Synergistic coordination of polyethylene glycol with ClpB/DnaKJE bichaperone for refolding of heat-denatured malate dehydrogenase.
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.
Nian
Rui
R
Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore.
Kim
Duck Sang
DS
Tan
Lihan
L
Kim
Chan-Wha
CW
Choe
Woo-Seok
WS
D000818:Animals; D004258:DNA Polymerase III / Q000235:genetics / Q000378:metabolism*; D049071:Endopeptidase Clp; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D050956:HSP40 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D008291:Malate Dehydrogenase / Q000737:chemistry*; D018832:Molecular Chaperones / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry*; D017510:Protein Folding; D020673:Protein Renaturation; D013552:Swine
D029968:Escherichia coli Proteins; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D011092:Polyethylene Glycols; D008291:Malate Dehydrogenase; C074904:DNA polymerase III, alpha subunit; D004258:DNA Polymerase III; D049071:Endopeptidase Clp; C063842:dnaK protein, E coli; C069775:ClpB protein, E coli