Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression system

"This is the peer reviewed version of the following article: Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression system, which has been published in final form at https://doi.org/10.1002/bit.24723. This article may...

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Autores: Tamayo Ramos, Juan Antonio, Barends, Sharief, de Lange, Dennis, de Jel, Annemarie, Verhaert, Raymond, de Graaff, Leo
Tipo de recurso: artículo
Estado:Versión aceptada para publicación
Fecha de publicación:2012
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/414751
Acceso en línea:http://hdl.handle.net/10261/414751
https://api.elsevier.com/content/abstract/scopus_id/84871722011
Access Level:acceso abierto
Palabra clave:Aspergillus niger
High-throughput screening
MRNA 5′UTR
Multicopper oxidase
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spelling Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression systemTamayo Ramos, Juan AntonioBarends, Shariefde Lange, Dennisde Jel, AnnemarieVerhaert, Raymondde Graaff, LeoAspergillus nigerHigh-throughput screeningMRNA 5′UTRMulticopper oxidaseAspergillus niger"This is the peer reviewed version of the following article: Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression system, which has been published in final form at https://doi.org/10.1002/bit.24723. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited."In filamentous fungi, most of the strategies used for the improvement of protein yields have been based on an increase in the transcript levels of a target gene. Strategies focusing at the translational level have been also described, but are far less explored. Here the 5' untranslated sequence of the glaA mRNA, a widely used expression system for the expression of recombinant proteins, was modified by the introduction of different nucleotide elements that have positive role in the translation process. Five Aspergillus niger laccase-like multicopper oxidases (MCOs) coding genes were fused to the native glaA 5'UTR and the three synthetic versions (sUTR1, sUTR2, and sUTR3) as well, and placed under the control of the glucoamylase gene promoter. Afterwards, a total of 20 fungal transformations were done using A. niger N593 as a recipient strain and 50 transformants per transformation were isolated and analyzed. The result of the incorporation of the synthetic 5'UTRs on the overall productivity of the transformants was assessed, on one hand by monitoring the laccase activity of all the isolated transformants, and on the other hand by quantifying and comparing the activity of those secreting the highest level of each MCO. For this purpose, a high-throughput method for the screening and selection of the best producers was developed. Once the best transformants producing the highest yield of McoA, McoB, McoC, McoD, and McoJ laccases were selected, their production level was quantified in supernatants of liquid cultures. The results obtained in this work indicate that modifications in the native glaA 5'UTR can lead to improvements in protein yields.This work has been carried out under the IBOS SME research project 053.63.322 granted by the ACTS national research program from the Netherlands. J.A.T.R. was recipient of a postdoctoral contract within research project 053.63.322.Peer reviewedJohn Wiley & Sons202620262012info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Postprintinfo:eu-repo/semantics/acceptedVersionapplication/pdfhttp://hdl.handle.net/10261/414751https://api.elsevier.com/content/abstract/scopus_id/84871722011reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)InglésBiotechnology and bioengineeringhttps://doi.org/10.1002/bit.24723Noinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/4147512026-05-22T06:33:51Z
dc.title.none.fl_str_mv Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression system
title Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression system
spellingShingle Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression system
Tamayo Ramos, Juan Antonio
Aspergillus niger
High-throughput screening
MRNA 5′UTR
Multicopper oxidase
Aspergillus niger
title_short Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression system
title_full Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression system
title_fullStr Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression system
title_full_unstemmed Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression system
title_sort Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression system
dc.creator.none.fl_str_mv Tamayo Ramos, Juan Antonio
Barends, Sharief
de Lange, Dennis
de Jel, Annemarie
Verhaert, Raymond
de Graaff, Leo
author Tamayo Ramos, Juan Antonio
author_facet Tamayo Ramos, Juan Antonio
Barends, Sharief
de Lange, Dennis
de Jel, Annemarie
Verhaert, Raymond
de Graaff, Leo
author_role author
author2 Barends, Sharief
de Lange, Dennis
de Jel, Annemarie
Verhaert, Raymond
de Graaff, Leo
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Aspergillus niger
High-throughput screening
MRNA 5′UTR
Multicopper oxidase
Aspergillus niger
topic Aspergillus niger
High-throughput screening
MRNA 5′UTR
Multicopper oxidase
Aspergillus niger
description "This is the peer reviewed version of the following article: Enhanced production of Aspergillus niger laccase-like multicopper oxidases through mRNA optimization of the glucoamylase expression system, which has been published in final form at https://doi.org/10.1002/bit.24723. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited."
publishDate 2012
dc.date.none.fl_str_mv 2012
2026
2026
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Postprint
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/414751
https://api.elsevier.com/content/abstract/scopus_id/84871722011
url http://hdl.handle.net/10261/414751
https://api.elsevier.com/content/abstract/scopus_id/84871722011
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Biotechnology and bioengineering
https://doi.org/10.1002/bit.24723
No
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv John Wiley & Sons
publisher.none.fl_str_mv John Wiley & Sons
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,81155

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