Purification and characterization of a family 5 endoglucanase from a moderately thermophilic strain of Bacillus licheniformis

被引:92
作者
Bischoff, Kenneth M. [1 ]
Rooney, Alejandro P. [1 ]
Li, Xin-ang Li [1 ]
Liu, Siqing [1 ]
Hughes, Stephen R. [1 ]
机构
[1] USDA ARS, Natl Ctr Agr Utilizat Res, Peoria, IL 61604 USA
关键词
Bacillus licheniformis; carboxymethylcellulase; endoglucanase; thermophilic;
D O I
10.1007/s10529-006-9153-0
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Strains of thermophilic bacilli were screened for cellulolytic activity by gel diffusion assay on selective medium at 55 degrees C. Strain B-41361, identified as a strain of Bacillus licheniformis, displayed activity against carboxymethylcellulose. Zymogram analysis demonstrated several catalytically active polypeptides with the most prominent species having a mass of 37 kDa. The enzyme was purified 60-fold with a 17% yield and specific activity of 183 U/mg. The amino terminal sequence was homologous to members of glycoside hydrolase family 5. Optimal temperature was 65 degrees C (measured over 30 min), but the enzyme was most stable at 60 degrees C, retaining greater than 90% activity after one hour. The enzyme had a broad pH range, with maximal activity at pH 6.0, 75% maximal activity at pH 4.5, and 40% at pH 10. The enzyme hydrolyzed p-nitrophenylcellobioside, barley beta-glucan, and lichenan, but no activity was detected against avicel or acid-swollen cellulose.
引用
收藏
页码:1761 / 1765
页数:5
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