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Isolation and Identification of Novel Bacillus tequilensis TB5 from Vegetable Waste and Analyze the Effect of Rudiment Compounds on Bio-Catalytic α-Amylase Production
Abstract
Abstract
Amylase is one of the basic components for leather, baking, textiles and paper industries. It hydrolyzes starch into simple productive sugar which reduces the cost of the final desired product. In this work, amylase producing novel Bacillus tequilensis TB5 was isolated from vegetable waste and different chemical compounds were added at different concentration in production medium to analyze its effect on amylase enzyme production. We observed that addition of weight/volume (w/v) of starch 1.5%, yeast extract 1%, ammonium nitrate 0.1%, calcium chloride 0.2% and triton x 100 0.2% in the production medium increases the production of enzyme up to 546.64±0.04 U/ml. Storage time of purified enzyme and activity yield was analysed and found to be 98.4% after 72 h, and reuse efficiency of immobilized enzyme was found to be >90% of efficiency after 5th cycle. Purification fold of enzyme shows increase from 1 to 1.93 and MW of purified enzyme was approximately 53 kDa. Enzyme kinetic studies reveal that the Michaelis constant (Km) was found to be 4.56 mg/ml and the reactions maximum velocity (Vmax) was 23.28 mg/ml/min. In summary, we demonstrate that some rudiment compounds present in production medium play a key role for the enhancement of enzyme production.
Keywords: α-amylase, enzyme production, Bacillus, media optimization, enzyme kinetics
Keywords
References
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