For sustainable chemical industry, it is important to produce bio-based chemicalcompounds. In this study, metabolically engineered Escherichia coli produced 1,3-diaminopropane (1,3-DAP), which is polyamide monomer. In silico flux analysis showedthat the C4 pathway employing Acinetobacter baumannii dat and ddc genes, encoding 2-ketoglutarate 4-aminotransferase and L-2,4-diaminobutanolate decarboxylase, is moreefficient than heterologous pathways of C4 and C5 for producing 1,3-DAP. Also, byapplying 128 of synthetic small RNAs, we found that pfkA gene knock out resulted inincreased 1,3-DAP production. After deleting pfkA gene, we overexpressed the ppc andaspC genes and it made 1,3-DAP production even higher. The final 1,3-DAP producingE. coli strain was went through fed-batch fermentation in a minimal medium withglucose producing 13 g/L of 1,3-DAP. [This work was supported by the TechnologyDevelopment Program to Solve Climate Changes on Systems Metabolic Engineering forBiorefineries from the Ministry of Science, ICT and Future Planning (MSIP) through theNational Research Foundation (NRF) of Korea.]