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首页> 外文期刊>Applied Microbiology >Characterization of Thermobifida fusca Cutinase-Carbohydrate-Binding Module Fusion Proteins and Their Potential Application in Bioscouring
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Characterization of Thermobifida fusca Cutinase-Carbohydrate-Binding Module Fusion Proteins and Their Potential Application in Bioscouring

机译：嗜热烟碱角质酶-碳水化合物结合模块融合蛋白的表征及其在生物精练中的潜在应用

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摘要

Cutinase from Thermobifida fusca is thermally stable and has potential application in the bioscouring of cotton in the textile industry. In the present study, the carbohydrate-binding modules (CBMs) from T. fusca cellulase Cel6A (CBM_(Cel6A)) and Cellulomonas fimi cellulase CenA (CBM_(CenA)) were fused, separately, to the carboxyl terminus of T. fusca cutinase. Both fusion enzymes, cutinase-CBM_(Cel6A) and cutinase-CBM_(CenA), were expressed in Escherichia coli and purified to homogeneity. Enzyme characterization showed that both displayed similar catalytic properties and pH stabilities in response to T. fusca cutinase. In addition, both fusion proteins displayed an activity half-life of 53 h at their optimal temperature of 50°C. Compared to T. fusca cutinase, in the absence of pectinase, the binding activity on cotton fiber was enhanced by 2% for cutinase-CBM_(Cel6A) and by 28% for cutinase-CBM_(CenA), whereas in the presence of pectinase, the binding activity was enhanced by 40% for the former and 45% for the latter. Notably, a dramatic increase of up to 3-fold was observed in the amount of released fatty acids from cotton fiber by both cutinase-CBM fusion proteins when acting in concert with pectinase. This is the first report of improving the scouring efficiency of cutinase by fusing it with CBM. The improvement in activity and the strong synergistic effect between the fusion proteins and pectinase suggest that they may have better applications in textile bioscouring than the native cutinase.
机译：来自Thermobifida fusca的角质酶是热稳定的，在纺织工业中对棉花的生物精练中具有潜在的应用。在本研究中，来自美国锥果纤维素酶Cel6A（CBM_（Cel6A））和纤维素单胞菌纤维素酶CenA（CBM_（CenA））的碳水化合物结合模块（CBMs）分别融合到美国锥果角质酶的羧基末端。角质酶-CBM_（Cel6A）和角质酶-CBM_（CenA）两种融合酶均在大肠杆菌中表达并纯化至同质。酶表征显示，响应于山毛榉角质酶，二者均显示出相似的催化性能和pH稳定性。此外，两种融合蛋白在最佳温度50°C下均显示53h的活性半衰期。与T. fusca角质酶相比，在不存在果胶酶的情况下，对角质酶CBM_（Cel6A）的棉纤维结合活性提高了2％，对角质酶CBM_（CenA）的增强了28％，而在存在果胶酶的情况下，前者的结合活性提高了40％，后者的结合活性提高了45％。值得注意的是，当与果胶酶协同作用时，两种角质酶-CBM融合蛋白从棉纤维中释放出的脂肪酸数量显着增加了多达3倍。这是通过将角质酶与CBM融合来提高角质酶精练效率的第一份报告。融合蛋白和果胶酶之间活性的增强和强大的协同作用表明，与天然角质酶相比，它们在纺织品生物精练中的应用可能更好。

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