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Inducer-independent production of pectinases in Aspergillus niger by overexpression of the D-galacturonic acid-responsive transcription factor gaaR

The transcription factor GaaR is needed for the expression of genes required for pectin degradation and transport and catabolism of the main degradation product, D-galacturonic acid (GA) in Aspergillus niger. In this study, we used the strong constitutive gpdA promoter of Aspergillus nidulans to ove...

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Autores principales: Alazi, Ebru, Knetsch, Tim, Di Falco, Marcos, Reid, Ian D., Arentshorst, Mark, Visser, Jaap, Tsang, Adrian, Ram, Arthur F. J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5847190/
https://www.ncbi.nlm.nih.gov/pubmed/29368217
http://dx.doi.org/10.1007/s00253-018-8753-7
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author Alazi, Ebru
Knetsch, Tim
Di Falco, Marcos
Reid, Ian D.
Arentshorst, Mark
Visser, Jaap
Tsang, Adrian
Ram, Arthur F. J.
author_facet Alazi, Ebru
Knetsch, Tim
Di Falco, Marcos
Reid, Ian D.
Arentshorst, Mark
Visser, Jaap
Tsang, Adrian
Ram, Arthur F. J.
author_sort Alazi, Ebru
collection PubMed
description The transcription factor GaaR is needed for the expression of genes required for pectin degradation and transport and catabolism of the main degradation product, D-galacturonic acid (GA) in Aspergillus niger. In this study, we used the strong constitutive gpdA promoter of Aspergillus nidulans to overexpress gaaR in A. niger. Overexpression of gaaR resulted in an increased transcription of the genes encoding pectinases, (putative) GA transporters, and catabolic pathway enzymes even under non-inducing conditions, i.e., in the absence of GA. Exoproteome analysis of a strain overexpressing gaaR showed that this strain secretes highly elevated levels of pectinases when grown in fructose. The genes encoding exo-polygalacturonases were found to be subjected to CreA-mediated carbon catabolite repression, even in the presence of fructose. Deletion of creA in the strain overexpressing gaaR resulted in a further increase in pectinase production in fructose. We showed that GaaR localizes mainly in the nucleus regardless of the presence of an inducer, and that overexpression of gaaR leads to an increased concentration of GaaR in the nucleus. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00253-018-8753-7) contains supplementary material, which is available to authorized users.
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spelling pubmed-58471902018-03-20 Inducer-independent production of pectinases in Aspergillus niger by overexpression of the D-galacturonic acid-responsive transcription factor gaaR Alazi, Ebru Knetsch, Tim Di Falco, Marcos Reid, Ian D. Arentshorst, Mark Visser, Jaap Tsang, Adrian Ram, Arthur F. J. Appl Microbiol Biotechnol Applied Genetics and Molecular Biotechnology The transcription factor GaaR is needed for the expression of genes required for pectin degradation and transport and catabolism of the main degradation product, D-galacturonic acid (GA) in Aspergillus niger. In this study, we used the strong constitutive gpdA promoter of Aspergillus nidulans to overexpress gaaR in A. niger. Overexpression of gaaR resulted in an increased transcription of the genes encoding pectinases, (putative) GA transporters, and catabolic pathway enzymes even under non-inducing conditions, i.e., in the absence of GA. Exoproteome analysis of a strain overexpressing gaaR showed that this strain secretes highly elevated levels of pectinases when grown in fructose. The genes encoding exo-polygalacturonases were found to be subjected to CreA-mediated carbon catabolite repression, even in the presence of fructose. Deletion of creA in the strain overexpressing gaaR resulted in a further increase in pectinase production in fructose. We showed that GaaR localizes mainly in the nucleus regardless of the presence of an inducer, and that overexpression of gaaR leads to an increased concentration of GaaR in the nucleus. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00253-018-8753-7) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2018-01-24 2018 /pmc/articles/PMC5847190/ /pubmed/29368217 http://dx.doi.org/10.1007/s00253-018-8753-7 Text en © The Author(s) 2018 Open Access This 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.
spellingShingle Applied Genetics and Molecular Biotechnology
Alazi, Ebru
Knetsch, Tim
Di Falco, Marcos
Reid, Ian D.
Arentshorst, Mark
Visser, Jaap
Tsang, Adrian
Ram, Arthur F. J.
Inducer-independent production of pectinases in Aspergillus niger by overexpression of the D-galacturonic acid-responsive transcription factor gaaR
title Inducer-independent production of pectinases in Aspergillus niger by overexpression of the D-galacturonic acid-responsive transcription factor gaaR
title_full Inducer-independent production of pectinases in Aspergillus niger by overexpression of the D-galacturonic acid-responsive transcription factor gaaR
title_fullStr Inducer-independent production of pectinases in Aspergillus niger by overexpression of the D-galacturonic acid-responsive transcription factor gaaR
title_full_unstemmed Inducer-independent production of pectinases in Aspergillus niger by overexpression of the D-galacturonic acid-responsive transcription factor gaaR
title_short Inducer-independent production of pectinases in Aspergillus niger by overexpression of the D-galacturonic acid-responsive transcription factor gaaR
title_sort inducer-independent production of pectinases in aspergillus niger by overexpression of the d-galacturonic acid-responsive transcription factor gaar
topic Applied Genetics and Molecular Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5847190/
https://www.ncbi.nlm.nih.gov/pubmed/29368217
http://dx.doi.org/10.1007/s00253-018-8753-7
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