Abstract：The effects of initial glucose concentration, agitation speed and aeration rate on hyaluronic acid (HA) production by Streptococcus zooepidemicus WSH24 were investigated in batch culture. The agitation speed had the most pronounced effect on HA yield and molecular weight. Under the condition of 200 r/min, the HA yield, YHA/C and molecular weight could reach 5.3 g/L, 0.13 g/g and 1.88×106 Da, respectively. When the initial glucose concentration was 65.8 g/L, the HA yield, YHA/C and molecular weight reached 5.9 g/L, 0.17 g/g and 1.90×106 Da, respectively. The aeration rate also affected HA fermentation. HA yield and molecular weight were higher at aeration rate of 1.2 L/(min·L) than those at 0.5 L/(min·L).