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Mechanistic platform knowledge of concomitant sugar uptake in Escherichia coli BL21(DE3) strains

When producing recombinant proteins, the use of Escherichia coli strain BL21(DE3) in combination with the T7-based pET-expression system is often the method of choice. In a recent study we introduced a mechanistic model describing the correlation of the specific glucose uptake rate (q(s,glu)) and th...

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Autores principales: Wurm, David J., Hausjell, Johanna, Ulonska, Sophia, Herwig, Christoph, Spadiut, Oliver
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5362885/
https://www.ncbi.nlm.nih.gov/pubmed/28332595
http://dx.doi.org/10.1038/srep45072
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author Wurm, David J.
Hausjell, Johanna
Ulonska, Sophia
Herwig, Christoph
Spadiut, Oliver
author_facet Wurm, David J.
Hausjell, Johanna
Ulonska, Sophia
Herwig, Christoph
Spadiut, Oliver
author_sort Wurm, David J.
collection PubMed
description When producing recombinant proteins, the use of Escherichia coli strain BL21(DE3) in combination with the T7-based pET-expression system is often the method of choice. In a recent study we introduced a mechanistic model describing the correlation of the specific glucose uptake rate (q(s,glu)) and the corresponding maximum specific lactose uptake rate (q(s,lac,max)) for a pET-based E. coli BL21(DE3) strain producing a single chain variable fragment (scFv). We showed the effect of q(s,lac,max) on productivity and product location underlining its importance for recombinant protein production. In the present study we investigated the mechanistic q(s,glu)/q(s,lac,max) correlation for four pET-based E. coli BL21(DE3) strains producing different recombinant products and thereby proved the mechanistic model to be platform knowledge for E. coli BL21(DE3). However, we found that the model parameters strongly depended on the recombinant product. Driven by this observation we tested different dynamic bioprocess strategies to allow a faster investigation of this mechanistic correlation. In fact, we succeeded and propose an experimental strategy comprising only one batch cultivation, one fed-batch cultivation as well as one dynamic experiment, to reliably determine the mechanistic model for q(s,glu)/q(s,lac,max) and get trustworthy model parameters for pET-based E. coli BL21(DE3) strains which are the basis for bioprocess development.
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spelling pubmed-53628852017-03-24 Mechanistic platform knowledge of concomitant sugar uptake in Escherichia coli BL21(DE3) strains Wurm, David J. Hausjell, Johanna Ulonska, Sophia Herwig, Christoph Spadiut, Oliver Sci Rep Article When producing recombinant proteins, the use of Escherichia coli strain BL21(DE3) in combination with the T7-based pET-expression system is often the method of choice. In a recent study we introduced a mechanistic model describing the correlation of the specific glucose uptake rate (q(s,glu)) and the corresponding maximum specific lactose uptake rate (q(s,lac,max)) for a pET-based E. coli BL21(DE3) strain producing a single chain variable fragment (scFv). We showed the effect of q(s,lac,max) on productivity and product location underlining its importance for recombinant protein production. In the present study we investigated the mechanistic q(s,glu)/q(s,lac,max) correlation for four pET-based E. coli BL21(DE3) strains producing different recombinant products and thereby proved the mechanistic model to be platform knowledge for E. coli BL21(DE3). However, we found that the model parameters strongly depended on the recombinant product. Driven by this observation we tested different dynamic bioprocess strategies to allow a faster investigation of this mechanistic correlation. In fact, we succeeded and propose an experimental strategy comprising only one batch cultivation, one fed-batch cultivation as well as one dynamic experiment, to reliably determine the mechanistic model for q(s,glu)/q(s,lac,max) and get trustworthy model parameters for pET-based E. coli BL21(DE3) strains which are the basis for bioprocess development. Nature Publishing Group 2017-03-23 /pmc/articles/PMC5362885/ /pubmed/28332595 http://dx.doi.org/10.1038/srep45072 Text en Copyright © 2017, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Wurm, David J.
Hausjell, Johanna
Ulonska, Sophia
Herwig, Christoph
Spadiut, Oliver
Mechanistic platform knowledge of concomitant sugar uptake in Escherichia coli BL21(DE3) strains
title Mechanistic platform knowledge of concomitant sugar uptake in Escherichia coli BL21(DE3) strains
title_full Mechanistic platform knowledge of concomitant sugar uptake in Escherichia coli BL21(DE3) strains
title_fullStr Mechanistic platform knowledge of concomitant sugar uptake in Escherichia coli BL21(DE3) strains
title_full_unstemmed Mechanistic platform knowledge of concomitant sugar uptake in Escherichia coli BL21(DE3) strains
title_short Mechanistic platform knowledge of concomitant sugar uptake in Escherichia coli BL21(DE3) strains
title_sort mechanistic platform knowledge of concomitant sugar uptake in escherichia coli bl21(de3) strains
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5362885/
https://www.ncbi.nlm.nih.gov/pubmed/28332595
http://dx.doi.org/10.1038/srep45072
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