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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/29058
Ver los metadatos del registro completo
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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 Técnicas2025-09-03T10:05:26Zoai: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:34982025-09-03 10:05:26.405CONICET 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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1842269910120529920 |
score |
13.13397 |