Abstract
The use of polyethylene glycol (PEG) as a refolding additive to a refolding cocktail comprising the molecular bichaperone ClpB and DnaKJE significantly enhances chaperone-mediated refolding of heat-denatured malate dehydrogenase (MDH). The critical factor to affect the refolding yield is the time point of introducing PEG to the refolding cocktail. The refolding efficiency reached approximately 90% only when PEG was added at the beginning of refolding reaction. The synergistic coordination of an inexpensive refolding additive PEG with the ClpB/DnaKJE bichaperone system may provide an economical route to further enhance the efficacy of ClpB/DnaKJE refolding cocktail approach, facilitating its implementation in large-scale refolding processes.
(c) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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DNA Polymerase III / genetics
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DNA Polymerase III / metabolism*
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Endopeptidase Clp
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Escherichia coli Proteins / genetics
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Escherichia coli Proteins / metabolism*
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HSP40 Heat-Shock Proteins / genetics
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HSP40 Heat-Shock Proteins / metabolism*
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HSP70 Heat-Shock Proteins / genetics
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HSP70 Heat-Shock Proteins / metabolism*
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Heat-Shock Proteins / genetics
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Heat-Shock Proteins / metabolism*
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Malate Dehydrogenase / chemistry*
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Molecular Chaperones / chemistry
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Polyethylene Glycols / chemistry*
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Protein Folding*
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Protein Renaturation
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Swine
Substances
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Escherichia coli Proteins
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HSP40 Heat-Shock Proteins
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HSP70 Heat-Shock Proteins
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Heat-Shock Proteins
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Molecular Chaperones
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Polyethylene Glycols
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Malate Dehydrogenase
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DNA polymerase III, alpha subunit
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DNA Polymerase III
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Endopeptidase Clp
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dnaK protein, E coli
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ClpB protein, E coli