Bacillus Thuringiensisfermentation of Wastewater and Wastewater Sludge – Presence and Characterization of Chitinases

This study investigated the presence of chitinases in Bacillus thuringiensis ssp kurstaki HD-1 (Bt) fermented broths of wastewater sludge (non-hydrolyzed and hydrolyzed); starch industry wastewater and soyameal. Chitinase activity was absent in soyameal and present in others. Chitinase demonstrated peaks at pH 4.0 and temperatures 40 and 50 degrees C with higher activity between pH 4-5 and 10-11. The chitinase band on SDS-PAGE was found to be between 36 and 45 kDa for non-hydrolyzed (NH) and hydrolyzed sludge (TH) and starch industry wastewater. The chitinase profile during fermentation showed peaks at 15 and 30 h for non-hydrolyzed and hydrolyzed sludge and 15 and 24 h for starch industry wastewater. Chitinase retained 96-99 % activity after two weeks incubation at room temperature and pH 4. Bioassays with supplementation of Bt chitinases showed 1.2 fold increase in entomotoxicity of wastewater sludge and a small increase in starch industry wastewater. This study sheds light on production of Bt chitinases in alternative media which will have a long term effect on entomotoxicity of these formulations.