Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system

Detalhes bibliográficos
Autor(a) principal: Maciel, Marília de Holanda Cavalcanti
Data de Publicação: 2014
Outros Autores: Ottoni, C. A., Herculano, Polyanna N., Porto, Tatiana Souza, Porto, Ana L. F., Santos, Cledir, Lima, Nelson, Moreira, Keila, Motta, Cristina Maria de Souza
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/1822/31601
Resumo: The partitioning and purification of polygalacturonases (PG) produced by Aspergillus niger URM 5162 were investigated in aqueous two-phase systems (ATPS), formed by polyethylene glycol and phosphate salts (PEG/phosphate). To evaluate the effect of the 4 independent variables – molar mass of polyethylene glycol (PEG) (400–8000 g/mol – MPEG), PEG concentration (12.5–17.5%, w/w – CPEG), phosphate concentration (15–25%, w/w, CPHOS) and pH (6.0–8.0) – on the 4 response variables: partition coefficient (K), activity yield (Y), purification factor (PF) and selectivity (S), a factorial design (24) was used. The endo-polygalacturonases (endo-PG) and exo-polygalacturonase (exo-PG) were preferentially partitioned in the top phase. For endo- and exo-PG, the highest values for the response variables K (1.23 and 2.40), Y (74.04% and 17.97%), PF (8.18 and 1.98) and S (24.68 and 48.07), respectively, were obtained for a CPEG of 12.5% (w/w), MPEG of 8000 g/mol, and CPHOS of 25% (w/w) at pH 6.0. These conditions were considered the most suitable for the purification of PG produced by A. niger URM 5162. Furthermore, the most important independent variables for endo- and exo-PG were CPHOS and MPEG, respectively. All independent variables studied and their interactions significantly influenced the response variables. According to these results, the PEG/phosphate system is a useful cost-effective alternative for purification of PG produced by A. niger URM 5162.
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spelling Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase systemAspergillus nigerPartitionPEG/phosphate ATPSPolygalacturonasesPurification factorScience & TechnologyThe partitioning and purification of polygalacturonases (PG) produced by Aspergillus niger URM 5162 were investigated in aqueous two-phase systems (ATPS), formed by polyethylene glycol and phosphate salts (PEG/phosphate). To evaluate the effect of the 4 independent variables – molar mass of polyethylene glycol (PEG) (400–8000 g/mol – MPEG), PEG concentration (12.5–17.5%, w/w – CPEG), phosphate concentration (15–25%, w/w, CPHOS) and pH (6.0–8.0) – on the 4 response variables: partition coefficient (K), activity yield (Y), purification factor (PF) and selectivity (S), a factorial design (24) was used. The endo-polygalacturonases (endo-PG) and exo-polygalacturonase (exo-PG) were preferentially partitioned in the top phase. For endo- and exo-PG, the highest values for the response variables K (1.23 and 2.40), Y (74.04% and 17.97%), PF (8.18 and 1.98) and S (24.68 and 48.07), respectively, were obtained for a CPEG of 12.5% (w/w), MPEG of 8000 g/mol, and CPHOS of 25% (w/w) at pH 6.0. These conditions were considered the most suitable for the purification of PG produced by A. niger URM 5162. Furthermore, the most important independent variables for endo- and exo-PG were CPHOS and MPEG, respectively. All independent variables studied and their interactions significantly influenced the response variables. According to these results, the PEG/phosphate system is a useful cost-effective alternative for purification of PG produced by A. niger URM 5162.M.H.C. Maciel thanks to Conselho Nacional de Desenvolvimento Cientlfico e Tecnologico (CNPq, Brazil), Fundacao de Amparo a Ciencia e Tecnologia do Estado de Pernambuco (FACEPE/APQ-0290-2.12/10), Financiadora de Estudos e Projetos (FINEP/ref. 2083/07) and Erasmus Mundus External Cooperation Window Lote 17. The authors thank the Laboratorio de Tecnologia de Bioativos (LABTECBIO), Federal Rural University of Pernambuco (Recife, Brazil). M.H.C. Maciel, C.A. Ottoni, C. Santos and N. Lima also thank the FCT Strategic Project PEst-OE/EQB/LA0023/2013 and the Project "BioEnv - Biotechnology and Bioengineering for a sustainable world", REF. NORTE-07-0124-FEDER-000048." Co-funded by the Programa Operacional Regional do Norte (ON.2-O Novo Norte), QREN, FEDER.Elsevier B.V.Universidade do MinhoMaciel, Marília de Holanda CavalcantiOttoni, C. A.Herculano, Polyanna N.Porto, Tatiana SouzaPorto, Ana L. F.Santos, CledirLima, NelsonMoreira, KeilaMotta, Cristina Maria de Souza20142014-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/31601engMaciel, M. H. C.; Ottoni, C. A.; Herculano, Polyanna N.; Porto, Tatiana S.; Porto, Ana L. F.; Santos, C.; Lima, Nelson; Moreira, Keila A.; Souza-Motta, Cristina, Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system. Fluid Phase Equilibria, 371, 125-130, 20140378-38120378-381210.1016/j.fluid.2014.03.018info: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:RCAAP2023-07-21T12:00:43Zoai:repositorium.sdum.uminho.pt:1822/31601Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:50:34.931553Repositório 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 Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system
title Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system
spellingShingle Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system
Maciel, Marília de Holanda Cavalcanti
Aspergillus niger
Partition
PEG/phosphate ATPS
Polygalacturonases
Purification factor
Science & Technology
title_short Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system
title_full Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system
title_fullStr Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system
title_full_unstemmed Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system
title_sort Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system
author Maciel, Marília de Holanda Cavalcanti
author_facet Maciel, Marília de Holanda Cavalcanti
Ottoni, C. A.
