固定化酶解猪小肠黏膜制备肝素的工艺研究

Preparation of Heparin from Porcine Small Intestine Mucosa by Immobilized Enzyme Hydrolysis

本文以肝素效价为指标,探索了固定化胰蛋白酶对小肠黏膜的酶解作用。在单因素试验基础上采用Box-Behnken中心组合试验设计和响应面(RSM)分析法,以肝素效价为响应值,通过考察p H值、加酶量、酶解温度及料液比,优化了固定化胰蛋白酶酶解制备肝素的工艺,建立了二次回归方程。结果表明,p H值、酶解温度、加酶量、及料液比均对肝素提取具有显著影响。固定化酶制备肝素最佳工艺条件为:p H为11,加酶质量分数为0.24%,酶解温度为46℃,料液比为1∶3.3(g/m L),酶解时间为5 h,在此条件下获得肝素酶解液效价为15.28 U。

In the present study,the effects of immobilized trypsin on small intestinal mucosa were investigated based on the heparin titer. Using heparin titer as the response value, and on the basis of the single factor experimental results, onefactor-at-a-time tests followed by RSM analysis was carried out to achieve the optimal extraction conditions of heparin from porcine intestinal mueosa. The results showed that heparin extraction was significantly affected by pH value,temperature ,enzyme dosage, and solid-llquid ratio. The optimal conditions was as follows： enzymatic hydrolysis temperature of 46 ℃, solid-liquid ratio of 1 ： 3.3 （g/mL） , 0.24% concentration of immobilized enzyme, pH 11 and 5 hr hydrolysis time, respectively. The heparin titer was reached to 15.28 U under the optimal conditions.