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OALib Journal期刊
ISSN: 2333-9721
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耐酸耐温α-淀粉酶基因在大肠杆菌中的功能表达
DOI: 10.3969/j.issn.1671-7627.2012.03.004, PP. 20-26
Keywords: 淀粉酶,密码子偏好,耐酸,耐温,表达
Abstract:
兼顾大肠杆菌与枯草芽孢杆菌密码子偏好性,优化获得了1个耐酸耐温淀粉酶的基因amycn1,并将其在大肠杆菌中实现了功能表达。纯化后的重组α-淀粉酶(amy1)表征结果表明:在最适ph5.5和最适温度75℃条件下表观米氏常数(km)值和催化效率(kcat/km)值分别为20.93g/l和98.20l/(g·s);低浓度的co2+(1mmol/l)可以提高30%的淀粉酶活力,mn2+抑制了大部分活力,包括ca2+在内的其他金属离子影响不显著;高浓度的edta(≥50mmol/l)抑制其活力;生物信息学分析表明,amy1具有α-淀粉酶家族典型的3-结构域分布,具有1个ca2+结合位点和5个zn2+结合位点,活性中心催化氨基酸残基d198和e222位于(β/α)8桶中β折叠片c端一侧。amy1优良的低ph耐受性和耐温性,有助于木薯淀粉生产燃料乙醇工业液化与糖化同步发酵工艺的实现。
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