Cargando…

Xpp1 regulates the expression of xylanases, but not of cellulases in Trichoderma reesei

BACKGROUND: The ascomycete Trichoderma reesei is industrially used for the production of cellulases. During the production process xylanases are co-secreted, which uses energy and nutrients. Cellulases and xylanases share the same main regulators, which makes a knowledge-based strain design difficul...

Descripción completa

Detalles Bibliográficos
Autores principales: Derntl, Christian, Rassinger, Alice, Srebotnik, Ewald, Mach, Robert L, Mach-Aigner, Astrid R
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4526299/
https://www.ncbi.nlm.nih.gov/pubmed/26246855
http://dx.doi.org/10.1186/s13068-015-0298-8
_version_ 1782384403833421824
author Derntl, Christian
Rassinger, Alice
Srebotnik, Ewald
Mach, Robert L
Mach-Aigner, Astrid R
author_facet Derntl, Christian
Rassinger, Alice
Srebotnik, Ewald
Mach, Robert L
Mach-Aigner, Astrid R
author_sort Derntl, Christian
collection PubMed
description BACKGROUND: The ascomycete Trichoderma reesei is industrially used for the production of cellulases. During the production process xylanases are co-secreted, which uses energy and nutrients. Cellulases and xylanases share the same main regulators, which makes a knowledge-based strain design difficult. However, previously a cis-element in the promoter of the main xylanase-encoding gene was identified as binding site for a putative repressor. Subsequently, three candidate repressors were identified in a pull-down approach. The expression of the most promising candidate, Xpp1 (Xylanase promoter-binding protein 1), was reported to be up-regulated on the repressing carbon source d-glucose and to bind the cis-element in vitro. RESULTS: In this study, Xpp1 was deleted and over-expressed in T. reesei. An in vivo DNA-footprint assay indicated that Xpp1 binds a palindromic sequence in the xyn2 promoter. Comparison of the deletion, the over-expression, and the parent strain demonstrated that Xpp1 regulates gene expression of xylanolytic enzymes at later cultivation stages. Xpp1 expression was found to be up-regulated, additionally to d-glucose, by high d-xylose availability. These findings together with the observed xyn2 transcript levels during growth on xylan suggest that Xpp1 is the mediator of a feedback mechanism. Notably, Xpp1 has neither influence on the d-xylose metabolism nor on the expression of cellulases. CONCLUSIONS: Xpp1 as regulator acting on the expression of xylanases, but not cellulases, is a highly promising candidate for knowledge-based strain design to improve the cellulases-to-xylanases ratio during industrial cellulase production. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-015-0298-8) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-4526299
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-45262992015-08-06 Xpp1 regulates the expression of xylanases, but not of cellulases in Trichoderma reesei Derntl, Christian Rassinger, Alice Srebotnik, Ewald Mach, Robert L Mach-Aigner, Astrid R Biotechnol Biofuels Research Article BACKGROUND: The ascomycete Trichoderma reesei is industrially used for the production of cellulases. During the production process xylanases are co-secreted, which uses energy and nutrients. Cellulases and xylanases share the same main regulators, which makes a knowledge-based strain design difficult. However, previously a cis-element in the promoter of the main xylanase-encoding gene was identified as binding site for a putative repressor. Subsequently, three candidate repressors were identified in a pull-down approach. The expression of the most promising candidate, Xpp1 (Xylanase promoter-binding protein 1), was reported to be up-regulated on the repressing carbon source d-glucose and to bind the cis-element in vitro. RESULTS: In this study, Xpp1 was deleted and over-expressed in T. reesei. An in vivo DNA-footprint assay indicated that Xpp1 binds a palindromic sequence in the xyn2 promoter. Comparison of the deletion, the over-expression, and the parent strain demonstrated that Xpp1 regulates gene expression of xylanolytic enzymes at later cultivation stages. Xpp1 expression was found to be up-regulated, additionally to d-glucose, by high d-xylose availability. These findings together with the observed xyn2 transcript levels during growth on xylan suggest that Xpp1 is the mediator of a feedback mechanism. Notably, Xpp1 has neither influence on the d-xylose metabolism nor on the expression of cellulases. CONCLUSIONS: Xpp1 as regulator acting on the expression of xylanases, but not cellulases, is a highly promising candidate for knowledge-based strain design to improve the cellulases-to-xylanases ratio during industrial cellulase production. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13068-015-0298-8) contains supplementary material, which is available to authorized users. BioMed Central 2015-08-06 /pmc/articles/PMC4526299/ /pubmed/26246855 http://dx.doi.org/10.1186/s13068-015-0298-8 Text en © Derntl et al. 2015 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 Article
Derntl, Christian
Rassinger, Alice
Srebotnik, Ewald
Mach, Robert L
Mach-Aigner, Astrid R
Xpp1 regulates the expression of xylanases, but not of cellulases in Trichoderma reesei
title Xpp1 regulates the expression of xylanases, but not of cellulases in Trichoderma reesei
title_full Xpp1 regulates the expression of xylanases, but not of cellulases in Trichoderma reesei
title_fullStr Xpp1 regulates the expression of xylanases, but not of cellulases in Trichoderma reesei
title_full_unstemmed Xpp1 regulates the expression of xylanases, but not of cellulases in Trichoderma reesei
title_short Xpp1 regulates the expression of xylanases, but not of cellulases in Trichoderma reesei
title_sort xpp1 regulates the expression of xylanases, but not of cellulases in trichoderma reesei
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4526299/
https://www.ncbi.nlm.nih.gov/pubmed/26246855
http://dx.doi.org/10.1186/s13068-015-0298-8
work_keys_str_mv AT derntlchristian xpp1regulatestheexpressionofxylanasesbutnotofcellulasesintrichodermareesei
AT rassingeralice xpp1regulatestheexpressionofxylanasesbutnotofcellulasesintrichodermareesei
AT srebotnikewald xpp1regulatestheexpressionofxylanasesbutnotofcellulasesintrichodermareesei
AT machrobertl xpp1regulatestheexpressionofxylanasesbutnotofcellulasesintrichodermareesei
AT machaignerastridr xpp1regulatestheexpressionofxylanasesbutnotofcellulasesintrichodermareesei