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Identification and characterization of putative xylose and cellobiose transporters in Aspergillus nidulans

BACKGROUND: The conversion of lignocellulosic biomass to biofuels (second-generation biofuel production) is an environmentally friendlier alternative to petroleum-based energy sources. Enzymatic deconstruction of lignocellulose, catalyzed by filamentous fungi such as Aspergillus nidulans, releases a...

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Autores principales: dos Reis, Thaila Fernanda, de Lima, Pollyne Borborema Almeida, Parachin, Nádia Skorupa, Mingossi, Fabiana Bombonato, de Castro Oliveira, Juliana Velasco, Ries, Laure Nicolas Annick, Goldman, Gustavo Henrique
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5037631/
https://www.ncbi.nlm.nih.gov/pubmed/27708711
http://dx.doi.org/10.1186/s13068-016-0611-1
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author dos Reis, Thaila Fernanda
de Lima, Pollyne Borborema Almeida
Parachin, Nádia Skorupa
Mingossi, Fabiana Bombonato
de Castro Oliveira, Juliana Velasco
Ries, Laure Nicolas Annick
Goldman, Gustavo Henrique
author_facet dos Reis, Thaila Fernanda
de Lima, Pollyne Borborema Almeida
Parachin, Nádia Skorupa
Mingossi, Fabiana Bombonato
de Castro Oliveira, Juliana Velasco
Ries, Laure Nicolas Annick
Goldman, Gustavo Henrique
author_sort dos Reis, Thaila Fernanda
collection PubMed
description BACKGROUND: The conversion of lignocellulosic biomass to biofuels (second-generation biofuel production) is an environmentally friendlier alternative to petroleum-based energy sources. Enzymatic deconstruction of lignocellulose, catalyzed by filamentous fungi such as Aspergillus nidulans, releases a mixture of mono- and polysaccharides, including hexose (glucose) and pentose (xylose) sugars, cellodextrins (cellobiose), and xylooligosaccharides (xylobiose). These sugars can subsequently be fermented by yeast cells to ethanol. One of the major drawbacks in this process lies in the inability of yeast, such as Saccharomyces cerevisiae, to successfully internalize sugars other than glucose. The aim of this study was, therefore, to screen the genome of A. nidulans, which encodes a multitude of sugar transporters, for transporters able to internalize non-glucose sugars and characterize them when introduced into S. cerevisiae. RESULTS: This work identified two proteins in A. nidulans, CltA and CltB, with roles in cellobiose transport and cellulose signaling, respectively. CltA, when introduced into S. cerevisiae, conferred growth on low and high concentrations of cellobiose. Deletion of cltB resulted in reduced growth and extracellular cellulase activity in A. nidulans in the presence of cellobiose. CltB, when introduced into S. cerevisiae, was not able to confer growth on cellobiose, suggesting that this protein is a sensor rather than a transporter. However, we have shown that the introduction of additional functional copies of CltB increases the growth in the presence of low concentrations of cellobiose, strongly indicating CltB is able to transport cellobiose. Furthermore, a previously identified glucose transporter, HxtB, was also found to be a major xylose transporter in A. nidulans. In S. cerevisiae, HxtB conferred growth on xylose which was accompanied by ethanol production. CONCLUSIONS: This work identified a cellobiose transporter, a xylose transporter, and a putative cellulose transceptor in A. nidulans. This is the first time that a sensor role for a protein in A. nidulans has been proposed. Both transporters are also able to transport glucose, highlighting the preference of A. nidulans for this carbon source. This work provides a basis for future studies which aim at characterizing and/or genetically engineering Aspergillus spp. transporters, which, in addition to glucose, can also internalize other carbon sources, to improve transport and fermentation of non-glucose sugars in S. cerevisiae. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0611-1) contains supplementary material, which is available to authorized users.
