Parameter's optimization and kinetics study of α-amylase enzyme of Bacillus sp. MB6 isolated from vegetable waste

- Isolation of bacteria from vegetable waste for amylase enzyme production.
- Optimization of enzyme production and activity condition.
- Immobilization and reuse of immobilized enzyme can reduce cost of the product.
- Purified enzyme showed very high specific activity against substrate.
- Enzyme kinetics was studied to check rate of reaction against substrate.

α-Amylase, a very critical enzyme for hydrolysis of starch into simple sugar and it has various applications in industrial settings. This study reports the identification of Bacillus sp. MB6 which produces increased amount of enzyme from less required resources. To optimize the yield of enzyme, we used various combinations of parameters. The most optimized conditions for production of amylase enzyme from the bacterium Bacillus sp. MB6 are pH of 6, temperature of 37 °C, and incubation period of 48 h. Condition of enzymatic activity were also examined and the results show that pH of 6, a temperature of 55 °C, and a reaction time of 30 min are the best available conditions for its activity. Purification of enzyme by 1.63 fold enhanced the specific activity of enzyme based upon its activity analysis as compared with unpurified enzyme. Enzyme kinetics studies show the Michaelis constant (Km) to be 5.45 mg/ml and maximum velocity of the reaction (Vmax) to be 24.15 mg/ml/min. In conclusion, we report enzyme production and purification methodology that exhibit better yield of alpha-amylase for commercial applications.

In this work, after purification, enzyme was highly purified, which clearly showed in band intensity that was nearly same as commercial enzyme.117