溶胶—凝胶法固定木瓜蛋白酶条件的优化

Optimization on the Conditions for Immobilizing Papain by Sol-gel Method

[目的]探索正硅酸乙酯包埋木瓜蛋白酶的过程。[方法]以正硅酸乙酯为前驱体,采用溶胶-凝胶法制备固定化木瓜蛋白酶并优化试验条件。[结果]在固定化木瓜蛋白酶的制备过程中,卵胆比、酶原溶液含水量、酶原溶液加入量、胆固醇含量、木糖含量的极差分别为39.318、13.280、15.350、9.810和2.210。卵胆比对木瓜蛋白酶固定化的影响最大,酶原溶液加入量次之。制备固定化木瓜蛋白酶的最佳条件为:酶原溶液含水量为90μlH2O/mg木瓜蛋白酶,木糖加入量为15 mg木糖/mg木瓜蛋白酶,正癸烷溶液中胆固醇含量为1.5%,卵磷脂与胆固醇的比值为4.5∶1.0(w/w),固定化过程中酶原溶液、正癸烷溶液和前驱体溶液的加入量分别为80、500和600μ。l[结论]该研究为固定化木瓜蛋白酶的制备奠定了试验基础。

[ Objective ] The research aimed to explore the process of embedding papain with ethyl silicate. [ Method ] With ethyl silicate as precursor, the immobilized papain was prepared by sol-gel method and the experimental conditions were optimized. [ Result] In the preparation process of immobihzed papain, the ranges of ratio of lecithin to cholesterol, water content in proenzyme solution, addition amount of proenzyme solution, cholesterol content and xylose content were 39. 318, 13. 280, 15. 350, 9. 810 and 2. 210 resp. The influence of ratio of lecithin to cholesterol on the immobilization of papain was greatest and that the addition amount of proenzyme solution was secondary. The optimum conditions for preparing immobilized papain were as follows： the water content in proenzyme solution was 90μl H2O/mg papain, the addition amount of xylose was 15 mg xylose/mg papain, the cholesterol content in n-decane solution was 1.5% , the ratio of lecithin to cholesterol was 4.5：1.0 （w/w） and the addition amounts of proenzyme solution, n-decane solution and precursor solution were 80, 500 and 600 μl resp. in immobilization process. [ Conclusion ] This research laid an experimental foundation for preparing immobilized papain.