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通过荧光光谱研究了枯草芽孢杆菌超氧化物歧化酶(SOD)的分子构象在不同浓度盐酸胍、尿素以及不同pH条件下的变化及其与功能的关系。结果表明,SOD构象与功能的变化与其荧光光谱的变化有明显的对应关系。随着盐酸胍浓度的增加,SOD发射峰红移,荧光强度不断下降,且这两种变化同步发生;酶则出现快失活慢变构现象。而利用尿素变性SOD时,SOD先是荧光强度增强,然后发射峰红移,酶活力的下降与构象变化同步进行。尿素和盐酸胍使SOD变性的机理明显不同。处于不同酸碱度时,SOD活力的变化与荧光光谱所表现出的构象变化几乎一致。通过荧光分析法是可以对不同条件下SOD分子的构象变化及其酶活进行快速测定的。
The change of molecular conformation of Bacillus subtilis superoxide dismutase (SOD) under different concentrations of guanidine hydrochloride, urea and different pH and its relationship with function were studied by fluorescence spectroscopy. The results showed that the change of conformation and function of SOD had obvious corresponding relation with the change of fluorescence spectrum. With the increase of the concentration of guanidine hydrochloride, SOD emission peak red-shift, fluorescence intensity decreased, and the two changes occur simultaneously; enzyme deactivation occurs slowly slow phenomenon. However, using urea-denatured SOD, the fluorescence intensity of SOD increases first and then the emission peak shifts red. The decrease of enzyme activity is in parallel with the conformational change. The mechanism of degeneration of SOD by urea and guanidine hydrochloride is obviously different. At different acidity and alkalinity, the change of SOD activity is almost the same as that of the fluorescence spectrum. Fluorescence analysis method is able to quickly determine the conformational changes of SOD molecules and their enzymatic activities under different conditions.