A pulsed electric field (PEF) treatment exhibits different effects on trypsin; however, the mechanism of enzyme activation remains unclear. Herein, chemical experiments combined with molecular dynamics simulations revealed the mechanism of trypsin activation by PEF treatment at the molecular level. The results indicated that compared with the values at 0 kV/cm, the enzyme activity, Vmax, and Kcat at 20 kV/cm increased by 9.30%, 4.74%, and 4.30%, respectively, and Km decreased by 11.14%, indicating an improved interaction between the enzyme and substrate. The simulation results revealed that PEF treatment increased the number of molecular hydrogen bonds and the solvent-accessible surface area, while decreasing the rotation radius and random coil content by 5.00% and 3.37%, respectively. Molecular docking indicated that PEF treatment altered the active center and increased the affinity between the enzyme and substrate. The simulation results were consistent with those of the spectroscopic experiments conducted on trypsin after PEF treatment.

脉冲电场 (PEF) 处理对胰蛋白酶表现出不同的影响；然而，酶激活的机制仍不清楚。在此，化学实验结合分子动力学模拟揭示了PEF处理在分子水平上激活胰蛋白酶的机制。结果表明，与0 kV/cm时的值相比，20 kV/cm时的酶活性、Vmax和Kcat分别提高了9.30%、4.74%和4.30%，Km降低了11.14%，表明改善酶和底物之间的相互作用。模拟结果表明，PEF处理增加了分子氢键的数量和溶剂可及的表面积，而旋转半径和无规卷曲含量分别降低了5.00%和3.37%。分子对接表明PEF处理改变了活性中心并增加了酶和底物之间的亲和力。模拟结果与PEF处理后对胰蛋白酶进行的光谱实验结果一致。