双功能化无机介孔材料对猪胰蛋白酶催化性能的影响

Effect of bifunctional mesoporous material on Porcine trypsin catalytic properties

通过在SBA-15介孔材料表面引入带有不同官能团及不同亲疏水性质的修饰剂(-SH,-Cl,-NH_2,-C_1,-C_3,-C_8及-C_(16)),改善介孔材料与猪胰蛋白酶的结合力及蛋白酶所处微环境,从而提高酶固定化效率,活性及其稳定性。结果表明,官能团修饰后载体固定化酶PT-NH2/C8-SBA活性达到255 U/mg胰蛋白,较之未修饰载体固定化酶PT-SBA活性(195 U/mg胰蛋白)有大幅度的提高;并且重复使用五次及在室温下保持5 d后,PT-NH_2/C_8-SBA活力仍然能保持初始活力的85%及81%。而PT-SBA仅仅保留了初始活性的51%及67%。固定化酶的酶学性质和催化反应动力学研究表明,底物与固定化酶PT-NH_2/C_8-SBA具有更好的亲和力,固定化酶PT-NH_2/C_8-SBA有较高的活力和稳定性。

In this paper, binding force between mesoporous materials and trypsin was improved by introducing different modifiers functional groups on the surface of SBA-15 （-SH,-Cl,-NH2,-C1,-C3.-C8 and -C16）in order to increase the efficiency of enzyme loading,activity and stability.Activity of PT-NH2/Cs-SBA could be found up to 255 U/mg trypsin,the activity of PT-SBA was just 195 U/mg trypsin.After the fifth recycle and storage at room temprature of 5 d, PT-NH2/C,-SBA was still able to maintain 85% and 81% of initial activity, while the PT-SBA retained only 51% and 67% of the initial activity.Kinetic studies showed that immobilized enzyme PT-NH2/C,-SBA has better affinity with substrate.