丙氨酸依赖型转氨酶高通量快速检测方法的建立  

Rapid and sensitive high-throughput determination method for alanine-dependent transaminase

作　　者：刘欢 吴旭日 陈依军 LIU Huan;WU Xuri;CHEN Yijun(Laboratory of Chemical Biology School of Life Science and Technology,China Pharmaceutical University,Nanjing 210009,China)

机构地区：[1]中国药科大学生命科学与技术学院化学生物学研究室,南京210009

出　　处：《中国药科大学学报》2018年第4期490-495,共6页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.21778076);江苏高校"青蓝工程"资助项目~~

摘　　要：本研究以来源自Vibrio fluvialis JS17的丙氨酸依赖型转氨酶VfTA为研究对象,偶联丙酮酸氧化酶和辣根过氧化物酶,建立了一种基于反应液颜色变化的转氨酶活性测定新方法,反应条件优化后,考察了VfTA活力单位数与反应液400 nm处吸收度的线性关系,并将此法用于商业化转氨酶ATA117活力的研究。结果显示:该法对转氨酶Vf TA的活力检测限可达0.45 U/mL,对ATA117的活力检测限可达0.5 U/mL,并且可根据反应液颜色深浅初步判断酶活力情况。A new method for determining transaminase activity based on the color change of the reaction solution was established,by using alanine-dependent transaminase Vf TA from Vibrio fluvialis JS17 as the research object coupled with pyruvate oxidase and horseradish peroxidase.After the optimization of the conditions,the linear relationship between Vf TA activity units and the absorbance at 400 nm was investigated.This method was also applied to determine the activity of commercial transaminase ATA117.The results showed that the detection limit of transaminase Vf TA activity was up to 0.45 U/mL and the detection limit of ATA117 activity was up to 0.5 U/mL.The transaminase activity could be quickly judged according to the color depth of the reaction solution.

关 键 词：丙氨酸依赖型转氨酶 丙酮酸氧化酶 辣根过氧化物酶 颜色反应 转氨酶活性测定 

分 类 号：TQ460[化学工程—制药化工]