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Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase

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Resumo:Aqueous two-phase systems (ATPS) based on tetrahydrofuran (THF) + potassium phosphate buffer (pH 7) were used in this work for the purification of lipases. Binodal curve, tie lines and critical point were obtained for the new THF-salt ATPS at 25 degrees C and the binodal curve were successfully correlated with the Merchuck and Hu equations. To optimize the extraction capability of this ATPS the effects of the concentration of components and temperature of equilibrium on the partition coefficients and extraction efficiencies were investigated using lipase from Burkholderia cepacia (commercially obtained) as a model compound. The optimum conditions for the purification of an extracellular lipase obtained by submerged fermentation were established through a surface response analysis by central composite rotational design applied allowing a purification factor (PF) of 103.9 +/- 0.9 and an enzyme recovery of 96.4 +/- 1.1 achieved using this process. Moreover, a commercial lipase by Candida antarctica B recombinant in Aspergillus niger was purified (PF= 4.84 +/- 0.24), confirming the potential of this new THF-based ATPS for purifying lipases. (C) 2015 Elsevier Ltd. All rights reserved.
Autores principais:Souza, Ranyere L.
Outros Autores:Lima, Rafaella A.; Coutinho, Joao A. P.; Soares, Cleide M. F.; Lima, Alvaro S.
Assunto:PORCINE PANCREATIC LIPASE POLYETHYLENE-GLYCOL PHASE-SEPARATION IONIC LIQUIDS INDUSTRIAL APPLICATIONS PROTEIN SEPARATION BIPHASIC SYSTEMS TOLERANT LIPASE PLUS WATER EXTRACTION
Ano:2015
País:Portugal
Tipo de documento:artigo
Tipo de acesso:acesso restrito
Instituição associada:Universidade de Aveiro
Idioma:inglês
Origem:RIA - Repositório Institucional da Universidade de Aveiro
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author Souza, Ranyere L.
author2 Lima, Rafaella A.
Coutinho, Joao A. P.
Soares, Cleide M. F.
Lima, Alvaro S.
author2_role author
author
author
author
author_facet Souza, Ranyere L.
Lima, Rafaella A.
Coutinho, Joao A. P.
Soares, Cleide M. F.
Lima, Alvaro S.
author_role author
country_str PT
creators_json_txt [{\"Person.name\":\"Souza, Ranyere L.\"},{\"Person.name\":\"Lima, Rafaella A.\"},{\"Person.name\":\"Coutinho, Joao A. P.\"},{\"Person.name\":\"Soares, Cleide M. F.\"},{\"Person.name\":\"Lima, Alvaro S.\"}]
datacite.creators.creator.creatorName.fl_str_mv Souza, Ranyere L.
Lima, Rafaella A.
Coutinho, Joao A. P.
Soares, Cleide M. F.
Lima, Alvaro S.
datacite.date.Accepted.fl_str_mv 2015-01-01T00:00:00Z
datacite.date.embargoed.fl_str_mv 10000-01-01T00:00:00Z
datacite.rights.fl_str_mv http://purl.org/coar/access_right/c_16ec
datacite.subjects.subject.fl_str_mv PORCINE PANCREATIC LIPASE
POLYETHYLENE-GLYCOL
PHASE-SEPARATION
IONIC LIQUIDS
INDUSTRIAL APPLICATIONS
PROTEIN SEPARATION
BIPHASIC SYSTEMS
TOLERANT LIPASE
PLUS WATER
EXTRACTION
datacite.titles.title.fl_str_mv Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase
dc.creator.none.fl_str_mv Souza, Ranyere L.
Lima, Rafaella A.
Coutinho, Joao A. P.
Soares, Cleide M. F.
Lima, Alvaro S.
dc.date.Accepted.fl_str_mv 2015-01-01T00:00:00Z
dc.date.embargoed.fl_str_mv 10000-01-01T00:00:00Z
dc.format.none.fl_str_mv application/pdf
dc.identifier.none.fl_str_mv http://hdl.handle.net/10773/20770
dc.language.none.fl_str_mv eng
dc.publisher.none.fl_str_mv ELSEVIER SCI LTD
dc.rights.none.fl_str_mv http://purl.org/coar/access_right/c_16ec
dc.subject.none.fl_str_mv PORCINE PANCREATIC LIPASE
POLYETHYLENE-GLYCOL
PHASE-SEPARATION
IONIC LIQUIDS
INDUSTRIAL APPLICATIONS
PROTEIN SEPARATION
BIPHASIC SYSTEMS
TOLERANT LIPASE
PLUS WATER
EXTRACTION
dc.title.fl_str_mv Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase
dc.type.none.fl_str_mv http://purl.org/coar/resource_type/c_6501
description Aqueous two-phase systems (ATPS) based on tetrahydrofuran (THF) + potassium phosphate buffer (pH 7) were used in this work for the purification of lipases. Binodal curve, tie lines and critical point were obtained for the new THF-salt ATPS at 25 degrees C and the binodal curve were successfully correlated with the Merchuck and Hu equations. To optimize the extraction capability of this ATPS the effects of the concentration of components and temperature of equilibrium on the partition coefficients and extraction efficiencies were investigated using lipase from Burkholderia cepacia (commercially obtained) as a model compound. The optimum conditions for the purification of an extracellular lipase obtained by submerged fermentation were established through a surface response analysis by central composite rotational design applied allowing a purification factor (PF) of 103.9 +/- 0.9 and an enzyme recovery of 96.4 +/- 1.1 achieved using this process. Moreover, a commercial lipase by Candida antarctica B recombinant in Aspergillus niger was purified (PF= 4.84 +/- 0.24), confirming the potential of this new THF-based ATPS for purifying lipases. (C) 2015 Elsevier Ltd. All rights reserved.
