The increasing need for eco-friendly and cost-effective enzyme production has driven interest in using agricultural and biowaste materials as fermentation substrates. This study aimed to optimize amylase production from Bacillus subtilis using rice gruel water and corn cob hydrolysate as sustainable substrates. B. subtilis was isolated from rhizosphere soil samples through the serial dilution (spread plate) method and identified by morphological, biochemical and selective media tests. The isolates were screened for amylolytic activity on starch agar and the strain exhibiting the largest clear zone was selected for optimization studies. The effects of pH, incubation time, carbon and nitrogen sources on enzyme production were systematically evaluated to determine optimal conditions. Maximum amylase yield was achieved at pH 7, with an incubation period of 2 days, using sucrose as the carbon source and yeast extract as the nitrogen source. The findings demonstrate that biowaste substrates can effectively support industrial enzyme production, reducing production costs and environmental impact. This approach provides a sustainable model for large scale amylase production and encourages further exploration of waste based bioprocessing systems.
