变性剂对橡胶籽中-葡萄糖苷酶结构及酶活的影响.pdfVIP

现代食品科技 Modern Food Science and Technology 2016, Vol.32, No.12 变性剂对橡胶籽中 β-葡萄糖苷酶结构及酶活的影响 刘宇 ，刘畅，刘石生 （海南大学热带生物资源教育部重点实验室，海南大学食品学院，海南海口 570228） 摘要：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。 关键词：橡胶籽；β-葡萄糖苷酶；变性特点；尿素 文章篇号：1673-9078(2016)12-120-124 DOI: 10.13982/j.mfst.1673-9078.2016.12.019 Effect of Denaturants on the Structure and Enzyme Activity of β-Glucosidase from Rubber Seeds LIU Yu, LIU Chang, LIU Shi-sheng (Key Laboratory of Tropical Biological Resources of Ministry of Education, College of Food Science, Hainan University, Haikou 570228, China) Abstract: β-Glucosidase extract was separated from a rubber seed protein solution to study the effect of different concentrations of denaturants on β-glucosidase activity, and laser particle size analysis and molecular fluorescence spectroscopy were used to study the effect of denaturants on the spatial structure of β-glucosidase. β-Glucosidase activity showed a declining trend with increased denaturant concentration, and increased temperature accelerated the pace of this decline. Analysis of the spatial structure of β-glucosidase showed that the molecular parti