Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalus

Autores
Martos, María Alicia; Butiuk, Ana Paula; Rojas, Natalia Lorena; Hours, Roque Alberto
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The aim of this work was to study the purification and physicochemical properties of an endo-polygalacturonase(PG) produced by Wickerhamomyces anomalus isolated from the citrus fruit peels. The enzyme was purified to homogeneity from the culture filtrate of W. anomalus grown on the yeast nitrogen base medium with glucose as carbon and energy source and citrus pectin as inductor. After anion-exchange chromatography and gel filtration chromatography, PG activity was eluted as a single peak, yielding 21% of the original activity. After dialysis and cation-exchange chromatography, only one fraction with PG activity was obtained, recovering 56% of initial enzyme activity and 1.3-fold increase in specific activity. The molecular weight of the enzyme was estimated as 43 kDa by the SDS-PAGE. The enzyme exhibited maximal activity at pH 4.2 and was stable over a pH range from 3.5 to 6.0 and up to 49ºC for 10 h. The Vmax and Km values with polygalacturonic acid as substrate were 0.26 mmol/L.min and 0.173 mg/mL, respectively. Cations such as Cu+2, Fe+3, Mg+2, Mn+2 and Zn+2 did not show any significant effect on PG activity but K+ and Ca+2 reduced it. The purified PG was able to macerate cassava tissues.
Fil: Martos, María Alicia. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; Argentina
Fil: Butiuk, Ana Paula. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Rojas, Natalia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina
Fil: Hours, Roque Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina
Materia
WICKERHAMOMYCES ANOMALUS
POLYGALACTURONASE
PURIFICATION
CHARACTERIZATION
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/29058
Acceder
id CONICETDig_b7941a99eae2d82de989c7133ce69c90
oai_identifier_str oai:ri.conicet.gov.ar:11336/29058
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalusMartos, María AliciaButiuk, Ana PaulaRojas, Natalia LorenaHours, Roque AlbertoWICKERHAMOMYCES ANOMALUSPOLYGALACTURONASEPURIFICATIONCHARACTERIZATIONhttps://purl.org/becyt/ford/2.9https://purl.org/becyt/ford/2The aim of this work was to study the purification and physicochemical properties of an endo-polygalacturonase(PG) produced by Wickerhamomyces anomalus isolated from the citrus fruit peels. The enzyme was purified to homogeneity from the culture filtrate of W. anomalus grown on the yeast nitrogen base medium with glucose as carbon and energy source and citrus pectin as inductor. After anion-exchange chromatography and gel filtration chromatography, PG activity was eluted as a single peak, yielding 21% of the original activity. After dialysis and cation-exchange chromatography, only one fraction with PG activity was obtained, recovering 56% of initial enzyme activity and 1.3-fold increase in specific activity. The molecular weight of the enzyme was estimated as 43 kDa by the SDS-PAGE. The enzyme exhibited maximal activity at pH 4.2 and was stable over a pH range from 3.5 to 6.0 and up to 49ºC for 10 h. The Vmax and Km values with polygalacturonic acid as substrate were 0.26 mmol/L.min and 0.173 mg/mL, respectively. Cations such as Cu+2, Fe+3, Mg+2, Mn+2 and Zn+2 did not show any significant effect on PG activity but K+ and Ca+2 reduced it. The purified PG was able to macerate cassava tissues.Fil: Martos, María Alicia. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; ArgentinaFil: Butiuk, Ana Paula. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rojas, Natalia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; ArgentinaFil: Hours, Roque Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; ArgentinaInst Tecnologia Parana2014-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/29058Martos, María Alicia; Butiuk, Ana Paula; Rojas, Natalia Lorena; Hours, Roque Alberto; Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalus; Inst Tecnologia Parana; Brazilian Archives Of Biology And Technology; 57; 4; 7-2014; 587-5941516-89131678-4324CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1590/S1516-8913201402214info:eu-repo/semantics/altIdentifier/url/http://ref.scielo.org/9jwt99info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:26:28Zoai:ri.conicet.gov.ar:11336/29058instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:26:29.03CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalus
title Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalus
spellingShingle Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalus
Martos, María Alicia
WICKERHAMOMYCES ANOMALUS
POLYGALACTURONASE
PURIFICATION
CHARACTERIZATION
title_short Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalus
