interference (CRISPRi) and CRISPR activation (CRISPRa) capabilities
in C. glutamicum by altering the binding
sites of crRNAs. The bifunctional regulation system was used to fine-tune
metabolic flux from shikimic acid (SA) and l-serine biosynthesis,
resulting in 27-fold and 10-fold increases in SA and l-serine
production, respectively, compared to the original strain. These findings
highlight the potential of the CRISPR/dCpf1-based bifunctional regulation
system in effectively enhancing the yield of target products in C. glutamicum.