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  • 标题:Production, purification and characterization of thermostable α-amylase from soil isolate Bacillus sp . strain B-10
  • 本地全文:下载
  • 作者:RAVINDRA NATH SINGH ; ASHUTOSH BAHUGUNA ; PEEYUSH CHAUHAN
  • 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
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