Fe3O4@mSiO2@nSiO2 can be modified with γ-aminopropyl triethoxysilane (APTES) for the immobilization of trypsin after glutaraldehyde cross-linking. The optimum conditions of the concentration of glutaral-dehyde, the given amount of enzyme and reaction time are as following; 1. 875% glutaraldehyde( volume rati-o) ,0. 3 mg/mL trypsin solution, 4 h reaction time. Under these optimum conditions, the immobilizing rate and the activity retention rate are as high as 50. 5% and 77. 3% respectively. The immobilizing trypsin has a high efficiency on microwave assisted BSA digestion at 40 ℃. The results show that in hydrolyzate there is a variation peak absorb at A = 280 nm. A significant increasing absorbs of peptide fragments and some new pep-tide fragments can be observed by HPLC analysis.%以Fe3O4@mSiO2@nSiO2介孔材料为载体,γ-氨丙基三乙氧基硅烷接枝,戊二醛交联,研究胰蛋白酶的固定化及其在蛋白酶解中的应用.对蛋白酶固定化条件的优化研究表明,φ(戊二醛)＝1.875％、给酶量为0.3 mg/mL、反应时间为4h时酶的固定化效率可达50.5％,且保持77.3％相同含量酶的活性.对牛血清蛋白的酶解实验表明,在微波辅助作用下,磁性微球材料固定化酶可快速高效地酶解牛血清蛋白,酶解液在λ=280 nm处有较明显的吸收信号变化,HPLC分析表明酶解液中多肽片段吸收值变化明显,同时有新的多肽片段生成.
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