Open AccessPurification of inclusion body—forming peptides and proteins in soluble form by fusion to Escherichia coli thermostable proteins
Author(s) -
Arjun Thapa,
Md. Shahnawaz,
Pratap Karki,
Giri Raj Dahal,
Golam Sharoar,
Song Yub Shin,
Jung Sup Lee,
Byungyun Cho,
IlSeon Park
Publication year - 2008
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000112728
Subject(s) - inclusion bodies , escherichia coli , fusion protein , biochemistry , maltose binding protein , biology , solubilization , chemistry , recombinant dna , gene
Proteins and peptides expressed in the prokaryotic system often form inclusion bodies. Solubilization and refolding procedures can be used for their recovery, but this process remains difficult. One strategy for improving the solubility of a protein of interest is to fuse it to a highly soluble protein. To select a suitable fusion partner capable of solubilizing the aggregation-prone (inclusion body–forming) proteins and peptides, Escherichia coli thermostable proteins were identified and tested. Among them, trigger factor (TF) protein was selected because of its high expression and stability. Using an expression system based on fusion to TF, selected proteins and peptides that otherwise form inclusion bodies were expressed in soluble state and were purified like other soluble proteins. This system provides a convenient method for production of aggregation-prone proteins and peptides.