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本文研究了染料木素与β-葡萄糖苷酶的相互作用。采用荧光光谱法研究了染料木素与β-葡萄糖苷酶的结合反应,二者的相互作用使β-葡萄糖苷酶发生内源荧光猝灭,属于静态猝灭机制。通过计算得到染料木素与β-葡萄糖苷酶在17、27和37℃下的结合常数分别为3.69×104、3.06×104和2.36×104 L·mol-1。同步荧光研究了染料木素对β-葡萄糖苷酶结构的影响,染料木素可使β-葡萄糖苷酶的构象发生变化、色氨酸残基所处微环境的疏水性降低。染料木素对酶活性的抑制试验结果表明其对酶的活性具有明显的抑制作用,双分子结合速率常数为1.2×103(mol·L-1)-1.min-1。结合AutoDock 4.2分子对接软件模拟研究了二者的键合模式和键合机制,表明氢键和疏水作用对染料木素与β-葡萄糖苷酶的结合起重要作用,同时还存在静电作用。
This article investigates the interaction of genistein with β-glucosidase. The binding reaction of genistein and β-glucosidase was studied by fluorescence spectroscopy. The interaction between them led to the endogenous fluorescence quenching of β-glucosidase, which belongs to the static quenching mechanism. The binding constants of genistein and β-glucosidase at 17,27 and 37 ℃ were calculated to be 3.69 × 104, 3.06 × 104 and 2.36 × 104 L · mol-1, respectively. The effect of genistein on the structure of β-glucosidase was studied by synchronous fluorescence. The genistein could change the conformation of β-glucosidase, and the hydrophobicity of tryptophan residues in the microenvironment decreased. The results of the inhibition test of genistein on the enzyme activity showed that the activity of the genistein was significantly inhibited, and the bimolecular binding rate constant was 1.2 × 103 (mol·L -1) -1.min -1. Combined with AutoDock 4.2 molecular docking software simulation study of the two bonding modes and bonding mechanism, indicating that hydrogen bonding and hydrophobic interaction on genistein and β-glucosidase play an important role, but also the existence of electrostatic interactions.