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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2017
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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. |
format | Online Article Text |
id | pubmed-5362885 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
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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