固定化乳酸脱氢酶活性的X射线微区分析  

X-ray Micro-area Analysis of Activity of Immobilized Lactate Dehydrogenase

作　　者：丁良[1] 姚子华[1] 李彤[1] 

机构地区：[1]河北大学化学与环境科学学院理化分析中心

出　　处：《分析测试学报》2003年第2期56-59,共4页Journal of Instrumental Analysis

摘　　要：利用X射线微区分析 ,对吸附法得到的固定化乳酸脱氢酶的活性进行了分析;以乳酸为底物,铁氰化钾及铜离子作为捕捉剂,底物经乳酸脱氢酶及氧化型辅酶NAD +的作用下催化产生丙酮酸,反应生成的还原性辅酶NADH使铁氰化物还原为亚铁氰化物,后者和铜离子反应产生具有高电子密度的亚铁氰化铜沉淀,可以确定固定化乳酸脱氢酶的催化活性部位 ;结果表明 ,以合成出的大孔吸附树脂为固定化酶载体 ,其酶活性较高 ,活性乳酸脱氢酶分布较均匀 ,得到了固定化乳酸脱氢酶的活性定位的最佳条件 ,并对不同结构载体固定化酶活性进行了研究。The activity of immobilized lactate dehydrogenase(LHD)prepared via absorption method was studied by X-ray micro-area analysis using lactate as a substrate,K 3 and copper ion as trapping agent.The substrate was catalysed by immobilized LHD and coenzyme NAD + to produce pyruvatic acid and coenzyme NADH.K 3 was reduced to K 4 by NADH.The resulting K 4 was captured by CuSO 4 to form Cu 2 precipitate having a high electron density.The precipitate deposited on active sites of imˉmobilized lactate dehydrogenase.Thus active sites of immobilized lactate dehydrogenase can be determined.The results showed that the LHD immobilized on the self-synthesized macroporous resin had greater activity,and the more uniform distribution.Most of activated enzyme was immobilized on the macroporous resin.The relationship between the location condition on macroporous resin and the activity of immobilized LHD was discussed.

关 键 词：扫描电子显微法 X射线能谱法 乳酸脱氢酶 大孔树脂 酶活性 

分 类 号：O657.99[理学—分析化学] Q554.9[理学—化学]