Ammonium limitation did not promote ply(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) production of Azotobacter vinelandii UWD. In acid phase, ammonium limitation during utilization of propionic acid and butyric acid led to 35% decrease in product yield. In glucose phase, both biomass yield and polymer yield decreased about 22% under ammonium limitation. However, in nitrogen-fixing culture glucose was consumed 25% faster and the final PHBV wt% decreased slightly. Under nitrogen limitation a portion of the carbon sources was used fro nitrogen fixation rather than biomass and polymer formation, resulting in a decrease in biomass yield and polymer yield.