响应面法优化蛋壳内膜酶解制备抑菌活性物质的工艺条件

摘要

T he bacteriostatic activity of protease enzymolysis of eggshell membrane was tested.The factors of alkaline pretreatment,digestion temperature,digestion time,quantity of enzyme,enzyme solution pH on bacteriostatic effect of eggshell membrane enzymatic hydrolysate were analyzed.On the basis of single factor experiment,the Design Expert 8.0.6 analysis software,the Box-Behnken central composite design and response surface method(RSM)were used to optimize the enzymolysis technology.The best process conditions for eggshell membrane enzymolysis were:45 ℃ for enzyme solution temperature,2 h for enzymolysis time, 37.51 U/mg of enzyme dosage,pH 10.14 for enzyme solution.Under this condition,the bacteriostatic circle diameter of eggshell membrane enzymatic hydrolysis on chicken S.enteritidis was(17.927±0.19)mm,close to the theory optimal value of (17.941 ±0.21)mm,bacteriostatic effect was improved 10.52% than before optimization(16.22 mm).The results showed that the fitted equation was consistent with the actual and the result was accurate and reliable.%本试验以鸡蛋壳内膜为试验材料,用蛋白酶进行酶解,测定酶解液对微生物的抑菌活性,分析碱性预处理、酶解温度、酶解时间、加酶量、酶解pH对鸡蛋壳内膜酶解液抑菌效果的影响,并在单因素试验基础上,利用De-sign Expert 8.0.6分析软件,采用Box-Behnken中心组合设计法及响应面分析法(RSM)优化酶解工艺条件,得到鸡蛋壳内膜酶解液抑菌效果最佳的酶解工艺条件为:酶解温度45℃,酶解时间2 h,酶用量37.51 U/mg,酶解pH 10.14.该工艺条件下,鸡蛋壳内膜酶解液对S.enteritidis的抑菌圈直径为(17.927±0.19)mm,与理论最优值(17.941±0.21)mm接近,抑菌效果比优化前提高了10.52%,说明拟合方程与实际相符合,所得结果准确可靠.