Large-scale production of a therapeutic protein in transgenic tobacco plants: effect of subcellular targeting on quality of a recombinant dog gastric lipase
V Gruber, PP Berna, T Arnaud, P Bournat, C Clément… - Molecular …, 2001 - Springer
V Gruber, PP Berna, T Arnaud, P Bournat, C Clément, D Mison, B Olagnier, L Philippe…
Molecular Breeding, 2001•SpringerA 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 …
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 …
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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