HA batch fermentation process using Streptococcus zooepidemicus was discussed under different temperatures which were varied from 30 ℃ to 40℃,the results show that the optimal bacterial growth temperature is 34℃ and the highest yield up to 82mg/100mL at 38℃.Application of a two-stage temperature control strategy was developed base on those studies.The orthogonal of L18(37)indicated that when pre-fermentation temperature was controlled at 34℃ and changed to 40℃ in product synthesis phase,the yield of HA increased to 105.30 mg/100 mL,which is 28.4% higher compared to that of fermentation at single temperature of 38℃.