In this present study, bagasse based pulp and paper industry wastewater was treated under different operating conditions such as initial pH (6-8), temperature (25-35 o C) and contact time (3-7 days) by using Bacillus subtilis. Response surface methodology (RSM) coupled with Box-Behnken response surface design (BBD) was employed to investigate the effect of process variables on the responses such as turbidity, biological oxygen demand (BOD) and chemical oxygen demand (COD) removal. The experimental data were analyzed by Pareto analysis of variance (ANOVA) and the second order polynomial models were developed. Interactive effects of the process variables on the responses were studied using plotting 3D response surface contour graph and the optimum process conditions were found to be: initial pH of 7, temperature of 30 o C and contact time of 5 days. Under these conditions, removal efficiencies of turbidity, BOD and COD were found to be 85%, 93% and 80% respectively which are close agreement with real experiments. These results indicate that the treatment of bagasse based pulp and paper industry wastewater using Bacillus subtilis is an effective and novel technique.