牛胰脂肪酶交连酶聚体的制备及性能研究

Preparation and properties of cross-linked enzyme aggregates of cattle pancreatic lipase

以牛胰脂肪酶(cattle pancreatic lipase,CPL)为研究对象,采用交联酶聚体技术,研究交联酶聚体脂肪酶(CPL-CLEAs)的制备条件、催化性能和酶促反应动力学参数。结果表明,交联酶聚体脂肪酶制备的最优制备条件为:硫酸铵饱和度80%,戊二醛终浓度1.5%,交联时间1 h,在此条件下,所得CPL-CLEAs最高酶活回收率为90%。CPL-CLEAs的最适催化温度是60℃,与游离脂肪酶相比,CPL-CLEAs的最适催化温度提高了10℃,且温度稳定性、储存稳定性和操作稳定性均有所提高。重复使用7次后,CPL-CLEAs仍能保持初始酶活的43%。

In this study,cross-linked enzyme aggregates（CLEAs） of cattle pancreatic lipase（CPL-CLEAs） was prepared by cross-linked enzyme aggregates technology,the preparation conditions,catalysis properties,and dynamics parameters of CPL-CLEAs were investigated. The results showed that the optimum conditions of the preparation of CPL- CLEAs were as followed ： final saturability of the ammonium sulfate was 80 %, final concentration of glutaraldehyde was 1. 5 %, and corss- link time was 1 h. Under this condition, the highest activity recovery reached 90%. In addition,compared to free lipase,the optimum temperature of CPL-CLEAs was 60 ℃,which was increased about 10 ℃. The thermal stability,operational stability,and storage stablility of CPL-CLEAs were significantly enhanced compared with free lipase. Furthermore,the CPL-CLEAs still retained 43% of its initial activity after consecutive 7 cycles.