Fraunhofer scientists report expression in plants of an active bacterial PNGase F and the production of recombinant proteins of interest in a non-glycosylated form

Dr. Tarlan Mamedov and his colleagues from Fraunhofer USA Center for Molecular Biotechnology report their work on the production of non-glycosylated proteins in the Plant Biotechnology Journal. Tarlan and his colleagues use in-vivo deglycosylation of recombinant N glycosylated proteins as a result of their co-expression with bacterial PNGase F.

Plasmodium parasites may have complex post-translationally modified proteins such as Pfs48/45 that do not carry N-linked glycans (Exp. Parasitol. 1998, 90, 165) but contain potential N-linked glycosylation sites that can be aberrantly glycosylated during expression in mammalian and plant systems.  Therefore, it is important to develop strategies for producing non-glycosylated forms of these targets to preserve biological activity and native conformation.

In addition to demonstrating in-vivo deglycosylation they show that the recognition of an in-vivo deglycosylated plant produced malaria vaccine candidate, Pfs48F1, by monoclonal antibodies I, III and V raised against various epitopes (I, III and V) of native Pfs48/45 of Plasmodium falciparium, was significantly stronger compared to that of the glycosylated form of plant produced Pfs48F1.

MS Bioworks mass spectrometry services were used to identify N-linked glycosylation sites on Pfs48F1.  Congratulations to Tarlan and the team for this great example of innovation in the field of biotechnology.

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