Application of Response Surface Methodology to Optimize Medium Components for Production of Alkaline Protease by Bacillus subtilis IC-5

Abstract

Optimization of fermentation medium for production of alkaline protease by using Bacillus subtilis IC-5 was carried out. The concentration of agricultural by product (soybean meal). carbon source (glucose) and nitrogen source (sodium nitrate) were selected for optimization. A central composite design and response surface methodology were used in designing of experiment and statistical analysis of results. This procedure limited the number of actual experiments to 20 and allowed for possible interaction between the three ingredients. The optimum concentrations of the ingredients (g/l) were soybean meal 1.80, glucose 2.1 land sodium nitrate 0.29 were predicted for production of 3468.94 units per ml alkaline protease by Bacillus subtilis IC-5. The verification run confirm the predicted optimized concentration of all the three ingredients tor maximum production of alkaline protease. These studies successfully demonstrated the use of central composite experimental design for the determination of optimum nutrient concentrations for maximum production of alkaline protease. Results also revealed that smaller and less time consuming experimental designs could be sufficient for optimization of fermentation processes.