Study On The Enzyme Catalytic Reaction And Interaction Between Proteins

As a biological macromolecule with biological catalytic function, enzyme is a kind of special protein with substrate specificity and catalytic activity, as we all know, there are a wide variety of proteins in cells, most researchers focus on static proteins to study the interaction between proteins and conduct theoretical research using traditional methods, while as an important participant in life activity, enzyme is seldom applied to the predecessors’ study. Therefore, this study aims at exploring the effects of different charged proteins on enzyme and enzymatic reaction to analyze the enzymatic reaction mechanism, then further explore the nature of the interaction between proteins and calculate the force who maintains the protein interaction.In the study, in order to investigate the effects of charged proteins on the enzymatic reaction, Zeta potential of charged proteins were measured, and α-amylase solution and Invertase solution were added into reduced glutathione(GSH), bovine serum albumin(BSA) and the two compounds with various concentrations for a known period of time, then the activity, kinetic parameters, and the activation energy were investigated. The results are as follows: the effects of charged proteins on different enzymes and enzymatic reactions are quite different. For α-amylase, under a certain concentration of these three kinds of proteins, such as 0.6 mmol/L, the activity of α-amylase was increased by 7.9%, 8.7%, 11.1% compared with the control. It was also found that both kinetic parameters Km and Vm could be decreased, Km decreased from 129.7 g/L to 116, 104.2, 85.1 g/L, respectively, whereas Vm decreased from 32 g/(L·min-1) to 30.5, 28.1, 23.7 g/(L·min-1). At the same time, the activation energy also decreased from 13.91 kJ/mol to 10.0, 9.91 and 9.88 kJ/mol. For Invertase, under a certain concentration of these three kinds of proteins, the activity of Invertase was increased by 7.35%, 7.74%, 8.64% compared with the control. It was also found that both kinetic parameters Km and Vm could be decreased, Km decreased from 44.51 g/L to 41.76, 39.76, 37.86 g/L, respectively, whereas Vm decreased from 24.1 g/(L·min-1) to 24.04, 23.7, 23.36 g/(L·min-1). At the same time, the activation energy also decreased from 11.9 kJ/mol to 11.58, 11.41 and 11.35 kJ/mol. In addition, this change in α-amylase was associated with the concentration of proteins, and the linear correlation was very good. Different charged proteins have different effects on the enzyme, and the mixture have the largest impact on enzyme, followed by BSA, and GSH. The results showed that owning to the existence of different charged proteins, on one hand, enzymatic activity and the affinity between enzyme and the substrate were increased, on the other hand, the rate of the reaction is faster, which is more efficient for enzyme to play its role in the reaction.From the perspective of the Zeta potential of protein and enzyme, the distance between proteins and enzymes and electric field force were calculated to have a deep discussion on the mechanism of influence charged proteins have on enzyme, it was thought that there was a certain distance formed between proteins and enzyme who were in the same potential, which created weak repulsion to enzyme, thus making the enzyme catalytic activity be better played. Based on this theory, the results of the experiment were analyzed, so as to lay a theoretical foundation in aspects such as improving the enzyme activity, synergy of multiple enzyme system in a variety of enzymes, and the interaction between biological macromolecules.