Application of surface response analysis to the optimization of penicillin acylase purification in aqueous two-phase systems
Autor(a) principal: | |
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Data de Publicação: | 2002 |
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/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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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) 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 |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799132679655915520 |