In this study, the influence of substrate-limited media (nitrogen or phosphorus) on Ralstonia eutropha (ATCC 17699) growth and poly-hydroxybutyrate (PHB) production in a batch fermenter at various temperatures (26, 30 and 35 degrees C) was explored. The intracellular PHB yields of bacteria using different limiting substrates were compared. The Ralstonia eutropha had a significantly shortened generation time at higher temperatures (between 26 and 35 degrees C), regardless of the limiting media used. In both limiting media, the PHB accumulation was the highest when the temperature was 35 degrees C. The highest ratios of PHB to biomass in the nitrogen-limiting medium and in the phosphorus-limiting medium were 66.1% and 43.1%, respectively.