Affinity precipitation of Aspergillus niger pectinase by microwave-treated alginate

被引：18
|
作者：
Mondal, K [1 ]
Mehta, P [1 ]
Gupta, MN [1 ]
机构：
[1] Indian Inst Technol, Dept Chem, New Delhi 110016, India
来源：
PROTEIN EXPRESSION AND PURIFICATION | 2004年 / 33卷 / 01期
关键词：
affinity precipitation; affinity ligands; alginate; microwave pretreatment; pectinase; protein purification;
D O I：
10.1016/j.pep.2003.08.013
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Affinity precipitation is a simple, single plate separation process in which the complex of a smart macroaffinity ligand with the target protein (from a crude broth) can be selectively precipitated by application of a suitable stimulus. Alginate is a copolymer of guluronic acid and mannuronic acid residues and precipitates with Ca2+ ions. It was found to bind to pectinase present in a commercial preparation of Aspergillus niger, Pectinex Ultra-SPL. Microwave pretreatment of alginate at 75 degreesC was found to enhance the selectivity of the affinity precipitation. Using microwave-treated alginate, 83% of the enzyme activity with 20-fold purification could be recovered. SDS-PAGE upon silver staining confirmed the enhanced selectivity of affinity precipitation when microwave-treated alginate was used. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码：104 / 109
页数：6
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