Application of surface response analysis to the optimization of penicillin acylase purification in aqueous two-phase systems

Detalhes bibliográficos
Autor(a) principal: Marcos, João Carlos
Data de Publicação: 2002
Outros Autores: Fonseca, Luís Pina, Ramalho, Maria Teresa, Cabral, J. M. S.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/1822/2007
Resumo: Penicillin acylase purification from an Escherichia coli crude extract using PEG 3350 – sodium citrate aqueous two phase systems was optimized. An experimental design was used to evaluate the influence of PEG, sodium citrate and sodium chloride on the purification parameters. A central composite design was defined centred on the previously found conditions for highest purification from an osmotic shock extract. Mathematical models for the partition coefficient of protein and enzyme, balance of protein and enzyme, yield and purification were calculated and statistically validated. Analysis of the contours of constant response as a function of PEG and sodium citrate concentrations for three different concentrations of NaCl revealed different effects of the three factors on the studied parameters. A maximum purification factor of 6.5 was predicted for PEG 3350, Sodium Citrate and NaCl concentrations of 15.1%, 11.0% and 8.52% respectively. However under these conditions the predicted yield was 61%. A better compromise between these two parameters can be found by superimposing the contour plots of the purification factor and yield for 10.3% NaCl. A region in the experimental space can be defined where the purification factor is always higher than 5.5 with yields exceeding 80%.
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spelling Application of surface response analysis to the optimization of penicillin acylase purification in aqueous two-phase systemsPenicillin acylasePurificationAqueous two-phase systemsExperimental designaqueous two-phase systems (ATPS)Science & TechnologyPenicillin acylase purification from an Escherichia coli crude extract using PEG 3350 – sodium citrate aqueous two phase systems was optimized. An experimental design was used to evaluate the influence of PEG, sodium citrate and sodium chloride on the purification parameters. A central composite design was defined centred on the previously found conditions for highest purification from an osmotic shock extract. Mathematical models for the partition coefficient of protein and enzyme, balance of protein and enzyme, yield and purification were calculated and statistically validated. Analysis of the contours of constant response as a function of PEG and sodium citrate concentrations for three different concentrations of NaCl revealed different effects of the three factors on the studied parameters. A maximum purification factor of 6.5 was predicted for PEG 3350, Sodium Citrate and NaCl concentrations of 15.1%, 11.0% and 8.52% respectively. However under these conditions the predicted yield was 61%. A better compromise between these two parameters can be found by superimposing the contour plots of the purification factor and yield for 10.3% NaCl. A region in the experimental space can be defined where the purification factor is always higher than 5.5 with yields exceeding 80%.ElsevierUniversidade do MinhoMarcos, João CarlosFonseca, Luís PinaRamalho, Maria TeresaCabral, J. M. S.20022002-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/2007eng"Enzyme and microbial technology". 31 (2002) 1006-1014.0141-022910.1016/S0141-0229(02)00230-2info: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:RCAAP2023-07-21T12:26:48Zoai:repositorium.sdum.uminho.pt:1822/2007Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:21:17.715125Repositório 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 Application of surface response analysis to the optimization of penicillin acylase purification in aqueous two-phase systems
title Application of surface response analysis to the optimization of penicillin acylase purification in aqueous two-phase systems
spellingShingle Application of surface response analysis to the optimization of penicillin acylase purification in aqueous two-phase systems
Marcos, João Carlos
Penicillin acylase
Purification
Aqueous two-phase systems
Experimental design
aqueous two-phase systems (ATPS)
Science & Technology
title_short Application of surface response analysis to the optimization of penicillin acylase purification in aqueous two-phase systems
title_full Application of surface response analysis to the optimization of penicillin acylase purification in aqueous two-phase systems
title_fullStr Application of surface response analysis to the optimization of penicillin acylase purification in aqueous two-phase systems
title_full_unstemmed Application of surface response analysis to the optimization of penicillin acylase purification in aqueous two-phase systems
title_sort Application of surface response analysis to the optimization of penicillin acylase purification in aqueous two-phase systems
author Marcos, João Carlos
author_facet Marcos, João Carlos
Fonseca, Luís Pina
Ramalho, Maria Teresa
Cabral, J. M. S.
author_role author
author2 Fonseca, Luís Pina
Ramalho, Maria Teresa
Cabral, J. M. S.
author2_role author
author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Marcos, João Carlos
Fonseca, Luís Pina
Ramalho, Maria Teresa
Cabral, J. M. S.
dc.subject.por.fl_str_mv Penicillin acylase
Purification
Aqueous two-phase systems
Experimental design
aqueous two-phase systems (ATPS)
Science & Technology
topic Penicillin acylase
Purification
Aqueous two-phase systems
Experimental design
aqueous two-phase systems (ATPS)
Science & Technology
description Penicillin acylase purification from an Escherichia coli crude extract using PEG 3350 – sodium citrate aqueous two phase systems was optimized. An experimental design was used to evaluate the influence of PEG, sodium citrate and sodium chloride on the purification parameters. A central composite design was defined centred on the previously found conditions for highest purification from an osmotic shock extract. Mathematical models for the partition coefficient of protein and enzyme, balance of protein and enzyme, yield and purification were calculated and statistically validated. Analysis of the contours of constant response as a function of PEG and sodium citrate concentrations for three different concentrations of NaCl revealed different effects of the three factors on the studied parameters. A maximum purification factor of 6.5 was predicted for PEG 3350, Sodium Citrate and NaCl concentrations of 15.1%, 11.0% and 8.52% respectively. However under these conditions the predicted yield was 61%. A better compromise between these two parameters can be found by superimposing the contour plots of the purification factor and yield for 10.3% NaCl. A region in the experimental space can be defined where the purification factor is always higher than 5.5 with yields exceeding 80%.
publishDate 2002
dc.date.none.fl_str_mv 2002
2002-01-01T00:00:00Z
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/1822/2007
url http://hdl.handle.net/1822/2007
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv "Enzyme and microbial technology". 31 (2002) 1006-1014.
0141-0229
10.1016/S0141-0229(02)00230-2
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 Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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