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Large-scale production of a therapeutic protein in transgenic tobacco plants: effect of subcellular targeting on quality of a recombinant dog gastric lipase

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Abstract

A recombinant dog gastric lipase with therapeutic potential for the treatment of exocrine pancreatic insufficiency was expressed in transgenic tobacco plants. We targeted the protein using two different signal sequences for either vacuolar retention or secretion. In both cases, an active glycosylated recombinant protein was obtained. The recombinant enzymes and the native enzyme displayed similar properties including acid resistance and acidic optimum pH. The proteolytic maturation and the specific activity of the recombinant proteins, however, were found to be dependent on subcellular compartmentalization. Expression levels of recombinant dog gastric lipase were about 5% and 7% of acid extractable plant proteins for vacuolar retention and secretion respectively. This expression system already has allowed the production of tens of grams of purified lipase through open-field culture of transgenic tobacco plants.

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Authors and Affiliations

  1. Meristem Therapeutics, 8 rue des Frères Lumière, Z. I. du Brézet, 63100, Clermont-Ferrand, France

    Véronique Gruber, Thierry Arnaud, Philippe Bournat, Christèle Clément, Dominique Mison, Béatrice Olagnier, Manfred Theisen, Sylvie Baudino, Nathalie Duchateau, Catherine Gueguen, Séverine Jacquet, Catherine Ollivo, Christine Poncetta & Bertrand Mérot

  2. Institut de Recherche Jouveinal/Parke-Davis, 3–9 rue de la loge, BP 100, 94265, Fresnes Cedex, France

    Patrick P. Berna, Laurence Philippe, Claude Bénicourt, Claire Cudrey, Carole Bloës, Sylvie Dufour, Annette Doherty & Charles Danzin

  3. CNRS UMR 7509, Université Louis Pasteur de Strasbourg, 1 rue B. Pascal, 67008, Strasbourg, France

    Nathalie Zorn & Alain Van Dorsselaer

  4. CSIC, Jordi Girona, 18-26, 08034, Barcelona, Spain

    Dolores Ludevid

  5. Laboratoire de lipolyse enzymatique du CNRS UPR 9025, 31 chemin Joseph Aiguier, BP 71, 13402, Marseille, France

    Robert Verger

Authors
  1. Véronique Gruber
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  2. Patrick P. Berna
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  6. Dominique Mison
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  7. Béatrice Olagnier
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  14. Nathalie Duchateau
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  15. Sylvie Dufour
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  16. Catherine Gueguen
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  17. Séverine Jacquet
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  18. Catherine Ollivo
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  19. Christine Poncetta
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  20. Nathalie Zorn
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  21. Dolores Ludevid
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  22. Alain Van Dorsselaer
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  23. Robert Verger
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  24. Annette Doherty
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  25. Bertrand Mérot
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Gruber, V., Berna, P.P., Arnaud, T. et al. Large-scale production of a therapeutic protein in transgenic tobacco plants: effect of subcellular targeting on quality of a recombinant dog gastric lipase. Molecular Breeding 7, 329–340 (2001). https://doi.org/10.1023/A:1011653220724

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