Organophosphate hydrolase enzyme has been reported to efficiently degrade organophosphate compounds. Thus its increased demand, has led us to improve its production by process optimization. Statistical improvement for OPH production by Response Surface Methodology (RSM) was performed by using the bacterial strain Brevibacillus parabrevis SR2729. Glucose and NaNO3 were observed as significant carbon and nitrogen sources, respectively. After RSM, the optimum activity of OPH was observed to be 365 U mL-1 at pH 4, temperature 52.5°C, incubation time 40.5h, carbon and nitrogen sources 0.5%, and pesticide concentration 6%. The results showed that at 6% of pesticide, low nutrients concentration, and short incubation time, the enzyme production was maximum, which could be a breakthrough for the degradation of pesticide. Such an optimized production of OPH can be practiced for the enhanced pesticide determination and degradation settings.