嗜热细菌Clostridium thermocellum阿魏酸酯酶基因的克

发布时间：2018-02-08 22:59

本文关键词： Clostridium thermocellum 嗜热阿魏酸酯酶碳水化合物结合域异源表达酶学特性　出处：《微生物学通报》2017年06期 　论文类型：期刊论文

【摘要】：【目的】阐明嗜热细菌Clostridium thermocellum Xyn Z蛋白的阿魏酸酯酶催化域的酶学特性,为其在生物质能源及其它发酵工业中的应用奠定基础。【方法】分别构建了C.thermocellum Xyn Z的阿魏酸酯酶催化域(FAE)及该阿魏酸酯酶催化域和碳水化合物结合域(FAE-CBM6)编码基因的原核表达载体,并在大肠杆菌菌株BL21(DE3)中异源表达,在此基础上分析比较了温度、pH、底物、金属离子及CBM6结合域对阿魏酸酯酶活性的影响。【结果】重组FAE酶及FAE-CBM6酶发挥催化活性的适宜pH值为5.0-9.0,适宜温度为50-70°C,它们对不同金属离子的响应有差异。【结论】在同一反应条件下,FAE-CBM6酶的酶活均比FAE高,说明CBM6结合域的存在对于阿魏酸酯酶活性有促进作用。
[Abstract]:[objective] to elucidate the enzymatic properties of ferulic esterase domain of Clostridium thermocellum Xyn Z protein. [methods] the prokaryotic expression vectors of ferulic acid esterase catalytic domain (FAE) of C. thermocellum Xyn Z and its carbohydrate binding domain (FAE-CBM6) and ferulic esterase catalytic domain (FAE) were constructed, respectively. On the basis of the heterologous expression of Escherichia coli strain BL21 (DE3), the pH of temperature and substrate were analyzed and compared. The effect of metal ions and CBM6 binding domain on ferulic acid esterase activity. [results] the optimum pH value of recombination FAE enzyme and FAE-CBM6 enzyme was 5.0-9.0, and the suitable temperature was 50-70 掳C. [conclusion] their responses to different metal ions were different. The enzyme activity of FAE-CBM6 was higher than that of FAE under the same reaction condition. It indicated that the existence of CBM6 binding domain could promote ferulic esterase activity.
【作者单位】：江苏大学环境与安全工程学院生物质能源研究所国家能源非粮生物质原料研发中心华东分中心;
【基金】：国家自然科学基金项目(No.31671754) 江苏大学高级人才启动基金项目(No.15JDG012)~~
【分类号】：Q78;Q936

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