Butovich I A, Tertykh V A
Ukr Biokhim Zh (1978). 1984 Jul-Aug;56(4):437-41.
A new simple and sensitive spectrophotometric method is suggested for determining the catecholase activity of diphenoloxidase. The method is based on the enzymatic oxidation of pyrocatechol to 1,2-benzoquinone (BQ) in the presence of the excess of ethylenediamine sulphate (EDA). The condensation product (products) of BQ and EDA (P365) is stable in the solution and possesses strong absorption in the range of 365 nm. The molar absorption factor, E365 (under condition that the molar reaction ratio of catechol to P365 is 1:1) is 15500 M-1 cm-1 on the average. Optimal reaction conditions (pH 7.0, T=25-30 degrees C, [EDA]o = 5 mg/ml) are determined. The advantages and restrictions of the suggested technique in comparison with the methods described earlier are discussed.
提出了一种用于测定二酚氧化酶的儿茶酚酶活性的简单灵敏的分光光度法。该方法基于在过量硫酸乙二胺（EDA）存在下，邻苯二酚被酶氧化为1,2 - 苯醌（BQ）。BQ与EDA的缩合产物（P365）在溶液中稳定，并且在365nm范围内具有强吸收。平均而言，摩尔吸收系数E365（在邻苯二酚与P365的摩尔反应比为1:1的条件下）为15500 M-1 cm-1。确定了最佳反应条件（pH 7.0，T = 25 - 30℃，[EDA]o = 5 mg/ml）。讨论了所建议技术与先前描述的方法相比的优点和局限性。
