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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2019
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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. |
format | Online Article Text |
id | pubmed-6801129 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
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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