Development of screening methods for functional characterization of UGTs from Stevia rebaudiana

Autor:	Veronica Vallejo, Laurent Camborde, Monique Berger, Jean Daydé, Eva Petit, Alban Jacques
Přispěvatelé:	Ecole d'Ingénieurs de Purpan (INPT - EI Purpan), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Interactions Microbiennes dans la Rhizosphère et les Racines, Laboratoire de Recherche en Sciences Végétales (LRSV), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), PepsiCo, PepsiCo Global R&D
Rok vydání:	2020
Předmět:	0106 biological sciences 0301 basic medicine Agroinfiltration Glycosylation MESH: Glycosyltransferases / metabolism Nicotiana benthamiana lcsh:Medicine MESH: Glycosyltransferases / genetics Steviol MESH: Biosynthetic Pathways MESH: Stevia / enzymology 01 natural sciences Biochemistry chemistry.chemical_compound Glucosides Stevia lcsh:Science MESH: Glucosides / metabolism Plant Proteins chemistry.chemical_classification Multidisciplinary Expression vector biology Chemistry Biological techniques MESH: Glycosylation Recombinant Proteins MESH: Plant Proteins / metabolism Diterpenes Kaurane Biotechnology digestive system Article Uridine Diphosphate 03 medical and health sciences Glycosyltransferase [SDV.BV]Life Sciences [q-bio]/Vegetal Biology MESH: Recombinant Proteins / isolation & purification lcsh:R fungi Glycoside Glycosyltransferases biology.organism_classification MESH: Plant Proteins / genetics Biosynthetic Pathways Stevia rebaudiana MESH: Diterpenes Kaurane / metabolism 030104 developmental biology MESH: Uridine Diphosphate / metabolism biology.protein lcsh:Q MESH: Recombinant Proteins / metabolism Plant sciences MESH: Recombinant Proteins / genetics 010606 plant biology & botany
Zdroj:	Scientific Reports Scientific Reports, Nature Publishing Group, 2020, 10 (1), pp.15137. ⟨10.1038/s41598-020-71746-9⟩ Scientific Reports, Vol 10, Iss 1, Pp 1-10 (2020)
ISSN:	2045-2322
Popis:	Glycosylation is a key modification that contributes to determine bioactivity and bioavailability of plant natural products, including that of terpenoids and steviol glycosides (SVglys). It is mediated by uridine-diphosphate glycosyltransferases (UGTs), that achieve their activity by transferring sugars on small molecules. Thus, the diversity of SVglys is due to the number, the position and the nature of glycosylations on the hydroxyl groups in C-13 and C-19 of steviol. Despite the intense sweetener property of SVglys and the numerous studies conducted, the SVglys biosynthetic pathway remains largely unknown. More than 60 SVglys and 68 putative UGTs have been identified in Stevia rebaudiana. This study aims to provide methods to characterize UGTs putatively involved in SVglys biosynthesis. After agroinfiltration-based transient gene expression in Nicotiana benthamiana, functionality of the recombinant UGT can be tested simply and directly in plants expressing it or from a crude extract. The combined use of binary vectors from pGWBs series to produce expression vectors containing the stevia's UGT, enables functionality testing with many substrates as well as other applications for further analysis, including subcellular localization.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::886e9ca646dc72fd0c8c449f8fb16caa https://pubmed.ncbi.nlm.nih.gov/32934264 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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