文章基本信息

标题：Production, purification and characterization of thermostable α-amylase from soil isolate Bacillus sp . strain B-10
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作者：RAVINDRA NATH SINGH ; ASHUTOSH BAHUGUNA ; PEEYUSH CHAUHAN 等
期刊名称：Journal of BioScience and Biotechnology
印刷版ISSN：1314-6238
电子版ISSN：1314-6246
出版年度：2016
卷号：5
期号：1
页码：37-43
出版社：Plovdiv University Press
摘要：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 Bacillus sp. Alpha-amylase produced by this isolate was purified by (NH4)2SO4 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. Km and Vmax were found to be 1.4 mg/mL and 6.2 U/mL, respectively.
关键词：Bacillus; α-amylase; SDS-PAGE; ion exchange chromatography; Michaelis constant; Km