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    • 2. 发明授权
    • Method for assay of peroxidase enzyme or reducing substrate activity
    • 测定过氧化物酶或降低底物活性的方法
    • US4780281A
    • 1988-10-25
    • US744673
    • 1985-06-14
    • Lawrence J. MarnettPaul E. Weller
    • Lawrence J. MarnettPaul E. Weller
    • C12Q1/28
    • C12Q1/28C12Q2326/00Y10S435/803Y10S435/81
    • A preferred method wherein 5-phenyl-4-pentenyl-hydroperoxide (PPHP) is reduced to 5-phenyl-4-pentenyl-alcohol (PPA) by plant and animal peroxidases in the presence of reducing substrates is described. The method also uses related homologs containing 3 to 8 carbon atoms. PHP and PPA are rapidly isolated with solid phase extraction, separated by isocrated reverse phase high performance liquid chromatography, and quantitated with a fixed-wavelength ultraviolet detector. The procedure described in suitable for detecting peroxide reducing enzymes, determining the kinetic properties of heme- and non-heme-containing peroxidases, and evaluating oxidizable compounds as reducing substrates for peroxidases. The method identifies compounds which are reducing substrates and also ranks them for relative activity. The method can be used to identify active antithrombotic, antimetastatic, or anti-inflammatory drugs as substrates as well as detect and characterize mammalian peroxidases.
    • 描述了在还原底物存在下,通过植物和动物过氧化物酶将5-苯基-4-戊烯基 - 过氧化氢(PPHP)还原为5-苯基-4-戊烯醇(PPA)的优选方法。 该方法还使用含有3至8个碳原子的相关同系物。 通过固相萃取快速分离出PHP和PPA,用反相高效液相色谱法分离,用固定波长紫外检测器定量。 适用于检测过氧化物还原酶,确定含血红素和非含血红素的过氧化物酶的动力学性质以及评价可氧化化合物作为过氧化物酶底物的方法。 该方法鉴定了减少底物的化合物,并将它们排列成相对活性。 该方法可用于鉴定活性抗血栓形成,抗转移或抗炎药物作为底物,以及检测和表征哺乳动物过氧化物酶。