| 金璐 王彦 薛芸 杨婵 阎超△.植物源β- 葡萄糖苷酶活性检测方法学研究[J].,2016,16(26):5032-5037 |
| 植物源β- 葡萄糖苷酶活性检测方法学研究 |
| Methodological Research on the Activityof β-Glycosidase fromPlant Origin |
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| DOI： |
| 中文关键词: β- 葡萄糖苷酶 酶活性 方法学研究 |
| 英文关键词: β-Glycosidase Enzyme activity Methodological research |
| 基金项目:国家自然科学基金项目（21175092）；上海市科委科研计划项目（15142200200） |
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| 中文摘要: |
| 目的：来源于植物β-葡萄糖苷酶活性检测的方法学研究。方法：以对硝基苯基β-D- 葡萄糖苷（pNPG）为底物，β-葡萄糖苷酶
为催化剂催化底物生成对硝基苯酚，用紫外分光光度法测定对硝基苯酚的含量，根据生成的对硝基苯酚含量计算酶活性，并对此
进行方法学研究。结果：对硝基苯酚标准曲线的相关系数为0.999；催化反应的线性相关系数为0.999；反应精密度的相对标准偏
差为0.05%；方法重复性的相对标准偏差为2.8%；方法耐用性考察中，酶活性均大于1200 U/g，九个不同方法参数的酶活性相对
标准偏差为4.6%；中间精密度实验中，十二份样品酶活性的相对标准偏差为6.6%；稳定性实验中，β- 葡萄糖苷酶与底物反应后，
放置0 小时、16小时、22 小时后，测定的酶活性均大于1200 U/g，相对标准偏差为1.1%。结论：此方法线性较好，有良好的方法重
复性、精密度、中间精密度、耐用性和溶液稳定性，适用于来源于植物的β- 葡萄糖苷酶活性检测。 |
| 英文摘要: |
| Objective:Methodology research for the activity test of β-glycosidase from plants was conducted.Methods:The
substrate of 4'-Nitrophenyl-β-D-glucopyranoside (pNPG) was catalyzed by β-glycosidase to generate 4-nitrophenol. The enzymatic
activity of β- glycosidase was determined through assay the concentration of 4-nitrophenol by the mean of ultraviolet spectrophotometry,
and the methodology research for this method was fully evaluated.Results:The correlation coefficient of standard curve for 4-nitrophenol
was 0.999; The linear correlation coefficient for catalytic reaction was 0.999; The relative standard deviation for precision was 0.05%;
The relative standard deviation for method reproducibility was 2.8%; The enzyme activity was more than 1200 U/g and the relative
standard deviation was 4.6% for nine different method parameters in the observation of method for durability; The relative standard
deviation of enzyme activity for 12 samples was 6.6%, respectively, in the experiment of intermediate precision; For the stability
experiment, three time points (0 hour, 16 hour, and 22 hour after the reaction ) were measured. The result shows that the activity of
β-glycosidase is above 1200 unit/g after the reaction with the substrate (0 hour), 16 hours later, as well as 22 hours later, where the RSD
is 1.1%.Conclusion:The developed analytical method showed good linearity, repeatability, precision, intermediate precision, stability
and durability. It can be applied in the determination of the activity for β-glycosidase fromplants origin. |
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