To effectively use methanol derived from CO2 or methane in E. coli, it is crucial to develop a fast-growing strain that can efficiently process methanol. We successfully developed a methylotrophic E. coli strain, SM6, which has a doubling time of 3.5 hours, surpassing the growth rates of typical native methylotrophs. This was achieved through evolutionary engineering that utilized dynamic copy number variation (CNV). The SM6 strain has demonstrated the ability to overcome the issue of formaldehyde-induced DNA-protein cross-linking (DPC) during its evolution, enabling rapid growth in methanol medium. Proteomics are conducted to understand the underlying metabolism / regulation change for the improved growth phenotype.