双功能化无机介孔材料对猪胰蛋白酶催化性能的影响

邹彬; 邓甜; 韦诗宇; 夏姣姣; 霍书豪

doi:10.13386/j.issn1002-0306.2016.07.015

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

摘要

摘要: 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,-NH₂,-C₁,-C₃,-C₈及-C₁₆),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-NH₂/C₈-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-NH₂/C₈-SBA具有更好的亲和力,固定化酶PT-NH₂/C₈-SBA有较高的活力和稳定性。

Abstract: 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/ C₈- SBA could be found up to255 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/ C₈- 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/ C₈- SBA has better affinity with substrate.