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Proteolysin, a Novel Highly Thermostable and Cosolvent-Compatible Protease from the Thermophilic Bacterium Coprothermobacter proteolyticus

机译:蛋白溶素,一种来自嗜热细菌蛋白水解副杆菌的高度热稳定且与溶剂兼容的新型蛋白酶

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摘要

Through genome mining, we identified a gene encoding a putative serine protease of the thermitase subgroup of subtilases (EC 3.4.21.66) in the thermophilic bacterium Coprothermobacter proteolyticus. The gene was functionally expressed in Escherichia coli, and the enzyme, which we called proteolysin, was purified to near homogeneity from crude cell lysate by a single heat treatment step. Proteolysin has a broad pH tolerance and is active at temperatures of up to 80°C. In addition, the enzyme shows good activity and stability in the presence of organic solvents, detergents, and dithiothreitol, and it remains active in 6 M guanidinium hydrochloride. Based on its stability and activity profile, proteolysin can be an excellent candidate for applications where resistance to harsh process conditions is required.
机译:通过基因组挖掘,我们在嗜热菌蛋白水解副杆菌中鉴定了一个编码枯草杆菌酶的热酶亚组推定丝氨酸蛋白酶的基因(EC 3.4.21.66)。该基因在大肠杆菌中进行了功能表达,并且通过单步热处理从粗细胞裂解物中将酶(我们称为蛋白水解酶)纯化至接近同质。蛋白溶素具有广泛的pH耐受性,并在高达80°C的温度下具有活性。此外,该酶在有机溶剂,去污剂和二硫苏糖醇的存在下显示出良好的活性和稳定性,并且在6 M胍盐酸盐中仍然具有活性。基于其稳定性和活性特征,蛋白溶素可以成为需要耐受苛刻工艺条件的应用的理想选择。

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