Cloning, gene mapping, and functional analysis of a fructan 1-exohydrolase (1-FEH) from Lolium perenne implicated in fructan synthesis rather than in fructan mobilization
| Autor: | Wim Van den Ende, André Van Laere, Annette Morvan-Bertrand, Katrien Le Roy, Jérémy Lothier, Philippe Barre, Bertrand Lasseur, Marie-Pascale Prud'homme |
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| Přispěvatelé: | Ecophysiologie Végétale, Agronomie et Nutritions (EVA), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Recherche Agronomique (INRA), Université Catholique de Louvain = Catholic University of Louvain (UCL), Unité de recherche d'Écophysiologie des Plantes Fourragères (UEPF), Institut National de la Recherche Agronomique (INRA) |
| Rok vydání: | 2007 |
| Předmět: |
0106 biological sciences
Glycoside Hydrolases Physiology Recombinant Fusion Proteins PICHIA PASTORIS Molecular Sequence Data Inulin GENE MAPPING Plant Science 01 natural sciences Pichia pastoris 03 medical and health sciences chemistry.chemical_compound Fructan Complementary DNA Lolium Glycoside hydrolase Amino Acid Sequence RNA Messenger Cloning Molecular Peptide sequence Phylogeny Plant Proteins 030304 developmental biology 2. Zero hunger 0303 health sciences RAY GRASS ANGLAIS Base Sequence FRUCTAN EXOHYDROLASE biology cDNA library Chromosome Mapping Biological Transport [SDV.BV.BOT]Life Sciences [q-bio]/Vegetal Biology/Botanics biology.organism_classification Fructans Invertase chemistry Biochemistry FRUCTAN BIOSYNTHESIS Sequence Alignment 010606 plant biology & botany |
| Zdroj: | Journal of Experimental Botany Journal of Experimental Botany, Oxford University Press (OUP), 2007, 58, pp.1969-1983. ⟨10.1093/jxb/erm053⟩ |
| ISSN: | 1460-2431 0022-0957 |
| DOI: | 10.1093/jxb/erm053 |
| Popis: | International audience; Fructans, which are beta-(2,1) and/or beta-(2,6) linked polymers of fructose, are important storage carbohydrates in many plants. They are mobilized via fructan exohydrolases (FEHs). The cloning, mapping, and functional analysis of the first 1-FEH (EC 3.2.1.153) from Lolium perenne L. var. Bravo is described here. By screening a perennial ryegrass cDNA library, a 1-FEH cDNA named Lp1-FEHa was cloned. The Lp1-FEHa deduced protein has a low iso-electric point (5.22) and it groups together with plant FEHs and cell-wall type invertases. The deduced amino acid sequence shows 75% identity to wheat 1-FEH w2. The Lpl-FEHa gene was mapped at a distal position on the linkage group 3 (LG3). Functional characterization of the recombinant protein in Pichia pastoris demonstrated that it had high FEH activity towards 1-kestotriose, 1,1-kestotetraose, and inulin, but low activity against 6-kestotriose and levan. Like other fructan-plant FEHs, no hydrolase activity could be detected towards sucrose, convincingly demonstrating that the enzyme is not a classic invertase. The expression pattern analysis of Lpl-FEHa revealed transcript accumulation in leaf tissues accumulating fructans while transcript level was low in the photosynthetic tissues. The high expression level of this 1-FEH in conditions of active fructan synthesis, together with its low expression level when fructan contents are low, suggest that it might play a role as a beta-(2,1) trimming enzyme acting during fructan synthesis in concert with fructan synthesis enzymes. |
| Databáze: | OpenAIRE |
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