多孔壳聚糖微球固定化碱性蛋白酶的研究

Study on immobilization of alkaline proteinase on porous chitosan microspheres

采用自制的多孔壳聚糖微球固定化碱性蛋白酶,以酶活回收率为参考指标,分别对酶浓度、酶与载体用量比、戊二醛浓度、吸附时间、交联时间、BSA浓度等进行了单因素试验,考察了其对碱性蛋白酶固定化的影响,确定了较好的固定化碱性蛋白酶的工艺条件。结果表明,碱性蛋白酶固定化的最佳工艺条件为:酶与载体用量比315 U/g、酶浓度45 mg/mL、固定化温度4℃、固定化pH7.2、吸附时间48 h、交联时间8 h、戊二醛浓度为1%、BSA浓度4.5 mg/mL,此时RRA达到65.46%,固定化碱性蛋白酶也具有较好的理化性质。

Alkaline protease was immobilized by using porous chitosan microspheres which were prepared by ourselves, the proportion of enzyme to carriers, the concentrations of alkaline protease, glutaraldehyde and BSA, pH, temperature, absorption and crosslinking time were studied by using recycling rate of activity as index and the better method was found. The test indicated that the optimum technics were as follows： the proportion of enzyme to carriers was 315 U/g, the concentration of alkaline protease, glutaraldehyde and BSA were 45 mg/ mL, 1%, 4.5 mg/mL respectively, pH was 7.2, temperature was 4 ℃, absorption and crosslinking time were 48 hours and 8 hours, the recycling rate of activity reached 65.46%, the alkaline protease has good physical and chemical properties.