Liquid-liquid extraction of protease from cold-adapted yeast Rhodotorula mucilaginosa L7 using biocompatible and biodegradable aqueous two-phase systems
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1080/01496395.2015.1080276 http://hdl.handle.net/11449/172479 |
Resumo: | This work aimed to optimize the extraction of an extracellular protease produced by the cold-adapted yeast Rhodotorula mucilaginosa L7 using aqueous two-phase systems (ATPS) comprising polyethylene glycol (PEG) and sodium citrate or sodium tartrate. First, the biocompatibility of the phase forming agents was assessed. The results obtained with PEG-2000, PEG-4000, and PEG-6000 demonstrated that even at large PEG concentrations (32 wt%) the protease maintains its activity after 3 h of reaction, whereas an increase in salt concentration provokes a gradual decrease in protease stability. Subsequently, the partitioning of the protease in both types of ATPS was assessed, evaluating the effect of temperature, molecular weight, and concentration of PEG on protease purification, using two 23-full factorial designs. The best partitioning conditions were obtained in PEG-6000/sodium tartrate-based ATPS, at 30ºC (with a yield of 81.09 ± 0.66% and a purification factor of 2.51 ± 0.03). Thus, considering the biodegradable characteristics of the system, the PEG/sodium tartrate ATPS is a viable and economic low-resolution step in protease purification, with a strong potential for future industrial application. |
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Liquid-liquid extraction of protease from cold-adapted yeast Rhodotorula mucilaginosa L7 using biocompatible and biodegradable aqueous two-phase systemsaqueous two phase systemscold-adapted microorganismsliquid-liquid extractionProteasepurificationThis work aimed to optimize the extraction of an extracellular protease produced by the cold-adapted yeast Rhodotorula mucilaginosa L7 using aqueous two-phase systems (ATPS) comprising polyethylene glycol (PEG) and sodium citrate or sodium tartrate. First, the biocompatibility of the phase forming agents was assessed. The results obtained with PEG-2000, PEG-4000, and PEG-6000 demonstrated that even at large PEG concentrations (32 wt%) the protease maintains its activity after 3 h of reaction, whereas an increase in salt concentration provokes a gradual decrease in protease stability. Subsequently, the partitioning of the protease in both types of ATPS was assessed, evaluating the effect of temperature, molecular weight, and concentration of PEG on protease purification, using two 23-full factorial designs. The best partitioning conditions were obtained in PEG-6000/sodium tartrate-based ATPS, at 30ºC (with a yield of 81.09 ± 0.66% and a purification factor of 2.51 ± 0.03). Thus, considering the biodegradable characteristics of the system, the PEG/sodium tartrate ATPS is a viable and economic low-resolution step in protease purification, with a strong potential for future industrial application.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Department of Biochemical and Pharmaceutical Technology University of São PauloDepartamento de Química-Física Facultad de Ciencias Bioquímicas y Farmacéuticas Universidad Nacional de RosarioDepartment of Bioprocess and Biotechnology School of Pharmaceutical Sciences UNESP – Univ Estadual PaulistaDepartment of Biochemistry and Microbiology Institute of Biosciences University of São Paulo (UNESP)Department of Bioprocess and Biotechnology School of Pharmaceutical Sciences UNESP – Univ Estadual PaulistaDepartment of Biochemistry and Microbiology Institute of Biosciences University of São Paulo (UNESP)FAPESP: 2011/20521-0FAPESP: 2012/23726-4FAPESP: 2013/19486-0Universidade de São Paulo (USP)Universidad Nacional de RosarioUniversidade Estadual Paulista (Unesp)Lario, Luciana DanielaMalpiedi, Luciana PellegriniPereira, Jorge Fernando Brandão [UNESP]Sette, Lara Durães [UNESP]Pessoa-Junior, Adalberto2018-12-11T17:00:34Z2018-12-11T17:00:34Z2016-01-02info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article57-67application/pdfhttp://dx.doi.org/10.1080/01496395.2015.1080276Separation Science and Technology (Philadelphia), v. 51, n. 1, p. 57-67, 2016.1520-57540149-6395http://hdl.handle.net/11449/17247910.1080/01496395.2015.10802762-s2.0-849566212182-s2.0-84956621218.pdf5969653098289575Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengSeparation Science and Technology (Philadelphia)0,3720,372info:eu-repo/semantics/openAccess2023-10-08T06:03:38Zoai:repositorio.unesp.br:11449/172479Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-10-08T06:03:38Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Liquid-liquid extraction of protease from cold-adapted yeast Rhodotorula mucilaginosa L7 using biocompatible and biodegradable aqueous two-phase systems |