Herculano, Polyanna N.
Porto, Tatiana Souza
Porto, Ana L. F.
Santos, Cledir
Lima, Nelson
Moreira, Keila
Motta, Cristina Maria de Souza
author_role author
author2 Ottoni, C. A.
Herculano, Polyanna N.
Porto, Tatiana Souza
Porto, Ana L. F.
Santos, Cledir
Lima, Nelson
Moreira, Keila
Motta, Cristina Maria de Souza
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Maciel, Marília de Holanda Cavalcanti
Ottoni, C. A.
Herculano, Polyanna N.
Porto, Tatiana Souza
Porto, Ana L. F.
Santos, Cledir
Lima, Nelson
Moreira, Keila
Motta, Cristina Maria de Souza
dc.subject.por.fl_str_mv Aspergillus niger
Partition
PEG/phosphate ATPS
Polygalacturonases
Purification factor
Science & Technology
topic Aspergillus niger
Partition
PEG/phosphate ATPS
Polygalacturonases
Purification factor
Science & Technology
description The partitioning and purification of polygalacturonases (PG) produced by Aspergillus niger URM 5162 were investigated in aqueous two-phase systems (ATPS), formed by polyethylene glycol and phosphate salts (PEG/phosphate). To evaluate the effect of the 4 independent variables – molar mass of polyethylene glycol (PEG) (400–8000 g/mol – MPEG), PEG concentration (12.5–17.5%, w/w – CPEG), phosphate concentration (15–25%, w/w, CPHOS) and pH (6.0–8.0) – on the 4 response variables: partition coefficient (K), activity yield (Y), purification factor (PF) and selectivity (S), a factorial design (24) was used. The endo-polygalacturonases (endo-PG) and exo-polygalacturonase (exo-PG) were preferentially partitioned in the top phase. For endo- and exo-PG, the highest values for the response variables K (1.23 and 2.40), Y (74.04% and 17.97%), PF (8.18 and 1.98) and S (24.68 and 48.07), respectively, were obtained for a CPEG of 12.5% (w/w), MPEG of 8000 g/mol, and CPHOS of 25% (w/w) at pH 6.0. These conditions were considered the most suitable for the purification of PG produced by A. niger URM 5162. Furthermore, the most important independent variables for endo- and exo-PG were CPHOS and MPEG, respectively. All independent variables studied and their interactions significantly influenced the response variables. According to these results, the PEG/phosphate system is a useful cost-effective alternative for purification of PG produced by A. niger URM 5162.
publishDate 2014
dc.date.none.fl_str_mv 2014
2014-01-01T00:00:00Z
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/1822/31601
url http://hdl.handle.net/1822/31601
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Maciel, M. H. C.; Ottoni, C. A.; Herculano, Polyanna N.; Porto, Tatiana S.; Porto, Ana L. F.; Santos, C.; Lima, Nelson; Moreira, Keila A.; Souza-Motta, Cristina, Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system. Fluid Phase Equilibria, 371, 125-130, 2014
0378-3812
0378-3812
10.1016/j.fluid.2014.03.018
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 B.V.
publisher.none.fl_str_mv Elsevier B.V.
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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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
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