Abstract: In this study,Streptococcus zooepidemicus( Streptococcus zooepidemicus) as the starting strain,the effect of glucose concentration and feeding methods on Hyaluronic acid fermentation was studied in 10 L automatic fermentation tank.The results showed that the total concentration of glucose should be controlled at 80 g / L,and both of the initial and supplemented glucose concentration was 40 g / L. The cell amount and production of hyaluronic acid were increased by 13.5% and 28.5% compared with the batch culture while the accumulation of lactic acid was reduced by 7.89%,the relative yield of hyaluronic acid was 10.02%,and average molecular weight was 1.56 × 10~6 u.Thus,the production of hyaluronic acid fermentation can be used single carbohydrate supplement cultured ways to enhance the intensity of production and product quality.