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Real-Time Monitoring of the Yeast Intracellular State During Bioprocesses With a Toolbox of Biosensors
Industrial fermentation processes strive for high robustness to ensure optimal and consistent performance. Medium components, fermentation products, and physical perturbations may cause stress and lower performance. Cellular stress elicits a range of responses, whose extracellular manifestations hav...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8776715/ https://www.ncbi.nlm.nih.gov/pubmed/35069506 http://dx.doi.org/10.3389/fmicb.2021.802169 |
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author | Torello Pianale, Luca Rugbjerg, Peter Olsson, Lisbeth |
author_facet | Torello Pianale, Luca Rugbjerg, Peter Olsson, Lisbeth |
author_sort | Torello Pianale, Luca |
collection | PubMed |
description | Industrial fermentation processes strive for high robustness to ensure optimal and consistent performance. Medium components, fermentation products, and physical perturbations may cause stress and lower performance. Cellular stress elicits a range of responses, whose extracellular manifestations have been extensively studied; whereas intracellular aspects remain poorly known due to lack of tools for real-time monitoring. Genetically encoded biosensors have emerged as promising tools and have been used to improve microbial productivity and tolerance toward industrially relevant stresses. Here, fluorescent biosensors able to sense the yeast intracellular environment (pH, ATP levels, oxidative stress, glycolytic flux, and ribosome production) were implemented into a versatile and easy-to-use toolbox. Marker-free and efficient genome integration at a conserved site on chromosome X of Saccharomyces cerevisiae strains and a commercial Saccharomyces boulardii strain was developed. Moreover, multiple biosensors were used to simultaneously monitor different intracellular parameters in a single cell. Even when combined together, the biosensors did not significantly affect key physiological parameters, such as specific growth rate and product yields. Activation and response of each biosensor and their interconnection were assessed using an advanced micro-cultivation system. Finally, the toolbox was used to screen cell behavior in a synthetic lignocellulosic hydrolysate that mimicked harsh industrial substrates, revealing differences in the oxidative stress response between laboratory (CEN.PK113-7D) and industrial (Ethanol Red) S. cerevisiae strains. In summary, the toolbox will allow both the exploration of yeast diversity and physiological responses in natural and complex industrial conditions, as well as the possibility to monitor production processes. |
format | Online Article Text |
id | pubmed-8776715 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-87767152022-01-22 Real-Time Monitoring of the Yeast Intracellular State During Bioprocesses With a Toolbox of Biosensors Torello Pianale, Luca Rugbjerg, Peter Olsson, Lisbeth Front Microbiol Microbiology Industrial fermentation processes strive for high robustness to ensure optimal and consistent performance. Medium components, fermentation products, and physical perturbations may cause stress and lower performance. Cellular stress elicits a range of responses, whose extracellular manifestations have been extensively studied; whereas intracellular aspects remain poorly known due to lack of tools for real-time monitoring. Genetically encoded biosensors have emerged as promising tools and have been used to improve microbial productivity and tolerance toward industrially relevant stresses. Here, fluorescent biosensors able to sense the yeast intracellular environment (pH, ATP levels, oxidative stress, glycolytic flux, and ribosome production) were implemented into a versatile and easy-to-use toolbox. Marker-free and efficient genome integration at a conserved site on chromosome X of Saccharomyces cerevisiae strains and a commercial Saccharomyces boulardii strain was developed. Moreover, multiple biosensors were used to simultaneously monitor different intracellular parameters in a single cell. Even when combined together, the biosensors did not significantly affect key physiological parameters, such as specific growth rate and product yields. Activation and response of each biosensor and their interconnection were assessed using an advanced micro-cultivation system. Finally, the toolbox was used to screen cell behavior in a synthetic lignocellulosic hydrolysate that mimicked harsh industrial substrates, revealing differences in the oxidative stress response between laboratory (CEN.PK113-7D) and industrial (Ethanol Red) S. cerevisiae strains. In summary, the toolbox will allow both the exploration of yeast diversity and physiological responses in natural and complex industrial conditions, as well as the possibility to monitor production processes. Frontiers Media S.A. 2022-01-07 /pmc/articles/PMC8776715/ /pubmed/35069506 http://dx.doi.org/10.3389/fmicb.2021.802169 Text en Copyright © 2022 Torello Pianale, Rugbjerg and Olsson. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Torello Pianale, Luca Rugbjerg, Peter Olsson, Lisbeth Real-Time Monitoring of the Yeast Intracellular State During Bioprocesses With a Toolbox of Biosensors |
title | Real-Time Monitoring of the Yeast Intracellular State During Bioprocesses With a Toolbox of Biosensors |
title_full | Real-Time Monitoring of the Yeast Intracellular State During Bioprocesses With a Toolbox of Biosensors |
title_fullStr | Real-Time Monitoring of the Yeast Intracellular State During Bioprocesses With a Toolbox of Biosensors |
title_full_unstemmed | Real-Time Monitoring of the Yeast Intracellular State During Bioprocesses With a Toolbox of Biosensors |
title_short | Real-Time Monitoring of the Yeast Intracellular State During Bioprocesses With a Toolbox of Biosensors |
title_sort | real-time monitoring of the yeast intracellular state during bioprocesses with a toolbox of biosensors |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8776715/ https://www.ncbi.nlm.nih.gov/pubmed/35069506 http://dx.doi.org/10.3389/fmicb.2021.802169 |
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