Induction temperature impacts the structure of recombinant HuGM-CSF inclusion bodies in thermoinducible E. coli
Background: The temperature upshift has been widely used as an induction system to produce recombinant proteins (RPs). However, thermoinduction could affect bacterial metabolism, RP production, and RP aggregation. Understanding the structure and functionality of those aggregates, known as inclusion...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-09-01
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| Series: | Electronic Journal of Biotechnology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0717345822000343 |
| _version_ | 1818065755070005248 |
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| author | Sara Restrepo-Pineda Diego Rosiles-Becerril Angélica B. Vargas-Castillo Luis P. Ávila-Barrientos Axel Luviano Nuria Sánchez-Puig Enrique García-Hernández Nestor O. Pérez Mauricio A. Trujillo-Roldán Norma A. Valdez-Cruz |
| author_facet | Sara Restrepo-Pineda Diego Rosiles-Becerril Angélica B. Vargas-Castillo Luis P. Ávila-Barrientos Axel Luviano Nuria Sánchez-Puig Enrique García-Hernández Nestor O. Pérez Mauricio A. Trujillo-Roldán Norma A. Valdez-Cruz |
| author_sort | Sara Restrepo-Pineda |
| collection | DOAJ |
| description | Background: The temperature upshift has been widely used as an induction system to produce recombinant proteins (RPs). However, thermoinduction could affect bacterial metabolism, RP production, and RP aggregation. Understanding the structure and functionality of those aggregates, known as inclusion bodies (IBs), is a research area of interest in bioprocesses being scarcely studied under thermoinduction. Here, we describe the effect of the thermoinduction (39°C or 42°C) on the production of the recombinant human granulocyte–macrophage colony-stimulating factor (rHuGM-CSF) using Escherichia coli W3110 under the system λpL/cI857. Results: Results indicated that at 39°C, the production of biomass was almost doubled as well as the acetate accumulation compared to 42°C. Cultures thermoinduced at 42°C improved 1.5-fold the total protein over biomass yield and 1.25-fold the RP over total protein yield. Furthermore, 42°C accelerated the onset of IB formation, changing its architecture. Additionally, IBs formed at 42°C were less soluble and presented higher disorderly structures compared with IBs formed at 39°C, enriched in α-helix and amyloidal-like structures. Conclusions: This study highlights the observation that IBs attain different architecture in response to small changes in environmental conditions, such as the induction temperature, being this helpful information to improve thermoinduced bioprocesses.How to cite: Restrepo-Pineda S, Rosiles-Becerril D, Vargas-Castillo AB, et al. Induction temperature impacts the structure of recombinant HuGM-CSF inclusion bodies in thermoinducible E. coli. Electron J Biotechnol 2022;59. https://doi.org/10.1016/j.ejbt.2022.08.004. |
| first_indexed | 2024-12-10T14:56:56Z |
| format | Article |
| id | doaj.art-25c75a7d5a5141f4a70b07f15ae552be |
| institution | Directory Open Access Journal |
| issn | 0717-3458 |
| language | English |
| last_indexed | 2024-12-10T14:56:56Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Electronic Journal of Biotechnology |
| spelling | doaj.art-25c75a7d5a5141f4a70b07f15ae552be2022-12-22T01:44:17ZengElsevierElectronic Journal of Biotechnology0717-34582022-09-015994106Induction temperature impacts the structure of recombinant HuGM-CSF inclusion bodies in thermoinducible E. coliSara Restrepo-Pineda0Diego Rosiles-Becerril1Angélica B. Vargas-Castillo2Luis P. Ávila-Barrientos3Axel Luviano4Nuria Sánchez-Puig5Enrique García-Hernández6Nestor O. Pérez7Mauricio A. Trujillo-Roldán8Norma A. Valdez-Cruz9Programa de Investigación de Producción de Biomoléculas, Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, CP 04510 Ciudad de México, MexicoPrograma de Investigación de Producción de Biomoléculas, Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, CP 04510 Ciudad de México, MexicoPrograma de Investigación de Producción de Biomoléculas, Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, CP 04510 Ciudad de México, Mexico; INSERM UMR-S-MD 1197, Hôpital Paul Brousse, Villejuif, FrancePrograma de Investigación de Producción de Biomoléculas, Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, CP 