Purification and characterization of a polygalacturonase produced by Wickerhamomyces anomalus

Autores
Martos, M. A.; Butiuk, A. P.; 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 49°C 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.
Centro de Investigación y Desarrollo en Fermentaciones Industriales
Materia
Ciencias Exactas
Characterization
Polygalacturonase
Purification
Wickerhamomyces anomalus
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
SEDICI (UNLP)
Institución
Universidad Nacional de La Plata
OAI Identificador
oai:sedici.unlp.edu.ar:10915/85644

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network_name_str SEDICI (UNLP)
spelling Purification and characterization of a polygalacturonase produced by Wickerhamomyces anomalusMartos, M. A.Butiuk, A. P.Rojas, Natalia LorenaHours, Roque AlbertoCiencias ExactasCharacterizationPolygalacturonasePurificationWickerhamomyces anomalusThe 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 49°C 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.Centro de Investigación y Desarrollo en Fermentaciones Industriales2014info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdf587-594http://sedici.unlp.edu.ar/handle/10915/85644enginfo:eu-repo/semantics/altIdentifier/issn/1516-8913info:eu-repo/semantics/altIdentifier/doi/10.1590/S1516-8913201402214info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2025-09-03T10:48:41Zoai:sedici.unlp.edu.ar:10915/85644Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-03 10:48:42.073SEDICI (UNLP) - Universidad Nacional de La Platafalse
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, M. A.
Ciencias Exactas
Characterization
Polygalacturonase
Purification
Wickerhamomyces anomalus
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, M. A.
Butiuk, A. P.
Rojas, Natalia Lorena
Hours, Roque Alberto
author Martos, M. A.
author_facet Martos, M. A.
Butiuk, A. P.
Rojas, Natalia Lorena
Hours, Roque Alberto
author_role author
author2 Butiuk, A. P.
Rojas, Natalia Lorena
Hours, Roque Alberto
author2_role author
author
author
dc.subject.none.fl_str_mv Ciencias Exactas
Characterization
Polygalacturonase
Purification
Wickerhamomyces anomalus
topic Ciencias Exactas
Characterization
Polygalacturonase
Purification
Wickerhamomyces anomalus
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 49°C 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.
Centro de Investigación y Desarrollo en Fermentaciones Industriales
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 49°C 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
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
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://sedici.unlp.edu.ar/handle/10915/85644
url http://sedici.unlp.edu.ar/handle/10915/85644
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/issn/1516-8913
info:eu-repo/semantics/altIdentifier/doi/10.1590/S1516-8913201402214
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
587-594
dc.source.none.fl_str_mv reponame:SEDICI (UNLP)
instname:Universidad Nacional de La Plata
instacron:UNLP
reponame_str SEDICI (UNLP)
collection SEDICI (UNLP)
instname_str Universidad Nacional de La Plata
instacron_str UNLP
institution UNLP
repository.name.fl_str_mv SEDICI (UNLP) - Universidad Nacional de La Plata
repository.mail.fl_str_mv alira@sedici.unlp.edu.ar
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