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
Nepřihlášeným uživatelům se plný text nezobrazuje