As methanol can be derived from either CO2 or methane, methanol economy may play a role in combating climate change. In this scenario, rapid utilization of methanol by an industrial microorganism is the first and crucial step for efficient utilization of the C1 feedstock chemical. Here, we report the development of a methylotrophic E. coli strain (SM6) with a doubling time of 3.5 hours, outpacing that of common native methylotrophs. We accomplish this using evolution engineering with dynamic copy number variation (CNV). We developed a bacterial artificial chromosome (BAC) with dynamic CNV to facilitate overcoming the formaldehyde-induced DNA-protein cross-linking (DPC) problem in the evolution process. The growth rate of the organism in methanol minimal medium improved significantly after it acquired a loss-of-function mutation in mutS. We tracked the genome variations of 72 cultures along the evolution process by next-generation sequencing, and identified the metabolic features of the fast-growing strain. This study illustrates the potential of dynamic CNV as an evolution tool and synthetic methylotrophs as a platform for sustainable biotechnological applications.

中文翻译：

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甲基营养型大肠杆菌的进化工程能够比天然甲基营养型大肠杆菌更快地生长

由于甲醇可以从二氧化碳或甲烷中提取，甲醇经济可以在应对气候变化方面发挥作用。在这种情况下，工业微生物对甲醇的快速利用是高效利用 C1 原料化学品的第一步，也是关键的一步。在这里，我们报告了甲基营养型大肠杆菌菌株 (SM6) 的发展，其倍增时间为 3.5 小时，超过了常见的天然甲基营养菌。我们使用具有动态拷贝数变异（CNV）的进化工程来实现这一目标。我们开发了一种具有动态CNV的细菌人工染色体（BAC），以帮助克服进化过程中甲醛诱导的DNA-蛋白质交联（DPC）问题。在获得mutS功能丧失突变后，该微生物在甲醇基本培养基中的生长速度显着提高。我们通过下一代测序追踪了 72 个培养物在进化过程中的基因组变异，并鉴定了快速生长菌株的代谢特征。这项研究说明了动态 CNV 作为进化工具和合成甲基营养菌作为可持续生物技术应用平台的潜力。