Medium optimization for the production of amylase by Bacillus subtilis RM16 in Shake-flask fermentation

This study was carried out in shaking incubator and covers the optimization of culture conditions of Bacillus subtilis for the maximum production of amylase. Optimal activity was found to be 350 U ml(-1) when soluble starch was used as a substrate. Parameters taken into consideration to observe their effect on the optimum production of amylase include incubation time, incubation temperature, pH, inoculum size, carbon source, nitrogen source and metallic ions. All parameters were monitored in order to obtain high level of the enzyme units in cell-free broth. The established optimized conditions for Bacillus subtiliss train RM16 were found to be: incubation time 24 hours, temperature 40°C and pH 8.0. Inoculum size was 5%, starch (1%) as a carbon source while yeast extract (1.5%) as a nitrogen source. Magnesium ions (0.1%) exerted maximum stimulating effect for the production of amylase which can be further used at large scale applications.