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One‐Step Purification of a Model Periplasmic Protein from Inclusion Bodies by Its Fusion to an Effective Metal‐Binding Peptide
Author(s) -
Beitle Robert R.,
Ataai Mohammad M.
Publication year - 1993
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp00019a009
Subject(s) - periplasmic space , inclusion bodies , fusion protein , affinity chromatography , chemistry , flag tag , recombinant dna , protein purification , escherichia coli , peptide , biochemistry , chromatography , metal , target protein , binding protein , amino acid , peptide sequence , enzyme , organic chemistry , gene
It has been demonstrated that the addition of a metal‐binding amino acid sequence to an exposed terminus of a protein can be useful for purification using immobilized metal affinity chromatography (IMAC). Polyhistidine extensions, wherein sequential histidyl residues are placed at the end of a protein, have been utilized for protein purification through IMAC. Natural metal‐binding peptides may also serve as starting points for the design of an affinity tail. As a model system, an octapeptide derived from angiotensin I was fused to TEM‐β‐lactamase. When the modified protein was expressed in Escherichia coli, a one‐step purification of this recombinant protein was accomplished from resolubilized inclusion body material.