棉织物对甲苯磺酰氯活化法固定化菊粉酶工艺的研究

Technology of Inulinase Immobilization on Tosylated Cotton Cloth

以4-甲苯磺酰氯活化的棉纤维为载体,对固定化菊粉酶的条件进行探讨。通过对不同纤维成分布料的比较,绒布为载体时得到了较高的固定化酶活。分别采用单因素试验和正交试验,得到优化的工艺参数:每克干基棉布加入10ml干吡啶,对甲苯磺酰氯与干吡啶的比例为1g:1ml,偶联pH值4.5,离子强度0.2mol/L,加酶量80U/g,菊粉酶酶活回收率达到83.60%。在优化的工艺条件下,对糖分组分进行HPLC分析,固定化酶与250g/L菊糖(5U/g菊糖)作用1.5h,低聚果糖含量达到28.21%。

Inulinase from Aspergillusficuum SK004 was immobilized on cotton cloth activated with p-toluenesulfonyl chloride （tosyl chloride）. The immobilization conditions were optimized and the results showed that the optimal amount of dry pyridine was 10ml to each gram of cotton and the ratio of tosyl （g）/pyridine （ml） was lg： lml. The optimal coupling pH was found to be 4.5 and ionic strength was 0.2mol/L. 80U enzyme was immobilized onto each gram of cotton cloth. The highest enzyme activity yield achieved 83.60% through orthogonal test. At these conditions, immobilized enzyme reacting with 250g/L inulin for 1.5h gained 28.21% FOS （fructose oligosaccharide） analyzed by HPLC. Among different fibrous materials tested, the charpie cotton cloth showed the highest immobilized enzyme activity.