1-propanol is considered as a promising new generation of biofuel that can substitute agasoline. In this study, we introduced a novel biosynthetic pathway toward 1-propanolusing L-threonine overproducing Escherichia coli as a base strain. The highest titer of1-propanol (3.5g/L) was achieved by Atsumi et al., using recombinant Escherichia coliharboring heterogenous gene encoding Methanococcus jannaschii citramalate synthase(CimA), which directly converts pyruvate into 2-ketobutyrate. The previously reportedE. coli TH20 which overproducing L-threonine was used as base strain in this study. Togenerate 1-propanol producing E. coli strain, competing pathways were deleted andcarbon source was optimized by in silico flux reponse analysis. Further metabolicengineering of the final strain improved the production rate moreover. [This work wassupported by the Advanced Biomass R&D Center of Korea through the Global Frontier Research Program of the Ministry of Education, Science andTechnology (MEST). Further supports by BioFuelChem, EEWS program of KAIST, andthe World Class University program of the MEST areappreciated.]