DnaK/DnaJ-assisted recombinant protein production in Trichoplusia ni larvae |
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Authors: | Mónica Martínez-Alonso Silvia Gómez-Sebastián José M Escribano Juan-Carlos Saiz Neus Ferrer-Miralles Antonio Villaverde |
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Institution: | 1. Institute for Biotechnology and Biomedicine, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain 2. CIBER de Bioingenieria, Biomateriales y Nanomedicina (CIBER-BBN), Bellaterra, 08193, Barcelona, Spain 5. Department of Genetics and Microbiology, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain 3. R&D Department, Alternative Gene Expression S.L. (ALGENEX), Centro empresarial Parque Científico y Tecnológico de la Universidad Politécnica de Madrid Campus de Montegancedo, Pozuelo de Alarcón, 28223, Madrid, Spain 4. Department of Biotechnology, INIA, Autovía A6 Km 7, 28040, Madrid, Spain
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Abstract: | The DnaK/DnaJ Escherichia coli chaperone pair, co-produced along with recombinant proteins, has been widely used to assist protein folding in bacterial
cells, although with poor consensus about the ultimate effect on protein quality and its general applicability. Here, we have
evaluated for the first time these bacterial proteins as folding modulators in a highly promising recombinant protein platform
based on insect larvae. Intriguingly, the bacterial chaperones enhanced the solubility of a reporter, misfolding-prone GFP,
doubling the yield of recombinant protein that can be recovered from the larvae extracts in a production process. This occurs
without negative effects on the yield of total protein (extractable plus insoluble), indicative of a proteolytic stability
of the chaperone substrate. It is in contrast with what has been observed in bacteria for the same reporter protein, which
is dramatically degraded in a DnaK-dependent manner. The reported data are discussed in the context of the biotechnological
potential and applicability of prokaryotic chaperones in complex, eukaryotic factories for recombinant protein production. |
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