α-淀粉酶固定化的研究

Research on the α-Amylase Immobilization

综述了固定化酶的优越性,酶的固定化的方法分类以及不同方法的优点和缺点。以甘蔗纤维素衍生物为载体,用共价键结合法固定α-淀粉酶。根据温度、pH值、α-淀粉酶的浓度以及α-淀粉酶与甘蔗纤维素衍生物载体的配比对α-淀粉酶固定的影响,通过正交试验得到最佳固定条件为:温度60℃,pH值为6.0,α-淀粉酶的浓度为60U/ml,α-淀粉酶与甘蔗纤维素衍生物载体的配比为50ml∶1g。缓冲溶液为柠檬酸-磷酸氢二钾缓冲液。通过吸光度法测定所得固定化酶的活力为34.77U/g固定剂。测得米氏常数为12.88g/L,半衰期为3.17h,固定化酶在使用过程中没有α-淀粉酶脱离在产品中,所以可以减少额外的加工费用,同时可以循环使用。

In this paper the advantage of the immobilized enzyme,the classification of immobilization,and the advantage and disadvantage of different methods were summarized.The sugarcane cellulose derivative was used as a carrier by covalent binding fixed-amylase.Depending on the temperature,pH,α-amylase concentration and-amylase and cane cellulose derivatives,the ratio of vector-amylase fixed,through the orthogonal fixed experimental the best condition was： temperature,60 ℃;pH=6.0;α-amylase concentration,60 U/ml;the ratio of α-amylase and cane cellulose derivatives vector,50 ml∶1 g and the buffer solution,citric acid-K2HPO4 buffer.The determination value of the immobilized enzyme activity was 34.77 U/g of fixatives.The Michaelis constant was 12.88 g/L.The half-life wasf 3.17 h,so that in the fixed-process there was no α-amylase from the products,thus,we can reduce the additional processing cost and recycle application at same time.