可见分光光度法测定α-淀粉酶活力  

Determination of the Activity of α-Amylase by Visible Spectrophotometry

作　　者：张雪娇[1] 田欢 刘春叶[1] 尤静[1] ZHANG Xuejiao;TIAN Huan;LIU Chunye;YOU Jing(School of Pharmacy,Xi′an Medical University,Xi′an,710021,China)

机构地区：[1]西安医学院药学院,陕西西安710021

出　　处：《化学与生物工程》2020年第3期65-68,共4页Chemistry & Bioengineering

基　　金：陕西省教育厅项目(16JK1652,18JK0665);西安医学院校级项目(2017PT05,2019YU032)。

摘　　要：利用I-3与淀粉络合所呈现的蓝色在一定波长下有特征吸收这一特性,采用可见分光光度法,通过测定添加α-淀粉酶前后发生水解的淀粉量的变化测定α-淀粉酶活力。结果表明,淀粉与I-3在pH值6.5的磷酸氢二钠-柠檬酸缓冲溶液中显色良好,并在580 nm处具有最大吸光度。加入一定量的淀粉,α-淀粉酶浓度(x)在0.001~0.008 mg·mL^-1范围内与酶活力(y)线性关系良好,标准曲线方程为y=375.75x+0.1208(R^2=0.9939)。因此,可以利用碘与淀粉的显色反应对α-淀粉酶活力进行测定。According to the characteristic absorption of complex formed by I-3 and starch,the activity ofα-amylase is determined by visible spectrophotometry,which is determined through the amount of starch hydrolyzed before and after addingα-amylase.From the experimental results,we find that the maximum absorbance of complex is at 580 nm,and the color is stability in Na2HPO4-citric acid buffer with pH value of 6.5.When a certain amount of starch is added,changing the concentration ofα-amylase from 0.001-0.008 mg·mL^-1,there is a good linear relationship between the concentration(x)and the activity(y)ofα-amylase,and the linear equation is y=375.75x+0.1208(R^2=0.9939).This method can be used to determine the activity ofα-amylase.

关 键 词：Α-淀粉酶 酶活力 可见分光光度法 

分 类 号：O657.7[理学—分析化学]