Choline-binding domain as a novel affinity tag for purification of fusion proteins produced in Pichia pastoris

被引：15

作者：

Caubín, J

Martín, H

Roa, A

Cosano, I

Pozuelo, M

de la Fuente, JM

Sánchez-Puelles, JM

Molina, M

Nombela, C

机构：

[1] Univ Complutense, Fac Farm, Dept Microbiol 2, E-28040 Madrid, Spain

[2] SmithKline Beecham SA, Ctr Invest Basica, Tres Cantos, Spain

来源：

BIOTECHNOLOGY AND BIOENGINEERING | 2001年 / 74卷 / 02期

关键词：

C-LYTA; affinity tag; heterologous expression; yeast;

D O I：

10.1002/bit.1106

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

The choline-binding domain (ChoBD) of the carboxy-terminal region of the Streptococcus pneumoniae amidase LYTA (C-LYTA) presents a strong affinity for tertiary amines. We report a method for single-step purification of proteins expressed in the methylotrophic yeast Pichia pastoris based on the fusion of C-LYTA to the protein of interest. We show that C-LYTA can be efficiently expressed and secreted in this host. Tagged proteins fused to this binding domain can be purified on inexpensive DEAE matrices. It therefore provides a useful system for the purification of recombinant proteins with high specificity suitable for industrial purposes. (C) 2001 John Wiley & Sons, Inc.

引用

页码：164 / 171

页数：8

共 20 条

[1] Design, total synthesis, and functional overexpression of the Candida rugosa lip1 gene coding for a major industrial lipase [J].