Integration of aqueous (micellar) two-phase systems on the proteins separation
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10773/30223 |
Resumo: | A two-step approach combining an aqueous two-phase system (ATPS) and an aqueous micellar two-phase system (AMTPS), both based on the thermo-responsive copolymer Pluronic L-35, is here proposed for the purification of proteins and tested on the sequential separation of three model proteins, cytochrome c, ovalbumin and azocasein. Phase diagrams were established for the ATPS, as well as co-existence curves for the AMTPS. Then, by scanning and choosing the most promising systems, the separation of the three model proteins was performed. The aqueous systems based on Pluronic L-35 and potassium phosphate buffer (pH = 6.6) proved to be the most selective platform to separate the proteins (SAzo/Cyt = 1667; SOva/Cyt = 5.33 e SAzo/Ova = 1676). The consecutive fractionation of these proteins as well as their isolation from the aqueous phases was proposed, envisaging the industrial application of this downstream strategy. The environmental impact of this downstream process was studied, considering the carbon footprint as the final output. The main contribution to the total carbon footprint comes from the ultrafiltration (~ 49%) and the acid precipitation (~ 33%) due to the energy consumption in the centrifugation. The ATPS step contributes to ~ 17% while the AMTPS only accounts for 0.30% of the total carbon footprint. |
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Integration of aqueous (micellar) two-phase systems on the proteins separationAqueous (micellar) two-phase systemsDownstream processThermo-responsive copolymersProteinsCarbon footprintA two-step approach combining an aqueous two-phase system (ATPS) and an aqueous micellar two-phase system (AMTPS), both based on the thermo-responsive copolymer Pluronic L-35, is here proposed for the purification of proteins and tested on the sequential separation of three model proteins, cytochrome c, ovalbumin and azocasein. Phase diagrams were established for the ATPS, as well as co-existence curves for the AMTPS. Then, by scanning and choosing the most promising systems, the separation of the three model proteins was performed. The aqueous systems based on Pluronic L-35 and potassium phosphate buffer (pH = 6.6) proved to be the most selective platform to separate the proteins (SAzo/Cyt = 1667; SOva/Cyt = 5.33 e SAzo/Ova = 1676). The consecutive fractionation of these proteins as well as their isolation from the aqueous phases was proposed, envisaging the industrial application of this downstream strategy. The environmental impact of this downstream process was studied, considering the carbon footprint as the final output. The main contribution to the total carbon footprint comes from the ultrafiltration (~ 49%) and the acid precipitation (~ 33%) due to the energy consumption in the centrifugation. The ATPS step contributes to ~ 17% while the AMTPS only accounts for 0.30% of the total carbon footprint.Springer Nature2021-01-05T15:48:47Z2019-01-30T00:00:00Z2019-01-30info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10773/30223eng2524-417510.1186/s42480-019-0004-xVicente, Filipa A.Santos, João H. P. M.Pereira, Inês M. M.Gonçalves, Cátia V. M.Dias, Ana C. R. V.Coutinho, João A. P.Ventura, Sónia P. M.info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-02-22T11:58:27Zoai:ria.ua.pt:10773/30223Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:02:23.049378Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Integration of aqueous (micellar) two-phase systems on the proteins separation |
title |
Integration of aqueous (micellar) two-phase systems on the proteins separation |
spellingShingle |
Integration of aqueous (micellar) two-phase systems on the proteins separation Vicente, Filipa A. Aqueous (micellar) two-phase systems Downstream process Thermo-responsive copolymers Proteins Carbon footprint |
title_short |
Integration of aqueous (micellar) two-phase systems on the proteins separation |
title_full |
Integration of aqueous (micellar) two-phase systems on the proteins separation |
title_fullStr |
Integration of aqueous (micellar) two-phase systems on the proteins separation |
title_full_unstemmed |
Integration of aqueous (micellar) two-phase systems on the proteins separation |
title_sort |
Integration of aqueous (micellar) two-phase systems on the proteins separation |
author |
Vicente, Filipa A. |
author_facet |
Vicente, Filipa A. Santos, João H. P. M. Pereira, Inês M. M. Gonçalves, Cátia V. M. Dias, Ana C. R. V. Coutinho, João A. P. Ventura, Sónia P. M. |
author_role |
author |
author2 |
Santos, João H. P. M. Pereira, Inês M. M. Gonçalves, Cátia V. M. Dias, Ana C. R. V. Coutinho, João A. P. Ventura, Sónia P. M. |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Vicente, Filipa A. Santos, João H. P. M. Pereira, Inês M. M. Gonçalves, Cátia V. M. Dias, Ana C. R. V. Coutinho, João A. P. Ventura, Sónia P. M. |
dc.subject.por.fl_str_mv |
Aqueous (micellar) two-phase systems Downstream process Thermo-responsive copolymers Proteins Carbon footprint |
topic |
Aqueous (micellar) two-phase systems Downstream process Thermo-responsive copolymers Proteins Carbon footprint |
description |
A two-step approach combining an aqueous two-phase system (ATPS) and an aqueous micellar two-phase system (AMTPS), both based on the thermo-responsive copolymer Pluronic L-35, is here proposed for the purification of proteins and tested on the sequential separation of three model proteins, cytochrome c, ovalbumin and azocasein. Phase diagrams were established for the ATPS, as well as co-existence curves for the AMTPS. Then, by scanning and choosing the most promising systems, the separation of the three model proteins was performed. The aqueous systems based on Pluronic L-35 and potassium phosphate buffer (pH = 6.6) proved to be the most selective platform to separate the proteins (SAzo/Cyt = 1667; SOva/Cyt = 5.33 e SAzo/Ova = 1676). The consecutive fractionation of these proteins as well as their isolation from the aqueous phases was proposed, envisaging the industrial application of this downstream strategy. The environmental impact of this downstream process was studied, considering the carbon footprint as the final output. The main contribution to the total carbon footprint comes from the ultrafiltration (~ 49%) and the acid precipitation (~ 33%) due to the energy consumption in the centrifugation. The ATPS step contributes to ~ 17% while the AMTPS only accounts for 0.30% of the total carbon footprint. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-01-30T00:00:00Z 2019-01-30 2021-01-05T15:48:47Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10773/30223 |
url |
http://hdl.handle.net/10773/30223 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
2524-4175 10.1186/s42480-019-0004-x |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Springer Nature |
publisher.none.fl_str_mv |
Springer Nature |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
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1799137679355412480 |