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Functional characterization of a highly specific l-arabinose transporter from Trichoderma reesei

BACKGROUND: Lignocellulose biomass has been investigated as a feedstock for second generation biofuels and other value-added products. Some of the processes for biofuel production utilize cellulases and hemicellulases to convert the lignocellulosic biomass into a range of soluble sugars before ferme...

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Autores principales: Havukainen, Sami, Pujol-Giménez, Jonai, Valkonen, Mari, Hediger, Matthias A., Landowski, Christopher P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8425032/
https://www.ncbi.nlm.nih.gov/pubmed/34496831
http://dx.doi.org/10.1186/s12934-021-01666-4
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author Havukainen, Sami
Pujol-Giménez, Jonai
Valkonen, Mari
Hediger, Matthias A.
Landowski, Christopher P.
author_facet Havukainen, Sami
Pujol-Giménez, Jonai
Valkonen, Mari
Hediger, Matthias A.
Landowski, Christopher P.
author_sort Havukainen, Sami
collection PubMed
description BACKGROUND: Lignocellulose biomass has been investigated as a feedstock for second generation biofuels and other value-added products. Some of the processes for biofuel production utilize cellulases and hemicellulases to convert the lignocellulosic biomass into a range of soluble sugars before fermentation with microorganisms such as yeast Saccharomyces cerevisiae. One of these sugars is l-arabinose, which cannot be utilized naturally by yeast. The first step in l-arabinose catabolism is its transport into the cells, and yeast lacks a specific transporter, which could perform this task. RESULTS: We identified Trire2_104072 of Trichoderma reesei as a potential l-arabinose transporter based on its expression profile. This transporter was described already in 2007 as d-xylose transporter XLT1. Electrophysiology experiments with Xenopus laevis oocytes and heterologous expression in yeast revealed that Trire2_104072 is a high-affinity l-arabinose symporter with a K(m) value in the range of [Formula: see text]  0.1–0.2 mM. It can also transport d-xylose but with low affinity (K(m) [Formula: see text]  9 mM). In yeast, l-arabinose transport was inhibited slightly by d-xylose but not by d-glucose in an assay with fivefold excess of the inhibiting sugar. Comparison with known l-arabinose transporters revealed that the expression of Trire2_104072 enabled yeast to uptake l-arabinose at the highest rate in conditions with low extracellular l-arabinose concentration. Despite the high specificity of Trire2_104072 for l-arabinose, the growth of its T. reesei deletion mutant was only affected at low l-arabinose concentrations. CONCLUSIONS: Due to its high affinity for l-arabinose and low inhibition by d-glucose or d-xylose, Trire2_104072 could serve as a good candidate for improving the existing pentose-utilizing yeast strains. The discovery of a highly specific l-arabinose transporter also adds to our knowledge of the primary metabolism of T. reesei. The phenotype of the deletion strain suggests the involvement of other transporters in l-arabinose transport in this species. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-021-01666-4.
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spelling pubmed-84250322021-09-10 Functional characterization of a highly specific l-arabinose transporter from Trichoderma reesei Havukainen, Sami Pujol-Giménez, Jonai Valkonen, Mari Hediger, Matthias A. Landowski, Christopher P. Microb Cell Fact Research BACKGROUND: Lignocellulose biomass has been investigated as a feedstock for second generation biofuels and other value-added products. Some of the processes for biofuel production utilize cellulases and hemicellulases to convert the lignocellulosic biomass into a range of soluble sugars before fermentation with microorganisms such as yeast Saccharomyces cerevisiae. One of these sugars is l-arabinose, which cannot be utilized naturally by yeast. The first step in l-arabinose catabolism is its transport into the cells, and yeast lacks a specific transporter, which could perform this task. RESULTS: We identified Trire2_104072 of Trichoderma reesei as a potential l-arabinose transporter based on its expression profile. This transporter was described already in 2007 as d-xylose transporter XLT1. Electrophysiology experiments with Xenopus laevis oocytes and heterologous expression in yeast revealed that Trire2_104072 is a high-affinity l-arabinose symporter with a K(m) value in the range of [Formula: see text]  0.1–0.2 mM. It can also transport d-xylose but with low affinity (K(m) [Formula: see text]  9 mM). In yeast, l-arabinose transport was inhibited slightly by d-xylose but not by d-glucose in an assay with fivefold excess of the inhibiting sugar. Comparison with known l-arabinose transporters revealed that the expression of Trire2_104072 enabled yeast to uptake l-arabinose at the highest rate in conditions with low extracellular l-arabinose concentration. Despite the high specificity of Trire2_104072 for l-arabinose, the growth of its T. reesei deletion mutant was only affected at low l-arabinose concentrations. CONCLUSIONS: Due to its high affinity for l-arabinose and low inhibition by d-glucose or d-xylose, Trire2_104072 could serve as a good candidate for improving the existing pentose-utilizing yeast strains. The discovery of a highly specific l-arabinose transporter also adds to our knowledge of the primary metabolism of T. reesei. The phenotype of the deletion strain suggests the involvement of other transporters in l-arabinose transport in this species. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-021-01666-4. BioMed Central 2021-09-08 /pmc/articles/PMC8425032/ /pubmed/34496831 http://dx.doi.org/10.1186/s12934-021-01666-4 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Havukainen, Sami
Pujol-Giménez, Jonai
Valkonen, Mari
Hediger, Matthias A.
Landowski, Christopher P.
Functional characterization of a highly specific l-arabinose transporter from Trichoderma reesei
title Functional characterization of a highly specific l-arabinose transporter from Trichoderma reesei
title_full Functional characterization of a highly specific l-arabinose transporter from Trichoderma reesei
title_fullStr Functional characterization of a highly specific l-arabinose transporter from Trichoderma reesei
title_full_unstemmed Functional characterization of a highly specific l-arabinose transporter from Trichoderma reesei
title_short Functional characterization of a highly specific l-arabinose transporter from Trichoderma reesei
title_sort functional characterization of a highly specific l-arabinose transporter from trichoderma reesei
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8425032/
https://www.ncbi.nlm.nih.gov/pubmed/34496831
http://dx.doi.org/10.1186/s12934-021-01666-4
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