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The conformational quality of insoluble recombinant proteins is enhanced at low growth temperatures
Author(s) -
Vera Andrea,
GonzálezMontalbán Nuria,
Arís Anna,
Villaverde Antonio
Publication year - 2006
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.21218
Subject(s) - recombinant dna , protein folding , green fluorescent protein , protein aggregation , escherichia coli , chemistry , inclusion bodies , folding (dsp implementation) , solubility , biophysics , biochemistry , protein quality , biology , gene , organic chemistry , engineering , electrical engineering
Protein aggregation is a major bottleneck during the bacterial production of recombinant proteins. In general, the induction of gene expression at sub‐optimal growth temperatures improves the solubility of aggregation‐prone polypeptides and minimizes inclusion body (IB) formation. However, the effect of low temperatures on the quality of the recombinant protein, especially within the insoluble cell fraction, has been hardly ever explored. In this work, we have examined the conformational status of a recombinant GFP protein when produced in Escherichia coli below 37°C. As expected, the fraction of aggregated protein largely decreased at lower temperatures, while the conformational quality of both soluble and aggregated GFP, as reflected by its specific fluorescence emission, progressively improved. This observation indicates that physicochemical conditions governing protein folding affect concurrently the quality of the soluble and the aggregated forms of a misfolding‐prone protein, and that protein misfolding and aggregation are clearly not coincident events. Biotechnol. Bioeng. 2007;96:1101–1106. © 2006 Wiley Periodicals, Inc.