用于高通量筛选蛋白酶的氨基酸间接荧光测定法  

Determination of amino acid by indirect fluorimetry for high flux screening of protease

作　　者：潘力[1] 李方方[1] 崔翠[1] 胡杰[1] 

机构地区：[1]华南理工大学生物科学与工程学院,广东广州510006

出　　处：《食品工业科技》2009年第6期338-340,共3页Science and Technology of Food Industry

基　　金：广东省科技计划项目(2007A010900001)

摘　　要：建立了一种用间接荧光分析快速测定氨基酸的新方法。Cu^2＋与钙黄绿素结合成钙黄绿素-Cu^2＋后发生荧光淬灭，氨基酸与Cu^2＋形成的配合物比钙黄绿素-Cu^2＋配合物更稳定，氨基酸与钙黄绿素-Cu^2＋反应使钙黄绿素游离释放出荧光．能灵敏、间接的测定氨基酸含量。对酪氨酸检测的线性范围为10×10^-3--100×10^-3g·L^-1，相关系数r=0．9991。结果表明，该方法可用于测定蛋白酶的水解活性以及实时监测释放游离氨基酸的反应过程。A novel indirect fluorescein-based method has been developed for measuring amino acids.Calcein forms a non-fluorescent complex with Cu^2＋.Since the amino acid-Cu^2＋ complex is more stable than the calcein-Cu^2＋ complex,the fluorescence of calcein would be released again, which corresponds to the concentration of amino acids.The linear range for tyrosine was 10×10^-3--100×10^-3g·L^-1, and the correlation coefficient was 0.9991. Meanwhile,the method can be used to detect hydrolytic activity of various proteases and to monitor reactions releasing free amino acids in real time.

关 键 词：荧光分析法 钙黄绿素 金属离子 氨基酸 蛋白酶 

分 类 号：TS207.3[轻工技术与工程—食品科学]