title_full Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalus
title_fullStr Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalus
title_full_unstemmed Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalus
title_sort Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalus
dc.creator.none.fl_str_mv Martos, María Alicia
Butiuk, Ana Paula
Rojas, Natalia Lorena
Hours, Roque Alberto
author Martos, María Alicia
author_facet Martos, María Alicia
Butiuk, Ana Paula
Rojas, Natalia Lorena
Hours, Roque Alberto
author_role author
author2 Butiuk, Ana Paula
Rojas, Natalia Lorena
Hours, Roque Alberto
author2_role author
author
author
dc.subject.none.fl_str_mv WICKERHAMOMYCES ANOMALUS
POLYGALACTURONASE
PURIFICATION
CHARACTERIZATION
topic WICKERHAMOMYCES ANOMALUS
POLYGALACTURONASE
PURIFICATION
CHARACTERIZATION
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.9
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv The aim of this work was to study the purification and physicochemical properties of an endo-polygalacturonase(PG) produced by Wickerhamomyces anomalus isolated from the citrus fruit peels. The enzyme was purified to homogeneity from the culture filtrate of W. anomalus grown on the yeast nitrogen base medium with glucose as carbon and energy source and citrus pectin as inductor. After anion-exchange chromatography and gel filtration chromatography, PG activity was eluted as a single peak, yielding 21% of the original activity. After dialysis and cation-exchange chromatography, only one fraction with PG activity was obtained, recovering 56% of initial enzyme activity and 1.3-fold increase in specific activity. The molecular weight of the enzyme was estimated as 43 kDa by the SDS-PAGE. The enzyme exhibited maximal activity at pH 4.2 and was stable over a pH range from 3.5 to 6.0 and up to 49ºC for 10 h. The Vmax and Km values with polygalacturonic acid as substrate were 0.26 mmol/L.min and 0.173 mg/mL, respectively. Cations such as Cu+2, Fe+3, Mg+2, Mn+2 and Zn+2 did not show any significant effect on PG activity but K+ and Ca+2 reduced it. The purified PG was able to macerate cassava tissues.
Fil: Martos, María Alicia. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; Argentina
Fil: Butiuk, Ana Paula. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Rojas, Natalia Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina
Fil: Hours, Roque Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; Argentina
description The aim of this work was to study the purification and physicochemical properties of an endo-polygalacturonase(PG) produced by Wickerhamomyces anomalus isolated from the citrus fruit peels. The enzyme was purified to homogeneity from the culture filtrate of W. anomalus grown on the yeast nitrogen base medium with glucose as carbon and energy source and citrus pectin as inductor. After anion-exchange chromatography and gel filtration chromatography, PG activity was eluted as a single peak, yielding 21% of the original activity. After dialysis and cation-exchange chromatography, only one fraction with PG activity was obtained, recovering 56% of initial enzyme activity and 1.3-fold increase in specific activity. The molecular weight of the enzyme was estimated as 43 kDa by the SDS-PAGE. The enzyme exhibited maximal activity at pH 4.2 and was stable over a pH range from 3.5 to 6.0 and up to 49ºC for 10 h. The Vmax and Km values with polygalacturonic acid as substrate were 0.26 mmol/L.min and 0.173 mg/mL, respectively. Cations such as Cu+2, Fe+3, Mg+2, Mn+2 and Zn+2 did not show any significant effect on PG activity but K+ and Ca+2 reduced it. The purified PG was able to macerate cassava tissues.
publishDate 2014
dc.date.none.fl_str_mv 2014-07
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/29058
Martos, María Alicia; Butiuk, Ana Paula; Rojas, Natalia Lorena; Hours, Roque Alberto; Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalus; Inst Tecnologia Parana; Brazilian Archives Of Biology And Technology; 57; 4; 7-2014; 587-594
1516-8913
1678-4324
CONICET Digital
CONICET
url http://hdl.handle.net/11336/29058
identifier_str_mv Martos, María Alicia; Butiuk, Ana Paula; Rojas, Natalia Lorena; Hours, Roque Alberto; Purification and Characterization of a Polygalacturonase Produced by Wickerhamomyces anomalus; Inst Tecnologia Parana; Brazilian Archives Of Biology And Technology; 57; 4; 7-2014; 587-594
1516-8913
1678-4324
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1590/S1516-8913201402214
info:eu-repo/semantics/altIdentifier/url/http://ref.scielo.org/9jwt99
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Inst Tecnologia Parana
publisher.none.fl_str_mv Inst Tecnologia Parana
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.069144

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