A comparative analysis of industrial Escherichia coli K-12 and B strains in high-glucose batch cultivations on process-, transcriptome- and proteome level.
Escherichia coli K-12 and B strains are among the most frequently used bacterial hosts for production of recombinant proteins on an industrial scale. To improve existing processes and to accelerate bioprocess development, we performed a detailed host analysis. We investigated the different behaviors...
Guardado en:
| Autores principales: | , , , , , , , |
|---|---|
| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
Public Library of Science (PLoS)
2013
|
| Materias: | |
| Acceso en línea: | https://doaj.org/article/d579f0860f3a467cb7e94f4ea14d9414 |
| Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
| id |
oai:doaj.org-article:d579f0860f3a467cb7e94f4ea14d9414 |
|---|---|
| record_format |
dspace |
| spelling |
oai:doaj.org-article:d579f0860f3a467cb7e94f4ea14d94142021-11-18T09:00:38ZA comparative analysis of industrial Escherichia coli K-12 and B strains in high-glucose batch cultivations on process-, transcriptome- and proteome level.1932-620310.1371/journal.pone.0070516https://doaj.org/article/d579f0860f3a467cb7e94f4ea14d94142013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23950949/?tool=EBIhttps://doaj.org/toc/1932-6203Escherichia coli K-12 and B strains are among the most frequently used bacterial hosts for production of recombinant proteins on an industrial scale. To improve existing processes and to accelerate bioprocess development, we performed a detailed host analysis. We investigated the different behaviors of the E. coli production strains BL21, RV308, and HMS174 in response to high-glucose concentrations. Tightly controlled cultivations were conducted under defined environmental conditions for the in-depth analysis of physiological behavior. In addition to acquisition of standard process parameters, we also used DNA microarray analysis and differential gel electrophoresis (Ettan(TM) DIGE). Batch cultivations showed different yields of the distinct strains for cell dry mass and growth rate, which were highest for BL21. In addition, production of acetate, triggered by excess glucose supply, was much higher for the K-12 strains compared to the B strain. Analysis of transcriptome data showed significant alteration in 347 of 3882 genes common among all three hosts. These differentially expressed genes included, for example, those involved in transport, iron acquisition, and motility. The investigation of proteome patterns additionally revealed a high number of differentially expressed proteins among the investigated hosts. The subsequently selected 38 spots included proteins involved in transport and motility. The results of this comprehensive analysis delivered a full genomic picture of the three investigated strains. Differentially expressed groups for targeted host modification were identified like glucose transport or iron acquisition, enabling potential optimization of strains to improve yield and process quality. Dissimilar growth profiles of the strains confirm different genotypes. Furthermore, distinct transcriptome patterns support differential regulation at the genome level. The identified proteins showed high agreement with the transcriptome data and suggest similar regulation within a host at both levels for the identified groups. Such host attributes need to be considered in future process design and operation.Karoline MarischKarl BayerTheresa ScharlJuergen MairhoferPeter M KremplKarin HummelEbrahim Razzazi-FazeliGerald StriednerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 8, p e70516 (2013) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
Medicine R Science Q |
| spellingShingle |
Medicine R Science Q Karoline Marisch Karl Bayer Theresa Scharl Juergen Mairhofer Peter M Krempl Karin Hummel Ebrahim Razzazi-Fazeli Gerald Striedner A comparative analysis of industrial Escherichia coli K-12 and B strains in high-glucose batch cultivations on process-, transcriptome- and proteome level. |
| description |
Escherichia coli K-12 and B strains are among the most frequently used bacterial hosts for production of recombinant proteins on an industrial scale. To improve existing processes and to accelerate bioprocess development, we performed a detailed host analysis. We investigated the different behaviors of the E. coli production strains BL21, RV308, and HMS174 in response to high-glucose concentrations. Tightly controlled cultivations were conducted under defined environmental conditions for the in-depth analysis of physiological behavior. In addition to acquisition of standard process parameters, we also used DNA microarray analysis and differential gel electrophoresis (Ettan(TM) DIGE). Batch cultivations showed different yields of the distinct strains for cell dry mass and growth rate, which were highest for BL21. In addition, production of acetate, triggered by excess glucose supply, was much higher for the K-12 strains compared to the B strain. Analysis of transcriptome data showed significant alteration in 347 of 3882 genes common among all three hosts. These differentially expressed genes included, for example, those involved in transport, iron acquisition, and motility. The investigation of proteome patterns additionally revealed a high number of differentially expressed proteins among the investigated hosts. The subsequently selected 38 spots included proteins involved in transport and motility. The results of this comprehensive analysis delivered a full genomic picture of the three investigated strains. Differentially expressed groups for targeted host modification were identified like glucose transport or iron acquisition, enabling potential optimization of strains to improve yield and process quality. Dissimilar growth profiles of the strains confirm different genotypes. Furthermore, distinct transcriptome patterns support differential regulation at the genome level. The identified proteins showed high agreement with the transcriptome data and suggest similar regulation within a host at both levels for the identified groups. Such host attributes need to be considered in future process design and operation. |
| format |
article |
| author |
Karoline Marisch Karl Bayer Theresa Scharl Juergen Mairhofer Peter M Krempl Karin Hummel Ebrahim Razzazi-Fazeli Gerald Striedner |
| author_facet |
Karoline Marisch Karl Bayer Theresa Scharl Juergen Mairhofer Peter M Krempl Karin Hummel Ebrahim Razzazi-Fazeli Gerald Striedner |
| author_sort |
Karoline Marisch |
| title |
A comparative analysis of industrial Escherichia coli K-12 and B strains in high-glucose batch cultivations on process-, transcriptome- and proteome level. |
| title_short |
A comparative analysis of industrial Escherichia coli K-12 and B strains in high-glucose batch cultivations on process-, transcriptome- and proteome level. |
| title_full |
A comparative analysis of industrial Escherichia coli K-12 and B strains in high-glucose batch cultivations on process-, transcriptome- and proteome level. |
| title_fullStr |
A comparative analysis of industrial Escherichia coli K-12 and B strains in high-glucose batch cultivations on process-, transcriptome- and proteome level. |
| title_full_unstemmed |
A comparative analysis of industrial Escherichia coli K-12 and B strains in high-glucose batch cultivations on process-, transcriptome- and proteome level. |
| title_sort |
comparative analysis of industrial escherichia coli k-12 and b strains in high-glucose batch cultivations on process-, transcriptome- and proteome level. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2013 |
| url |
https://doaj.org/article/d579f0860f3a467cb7e94f4ea14d9414 |
| work_keys_str_mv |
AT karolinemarisch acomparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT karlbayer acomparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT theresascharl acomparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT juergenmairhofer acomparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT petermkrempl acomparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT karinhummel acomparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT ebrahimrazzazifazeli acomparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT geraldstriedner acomparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT karolinemarisch comparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT karlbayer comparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT theresascharl comparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT juergenmairhofer comparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT petermkrempl comparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT karinhummel comparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT ebrahimrazzazifazeli comparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel AT geraldstriedner comparativeanalysisofindustrialescherichiacolik12andbstrainsinhighglucosebatchcultivationsonprocesstranscriptomeandproteomelevel |
| _version_ |
1718421052022849536 |