通过制备型聚丙烯酰胺凝胶电泳从马铃薯提取物中分离醛氧化酶（EC 1.2.3.1）。

Isolation of aldehyde oxidase (EC 1.2.3.1) from potato extracts by preparative page.

作者信息

De Pooter H L, Huang Q L, Schamp N M

机构信息

Laboratory of Organic Chemistry, Faculty of Agricultural Sciences, State University of Gent, Belgium.

出版信息

Prep Biochem. 1989;19(1):61-7. doi: 10.1080/10826068908544897.

DOI:10.1080/10826068908544897

PMID:2500646

Abstract

A method is proposed for the isolation (and purification) of enzymes, with retention of their activity, from solutions or gels of preparative PAGE runs. It is based on the inclusion of Sephadex G-25 as a supporting medium for a collector buffer in otherwise normal disc-PAGE gels. The collector buffer has a lower pH and higher concentration than the stacking gel buffer. This makes the proteins concentrate in a very narrow, slowly moving band in the Sephadex on electrophoresis, and makes their recovery easy. The method is illustrated by the isolation of aldehyde oxidase from potato extracts (which was unsuccessful by classical methods), and of one isoenzyme from commercial lipoxygenase after preparative PAGE. Recovery of chicken egg albumin after PAGE was over 90%.

摘要

本文提出了一种从制备性聚丙烯酰胺凝胶电泳（PAGE）的溶液或凝胶中分离（及纯化）酶并保留其活性的方法。该方法基于在常规圆盘PAGE凝胶中加入葡聚糖G - 25作为收集缓冲液的支持介质。收集缓冲液的pH值低于堆积凝胶缓冲液，浓度高于堆积凝胶缓冲液。这使得蛋白质在电泳时在葡聚糖中浓缩成一条非常窄且移动缓慢的带，便于回收。通过从马铃薯提取物中分离醛氧化酶（经典方法未成功）以及从制备性PAGE后的商业脂氧合酶中分离一种同工酶来说明该方法。PAGE后鸡卵清蛋白的回收率超过90%。