Methanol is a promising alternative feedstock for biomanufacturing. However, natural platform industrial microorganisms such as Escherichia coli and Corynebacterium glutamicum cannot assimilate methanol. Although some methanol assimilation pathways differ from the typical sugar metabolism by only a few enzymes, engineering platform microorganisms to efficiently utilize methanol is very challenging. Recent advances in creating the first methylotrophic E. coli growing solely on methanol have underscored the value of metabolic engineering-guided adaptive laboratory evolution (ME-ALE) in developing non-natural methanol utilizers. This chapter reviews the recent progress in engineering platform microorganisms to utilize methanol by ME-ALE and discusses the future challenges in developing superior synthetic methylotrophs for methanol-based biomanufacturing.