Engineering mammalian-specific post-translational modifications in plant-derived proteins: phosphorylation and Mucin-type O-glycosylation as a challenge.
Ramírez-Alanis, Israel A.
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Expression of economically relevant plant-derived recombinant proteins in alternative expression platforms, especially plant expression platforms, has gained significant interest in recent years, due to the possibility to reduce production costs, or because of product quality of production. Among the different qualities that plants can offer for the production of recombinant proteins, capability to perform post-translational modifications like protein glycosylation and phosphorylation are some of the crucial ones since it has an impact on pharmaceuticals functionality and/or stability or protein activity, respectively. In this dissertation, the pharmaceutical glycoprotein human Granulocyte-Colony Stimulating Factor is transiently expressed in N. benthamiana, as several protein versions targeted to different compartments (apoplast, cytoplasm and as protein bodies), offering an alternative for the consideration of production of this protein. Furthermore, the glycoprotein was subjected to the native GalNAc-O-glycosylation, by co-expressing the pharmaceutical, together with the enzymes responsible for such glycosylation. In the case of phosphoproteins, the bovine β- and κ-caseins and their specific kinase the bovine Fam20C were also expressed for the first time in N. benthamiana plants, to assess the feasibility of controlling their phosphorylation pattern, which could be considered for the generation of soybean transgenic lines, enriched with such nutraceutical and nutrimental phosphoproteins.
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