Plant biotech & Risk Assessment
Goulet, Charles , Anguenot, Raphaël , Benchabane, Meriem , Michaud, Dominique .
Tomato cathepsin D inhibitor; An in-built stabilizing agent for recombinant proteins in plant-based expression systems?
Protein content is a major issue in plant molecular farming, where final yields in recombinant protein have a direct impact on the actual potential of the production scheme adopted. Several strategies have been considered in recent years to optimize the accumulation of recombinant proteins in plant cells. Here we show that the expression of a cathepsin D inhibitor (CDI) from tomato in the cytosol of plant (potato) leaf cells positively alters the synthesis/degradation balance of most proteins in leaf tissues, with a resulting increase in protein levels compared to potato lines not expressing the tomato inhibitor. As characterized by two-dimensional gel electrophoresis, the expression of tomato CDI in potato leaves was correlated with a relative increase in the content of most proteins in vivo, with some proteins showing a more than two-fold increase on a fresh weight basis. Interestingly, CDI would also allow the protection of heterologous proteins in planta, as illustrated here with the alpha-1-antichymotrypsin, a human serine protease inhibitor of therapeutic value, transiently expressed in leaves of CDI-expressing lines. Overall, these observations suggest the potential of tomato CDI as an in-built stabilizing agent for the protection of heterologous proteins expressed in transgenic plant systems.
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1 - Université Laval, Nutraceuticals and Functional Foods Institute (INAF), 2440 Hochelaga, Pavillon de l'INAF, Université Laval, quebec, quebec, G1K 7P4, canada
2 - Université Laval
3 - Université Laval, Nutraceuticals and Functional Foods Institute (INAF
Increased protein content
Presentation Type: Plant Biology Abstract
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM