Abstract:Higher alcohols that contain more than two carbon atoms have better fuel properties than ethanol, making them important supplements and alternatives to fossil fuels. Using microbes to produce higher alcohols from renewable biomass can alleviate the current energy and environmental crises, and has become a major future direction for green biomanufacturing. Since natural microbes can only produce a few higher alcohols in small amounts, it is necessary to reconstruct the synthetic pathways for higher alcohols in model industrial strains through metabolic engineering and synthetic biology to overcome the metabolic bottlenecks. A series of milestones have been accomplished in past decades. The authors of this review have witnessed the entire journey of this field from its first success to the leaping development. On the 30th anniversary of the founding of the discipline of metabolic engineering, this review dates back to the great milestones in achieving heterologous production of higher alcohols in non-native strains. The design and optimization of high alcohol biosynthetic pathways, the expansion of feedstock, the engineering of host strains and the industrialization process are summarized. This review aims to draw further attention to microbial synthesis of higher alcohols, inspire the development of novel techniques and strategies of metabolic engineering, and promote the innovation and upgrade of China’s biofuel industry.