Improving endoglucanase activity by adding the carbohydrate-binding module from Corticium rolfsii.
Improving endoglucanase activity by adding the carbohydrate-binding module from Corticium rolfsii.
复制标题
DOI：
10.4014/jmb.1311.11007
复制
发表时间：
2014-04
影响因子：
2.8
通讯作者：
Zizhong Tang;Hui Chen;Lijiao Chen;San Liu;Xue-yi Han;Qi Wu
中科院分区：
文献类型：
--
作者：
Zizhong Tang;Hui Chen;Lijiao Chen;San Liu;Xue-yi Han;Qi Wu
碳水化合物结合模块(CBM)是大多数纤维素酶的重要结构域，在纤维素酶的降解过程中起着关键作用。重组了中性内切葡聚糖酶(EG1)基因。重新设计了一种内切葡聚糖酶，命名为EG2，用含有Cortisum rofsii连接子的CBM构建(GenBank登录号：D49448)。对重新设计的EG基因在大肠杆菌中进行了表达，并对其特性进行了讨论。结果表明，EG2对纤维素的降解率约为EG1的两倍。在优化条件下测定了EG1和EG2的比活力，在纤维素降解过程中，EG2对CMC-Na的活性(169.1±2.74U/mg)高于EG1(84.0±1.98)U/mg。EG1和EG2的最适pH和温度、pH稳定性和热稳定性相似。结果表明，煤层气在纤维素的降解过程中起着至关重要的作用。我们可以通过添加煤层气来提高EG的催化能力。
The carbohydrate-binding module (CBM) is an important domain of most cellulases that plays a key role in the hydrolysis of cellulose. The neutral endoglucanase (EG1) gene was reconstructed. A redesigned endoglucanase, named EG2, was constructed with a CBM containing a linker from Corticium rolfsii (GenBank Accession No. D49448). The redesigned EG genes were expressed in Escherichia coli, and their characteristics are discussed. Results showed that the degradation of cellulose by EG2 was about double that by EG1. The specific activities of EG1 and EG2 were tested under optimal conditions, and EG2 had higher activity (169.1 ± 2.74 U/mg) toward CMC-Na than did EG1 (84.0 ± 1.98) in the process of cellulose degradation. The optimal pH and temperature, pH stability, and heat stability of EG1 and EG2 were similar. Results indicated that the CBM plays an essential role in the hydrolysis of cellulose. We can improve EG's catalytic power by adding the CBM from Corticium rolfsii.