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Innovative microscale workflow from fungi cultures to Cell Wall‐Degrading Enzyme screening

This study aimed at developing a complete miniaturized high‐throughput screening workflow for the evaluation of the Cell Wall‐Degrading Enzyme (CWDE) activities produced by any fungal strain directly cultivated on raw feedstock in a submerged manner. In this study, wheat straw was selected as model...

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Autores principales: Raulo, Roxane, Heuson, Egon, Siah, Ali, Phalip, Vincent, Froidevaux, Renato
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6801129/
https://www.ncbi.nlm.nih.gov/pubmed/31006173
http://dx.doi.org/10.1111/1751-7915.13405
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author Raulo, Roxane
Heuson, Egon
Siah, Ali
Phalip, Vincent
Froidevaux, Renato
author_facet Raulo, Roxane
Heuson, Egon
Siah, Ali
Phalip, Vincent
Froidevaux, Renato
author_sort Raulo, Roxane
collection PubMed
description This study aimed at developing a complete miniaturized high‐throughput screening workflow for the evaluation of the Cell Wall‐Degrading Enzyme (CWDE) activities produced by any fungal strain directly cultivated on raw feedstock in a submerged manner. In this study, wheat straw was selected as model substrate as it represents an important carbon source but yet poorly valorised to yield high added value products. Fungi were grown in a microbioreactor in a high‐throughput (HT) way to replace the fastidious shaking flask cultivations. Both approaches were compared in order to validate our new methodology. The range of CWDE activities produced from the cultures was assayed using AZO‐died and pNP‐linked substrates in an SBS plate format using a Biomek FXp pipetting platform. As highlighted in this study, it was shown that the CWDE activities gathered from the microbioreactor cultivations were similar or higher to those obtained from shake flasks cultures, with a lower standard deviation on the measured values, making this new method much faster than the traditional one and suitable for HT CWDE production thanks to its pipetting platform compatibility. Also, the results showed that the enzymatic activities measured were the same when doing the assay manually or using the automated method.
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spelling pubmed-68011292019-10-22 Innovative microscale workflow from fungi cultures to Cell Wall‐Degrading Enzyme screening Raulo, Roxane Heuson, Egon Siah, Ali Phalip, Vincent Froidevaux, Renato Microb Biotechnol Brief Reports This study aimed at developing a complete miniaturized high‐throughput screening workflow for the evaluation of the Cell Wall‐Degrading Enzyme (CWDE) activities produced by any fungal strain directly cultivated on raw feedstock in a submerged manner. In this study, wheat straw was selected as model substrate as it represents an important carbon source but yet poorly valorised to yield high added value products. Fungi were grown in a microbioreactor in a high‐throughput (HT) way to replace the fastidious shaking flask cultivations. Both approaches were compared in order to validate our new methodology. The range of CWDE activities produced from the cultures was assayed using AZO‐died and pNP‐linked substrates in an SBS plate format using a Biomek FXp pipetting platform. As highlighted in this study, it was shown that the CWDE activities gathered from the microbioreactor cultivations were similar or higher to those obtained from shake flasks cultures, with a lower standard deviation on the measured values, making this new method much faster than the traditional one and suitable for HT CWDE production thanks to its pipetting platform compatibility. Also, the results showed that the enzymatic activities measured were the same when doing the assay manually or using the automated method. John Wiley and Sons Inc. 2019-04-20 /pmc/articles/PMC6801129/ /pubmed/31006173 http://dx.doi.org/10.1111/1751-7915.13405 Text en © 2019 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Brief Reports
Raulo, Roxane
Heuson, Egon
Siah, Ali
Phalip, Vincent
Froidevaux, Renato
Innovative microscale workflow from fungi cultures to Cell Wall‐Degrading Enzyme screening
title Innovative microscale workflow from fungi cultures to Cell Wall‐Degrading Enzyme screening
title_full Innovative microscale workflow from fungi cultures to Cell Wall‐Degrading Enzyme screening
title_fullStr Innovative microscale workflow from fungi cultures to Cell Wall‐Degrading Enzyme screening
title_full_unstemmed Innovative microscale workflow from fungi cultures to Cell Wall‐Degrading Enzyme screening
title_short Innovative microscale workflow from fungi cultures to Cell Wall‐Degrading Enzyme screening
title_sort innovative microscale workflow from fungi cultures to cell wall‐degrading enzyme screening
topic Brief Reports
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6801129/
https://www.ncbi.nlm.nih.gov/pubmed/31006173
http://dx.doi.org/10.1111/1751-7915.13405
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