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identifier.url.fl_str_mv http://hdl.handle.net/10773/20770
instacron_str ua
institution Universidade de Aveiro
instname_str Universidade de Aveiro
language eng
network_acronym_str ria
network_name_str RIA - Repositório Institucional da Universidade de Aveiro
oai_identifier_str oai:ria.ua.pt:10773/20770
organization_str_mv urn:organizationAcronym:ua
person_str_mv Souza, Ranyere L.
Lima, Rafaella A.
Coutinho, Joao A. P.
Soares, Cleide M. F.
Lima, Alvaro S.
publishDate 2015
publisher.none.fl_str_mv ELSEVIER SCI LTD
reponame_str RIA - Repositório Institucional da Universidade de Aveiro
repository_id_str urn:repositoryAcronym:ria
service_str_mv urn:repositoryAcronym:ria
spelling porAqueous two-phase systems (ATPS) based on tetrahydrofuran (THF) + potassium phosphate buffer (pH 7) were used in this work for the purification of lipases. Binodal curve, tie lines and critical point were obtained for the new THF-salt ATPS at 25 degrees C and the binodal curve were successfully correlated with the Merchuck and Hu equations. To optimize the extraction capability of this ATPS the effects of the concentration of components and temperature of equilibrium on the partition coefficients and extraction efficiencies were investigated using lipase from Burkholderia cepacia (commercially obtained) as a model compound. The optimum conditions for the purification of an extracellular lipase obtained by submerged fermentation were established through a surface response analysis by central composite rotational design applied allowing a purification factor (PF) of 103.9 +/- 0.9 and an enzyme recovery of 96.4 +/- 1.1 achieved using this process. Moreover, a commercial lipase by Candida antarctica B recombinant in Aspergillus niger was purified (PF= 4.84 +/- 0.24), confirming the potential of this new THF-based ATPS for purifying lipases. (C) 2015 Elsevier Ltd. All rights reserved.application/pdfengELSEVIER SCI LTDporNovel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipaseSouza, Ranyere L.Lima, Rafaella A.Coutinho, Joao A. P.Soares, Cleide M. F.Lima, Alvaro S.Handlehttp://hdl.handle.net/10773/20770ISSNIsPartOf1359-5113DOIIsPartOf10.1016/j.procbio.2015.05.01510000-01-01T00:00:00Z2015-01-01T00:00:00Z2015http://purl.org/coar/access_right/c_16ecrestricted accessporPORCINE PANCREATIC LIPASEporPOLYETHYLENE-GLYCOLporPHASE-SEPARATIONporIONIC LIQUIDSporINDUSTRIAL APPLICATIONSporPROTEIN SEPARATIONporBIPHASIC SYSTEMSporTOLERANT LIPASEporPLUS WATERporEXTRACTION1401526 byteshttp://purl.org/coar/access_right/c_16ecapplication/pdffulltexthttps://ria.ua.pt/bitstream/10773/20770/2/Novel%20aqueous%20two-phase%20systems%20based%20on%20tetrahydrofuran%20and%20potassium%20phosphate%20buffer%20for%20purification%20of%20lipase_10.1016j.procbio.2015.05.015.pdfliteraturehttp://purl.org/coar/resource_type/c_6501journal article
spellingShingle Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase
Souza, Ranyere L.
PORCINE PANCREATIC LIPASE
POLYETHYLENE-GLYCOL
PHASE-SEPARATION
IONIC LIQUIDS
INDUSTRIAL APPLICATIONS
PROTEIN SEPARATION
BIPHASIC SYSTEMS
TOLERANT LIPASE
PLUS WATER
EXTRACTION
status SINGLETON
subject.fl_str_mv PORCINE PANCREATIC LIPASE
POLYETHYLENE-GLYCOL
PHASE-SEPARATION
IONIC LIQUIDS
INDUSTRIAL APPLICATIONS
PROTEIN SEPARATION
BIPHASIC SYSTEMS
TOLERANT LIPASE
PLUS WATER
EXTRACTION
title Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase
title_full Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase
title_fullStr Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase
title_full_unstemmed Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase
title_short Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase
title_sort Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase
topic PORCINE PANCREATIC LIPASE
POLYETHYLENE-GLYCOL
PHASE-SEPARATION
IONIC LIQUIDS
INDUSTRIAL APPLICATIONS
PROTEIN SEPARATION
BIPHASIC SYSTEMS
TOLERANT LIPASE
PLUS WATER
EXTRACTION
topic_facet PORCINE PANCREATIC LIPASE
POLYETHYLENE-GLYCOL
PHASE-SEPARATION
IONIC LIQUIDS
INDUSTRIAL APPLICATIONS
PROTEIN SEPARATION
BIPHASIC SYSTEMS
TOLERANT LIPASE
PLUS WATER
EXTRACTION
url http://hdl.handle.net/10773/20770
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