Production, purification and characterization of ther[sub][/sub]mostable α-amylase from soil isolate [i]Bacillus sp. [/i]strain B-10
Journal Title: Journal of BioScience and Biotechnology - Year 2016, Vol 5, Issue 1
Abstract
A bacterial strain B-10 that produces α-amylase was isolated from compost and kitchen waste receiving agricultural soil. Based on microbiological and biochemical tests the isolate B-10 was identified as [i]Bacillus sp.[/i] Alpha-amylase produced by this isolate was purified by (NH4)[sub]2[/sub]SO[sub]4[/sub] precipitation and DEAE cellulose ion-exchange chromatography showing 15.91 and 48.21 fold purification, respectively. SDS-PAGE of the purified enzyme confirmed the purification and monomeric nature of the enzyme. The purified α-amylase showed maximum activity at pH 7 and temperature 50°C. The enzyme was significantly active in the temperature range of 30-60°C for the studied period of 2 h. During the incubation of purified enzyme at pH ranging from 5 to 10 for 24 h the maximum stability was observed at pH 7 followed by pH 8, whereas at extreme pH, the stability was very poor. K[sub]m[/sub] and V[sub]max[/sub] were found to be 1.4 mg/mL and 6.2 U/mL, respectively.
Authors and Affiliations
Ravindra Singh, Ashutosh Bahuguna, Peeyush Chauhan, Vijay Sharma, Satinder Kaur, Santosh Singh, Amir Khan
Keywords
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