用于纯化鸡蛋清卵黏蛋白、溶菌酶、转铁蛋白和卵清蛋白的两步色谱法及纯化蛋白的表征

Two-step chromatographic procedure for the purification of hen egg white ovomucin, lysozyme, ovotransferrin and ovalbumin and characterization of purified proteins.

作者信息

Awadé A C, Moreau S, Mollé D, Brulé G, Maubois J L

机构信息

Laboratoire de Recherches et de Technologie Laitière, Institut National de la Recherche Agronomique, Rennes, France.

出版信息

J Chromatogr A. 1994 Aug 19;677(2):279-88. doi: 10.1016/0021-9673(94)80156-8.

DOI:10.1016/0021-9673(94)80156-8

PMID:7921188

Abstract

An improved procedure is described involving gel permeation and anion-exchange chromatography for the purification of four major hen egg white proteins. The procedure involves a first-step purification of ovomucin and lysozyme by gel permeation on a Superose 6 Prep Grade column. In the second step, anion-exchange chromatography on Q Sepharose Fast Flow led to the isolation of ovotransferrin and ovalbumin from a gel permeation chromatographic peak. The purities were estimated as ca. 80, 100, 80 and 100% for ovomucin, lysozyme, ovotransferrin and ovalbumin, respectively. The purification yield was over 60% for each protein. Further characterization of purified lysozyme revealed that it was fully active and homogeneous in relation to the electrospray ionization mass spectrum. The electrospray ionization mass spectrum showed different ovotransferrin species. The amino acid composition of purified ovomucin was compared to those published previously.

摘要

本文描述了一种改进的方法，该方法涉及凝胶渗透色谱法和阴离子交换色谱法，用于纯化四种主要的蛋清蛋白。该方法包括第一步，通过在Superose 6制备级柱上进行凝胶渗透，纯化卵粘蛋白和溶菌酶。第二步，在Q Sepharose Fast Flow上进行阴离子交换色谱，从凝胶渗透色谱峰中分离出转铁蛋白和卵清蛋白。卵粘蛋白、溶菌酶、转铁蛋白和卵清蛋白的纯度估计分别约为80%、100%、80%和100%。每种蛋白质的纯化产率均超过60%。对纯化后的溶菌酶进行进一步表征，结果表明其在电喷雾电离质谱方面具有完全活性且均一。电喷雾电离质谱显示了不同的转铁蛋白种类。将纯化后的卵粘蛋白的氨基酸组成与先前发表的结果进行了比较。