PMID:15382106
| Citation |
Han, MJ, Park, SJ, Park, TJ and Lee, SY (2004) Roles and applications of small heat shock proteins in the production of recombinant proteins in Escherichia coli. Biotechnol. Bioeng. 88:426-36 |
|---|---|
| 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. |
| Links |
PubMed Online version:10.1002/bit.20227 |
| Keywords |
Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli Proteins/biosynthesis; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Gene Expression Regulation, Bacterial/physiology; Genetic Enhancement; Heat-Shock Proteins/biosynthesis; Heat-Shock Proteins/genetics; Heat-Shock Proteins/metabolism; Inclusion Bodies/metabolism; Models, Biological; Protein Engineering/methods; Recombinant Proteins/biosynthesis |
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