Evaluation of Industrial Wastewaters as Low-Cost Resources for Sustainable Enzyme Production by Bacillus Species

The increasing demand for industrial enzymes calls for cost-effective and sustainable production strategies. This study investigates the potential of industrial wastewater as an alternative fermentation medium for enzyme synthesis, aligning with the principles of the circular bioeconomy. Four wastewater types from Québec, Canada—beverage wastewater (BW), pulp and paper mill activated sludge (PPMS), food industry wastewater (FIW), and starch industry wastewater (SIW)—were evaluated for their potential to support protease, amylase, and lipase production using Bacillus licheniformis , Bacillus amyloliquefaciens , and Bacillus megaterium . Initial screening identified SIW as optimal for amylase production with B. amyloliquefaciens , and PPMS for protease production with B. megaterium . Optimization using the Box–Behnken design was then performed, followed by scale-up experiments in 5 L bioreactors. B. amyloliquefaciens achieved 5.73 ± 0.01 U/mL of amylase at 48 h under 40 g/L total solids, 30 °C, and a 2% inoculum size, while B. megaterium produced the highest protease of 55.41 ± 3.54 U/mL at 24 h. Lipase production remained negligible across all media and strains. These findings demonstrate the feasibility of the potential of wastewater-based enzyme production, reducing reliance on expensive synthetic substrates, mitigating environmental burdens, and contributing to the transition to a circular bioeconomy.