Background: Glycogen storage disease type III (GSDIII, Cori/Forbes disease) is a
metabolic disorder due to the deficiency of the Glycogen Debranching Enzyme (GDE), a large
monomeric protein (about 176 kDa) with two distinct enzymatic activities: 4-α-glucantransferase
and amylo-α-1,6-glucosidase. Several mutations along the amylo-alpha-1,6-glucosidase,4-alphaglucanotransferase
(Agl) gene are associated with loss of enzymatic activity. The unique treatment
for GSDIII, at the moment, is based on diet.
The potential of plants to manufacture exogenous engineered compounds for pharmaceutical
purposes, from small to complex protein molecules such as vaccines, antibodies and other
therapeutic/prophylactic entities, was shown by modern biotechnology through “Plant Molecular
Objective and Methods: In an attempt to develop novel protein-based therapeutics for GSDIII, the
Agl gene, encoding for the human GDE (hGDE) was engineered for expression as a histidinetagged
GDE protein both in Nicotiana benthamiana plants by a transient expression approach, and
in axenic hairy root in vitro cultures (HR) from Lycopersicum esculentum and Beta vulgaris.
Results: In both plant-based expression formats, the hGDE protein accumulated in the soluble
fraction of extracts. The plant-derived protein was purified by affinity chromatography in native
conditions showing glycogen debranching activity.
Conclusion: These investigations will be useful for the design of a new generation of
biopharmaceuticals based on recombinant GDE protein that might represent, in the future, a
possible therapeutic option for GSDIII.