Chuturgoon A A, Dutton M F, Berry R K
Department of Biochemistry, University of Natal, Pietermaritzburg, South Africa.
Biochem Biophys Res Commun. 1990 Jan 15;166(1):38-42. doi: 10.1016/0006-291x(90)91908-b.
An affinity matrix for the purification of norsolorinic acid dehydrogenase, an enzyme involved in aflatoxin biosynthesis, was prepared by coupling norsolorinic acid to an agarose gel. This matrix was found to be ineffective in isolating active enzyme, and was therefore modified by methylation, using diazomethane. The methylated matrix produced a one-step purification of the enzyme from a crude homogenate, resulting in a 138-fold purification. The active isolate was found to contain one major and two minor bands upon nondenaturing electrophoresis, and all the norsolorinic acid dehydrogenase activity was associated with the major band. It was concluded that the matrix exhibited true affinity for the enzyme, and that affinity chromatography was a valuable approach to isolating other secondary metabolic enzymes involved in the biosynthesis of the aflatoxins.
通过将降红霉内酯酸偶联到琼脂糖凝胶上，制备了用于纯化降红霉内酯酸脱氢酶（一种参与黄曲霉毒素生物合成的酶）的亲和基质。发现该基质在分离活性酶方面无效，因此使用重氮甲烷进行甲基化修饰。甲基化基质从粗匀浆中一步纯化该酶，得到了138倍的纯化倍数。在非变性电泳中，活性分离物显示有一条主带和两条次带，所有降红霉内酯酸脱氢酶活性都与主带相关。得出的结论是，该基质对该酶表现出真正的亲和力，并且亲和色谱法是分离参与黄曲霉毒素生物合成的其他次生代谢酶的一种有价值的方法。
