The influence of media components plays an important role on the enhanced production of products from microorganism. An approach was done to enhance the production of laccase from Pleurotus ostreatus – white rot fungi by manipulating the different concertation of variables using Response surface methodology. The Enzyme production was correlated with the biomass produced during the production process.6 g/L NaNO3, 1.5 g/L K2HPO4, 1 g/L Tryptone, 10 g/L Glucose, 0.5 g/L MgSO4 combination showed maximum enzyme activity of 96 IU/ml with an agreeable increase in biomass (5.2 g/L). 50 experiments runs were attained this optimum conditions for enzyme production and the coefficient of determination was 99.5%.