中性蛋白酶在海藻酸钠和壳聚糖中的固定化研究

Study on Immobilization of Neutral Protease in Sodium Alginate and Chitosan

以天然无毒海藻酸钠、壳聚糖为固定化载体材料,以戊二醛为交联剂,选用包埋交联法对中性蛋白酶进行固定化研究,并对固定化条件进行了优化,测定了固定化酶的部分酶学性质。通过单因素试验和正交试验确定了该固定化酶的最佳条件参数:壳聚糖浓度为2%,海藻酸钠浓度为2%,冰醋酸浓度为1%,CaCl2浓度为3%,戊二醛浓度为0.75%,固定化时间为18h,pH为8.0。,该条件下固定化酶酶活回收可达92.91%;固定化酶的最适pH为6.9,最适温度为51℃。相对于游离酶,固定化酶便于分离回收,同时增加了利用效率,有效的降低了生产成本,具备较好的经济效益和研究价值。

Natural non-toxic chitosan and sodium alginate were used as the immobilization materials, glutaraldehyde was chosen as cross-linking agent to immobilize the neutral protease with embedding cross-linking. The immobilization conditions and part of enzymatic properties of immobilized enzyme were studied in this paper. The best conditions for immobilized enzyme were determined by orthogonal and single factor experiments. The results were as follows： the chitosan concentration of 2%, 1% acetic acid, 2%sodium alginate, 3% CaC12, 0.75% glutaraldehyde, immobilized time 18h and pH 8.0. Using this method, 92.91% enzyme activity was recovered. The best enzymatic reaction conditions for immobilization enzyme were 51℃ and pH 6.9. Compared with free enzyme, the immobilized enzyme has better economic benefits and significant value because of easiness in separation, increasing the use efficiency and reducing the production cost effectively.