title |
Liquid-liquid extraction of protease from cold-adapted yeast Rhodotorula mucilaginosa L7 using biocompatible and biodegradable aqueous two-phase systems |
spellingShingle |
Liquid-liquid extraction of protease from cold-adapted yeast Rhodotorula mucilaginosa L7 using biocompatible and biodegradable aqueous two-phase systems Lario, Luciana Daniela aqueous two phase systems cold-adapted microorganisms liquid-liquid extraction Protease purification |
title_short |
Liquid-liquid extraction of protease from cold-adapted yeast Rhodotorula mucilaginosa L7 using biocompatible and biodegradable aqueous two-phase systems |
title_full |
Liquid-liquid extraction of protease from cold-adapted yeast Rhodotorula mucilaginosa L7 using biocompatible and biodegradable aqueous two-phase systems |
title_fullStr |
Liquid-liquid extraction of protease from cold-adapted yeast Rhodotorula mucilaginosa L7 using biocompatible and biodegradable aqueous two-phase systems |
title_full_unstemmed |
Liquid-liquid extraction of protease from cold-adapted yeast Rhodotorula mucilaginosa L7 using biocompatible and biodegradable aqueous two-phase systems |
title_sort |
Liquid-liquid extraction of protease from cold-adapted yeast Rhodotorula mucilaginosa L7 using biocompatible and biodegradable aqueous two-phase systems |
author |
Lario, Luciana Daniela |
author_facet |
Lario, Luciana Daniela Malpiedi, Luciana Pellegrini Pereira, Jorge Fernando Brandão [UNESP] Sette, Lara Durães [UNESP] Pessoa-Junior, Adalberto |
author_role |
author |
author2 |
Malpiedi, Luciana Pellegrini Pereira, Jorge Fernando Brandão [UNESP] Sette, Lara Durães [UNESP] Pessoa-Junior, Adalberto |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Universidade de São Paulo (USP) Universidad Nacional de Rosario Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Lario, Luciana Daniela Malpiedi, Luciana Pellegrini Pereira, Jorge Fernando Brandão [UNESP] Sette, Lara Durães [UNESP] Pessoa-Junior, Adalberto |
dc.subject.por.fl_str_mv |
aqueous two phase systems cold-adapted microorganisms liquid-liquid extraction Protease purification |
topic |
aqueous two phase systems cold-adapted microorganisms liquid-liquid extraction Protease purification |
description |
This work aimed to optimize the extraction of an extracellular protease produced by the cold-adapted yeast Rhodotorula mucilaginosa L7 using aqueous two-phase systems (ATPS) comprising polyethylene glycol (PEG) and sodium citrate or sodium tartrate. First, the biocompatibility of the phase forming agents was assessed. The results obtained with PEG-2000, PEG-4000, and PEG-6000 demonstrated that even at large PEG concentrations (32 wt%) the protease maintains its activity after 3 h of reaction, whereas an increase in salt concentration provokes a gradual decrease in protease stability. Subsequently, the partitioning of the protease in both types of ATPS was assessed, evaluating the effect of temperature, molecular weight, and concentration of PEG on protease purification, using two 23-full factorial designs. The best partitioning conditions were obtained in PEG-6000/sodium tartrate-based ATPS, at 30ºC (with a yield of 81.09 ± 0.66% and a purification factor of 2.51 ± 0.03). Thus, considering the biodegradable characteristics of the system, the PEG/sodium tartrate ATPS is a viable and economic low-resolution step in protease purification, with a strong potential for future industrial application. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-01-02 2018-12-11T17:00:34Z 2018-12-11T17:00:34Z |
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://dx.doi.org/10.1080/01496395.2015.1080276 Separation Science and Technology (Philadelphia), v. 51, n. 1, p. 57-67, 2016. 1520-5754 0149-6395 http://hdl.handle.net/11449/172479 10.1080/01496395.2015.1080276 2-s2.0-84956621218 2-s2.0-84956621218.pdf 5969653098289575 |
url |
http://dx.doi.org/10.1080/01496395.2015.1080276 http://hdl.handle.net/11449/172479 |
identifier_str_mv |
Separation Science and Technology (Philadelphia), v. 51, n. 1, p. 57-67, 2016. 1520-5754 0149-6395 10.1080/01496395.2015.1080276 2-s2.0-84956621218 2-s2.0-84956621218.pdf 5969653098289575 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Separation Science and Technology (Philadelphia) 0,372 0,372 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
57-67 application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1799964478961352704 |