Cloning and functional characterization of a fructan 1-exohydrolase (1-FEH) in the cold tolerant Patagonian species Bromus pictus

Fructans are fructose polymers synthesized in a wide range of species such as bacteria, fungi and plants. Fructans are synthesized by fructosyltransferases (FTs) and depolymerized by fructan exohydrolases (FEHs). Bromus pictus is a graminean decaploid species from the Patagonian region of Argentina,...

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Autores principales:	Del Viso, F., Puebla, A.F., Hopp, H.E., Heinz, R.A.
Formato:	JOUR
Materias:	Bromus pictus Fructan Fructan exohydrolase Low temperature Patagonia fructan fructan beta fructosidase fructan beta-fructosidase glycosidase glycosyltransferase inulosucrase recombinant protein adaptation amino acid sequence article Bromus chemistry cold enzymology gene expression profiling gene expression regulation genetics metabolism molecular cloning molecular genetics nucleotide sequence phylogeny Pichia plant gene plant leaf sequence alignment Southern blotting Adaptation, Physiological Amino Acid Sequence Blotting, Southern Cloning, Molecular Cold Temperature Fructans Gene Expression Profiling Gene Expression Regulation, Enzymologic Gene Expression Regulation, Plant Genes, Plant Glycoside Hydrolases Hexosyltransferases Molecular Sequence Data Phylogeny Plant Leaves Recombinant Proteins Sequence Alignment Bacteria (microorganisms) Fungi Graminea Pichia pastoris
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_00320935_v231_n1_p13_DelViso
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_00320935_v231_n1_p13_DelViso
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spelling	todo:paper_00320935_v231_n1_p13_DelViso2023-10-03T14:45:01Z Cloning and functional characterization of a fructan 1-exohydrolase (1-FEH) in the cold tolerant Patagonian species Bromus pictus Del Viso, F. Puebla, A.F. Hopp, H.E. Heinz, R.A. Bromus pictus Fructan Fructan exohydrolase Low temperature Patagonia fructan fructan beta fructosidase fructan beta-fructosidase glycosidase glycosyltransferase inulosucrase recombinant protein adaptation amino acid sequence article Bromus chemistry cold enzymology gene expression profiling gene expression regulation genetics metabolism molecular cloning molecular genetics nucleotide sequence phylogeny Pichia plant gene plant leaf sequence alignment Southern blotting Adaptation, Physiological Amino Acid Sequence Blotting, Southern Bromus Cloning, Molecular Cold Temperature Fructans Gene Expression Profiling Gene Expression Regulation, Enzymologic Gene Expression Regulation, Plant Genes, Plant Glycoside Hydrolases Hexosyltransferases Molecular Sequence Data Phylogeny Pichia Plant Leaves Recombinant Proteins Sequence Alignment Bacteria (microorganisms) Bromus Bromus pictus Fungi Graminea Pichia pastoris Fructans are fructose polymers synthesized in a wide range of species such as bacteria, fungi and plants. Fructans are synthesized by fructosyltransferases (FTs) and depolymerized by fructan exohydrolases (FEHs). Bromus pictus is a graminean decaploid species from the Patagonian region of Argentina, which accumulates large amounts of fructans even at temperate temperatures. The first gene isolated from B. pictus fructan metabolism was a putative sucrose:fructan 6-fructosyltransferase (6-SFT). Here, a complete cDNA of the first fructan exohydrolase (FEH) from B. pictus (Bp1-FEHa) was isolated using RT-PCR strategies. The Bp1-FEHa encoding gene is present as a single copy in B. pictus genome. Functional characterization in Pichia pastoris confirmed Bp1-FEHa is a fructan exohydrolase with predominant activity towards β-(2-1) linkages. Its expression was analyzed in different leaf sections, showing the highest expression levels in the second section of the sheath and the tip of the blade. Bp1-FEHa expression was studied along with FEH and FT activities and fructan accumulation profile in response to chilling conditions during a 7-day time course experiment. Bp1-FEHa expression and FEH activity followed a similar pattern in response to low temperatures, especially in basal sections of the sheaths. In these sections the FEH and FT activities were particularly high and they were significantly correlated to fructan accumulation profile, along with cold treatment. © 2009 Springer-Verlag. Fil:Del Viso, F. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Puebla, A.F. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Hopp, H.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Heinz, R.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00320935_v231_n1_p13_DelViso
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Bromus pictus Fructan Fructan exohydrolase Low temperature Patagonia fructan fructan beta fructosidase fructan beta-fructosidase glycosidase glycosyltransferase inulosucrase recombinant protein adaptation amino acid sequence article Bromus chemistry cold enzymology gene expression profiling gene expression regulation genetics metabolism molecular cloning molecular genetics nucleotide sequence phylogeny Pichia plant gene plant leaf sequence alignment Southern blotting Adaptation, Physiological Amino Acid Sequence Blotting, Southern Bromus Cloning, Molecular Cold Temperature Fructans Gene Expression Profiling Gene Expression Regulation, Enzymologic Gene Expression Regulation, Plant Genes, Plant Glycoside Hydrolases Hexosyltransferases Molecular Sequence Data Phylogeny Pichia Plant Leaves Recombinant Proteins Sequence Alignment Bacteria (microorganisms) Bromus Bromus pictus Fungi Graminea Pichia pastoris
