Design of a molecular chaperone-assisted protein folding bioreactor.
Escherichia coli molecular chaperone GroEL and co-chaperone GroES are well known to assist the folding/refolding of a diverse range of substrate proteins. Despite this, there have been relatively few reports of the GroEL/GroES molecular chaperone system being used as a biotechnology tool for protein folding/refolding. In this paper, a solution-phase protein folding bioreactor is described that involves the complete GroEL/GroES system. The main features of this bioreactor are the use of a stirred-cell concentrator fitted with a 100 kDa molecular weight cutoff membrane and an attached buffer reservoir. This bioreactor system was used successfully for assisted-batch refolding of guanidinium chloride (Gu-HCl) unfolded mitochondrial malate dehydrogenase (mMDH). We believe that protein folding bioreactor systems of this type could have wide potential utility for the folding/refolding of unfolded protein substrates.
Imperial College Genetic Therapies Centre, Department of Chemistry, Imperial College of Science, Technology & Medicine, South Kensington, London SW7 2AY, U.K.
R J
RJ
Kohler
M
M
Preuss
A D
AD
Miller
D000818:Animals; D019149:Bioreactors; D018835:Chaperonin 10 / Q000378:metabolism*; D018834:Chaperonin 60 / Q000378:metabolism*; D008291:Malate Dehydrogenase / Q000378:metabolism*; D008929:Mitochondria, Heart / Q000201:enzymology; D018832:Molecular Chaperones / Q000378:metabolism*; D017510:Protein Folding; D013552:Swine
D018835:Chaperonin 10; D018834:Chaperonin 60; D018832:Molecular Chaperones; D008291:Malate Dehydrogenase