Strategies for the production of difficult-to-express full-length eukaryotic proteins using microbial cell factories: production of human alpha-galactosidase A
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| 024 | 7 | 0 | |a 10.1007/s00253-014-6328-9 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-014-6328-9 | ||
| 245 | 0 | 0 | |a Strategies for the production of difficult-to-express full-length eukaryotic proteins using microbial cell factories: production of human alpha-galactosidase A |h [Elektronische Daten] |c [Ugutz Unzueta, Felicitas Vázquez, Giulia Accardi, Rosa Mendoza, Verónica Toledo-Rubio, Maria Giuliani, Filomena Sannino, Ermenegilda Parrilli, Ibane Abasolo, Simo Schwartz Jr., Maria Tutino, Antonio Villaverde, José Corchero, Neus Ferrer-Miralles] |
| 520 | 3 | |a Obtaining high levels of pure proteins remains the main bottleneck of many scientific and biotechnological studies. Among all the available recombinant expression systems, Escherichia coli facilitates gene expression by its relative simplicity, inexpensive and fast cultivation, well-known genetics and the large number of tools available for its biotechnological application. However, recombinant expression in E. coli is not always a straightforward procedure and major obstacles are encountered when producing many eukaryotic proteins and especially membrane proteins, linked to missing posttranslational modifications, proteolysis and aggregation. In this context, many conventional and unconventional eukaryotic hosts are under exploration and development, but in some cases linked to complex culture media or processes. In this context, alternative bacterial systems able to overcome some of the limitations posed by E. coli keeping the simplicity of prokaryotic manipulation are currently emerging as convenient hosts for protein production. We have comparatively produced a "difficult-to-express” human protein, the lysosomal enzyme alpha-galactosidase A (hGLA) in E. coli and in the psychrophilic bacterium Pseudoalteromonas haloplanktis TAC125 cells (P. haloplanktis TAC125). While in E. coli the production of active hGLA was unreachable due to proteolytic instability and/or protein misfolding, the expression of hGLA gene in P. haloplanktis TAC125 allows obtaining active enzyme. These results are discussed in the context of emerging bacterial systems for protein production that represent appealing alternatives to the regular use of E. coli and also of more complex eukaryotic systems. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2015 | ||
| 690 | 7 | |a Recombinant protein |2 nationallicence | |
| 690 | 7 | |a Expression systems |2 nationallicence | |
| 690 | 7 | |a Escherichia coli |2 nationallicence | |
| 690 | 7 | |a Pseudoalteromonas haloplanktis TAC125 |2 nationallicence | |
| 690 | 7 | |a Human alpha-galactosidase A |2 nationallicence | |
| 690 | 7 | |a Fabry's disease |2 nationallicence | |
| 700 | 1 | |a Unzueta |D Ugutz |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | |
| 700 | 1 | |a Vázquez |D Felicitas |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | |
| 700 | 1 | |a Accardi |D Giulia |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | |
| 700 | 1 | |a Mendoza |D Rosa |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | |
| 700 | 1 | |a Toledo-Rubio |D Verónica |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | |
| 700 | 1 | |a Giuliani |D Maria |u Dipartimento di Scienze Biochimiche, Facoltà di Medicina e Chirurgia, Università degli studi di Palermo, 90127, Palermo, Italy |4 aut | |
| 700 | 1 | |a Sannino |D Filomena |u Dipartimento di Scienze Biochimiche, Facoltà di Medicina e Chirurgia, Università degli studi di Palermo, 90127, Palermo, Italy |4 aut | |
| 700 | 1 | |a Parrilli |D Ermenegilda |u Dipartimento di Scienze Biochimiche, Facoltà di Medicina e Chirurgia, Università degli studi di Palermo, 90127, Palermo, Italy |4 aut | |
| 700 | 1 | |a Abasolo |D Ibane |u CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Barcelona, Spain |4 aut | |
| 700 | 1 | |a Schwartz Jr. |D Simo |u CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Barcelona, Spain |4 aut | |
| 700 | 1 | |a Tutino |D Maria |u Dipartimento di Scienze Biochimiche, Facoltà di Medicina e Chirurgia, Università degli studi di Palermo, 90127, Palermo, Italy |4 aut | |
| 700 | 1 | |a Villaverde |D Antonio |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | |
| 700 | 1 | |a Corchero |D José |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | |
| 700 | 1 | |a Ferrer-Miralles |D Neus |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/14(2015-07-01), 5863-5874 |x 0175-7598 |q 99:14<5863 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-014-6328-9 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-014-6328-9 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Unzueta |D Ugutz |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Vázquez |D Felicitas |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Accardi |D Giulia |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Mendoza |D Rosa |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Toledo-Rubio |D Verónica |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Giuliani |D Maria |u Dipartimento di Scienze Biochimiche, Facoltà di Medicina e Chirurgia, Università degli studi di Palermo, 90127, Palermo, Italy |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Sannino |D Filomena |u Dipartimento di Scienze Biochimiche, Facoltà di Medicina e Chirurgia, Università degli studi di Palermo, 90127, Palermo, Italy |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Parrilli |D Ermenegilda |u Dipartimento di Scienze Biochimiche, Facoltà di Medicina e Chirurgia, Università degli studi di Palermo, 90127, Palermo, Italy |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Abasolo |D Ibane |u CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Schwartz Jr |D Simo |u CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Tutino |D Maria |u Dipartimento di Scienze Biochimiche, Facoltà di Medicina e Chirurgia, Università degli studi di Palermo, 90127, Palermo, Italy |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Villaverde |D Antonio |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Corchero |D José |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Ferrer-Miralles |D Neus |u Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/14(2015-07-01), 5863-5874 |x 0175-7598 |q 99:14<5863 |1 2015 |2 99 |o 253 | ||