04510 Ciudad de México, Mexico; Instituto de Química, Universidad Nacional Autónoma de México, Ciudad Universitaria, Ciudad de México 04510, MexicoInstituto de Química, Universidad Nacional Autónoma de México, Ciudad Universitaria, Ciudad de México 04510, MexicoInstituto de Química, Universidad Nacional Autónoma de México, Ciudad Universitaria, Ciudad de México 04510, MexicoInstituto de Química, Universidad Nacional Autónoma de México, Ciudad Universitaria, Ciudad de México 04510, MexicoProbiomed S.A. de C.V. Planta Tenancingo, Cruce de Carreteras Acatzingo-Zumpahuacan SN, Tenancingo, Edo., C.P. 52400 de México, MexicoPrograma de Investigación de Producción de Biomoléculas, Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, CP 04510 Ciudad de México, Mexico; Unidad de Bioprocesos, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, CP 04510 Ciudad de México, MexicoPrograma de Investigación de Producción de Biomoléculas, Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, CP 04510 Ciudad de México, Mexico; Corresponding author.Background: The temperature upshift has been widely used as an induction system to produce recombinant proteins (RPs). However, thermoinduction could affect bacterial metabolism, RP production, and RP aggregation. Understanding the structure and functionality of those aggregates, known as inclusion bodies (IBs), is a research area of interest in bioprocesses being scarcely studied under thermoinduction. Here, we describe the effect of the thermoinduction (39°C or 42°C) on the production of the recombinant human granulocyte–macrophage colony-stimulating factor (rHuGM-CSF) using Escherichia coli W3110 under the system λpL/cI857. Results: Results indicated that at 39°C, the production of biomass was almost doubled as well as the acetate accumulation compared to 42°C. Cultures thermoinduced at 42°C improved 1.5-fold the total protein over biomass yield and 1.25-fold the RP over total protein yield. Furthermore, 42°C accelerated the onset of IB formation, changing its architecture. Additionally, IBs formed at 42°C were less soluble and presented higher disorderly structures compared with IBs formed at 39°C, enriched in α-helix and amyloidal-like structures. Conclusions: This study highlights the observation that IBs attain different architecture in response to small changes in environmental conditions, such as the induction temperature, being this helpful information to improve thermoinduced bioprocesses.How to cite: Restrepo-Pineda S, Rosiles-Becerril D, Vargas-Castillo AB, et al. Induction temperature impacts the structure of recombinant HuGM-CSF inclusion bodies in thermoinducible E. coli. Electron J Biotechnol 2022;59. https://doi.org/10.1016/j.ejbt.2022.08.004.http://www.sciencedirect.com/science/article/pii/S0717345822000343AcetateAggregationBiomassBioprocessesEscherichia coliInclusion bodies |
| spellingShingle | Sara Restrepo-Pineda Diego Rosiles-Becerril Angélica B. Vargas-Castillo Luis P. Ávila-Barrientos Axel Luviano Nuria Sánchez-Puig Enrique García-Hernández Nestor O. Pérez Mauricio A. Trujillo-Roldán Norma A. Valdez-Cruz Induction temperature impacts the structure of recombinant HuGM-CSF inclusion bodies in thermoinducible E. coli Electronic Journal of Biotechnology Acetate Aggregation Biomass Bioprocesses Escherichia coli Inclusion bodies |
| title | Induction temperature impacts the structure of recombinant HuGM-CSF inclusion bodies in thermoinducible E. coli |
| title_full | Induction temperature impacts the structure of recombinant HuGM-CSF inclusion bodies in thermoinducible E. coli |
| title_fullStr | Induction temperature impacts the structure of recombinant HuGM-CSF inclusion bodies in thermoinducible E. coli |
| title_full_unstemmed | Induction temperature impacts the structure of recombinant HuGM-CSF inclusion bodies in thermoinducible E. coli |
| title_short | Induction temperature impacts the structure of recombinant HuGM-CSF inclusion bodies in thermoinducible E. coli |
| title_sort | induction temperature impacts the structure of recombinant hugm csf inclusion bodies in thermoinducible e coli |
| topic | Acetate Aggregation Biomass Bioprocesses Escherichia coli Inclusion bodies |
| url | http://www.sciencedirect.com/science/article/pii/S0717345822000343 |
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