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Roles and applications of small heat shock proteins in the production of recombinant proteins in Escherichia coli
Author(s) -
Han MeeJung,
Park Si Jae,
Park Tae Jung,
Lee Sang Yup
Publication year - 2004
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20227
Subject(s) - recombinant dna , escherichia coli , green fluorescent protein , biology , heat shock protein , inclusion bodies , secretion , proteome , proteases , biochemistry , cytosol , mutant , microbiology and biotechnology , chemistry , gene , enzyme
Abstract Proteome profiling of the inclusion body (IB) fraction of recombinant proteins produced in Escherichia coli suggested that two small heat shock proteins, IbpA and IbpB, are the major proteins associated with IBs. In this study, we demonstrate that IbpA and IbpB facilitate the production of recombinant proteins in E. coli and play important roles in protecting recombinant proteins from degradation by cytoplasmic proteases. We examined the cytosolic production, and Tat‐ or Sec‐dependent secretion of the enhanced green fluorescent protein (EGFP) in wild type, ibpAB − mutant, and ibpAB ‐amplified E. coli strains. Analysis of fluorescence histograms and confocal microscopic imaging revealed that over‐expression of the ibpA and/or ibpB genes enhanced cytosolic EGFP production whereas knocking out the ibpAB genes enhanced secretory production. This strategy seems to be generally applicable as it was successfully employed for the enhanced cytosolic or secretory production of several other recombinant proteins in E. coli . © 2004 Wiley Periodicals, Inc.