Meng Xin, Hu Guipeng, Li Xiaomin, Gao Cong, Song Wei, Wei Wanqing, Wu Jing, Liu Liming
School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wux, China.
School of Life Sciences and Health Engineering, Jiangnan University, Wuxi, China.
Nat Commun. 2025 Jan 2;16(1):31. doi: 10.1038/s41467-024-55502-5.
Microbial utilization of methanol for valorization is an effective way to advance green bio-manufacturing technology. Although synthetic methylotrophs have been developed, strategies to enhance their cell growth rate and internal regulatory mechanism remain underexplored. In this study, we design a synthetic methanol assimilation (SMA) pathway containing only six enzymes linked to central carbon metabolism, which does not require energy and carbon emissions. Through rational design and laboratory evolution, E. coli harboring with the SMA pathway is converted into a synthetic methylotroph. By self-adjusting the expression of TOPAI (topoisomerase I inhibitor) to alleviate transcriptional-replication conflicts (TRCs), the doubling time of methylotrophic E. coli is reduced to 4.5 h, approaching that of natural methylotrophs. This work has the potential to overcome the growth limitation of C1-assimilating microbes and advance the development of a circular carbon economy.
微生物利用甲醇进行增值是推进绿色生物制造技术的有效途径。尽管已经开发出合成甲基营养菌，但提高其细胞生长速率的策略和内部调节机制仍未得到充分探索。在本研究中，我们设计了一条仅包含六种与中心碳代谢相关酶的合成甲醇同化（SMA）途径，该途径无需能量和碳排放。通过合理设计和实验室进化，携带SMA途径的大肠杆菌被转化为合成甲基营养菌。通过自我调节拓扑异构酶I抑制剂（TOPAI）的表达来缓解转录-复制冲突（TRC），甲基营养型大肠杆菌的倍增时间缩短至4.5小时，接近天然甲基营养菌。这项工作有可能克服C1同化微生物的生长限制，并推动循环碳经济的发展。
