TY - JOUR
T1 - Rehosting of bacterial chaperones for high-quality protein production
AU - Martínez-Alonso, Mónica
AU - Toledo-Rubio, Verónica
AU - Noad, Rob
AU - Unzueta, Ugutz
AU - Ferrer-Miralles, Neus
AU - Roy, Polly
AU - Villaverde, Antonio
PY - 2009/12/1
Y1 - 2009/12/1
N2 - Coproduction of DnaK/DnaJ in Escherichia coli enhances solubility but promotes proteolytic degradation of their substrates, minimizing the yield of unstable polypeptides. Higher eukaryotes have orthologs of DnaK/DnaJ but lack the linked bacterial proteolytic system. By coexpression of DnaK and DnaJ in insect cells with inherently misfolding-prone recombinant proteins, we demonstrate simultaneous improvement of soluble protein yield and quality and proteolytic stability. Thus, undesired side effects of bacterial folding modulators can be avoided by appropriate rehosting in heterologous cell expression systems. Copyright © 2009, American Society for Microbiology. All Rights Reserved.
AB - Coproduction of DnaK/DnaJ in Escherichia coli enhances solubility but promotes proteolytic degradation of their substrates, minimizing the yield of unstable polypeptides. Higher eukaryotes have orthologs of DnaK/DnaJ but lack the linked bacterial proteolytic system. By coexpression of DnaK and DnaJ in insect cells with inherently misfolding-prone recombinant proteins, we demonstrate simultaneous improvement of soluble protein yield and quality and proteolytic stability. Thus, undesired side effects of bacterial folding modulators can be avoided by appropriate rehosting in heterologous cell expression systems. Copyright © 2009, American Society for Microbiology. All Rights Reserved.
U2 - 10.1128/AEM.01532-09
DO - 10.1128/AEM.01532-09
M3 - Article
VL - 75
SP - 7850
EP - 7854
IS - 24
ER -