spellingShingle	Bromus pictus Fructan Fructan exohydrolase Low temperature Patagonia fructan fructan beta fructosidase fructan beta-fructosidase glycosidase glycosyltransferase inulosucrase recombinant protein adaptation amino acid sequence article Bromus chemistry cold enzymology gene expression profiling gene expression regulation genetics metabolism molecular cloning molecular genetics nucleotide sequence phylogeny Pichia plant gene plant leaf sequence alignment Southern blotting Adaptation, Physiological Amino Acid Sequence Blotting, Southern Bromus Cloning, Molecular Cold Temperature Fructans Gene Expression Profiling Gene Expression Regulation, Enzymologic Gene Expression Regulation, Plant Genes, Plant Glycoside Hydrolases Hexosyltransferases Molecular Sequence Data Phylogeny Pichia Plant Leaves Recombinant Proteins Sequence Alignment Bacteria (microorganisms) Bromus Bromus pictus Fungi Graminea Pichia pastoris Del Viso, F. Puebla, A.F. Hopp, H.E. Heinz, R.A. Cloning and functional characterization of a fructan 1-exohydrolase (1-FEH) in the cold tolerant Patagonian species Bromus pictus
topic_facet	Bromus pictus Fructan Fructan exohydrolase Low temperature Patagonia fructan fructan beta fructosidase fructan beta-fructosidase glycosidase glycosyltransferase inulosucrase recombinant protein adaptation amino acid sequence article Bromus chemistry cold enzymology gene expression profiling gene expression regulation genetics metabolism molecular cloning molecular genetics nucleotide sequence phylogeny Pichia plant gene plant leaf sequence alignment Southern blotting Adaptation, Physiological Amino Acid Sequence Blotting, Southern Bromus Cloning, Molecular Cold Temperature Fructans Gene Expression Profiling Gene Expression Regulation, Enzymologic Gene Expression Regulation, Plant Genes, Plant Glycoside Hydrolases Hexosyltransferases Molecular Sequence Data Phylogeny Pichia Plant Leaves Recombinant Proteins Sequence Alignment Bacteria (microorganisms) Bromus Bromus pictus Fungi Graminea Pichia pastoris
description	Fructans are fructose polymers synthesized in a wide range of species such as bacteria, fungi and plants. Fructans are synthesized by fructosyltransferases (FTs) and depolymerized by fructan exohydrolases (FEHs). Bromus pictus is a graminean decaploid species from the Patagonian region of Argentina, which accumulates large amounts of fructans even at temperate temperatures. The first gene isolated from B. pictus fructan metabolism was a putative sucrose:fructan 6-fructosyltransferase (6-SFT). Here, a complete cDNA of the first fructan exohydrolase (FEH) from B. pictus (Bp1-FEHa) was isolated using RT-PCR strategies. The Bp1-FEHa encoding gene is present as a single copy in B. pictus genome. Functional characterization in Pichia pastoris confirmed Bp1-FEHa is a fructan exohydrolase with predominant activity towards β-(2-1) linkages. Its expression was analyzed in different leaf sections, showing the highest expression levels in the second section of the sheath and the tip of the blade. Bp1-FEHa expression was studied along with FEH and FT activities and fructan accumulation profile in response to chilling conditions during a 7-day time course experiment. Bp1-FEHa expression and FEH activity followed a similar pattern in response to low temperatures, especially in basal sections of the sheaths. In these sections the FEH and FT activities were particularly high and they were significantly correlated to fructan accumulation profile, along with cold treatment. © 2009 Springer-Verlag.
format	JOUR
author	Del Viso, F. Puebla, A.F. Hopp, H.E. Heinz, R.A.
author_facet	Del Viso, F. Puebla, A.F. Hopp, H.E. Heinz, R.A.
author_sort	Del Viso, F.
title	Cloning and functional characterization of a fructan 1-exohydrolase (1-FEH) in the cold tolerant Patagonian species Bromus pictus
title_short	Cloning and functional characterization of a fructan 1-exohydrolase (1-FEH) in the cold tolerant Patagonian species Bromus pictus
title_full	Cloning and functional characterization of a fructan 1-exohydrolase (1-FEH) in the cold tolerant Patagonian species Bromus pictus
title_fullStr	Cloning and functional characterization of a fructan 1-exohydrolase (1-FEH) in the cold tolerant Patagonian species Bromus pictus
title_full_unstemmed	Cloning and functional characterization of a fructan 1-exohydrolase (1-FEH) in the cold tolerant Patagonian species Bromus pictus
title_sort	cloning and functional characterization of a fructan 1-exohydrolase (1-feh) in the cold tolerant patagonian species bromus pictus
url	http://hdl.handle.net/20.500.12110/paper_00320935_v231_n1_p13_DelViso
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Cloning and functional characterization of a fructan 1-exohydrolase (1-FEH) in the cold tolerant Patagonian species Bromus pictus

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