The hyaluronic acidproducing Streptococcus zooepidermicus ATCC 43079 was used to study the hyaluronic acid production. We used the Plackett-Burman design of optimum multifactorial experiments to estimate the level effects of different factors such as peptone, meat extract, yeast extract, glucose, tween 80, dipotassium phosphate, sodium acetate, ammonium citrate, magnesium sulphate and manganese sulphate on hyaluronic acid production. As a result, meat extract (0.2%), yeast extract (0.1%) and glucose (0.4%) gave the highest production in Streptococcus zooepidermicus at temperature (40oC). In order to confirm the estimated hyaluronic acid yield, the designated experiments were done. The maximum yield of hyaluronic acid in Streptococcus zooepidermicus ATCC 43079 was 42.38 mg/l while the predicted HA was 42.32 mg/l. The results also showed the Plackett - Burman can be used to predict the HA yield.The study was the first report of HA production affected by complex nutrient, temperature and salts.