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在模拟人体生理pH条件下,采用荧光光谱法和紫外-可见光谱法研究不同温度下(298 K、304 K和310 K)芦丁与溶菌酶相互作用的光谱特征,明确了芦丁对溶菌酶荧光猝灭的机理,确定了二者作用间结合位点及结合常数,测定了芦丁对溶菌酶活性的影响趋势。结果表明芦丁能与溶菌酶发生弱相互作用,该作用是由焓驱动的低温自发反应过程,芦丁通过静态猝灭机制使溶菌酶内源荧光产生猝灭。两者结合位点数接近于1,结合驱动力为氢键或范德华力,298 K时结合距离为4.02 nm。紫外吸收、同步荧光和三维荧光光谱均表明芦丁导致溶菌酶构象变得更加紧密。采用比浊法测定溶菌酶活性结果表明,芦丁可能影响溶菌酶活性位点Asp-52所处微环境极性,不利于Asp-52发挥氢键受体的催化作用,使溶菌酶溶菌活性降低。
Fluorescence spectroscopy and UV-Vis spectroscopy were used to study the spectral characteristics of interaction between rutin and lysozyme at different temperatures (298 K, 304 K and 310 K) under simulated physiological pH conditions. The effect of rutin on lysozyme Fluorescence quenching mechanism to determine the role of both binding sites and binding constants, determination of rutin lysozyme activity trends. The results showed that rutin could weakly interact with lysozyme, which is driven by the enthalpy low-temperature spontaneous reaction process. Rutin quenches endogenous fluorescence of lysozyme through static quenching mechanism. The number of binding sites between the two is close to 1, and the binding force is hydrogen bond or van der Waal’s force. The binding distance at 298 K is 4.02 nm. UV absorption, synchronous fluorescence and three-dimensional fluorescence spectra both showed that rutin led to a more compact conformation of lysozyme. The results of turbidimetric assay of lysozyme showed that rutin could affect the microenvironment polarity of lysozyme active site Asp-52, which is not conducive to the catalytic effect of Asp-52 on hydrogen bond acceptor, .