聚乙二醇化黑曲霉脂肪酶的性质分析
Study on the property of dimethoxy polyethylene glycol modified lipase from Aspergillus niger
作者：李林俐;李刚锐;范翔;孟延发;
Author：
收稿日期： 年卷（期）页码：2014,29(03):-282-285
期刊名称：华西药学杂志
Journal Name：WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字：黑曲霉;脂肪酶;聚乙二醇修饰;酶学性质
Key words：
基金项目：
中文摘要
目的用聚乙二醇对脂肪酶进行化学修饰,以提高酶的物化特性。方法选择20 kD(mPEG)2-Lys-NHS作为修饰剂,实验条件下与黑曲霉脂肪酶发生化学修饰反应;采用Sephacryl S-200凝胶层析法分离聚乙二醇化脂肪酶,利用SDSPAGE法测定分子量,采用常规酶学方法分析比较脂肪酶经聚乙二醇化前后的酶学动力学行为。结果经SDS-PAGE检测获得的脂肪酶为两条带:mPEG2-lipase-Ⅰ、mPEG2-lipase-Ⅱ(分子量分别约105.2、88.7 kD);修饰后脂肪酶的活性保留69%,修饰度50%。以p-NPP为底物,在pH6、50℃条件下比较修饰前后脂肪酶的酶学性质表明:lipase和mPEG2-lipase的Km值分别为21.6、24.8 mmol·L-1,最适反应温度均为50℃,mPEG2-lipase的最适反应pH5,比lipase向碱性方向移动了1个单位,修饰后热稳定性和pH稳定性有了大幅度提高;mPEG2-lipase在4℃下保留6个月,而未修饰酶的活性减少20%;修饰后脂肪酶在有机试剂中耐受性略微提高。结论研究确定了黑曲霉脂肪酶经过聚乙二醇修饰后部分酶学性质,优化了脂肪酶的物化性能,为脂肪酶在食品、化妆品、化工、医药等行业的应用提供了理论依据。
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