用固定化绿皮西葫芦抗坏血酸氧化酶测定抗坏血酸

Determination of ascorbic acid with immobilized green zucchini ascorbate oxidase.

作者信息

Stevanato R, Avigliano L, Finazzi-Agrò A, Rigo A

出版信息

Anal Biochem. 1985 Sep;149(2):537-42. doi: 10.1016/0003-2697(85)90610-4.

Abstract

Ascorbate oxidase from zucchini squash was immobilized onto CH-Sepharose via carbodiimide. The properties of the immobilized enzyme were found to be similar to those of the free ascorbate oxidase. The immobilized enzyme was utilized in a flow-through system equipped with a polarographic detector which monitors the oxygen depletion due to the reaction ascorbic acid + 1/2 O2----dehydroascorbic acid + H2O. This method, the response of which is linear between 3 X 10(-7) and 5 X 10(-4) M ascorbate, was utilized to measure the ascorbic acid in biological samples such as human plasma and fruit juices at a rate of about 60 determinations every hour with a standard deviation lower than 5%.

摘要

通过碳二亚胺将西葫芦中的抗坏血酸氧化酶固定在CH-琼脂糖上。发现固定化酶的性质与游离抗坏血酸氧化酶的性质相似。固定化酶用于配备极谱检测器的流通系统中，该检测器监测由于抗坏血酸+ 1/2 O2→脱氢抗坏血酸+ H2O反应导致的氧气消耗。该方法在3×10（-7）至5×10（-4）M抗坏血酸盐之间的响应呈线性，用于以每小时约60次测定的速率测量生物样品（如人血浆和果汁）中的抗坏血酸，标准偏差低于5%。