Open AccessRehosting of Bacterial Chaperones for High-Quality Protein Production
Author(s) -
Maira Martínez-Alonso,
Verónica Toledo-Rubio,
Rob Noad,
Ugutz Unzueta,
Neus Ferrer-Miralles,
Polly Roy,
Antonio Villaverde
Publication year - 2009
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.01532-09
Subject(s) - escherichia coli , proteolysis , protein folding , heterologous , biology , biochemistry , recombinant dna , escherichia coli proteins , chaperone (clinical) , protein stability , bacterial protein , microbiology and biotechnology , enzyme , gene , medicine , pathology
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.