Clone, expression, and purification of a hyperthermophilic endoglucanase gene cel12B from Thermotoga maritima

摘要

The thermostability and catalytic activity of endoglucanase from thermophilic Thermotoga maritima were improved by evolutionary molecular engineering. The cell2B gene from Thermotoga maritima was cloned and expressed, and the expressed Cell2B protein was purified by His-tag affinity chromatography. The constructed plasmids were screened on LB plate with ampicillin using Escherichia coli BL21 (DE3) as a host. The results showed that both the enzyme activity and yield of the Cell12B protein expressed in pET15b-cell2B were 2 times as that of the protein expressed in pET20b-cell2B.