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spelling pubmed-50376312016-10-05 Identification and characterization of putative xylose and cellobiose transporters in Aspergillus nidulans dos Reis, Thaila Fernanda de Lima, Pollyne Borborema Almeida Parachin, Nádia Skorupa Mingossi, Fabiana Bombonato de Castro Oliveira, Juliana Velasco Ries, Laure Nicolas Annick Goldman, Gustavo Henrique Biotechnol Biofuels Research BACKGROUND: The conversion of lignocellulosic biomass to biofuels (second-generation biofuel production) is an environmentally friendlier alternative to petroleum-based energy sources. Enzymatic deconstruction of lignocellulose, catalyzed by filamentous fungi such as Aspergillus nidulans, releases a mixture of mono- and polysaccharides, including hexose (glucose) and pentose (xylose) sugars, cellodextrins (cellobiose), and xylooligosaccharides (xylobiose). These sugars can subsequently be fermented by yeast cells to ethanol. One of the major drawbacks in this process lies in the inability of yeast, such as Saccharomyces cerevisiae, to successfully internalize sugars other than glucose. The aim of this study was, therefore, to screen the genome of A. nidulans, which encodes a multitude of sugar transporters, for transporters able to internalize non-glucose sugars and characterize them when introduced into S. cerevisiae. RESULTS: This work identified two proteins in A. nidulans, CltA and CltB, with roles in cellobiose transport and cellulose signaling, respectively. CltA, when introduced into S. cerevisiae, conferred growth on low and high concentrations of cellobiose. Deletion of cltB resulted in reduced growth and extracellular cellulase activity in A. nidulans in the presence of cellobiose. CltB, when introduced into S. cerevisiae, was not able to confer growth on cellobiose, suggesting that this protein is a sensor rather than a transporter. However, we have shown that the introduction of additional functional copies of CltB increases the growth in the presence of low concentrations of cellobiose, strongly indicating CltB is able to transport cellobiose. Furthermore, a previously identified glucose transporter, HxtB, was also found to be a major xylose transporter in A. nidulans. In S. cerevisiae, HxtB conferred growth on xylose which was accompanied by ethanol production. CONCLUSIONS: This work identified a cellobiose transporter, a xylose transporter, and a putative cellulose transceptor in A. nidulans. This is the first time that a sensor role for a protein in A. nidulans has been proposed. Both transporters are also able to transport glucose, highlighting the preference of A. nidulans for this carbon source. This work provides a basis for future studies which aim at characterizing and/or genetically engineering Aspergillus spp. transporters, which, in addition to glucose, can also internalize other carbon sources, to improve transport and fermentation of non-glucose sugars in S. cerevisiae. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-016-0611-1) contains supplementary material, which is available to authorized users. BioMed Central 2016-09-26 /pmc/articles/PMC5037631/ /pubmed/27708711 http://dx.doi.org/10.1186/s13068-016-0611-1 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
dos Reis, Thaila Fernanda
de Lima, Pollyne Borborema Almeida
Parachin, Nádia Skorupa
Mingossi, Fabiana Bombonato
de Castro Oliveira, Juliana Velasco
Ries, Laure Nicolas Annick
Goldman, Gustavo Henrique
Identification and characterization of putative xylose and cellobiose transporters in Aspergillus nidulans
title Identification and characterization of putative xylose and cellobiose transporters in Aspergillus nidulans
title_full Identification and characterization of putative xylose and cellobiose transporters in Aspergillus nidulans
title_fullStr Identification and characterization of putative xylose and cellobiose transporters in Aspergillus nidulans
title_full_unstemmed Identification and characterization of putative xylose and cellobiose transporters in Aspergillus nidulans
title_short Identification and characterization of putative xylose and cellobiose transporters in Aspergillus nidulans
title_sort identification and characterization of putative xylose and cellobiose transporters in aspergillus nidulans
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5037631/
https://www.ncbi.nlm.nih.gov/pubmed/27708711
http://dx.doi.org/10.1186/s13068-016-0611-1
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