谷氨酸棒杆菌
双功能
清脆的
焊剂(冶金)
CRISPR干扰
代谢工程
代谢途径
Cas9
化学
丝氨酸
生物化学
生物
计算生物学
酶
基因
有机化学
催化作用
作者
Lianghong Yin,Dandan Xi,Yuefeng Shen,Nana Ding,Qingsong Shao,Yongchang Qian,Yu Fang
标识
DOI:10.1021/acs.jafc.3c08529
摘要
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.
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