FURUYA A, HAYASHI J A
J Bacteriol. 1963 May;85(5):1124-31. doi: 10.1128/jb.85.5.1124-1131.1963.
Furuya, Akira (University of Illinois College of Medicine, Chicago) and James A. Hayashi. Glycolic acid oxidation by Escherichia coli adapted to glycolate. J. Bacteriol. 85:1124-1131. 1963.-A procedure is described for extraction and partial purification of glycolic acid oxidase from Escherichia coli adapted to grow on glycolate as the sole carbon source. Enzyme activity was assayed by oxygen uptake and by reduction of 2,6-dichlorophenol-indophenol. Glyoxylic acid was the product of glycolate oxidation by the enzyme. Enzyme activity, which diminishes rapidly on storage, shows a maximum at pH 6 to 7. We were unable to show any cofactor requirement. Compounds which inhibited glycolate oxidation and their order of inhibitory activity were: p-hydroxymercuribenzoate > sodium azide > iodoacetate and o-phenanthroline > ethylenediaminetetraacetic acid. Tests of enzyme specificity showed that the following compounds were oxidized, but at different rates: glycolate, d-lactate, l-lactate, dl-alpha-hydroxybutyrate, dl-malate, and dl-glycerate. Citrate, tartrate, and dl-beta-hydroxybutyrate were not oxidized. Potassium cyanide stimulated oxygen uptake when glycolate and lactate were oxidized. Whether the oxidations were due to different oxidases or to a single oxidase with a wide range of specificities was tested by observing the oxidation of glycolate, d-lactate, and l-lactate under various conditions. Ammonium sulfate fractionation of a crude extract did not change the relative ability to oxidize the three acids. However, the three oxidative capacities diminished at different rates during storage at 0 C for 6 days. The partially purified glycolic oxidase preparations were probably mixtures of several different oxidases.
古屋彰（伊利诺伊大学医学院，芝加哥）和詹姆斯·A·林。适应乙醇酸的大肠杆菌对乙醇酸的氧化作用。《细菌学杂志》85:1124 - 1131。1963年。——本文描述了一种从适应以乙醇酸作为唯一碳源生长的大肠杆菌中提取和部分纯化乙醇酸氧化酶的方法。通过氧气摄取和2,6 - 二氯酚靛酚的还原反应来测定酶活性。乙醛酸是该酶氧化乙醇酸的产物。酶活性在储存时迅速降低，在pH 6至7时达到最大值。我们未能证明其对任何辅助因子有需求。抑制乙醇酸氧化的化合物及其抑制活性顺序为：对羟基汞苯甲酸＞叠氮化钠＞碘乙酸和邻菲罗啉＞乙二胺四乙酸。酶特异性测试表明，以下化合物可被氧化，但速率不同：乙醇酸、d - 乳酸、l - 乳酸、dl - α - 羟基丁酸、dl - 苹果酸和dl - 甘油酸。柠檬酸、酒石酸和dl - β - 羟基丁酸未被氧化。当乙醇酸和乳酸被氧化时，氰化钾刺激氧气摄取。通过观察在各种条件下乙醇酸、d - 乳酸和l - 乳酸的氧化情况，测试了这些氧化反应是由不同的氧化酶引起还是由具有广泛特异性的单一氧化酶引起。粗提取物的硫酸铵分级分离并未改变氧化这三种酸的相对能力。然而，在0℃储存6天期间，这三种氧化能力以不同速率降低。部分纯化的乙醇酸氧化酶制剂可能是几种不同氧化酶的混合物。
