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Recombinant Peptide Production Softens Escherichia coli Cells and Increases Their Size during C-Limited Fed-Batch Cultivation

Stress-associated changes in the mechanical properties at the single-cell level of Escherichia coli (E. coli) cultures in bioreactors are still poorly investigated. In our study, we compared peptide-producing and non-producing BL21(DE3) cells in a fed-batch cultivation with tightly controlled proces...

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Autores principales: Weber, Andreas, Gibisch, Martin, Tyrakowski, Daniel, Cserjan-Puschmann, Monika, Toca-Herrera, José L., Striedner, Gerald
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9916741/
https://www.ncbi.nlm.nih.gov/pubmed/36768962
http://dx.doi.org/10.3390/ijms24032641
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author Weber, Andreas
Gibisch, Martin
Tyrakowski, Daniel
Cserjan-Puschmann, Monika
Toca-Herrera, José L.
Striedner, Gerald
author_facet Weber, Andreas
Gibisch, Martin
Tyrakowski, Daniel
Cserjan-Puschmann, Monika
Toca-Herrera, José L.
Striedner, Gerald
author_sort Weber, Andreas
collection PubMed
description Stress-associated changes in the mechanical properties at the single-cell level of Escherichia coli (E. coli) cultures in bioreactors are still poorly investigated. In our study, we compared peptide-producing and non-producing BL21(DE3) cells in a fed-batch cultivation with tightly controlled process parameters. The cell growth, peptide content, and cell lysis were analysed, and changes in the mechanical properties were investigated using atomic force microscopy. Recombinant-tagged somatostatin-28 was expressed as soluble up to 197 ± 11 mg g(−1). The length of both cultivated strains increased throughout the cultivation by up to 17.6%, with nearly constant diameters. The peptide-producing cells were significantly softer than the non-producers throughout the cultivation, and respective Young’s moduli decreased by up to 57% over time. A minimum Young’s modulus of 1.6 MPa was observed after 23 h of the fed-batch. Furthermore, an analysis of the viscoelastic properties revealed that peptide-producing BL21(DE3) appeared more fluid-like and softer than the non-producing reference. For the first time, we provide evidence that the physical properties (i.e., the mechanical properties) on the single-cell level are significantly influenced by the metabolic burden imposed by the recombinant peptide expression and C-limitation in bioreactors.
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spelling pubmed-99167412023-02-11 Recombinant Peptide Production Softens Escherichia coli Cells and Increases Their Size during C-Limited Fed-Batch Cultivation Weber, Andreas Gibisch, Martin Tyrakowski, Daniel Cserjan-Puschmann, Monika Toca-Herrera, José L. Striedner, Gerald Int J Mol Sci Article Stress-associated changes in the mechanical properties at the single-cell level of Escherichia coli (E. coli) cultures in bioreactors are still poorly investigated. In our study, we compared peptide-producing and non-producing BL21(DE3) cells in a fed-batch cultivation with tightly controlled process parameters. The cell growth, peptide content, and cell lysis were analysed, and changes in the mechanical properties were investigated using atomic force microscopy. Recombinant-tagged somatostatin-28 was expressed as soluble up to 197 ± 11 mg g(−1). The length of both cultivated strains increased throughout the cultivation by up to 17.6%, with nearly constant diameters. The peptide-producing cells were significantly softer than the non-producers throughout the cultivation, and respective Young’s moduli decreased by up to 57% over time. A minimum Young’s modulus of 1.6 MPa was observed after 23 h of the fed-batch. Furthermore, an analysis of the viscoelastic properties revealed that peptide-producing BL21(DE3) appeared more fluid-like and softer than the non-producing reference. For the first time, we provide evidence that the physical properties (i.e., the mechanical properties) on the single-cell level are significantly influenced by the metabolic burden imposed by the recombinant peptide expression and C-limitation in bioreactors. MDPI 2023-01-30 /pmc/articles/PMC9916741/ /pubmed/36768962 http://dx.doi.org/10.3390/ijms24032641 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Weber, Andreas
Gibisch, Martin
Tyrakowski, Daniel
Cserjan-Puschmann, Monika
Toca-Herrera, José L.
Striedner, Gerald
Recombinant Peptide Production Softens Escherichia coli Cells and Increases Their Size during C-Limited Fed-Batch Cultivation
title Recombinant Peptide Production Softens Escherichia coli Cells and Increases Their Size during C-Limited Fed-Batch Cultivation
title_full Recombinant Peptide Production Softens Escherichia coli Cells and Increases Their Size during C-Limited Fed-Batch Cultivation
title_fullStr Recombinant Peptide Production Softens Escherichia coli Cells and Increases Their Size during C-Limited Fed-Batch Cultivation
title_full_unstemmed Recombinant Peptide Production Softens Escherichia coli Cells and Increases Their Size during C-Limited Fed-Batch Cultivation
title_short Recombinant Peptide Production Softens Escherichia coli Cells and Increases Their Size during C-Limited Fed-Batch Cultivation
title_sort recombinant peptide production softens escherichia coli cells and increases their size during c-limited fed-batch cultivation
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9916741/
https://www.ncbi.nlm.nih.gov/pubmed/36768962
http://dx.doi.org/10.3390/ijms24032641
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