戊二醛交联法固定化β-D-呋喃果糖苷酶的研究

Study on Immobilized β-D-fructofuranosidase by Cross-linking with Glutaraldehyde

大豆低聚糖属于功能性低聚糖的一种,具有促进双歧杆菌生长繁殖等生理特性。采用戊二醛交联法固定β-D-呋喃果糖苷酶。研究固定化酶法生产大豆低聚糖过程中对酶固定化制备的条件、操作稳定性。采用戊二醛交联法固定β-D-呋喃果糖苷酶。结果表明:最佳固定化条件为:戊二醛浓度20 mmol.L-1,蛋白质浓度1.0%,酶与固定液之比1∶10,固定化时间2 h;此条件下制备的固定化酶平均酶活达340 U.g-1,酶活保留率80%以上。固定化酶在间歇反应器中具有较高的操作稳定性,固定化酶的半衰期为58 d,完全可以满足工业化生产的要求。

The soybean oligosaccharide is a kind of functional oligosaccharides, which can multiply the number of Bifidobacteria. A novel method to immobilize β- D- fructofuranosidase by cross- linking with glutaraldehyde was successfully constructed. The preparation condition and operation stability of immobilization enzyme were investigated during the process of soybean oligosaccharide production. A novel method to immobilize β-D-fructofuranosidase by cross-linking with glutaraldehyde was successfully constructed. The conditions of immobilization were investigated by orthogonal experiment method. The optimal conditions of immobilization investigated by orthogonal experiment method were as follows ： the concentration of glutaraldehyde and protein were 20 mmol · L^-1 and 1.0% , the ratio of enzyme and immobilization liquid was 1 ： 10, the immobilizing time was 2 h. Under the optimal condition the average activity of immobilized enzyme reached 340 U · g ^- 1 , after immobilization and the activity could retain more than 80%, The immobilized enzyme showed high operation stability in batch reactor,whose half-life was 58 d. The stability of immobilized enzyme could completely meet the need of industrial production.