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本文以橡胶籽蛋白溶液为原料,分离得到提取β-葡萄糖苷酶,研究了不同浓度的变性剂对β-葡萄糖苷酶酶活的影响,并通用激光粒径分析和分子荧光光谱分析等方法研究了变性剂对酶的空间结构的影响。结果表明:β-葡萄糖苷酶的酶活随变性剂浓度的增加大体上呈现下降趋势,温度的增加会加速下降的速度,通过对β-葡萄糖苷酶的空间结构分析发现尿素变性酶后,酶溶液的分子粒径变小,推断为β-葡萄糖苷酶的多聚体结构发生解聚。低浓度尿素在1 h时间内对β-葡萄糖苷酶有激活作用,酶活力明显提高,表明酶的空间结构变化与尿素浓度大小有关。通过分子荧光光谱分析发现随酶溶液中尿素浓度增加,荧光峰位红移,表明色氨酸残基不断暴露,蛋白质分子结构不断展开。本研究为进一步开发利用橡胶籽中β-葡萄糖苷酶提供了实验依据。
In this paper, β-glucosidase was extracted from rubber seed protein solution, and the effects of different denaturant denaturing agents on β-glucosidase enzyme activity were studied. The methods of universal laser particle size analysis and molecular fluorescence spectroscopy The effect of denaturant on the spatial structure of the enzyme. The results showed that the increase of the concentration of β-glucosidase enzyme activity showed a general downward trend, the increase of temperature will accelerate the rate of decline, through the spatial structure analysis of β-glucosidase urea denaturation enzyme, enzyme The molecular size of the solution became smaller, suggesting that the polymer structure of β-glucosidase was depolymerized. Low concentration urea activated β-glucosidase within 1 h, and the enzyme activity was significantly increased, indicating that the spatial structure of the enzyme was related to the urea concentration. By molecular fluorescence spectroscopy analysis found that with the increase of urea concentration in the enzyme solution, the fluorescence peak redshift, indicating that tryptophan residues continue to be exposed, the molecular structure of the protein continues to unfold. This study provides experimental evidence for the further development and utilization of β-glucosidase in rubber seeds.