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.
Copyright 2001 John Wiley & Sons, Inc.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Base Sequence
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Binding Sites
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Choline / metabolism*
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Chromatography, DEAE-Cellulose / methods*
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Molecular Sequence Data
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N-Acetylmuramoyl-L-alanine Amidase / genetics*
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N-Acetylmuramoyl-L-alanine Amidase / isolation & purification
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N-Acetylmuramoyl-L-alanine Amidase / metabolism
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Pichia / genetics*
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Pichia / metabolism
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / isolation & purification*
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Recombinant Fusion Proteins / metabolism*
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Streptococcus pneumoniae / enzymology
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Streptococcus pneumoniae / genetics
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beta-Lactamases / genetics
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beta-Lactamases / isolation & purification
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beta-Lactamases / metabolism
Substances
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Recombinant Fusion Proteins
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EJL amidase
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N-Acetylmuramoyl-L-alanine Amidase
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beta-Lactamases
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Choline