Purification and characterization of an acidothermophilic cellulase enzyme produced by Bacillus subtilis strain LFS3

被引：0
作者：
Rekha Rawat
Lakshmi Tewari
机构：
[1] G.B. Pant University of Agriculture and Technology,Department of Microbiology
来源：
Extremophiles | 2012年 / 16卷
关键词：
Carboxymethylcellulase; Acidophilic; Purification; Characterization;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
In the present investigation, a microorganism hydrolyzing carboxymethylcellulose (CMC) was isolated and identified as Bacillus subtilis strain LFS3 by 16S rDNA sequence analysis. The carboxymethylcellulase (CMCase) enzyme produced by the B. subtilis strain LFS3 was purified by (NH4)2SO4 precipitation, ion exchange and gel filtration chromatography, with an overall recovery of 15 %. Native-PAGE analysis of purified CMCase revealed the molecular weight of enzyme to be about 185 kDa. The activity profile of CMCase enzyme showed the optimum activity at temperature 60 °C and pH 4.0, respectively. The enzyme activity was induced by Na+, Mg2+, NH4+, and EDTA, whereas strongly inhibited by Hg2+ and Fe3+. The purified enzyme hydrolyzed CMC, filter paper, and xylan, but not p-nitrophenyl β-d-glucopyranoside and cellulose. Kinetic analysis of purified enzyme showed the Km value of 2.2 mg/ml. Thus, acidophilic as well as thermophilic nature makes this cellulase a suitable candidate for current mainstream biomass conversion into fuel and other industrial processes.
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页码：637 / 644
页数：7
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