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dc.contributor.author
Kangwa, Martin
dc.contributor.author
Yelemane, Vikas
dc.contributor.author
Polat, Ayse Nur
dc.contributor.author
Gorrepati, Kanaka Durga Devi
dc.contributor.author
Grasselli, Mariano
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dc.contributor.author
Fernández Lahore, Marcelo
dc.date.available
2018-08-02T20:27:32Z
dc.date.issued
2015-12
dc.identifier.citation
Kangwa, Martin; Yelemane, Vikas; Polat, Ayse Nur; Gorrepati, Kanaka Durga Devi; Grasselli, Mariano; et al.; High-level fed-batch fermentative expression of an engineered Staphylococcal protein A based ligand in E. coli: purification and characterization; Springer Verlag; AMB Express; 5; 1; 12-2015; 1-10
dc.identifier.issn
2191-0855
dc.identifier.uri
http://hdl.handle.net/11336/53975
dc.description.abstract
The major platform for high level recombinant protein production is based on genetically modified microorganisms like Escherichia coli (E. coli) due to its short dividing time, ability to use inexpensive substrates and additionally, its genetics is comparatively simple, well characterized and can be manipulated easily. Here, we investigated the possibilities of finding the best media for high cell density fermentation, by analyzing different media samples, focusing on improving fermentation techniques and recombinant protein production. Initial fermentation of E. coli BL21 DE3:pAV01 in baffled flasks showed that high cell density was achieved when using complex media, Luria–Bertani (LB) and Terrific medium broth (TB) (10 and 14 g/L wet weight, respectively), as compared to mineral media M9, modified minimal medium (MMM) and Riesenberg mineral medium (RM) (7, 8 and 7 g/L, respectively). However, in fed-batch fermentation processes when using MMM after 25 h cultivation, it was possible to yield an optical density (OD600) of 139 corresponding to 172 g/L of wet biomass was produced in a 30 L TV Techfors-S Infors HT fermenter, with a computer controlled nutrient supply (glucose as a carbon source) delivery system, indicating nearly 1.5 times that obtained from TB. Upon purification, a total of 1.65 mg/g of protein per gram cell biomass was obtained and the purified AviPure showed affinity for immunoglobulin. High cell density fed batch fermentation was achieved by selecting the best media and growth conditions, by utilizing a number of fermentation parameters like media, fermentation conditions, chemical concentrations, pO2 level, stirrer speed, pH level and feed media addition. It is possible to reach cell densities higher than shake flasks and stirred tank reactors with the improved oxygen transfer rate and feed.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Springer Verlag
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Avipure
dc.subject
E. Coli
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Fed-Batch Fermentation
dc.subject
High Cell Density
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Recombinant Protein A
dc.subject.classification
Biotecnología Industrial
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dc.subject.classification
Biotecnología Industrial
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dc.subject.classification
INGENIERÍAS Y TECNOLOGÍAS
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dc.title
High-level fed-batch fermentative expression of an engineered Staphylococcal protein A based ligand in E. coli: purification and characterization
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2018-08-02T15:31:05Z
dc.journal.volume
5
dc.journal.number
1
dc.journal.pagination
1-10
dc.journal.pais
Países Bajos
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dc.journal.ciudad
Heidelberg
dc.description.fil
Fil: Kangwa, Martin. Jacobs University; Alemania
dc.description.fil
Fil: Yelemane, Vikas. Jacobs University; Alemania
dc.description.fil
Fil: Polat, Ayse Nur. Jacobs University; Alemania
dc.description.fil
Fil: Gorrepati, Kanaka Durga Devi. Jacobs University; Alemania
dc.description.fil
Fil: Grasselli, Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes; Argentina
dc.description.fil
Fil: Fernández Lahore, Marcelo. Jacobs University; Alemania
dc.journal.title
AMB Express
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/https://dx.doi.org/10.1186/s13568-015-0155-y
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://amb-express.springeropen.com/articles/10.1186/s13568-015-0155-y
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