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

✍ Scribed by J. Caubín; H. Martín; A. Roa; I. Cosano; M. Pozuelo; J. M. de la Fuente; J. M. Sánchez-Puelles; M. Molina; C. Nombela

• Publisher: John Wiley and Sons
• Year: 2001
• Tongue: English
• Weight: 511 KB
• Volume: 74
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/bit.1106

✦ Synopsis

Abstract

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. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 74: 164–171, 2001.