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非水介质中辣根过氧化物酶(HRP)荧光活性测定法

Fluorometric Assays for the Activity of Horseradish Peroxidase in Nonaqueous Media

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【作者】 孙旭东马林刘伟黄仲立董爱武丁忠田唐玉秋孙天霄由德林程玉华

【Author】 Sun,Xu-dong;Ma,Lin; Liu,Wei; Huang,Zhong-li;Dong,Ai-wu;Ding,Zhong-tian; Tang,Yu-qiu; Sun,Tian-xiao; You,De-lin; Cheng,Yu-hua(The Lab of Enzymes Engineering,Jilin University,Chagchun 130023)

【机构】 吉林大学酶工程实验室

【摘要】 建立了一种分析HRP催化活力的新方法。该方法基于单体(底物)、聚合物(产物)的荧光发射光谱不重叠,使用荧光光谱仪,通过测量底物荧光淬灭来检测HRP在非水介质中(二氧六环-水、乙醇-水、丙酮-水体系)催化酚类、芳香胺类物质聚合的活力。此方法迅速、简便,结果是定量并可重复的,并能定量地计算底物转化率。

【Abstract】 A new method to assay horseradish peroxidase(HRP)catalytic activity of coupling phenols and aromatic amines in nonaqueous media with fluorescence spectrophotometer was developed based on that the fluorescence emission spectra of the substrates and corresponding product polymers were not overlapped.This method could detect the activity of HRP for different substrates in various nonqueous reaction systems(such as dioxane-water mixture,acetone-water mixture) by measuring the quenching of the substrate fluorescence.The present work offered a fast and convenient method for determining the activity of HRP in nonqueous media.This assay is qualitatively reproducible.It can also be used for quantitative calculation of the substrate conversion.

【基金】 国家自然科学基金
  • 【分类号】Q550.2
  • 【被引频次】1
  • 【下载频次】164
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