非水介质中辣根过氧化物酶(HRP)荧光活性测定法

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

中国生物化学与分子生物学报 ›› 1994, Vol. 10 ›› Issue (04) : 490-495.

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PDF(384 KB)
中国生物化学与分子生物学报 ›› 1994, Vol. 10 ›› Issue (04) : 490-495.
论文

非水介质中辣根过氧化物酶(HRP)荧光活性测定法

  • 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
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Fluorometric Assays for the Activity of Horseradish Peroxidase in Nonaqueous Media

  • 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
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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.

关键词

辣根过氧化物酶;非水相;活力测定;荧光

Key words

Horseradish peroxidase / Nonaqueous media / Fluorescence / Activity

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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. 非水介质中辣根过氧化物酶(HRP)荧光活性测定法[J]. 中国生物化学与分子生物学报, 1994, 10(04): 490-495
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. Fluorometric Assays for the Activity of Horseradish Peroxidase in Nonaqueous Media[J]. Chinese Journal of Biochemistry and Molecular Biology, 1994, 10(04): 490-495

参考文献

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