Biobutanol has been attracting much attention as a clean fuel and chemical due to that the use of fossil fuels lead to aggravation of global warming and energy crisis. Escherichia coli is an ideal candidate for butanol production because it is easy to manipulate genetically. Butanol toxicity has been a bottleneck for industrial-scale biobutanol production,so the improvement in butanol tolerance is essential for high titer butanol production. Butanol destroyed the barrier and transport functions of cell membrane, and cell produces physiological response,which is similar to that of heat shock,osmotic stress,etc. Cell regulates transcription and translation to resist butanol stress. In the light of the above points,the butanol tolerance mechanism of E. coli and recent advances in development of butanol-tolerant strains by rational design strategy are summarized in this review. Nevertheless,the mechanism has not been yet fully elucidated,which limits the use of rational design strategy. There is also concern about the application of inverse metabolic engineering in this area,which means that the butanol-tolerant strains are obtained through directed evolution and the functional genes are further revealed. In addition,the progress on application of the latest strategies for improving butanol tolerance,such as combined strategy,chemical modification,and propose the potential key points for enhancing butanol tolerance of E. coli were reviewed.