Calderón A A, Pedreño M A, Ros Barceló A, Muñoz R
Department of Plant Biology (Plant Physiology), University of Murcia, Spain.
J Biochem Biophys Methods. 1990 Mar;20(3):171-80. doi: 10.1016/0165-022x(90)90075-n.
A new and sensitive spectrophotometric assay has been developed to study and to characterize kinetically the oxidation of 4-hydroxystilbene (an analogue of the putative viniferin precursor, trans-resveratrol, directly involved in the resistance mechanism of the grapevine to fungal diseases) by peroxidase-H2O2 systems. The technique measures the overall increase in absorbance at 248 nm in the reaction media, probably due to the formation of a phenoxy radical of the 4-hydroxystilbene (4-HS). This technique was developed by using the purified isoenzyme C of horseradish peroxidase and all the validity criteria (sensitivity and reproducibility) were checked. The results show that it is especially suitable for low activity measurements. It was finally applied to the determination of the oxidation rate of 4-HS by peroxidases isolated from the media of suspension-cultured grapevine cells, at two different developmental stages.
已开发出一种新型灵敏的分光光度测定法，用于研究过氧化物酶-H₂O₂系统对4-羟基茋（一种假定的葡萄素前体反式白藜芦醇的类似物，直接参与葡萄对真菌病害的抗性机制）的氧化作用，并对其进行动力学表征。该技术通过测量反应介质在248nm处吸光度的总体增加来实现，这可能是由于形成了4-羟基茋（4-HS）的苯氧自由基。此技术是利用辣根过氧化物酶的纯化同工酶C开发的，并检查了所有有效性标准（灵敏度和重现性）。结果表明，它特别适用于低活性测量。最终，该技术被应用于测定从悬浮培养的葡萄细胞培养基中分离出的过氧化物酶在两个不同发育阶段对4-HS的氧化速率。
