Laccase enzyme has a range of biotechnological and industrial applications but its high cost hindered its use. Development of low-cost biotechnological process for laccase production has gained great attention. Present study is concerned with the enhanced production of laccase by Aspergillus flavus Maf 0139. Five different cultural media were tested and best one was selected for laccase production. Different cultural conditions such as inoculum size, pH, temperature, incubation period, moisture content and inducer were studied. Medium components were further optimized by Plackett-Burman and central composite designs. Results showed that maximum enzyme activity (297±0.1 U/ml/min) was obtained by using AFM02 medium containing g/L: sawdust, 5.0; glucose, 5.0; peptone, 6.0; MgSO4.7H2O, 0.25; KCl, 0.5; KH2PO4, 0.025. Optimized conditions for maximum enzyme yield were inoculum size 2.5%, Temperature 25ËšC, pH 4.5, incubation period 7 days and moisture content 60%. Eleven different inducers were evaluated but FeSO4.7H2O showed maximum enzyme production (2156±0.57 U/mL/min). By using these statistical techniques more than 13 folds increase in enzyme yield (4022±0.25 U/mL/min) was observed. It is concluded that cultural conditions have great influence on enzyme production and enzyme yield could be enhanced by using advanced statistical techniques.