复合保护剂对透明质酸酶的稳定性研究

以球形节杆菌(Arthrobacter globiformis)A152发酵生产的透明质酸酶(HAase)的酶活力为衡量指标,通过研究糖类、醇类、生物大分子和金属离子等保护剂对提高透明质酸酶热稳定性的作用,考察添加复合保护剂对透明质酸酶热稳定性和储存稳定性的影响。结果表明,山梨糖醇、蔗糖和可溶性淀粉作为保护剂,可提高HAase酶的热稳定性。通过单因素和正交实验优化后得到了效果最佳的复合保护剂配方,即山梨醇7.5%、蔗糖10%、可溶性淀粉0.2%。优化后添加保护剂的HAase在42℃保温2 h后,其酶活保留率为88.68%,较对照提高了64.18%,在37℃,通过添加复合保护剂,其半衰期提高了48%。在4℃储藏90 d,添加复合保护剂和防腐剂(0.1%的山梨酸钾和0.05%的苯甲酸钠)的HAase酶活保留率高达85.52%。

Study on Composite Protective Agent in the Stability of Hyaluronidase

The enzyme activity of hyaluronanlyase(HAase)produced by fermentation of Arthrobacter globiformis A152 was used as a measure,in order to enhance the thermostability of HAase,sugars,alcohols,biological macromolecules and metal ions were added into the preparation of HAase,and the effects of composite protective agent on the thermostability and storage stability of HAase were studied. The results indicated that sorbitol,sucrose and soluble starch as protective agents could improve the thermostability of HAase,significantly. The best composite protective agent(7.5%sorbitol,10%sucrose and 0.2%soluble starch)was obtained using the orthogonal experimental design. The residual activity of HAase with composite protective agent maintained 88.68% after incubating 2 h at 42℃,which was 64.18% higher than that of control without composite protective agent. Furthermore,the half-life of HAase with composite protective agent was 48% higher than that of control at 37℃. Finally the residual activity of HAase containing composite protective agent and preservatives(0.1% potassium sorbate,0. 05% sodium benzoate)reached 85.52% after storing at 4℃ for 90 days.