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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 \u201cPlant Molecular Farming\u201d.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective\/method:<\/jats:title><jats:p>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.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results:<\/jats:title><jats:p>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.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion:<\/jats:title><jats:p>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.<\/jats:p><\/jats:sec>","DOI":"10.2174\/0929866526666191014154047","type":"journal-article","created":{"date-parts":[[2019,10,14]],"date-time":"2019-10-14T14:25:48Z","timestamp":1571063148000},"page":"145-157","update-policy":"https:\/\/doi.org\/10.2174\/bsp_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Expression of a Functional Recombinant Human Glycogen Debranching Enzyme (hGDE) in N. benthamiana Plants and in Hairy Root Cultures"],"prefix":"10.2174","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5251-7154","authenticated-orcid":true,"given":"Meilyn","family":"Rodriguez-Hernandez","sequence":"first","affiliation":[{"name":"Center for Genetic Engineering and Biotechnology (CIGB), Direction of Agricultural Biotechnology, Havana, Cuba"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2326-1214","authenticated-orcid":true,"given":"Doriana","family":"Triggiani","sequence":"additional","affiliation":[{"name":"Italian Glycogen Storage Disease Association (AIG) NPO, Assago, Milan, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2087-6870","authenticated-orcid":true,"given":"Fiona","family":"Ivison","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Manchester University NHS Foundation Trust, Manchester, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3344-5754","authenticated-orcid":true,"given":"Olivia C.","family":"Demurtas","sequence":"additional","affiliation":[{"name":"Department of Sustainability (SSPT), Biotechnology Laboratory, ENEA, Italian National Agency for New Technologies, Energy and Sustainable Economic Development, Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8908-9755","authenticated-orcid":true,"given":"Elena","family":"Illiano","sequence":"additional","affiliation":[{"name":"Department of Sustainability (SSPT), Biomedical Technologies Laboratory, Italian National Agency for New Technologies, Energy and Sustainable Economic Development ENEA, Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carmela","family":"Marino","sequence":"additional","affiliation":[{"name":"Department of Sustainability (SSPT), Biomedical Technologies Laboratory, Italian National Agency for New Technologies, Energy and Sustainable Economic Development ENEA, Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4982-1646","authenticated-orcid":true,"given":"Rosella","family":"Franconi","sequence":"additional","affiliation":[{"name":"Department of Sustainability (SSPT), Biomedical Technologies Laboratory, Italian National Agency for New Technologies, Energy and Sustainable Economic Development ENEA, Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8429-4031","authenticated-orcid":true,"given":"Silvia","family":"Massa","sequence":"additional","affiliation":[{"name":"Department of Sustainability (SSPT), Biotechnology Laboratory, ENEA, Italian National Agency for New Technologies, Energy and Sustainable Economic Development, Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"965","reference":[{"key":"ref=1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.ymgme.2014.02.011","volume":"112","author":"Grabowski G.A.","year":"2014","unstructured":"Grabowski G.A.; Golembo M.; Shaaltiel Y.; Taliglucerase alfa: An enzyme replacement therapy using plant cell expression technology. 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