正丁醇和异丙醇对溶菌酶毒性效应及作用机理研究

发布时间：2018-01-11 05:04

本文关键词：正丁醇和异丙醇对溶菌酶毒性效应及作用机理研究　出处：《山东大学》2017年硕士论文　论文类型：学位论文

【摘要】：作为挥发性有机污染物(VOC)的其中成员,正丁醇和异丙醇广泛应用于各种生产活动,由此导致空气的污染并进一步危害人类健康。因此对二者进行环境毒理学研究势在必行。本文选取溶菌酶作为生物大分子的代表,以分子毒理学理论作为研究指导,采用多种光谱技术、酶活性检测和分子对接技术,研究正丁醇和异丙醇与溶菌酶相互作用时溶菌酶的构像、活性的变化、微环境以及两者的结合位点和结合模式等,并制定了有效的实验方法。荧光光谱结果显示正丁醇能够和溶菌酶分子结合并且使溶菌酶中的荧光基团远离猝灭剂(正丁醇)。同步荧光和三维荧光的结果表明正丁醇改变了色氨酸残基的微环境。紫外-可见光谱和圆二色光谱结果表明正丁醇改变了溶菌酶的二级结构,进而导致α-螺旋的含量增加和β-折叠的含量减少。分子对接研究结果展示了正丁醇和溶菌酶二者的结合位点和结合力。酶活性实验正丁醇对溶菌酶的酶活影响很小,表明正丁醇没有结合到溶菌酶的酶活中心。荧光光谱结果显示异丙醇导致溶菌酶的荧光发生猝灭,同步荧光结果表明色氨酸残基和酪氨酸残基的微环境变得更加疏水。紫外和圆二色光谱结果一致,显示异丙醇使溶菌酶的二级结构更加疏松。结合位点和正丁醇不一样但结合力都是氢键和疏水力。酶活实验结果也暗示了异丙醇没有结合到溶菌酶活性中心,所以对酶活影响不大。本研究不仅可以揭示正丁醇和异丙醇对溶菌酶的毒性作用及机理,而且为溶菌酶在医疗、食品防腐及生物工程上广泛应用以及为正丁醇和异丙醇作用的研究提供相应的资料支持。
[Abstract]:As a member of volatile organic pollutant (VOC), n-butanol and isopropanol are widely used in various production activities. As a result, air pollution and further harm to human health. Therefore, it is imperative to study the environmental toxicology of the two. In this paper, lysozyme is selected as the representative of biological macromolecules. Under the guidance of molecular toxicology theory, the conformation and activity of lysozyme in the interaction of n-butanol and isopropanol with lysozyme were studied by a variety of spectroscopic techniques, enzyme activity detection and molecular docking techniques. Microenvironment and their binding sites and binding patterns. The fluorescence spectra showed that n-butanol could bind to lysozyme molecule and make the fluorescence group in lysozyme away from quenching agent (n-butanol). The results of synchronous fluorescence and 3D fluorescence showed that n-butanol changed the microenvironment of tryptophan residues, and the UV-Vis spectra and circular dichroism spectra showed that n-butanol changed the secondary structure of lysozyme. The results of molecular docking show the binding sites and binding force of n-butanol and lysozyme. Enzyme activity experiment of n-butanol on lysozyme. It's a little loud. The fluorescence spectra showed that isopropanol caused fluorescence quenching of lysozyme. The results of synchronous fluorescence showed that the microenvironment of tryptophan residues and tyrosine residues became more hydrophobic. The results of UV and circular dichroism spectra were consistent. The results showed that isopropanol made the secondary structure of lysozyme more loose. The binding site was different from n-butanol but the binding force was hydrogen bond and hydrophobicity. The results of enzyme activity also suggested that isopropanol did not bind to the active center of lysozyme. This study can not only reveal the toxicity and mechanism of n-butanol and isopropanol to lysozyme, but also the lysozyme in medical treatment. Food anticorrosion and bioengineering are widely used, and the corresponding data support for the study of n-butanol and isopropanol is provided.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：X171.5

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