Among PHA copolymers, [P(3HB-co-3HV)] is one of the important copolymer. So far,adding of second auxiliary carbon source was needed for the production of P(3HB-co-3HV). However, due to the toxicity of auxiliary carbon source, maintaining the balancebetween cell growth and polymer production was hard. Thus, we developed the E. colican stably synthesize 3HB-CoA and 3HV-CoA in controlled ratio from glucose, and canefficiently synthesize P(3HB-co-3HV) without feeding auxiliary carbon source. [Thiswork was supported by the Technology Development Program to Solve ClimateChanges from National Research Foundation of Korea Development of systemsmetabolic engineering platform technologies for biorefineries.]