Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures

Autores
Rojas, Natalia Lorena; Ortiz, Gastón Ezequiel; Chesini, Mariana; Baruque, Diego Jorge; Cavalitto, Sebastian Fernando
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Polygalacturonases (PG; EC 3.2.1.15) catalyze the hydrolysis of pectin and/or pectic acid and are useful for industrial applications such as juice clarification and pectin extraction. Growth and heterologous expression of recombinant S. cerevisiae which expresses an acidic PG from Aspergillus kawachii was studied in batch and fed batch cultures. Kinetics and stoichiometric parameters of the recombinant yeast were determined in batch cultures in a synthetic medium. In these cultures, the total biomass concentration, protein concentration, and enzyme activity achieved were 2.2 g/l, 10 mg/l, and 3 U/ml, respectively, to give a productivity of 0.06 U/ml.h. In fed batch cultures, various strategies for galactose feeding were used: a) after a glucose growth phase, the addition of a single pulse of galactose which gave a productivity of 0.19 U/ml.h.; b) after a glucose growth phase, a double pulse of galactose at the same final concentration, resulting in a productivity of 0.21 U/ml.h.; c) a simultaneous feeding of glucose and galactose, yielding a productivity of 1.32 U/ml.h. Based on these results, the simultaneous feeding of glucose and galactose was by far the most suitable strategy for the production of this enzyme. Moreover, some biochemical characteristics of the recombinant enzyme such as a MW of ~60 kDa, an isoelectric point of 3.7 and its ability to hydrolyze polygalacturonic acid at pH 2.5 were determined.
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: Ortiz, Gastón Ezequiel. 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: Chesini, Mariana. 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: Baruque, Diego Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Cavalitto, Sebastian Fernando. 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
PG1
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:11336/184061

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spelling Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch culturesRojas, Natalia LorenaOrtiz, Gastón EzequielChesini, MarianaBaruque, Diego JorgeCavalitto, Sebastian FernandoPG1https://purl.org/becyt/ford/2.9https://purl.org/becyt/ford/2Polygalacturonases (PG; EC 3.2.1.15) catalyze the hydrolysis of pectin and/or pectic acid and are useful for industrial applications such as juice clarification and pectin extraction. Growth and heterologous expression of recombinant S. cerevisiae which expresses an acidic PG from Aspergillus kawachii was studied in batch and fed batch cultures. Kinetics and stoichiometric parameters of the recombinant yeast were determined in batch cultures in a synthetic medium. In these cultures, the total biomass concentration, protein concentration, and enzyme activity achieved were 2.2 g/l, 10 mg/l, and 3 U/ml, respectively, to give a productivity of 0.06 U/ml.h. In fed batch cultures, various strategies for galactose feeding were used: a) after a glucose growth phase, the addition of a single pulse of galactose which gave a productivity of 0.19 U/ml.h.; b) after a glucose growth phase, a double pulse of galactose at the same final concentration, resulting in a productivity of 0.21 U/ml.h.; c) a simultaneous feeding of glucose and galactose, yielding a productivity of 1.32 U/ml.h. Based on these results, the simultaneous feeding of glucose and galactose was by far the most suitable strategy for the production of this enzyme. Moreover, some biochemical characteristics of the recombinant enzyme such as a MW of ~60 kDa, an isoelectric point of 3.7 and its ability to hydrolyze polygalacturonic acid at pH 2.5 were determined.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; ArgentinaFil: Ortiz, Gastón Ezequiel. 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: Chesini, Mariana. 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: Baruque, Diego Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Cavalitto, Sebastian Fernando. 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; ArgentinaUniversity of Zagreb. Faculty of Food Technology and Biotechnology2011-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/184061Rojas, Natalia Lorena; Ortiz, Gastón Ezequiel; Chesini, Mariana; Baruque, Diego Jorge; Cavalitto, Sebastian Fernando; Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures; University of Zagreb. Faculty of Food Technology and Biotechnology; Food Technology and Biotechnology; 49; 2-2011; 316-3211330-98621334-2606CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.ftb.com.hr/index.php/component/content/article/56-alias/70-optimization-of-the-production-of-polygalacturonase-from-aspergillus-kawachii-cloned-in-saccharomyces-cerevisiae-in-batch-and-fed-batch-culturesinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:24:30Zoai:ri.conicet.gov.ar:11336/184061instacron: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:34982025-10-15 14:24:30.865CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures
title Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures
spellingShingle Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures
Rojas, Natalia Lorena
PG1
title_short Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures
title_full Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures
title_fullStr Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures
title_full_unstemmed Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures
title_sort Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures
dc.creator.none.fl_str_mv Rojas, Natalia Lorena
Ortiz, Gastón Ezequiel
Chesini, Mariana
Baruque, Diego Jorge
Cavalitto, Sebastian Fernando
author Rojas, Natalia Lorena
author_facet Rojas, Natalia Lorena
Ortiz, Gastón Ezequiel
Chesini, Mariana
Baruque, Diego Jorge
Cavalitto, Sebastian Fernando
author_role author
author2 Ortiz, Gastón Ezequiel
Chesini, Mariana
Baruque, Diego Jorge
Cavalitto, Sebastian Fernando
author2_role author
author
author
author
dc.subject.none.fl_str_mv PG1
topic PG1
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.9
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv Polygalacturonases (PG; EC 3.2.1.15) catalyze the hydrolysis of pectin and/or pectic acid and are useful for industrial applications such as juice clarification and pectin extraction. Growth and heterologous expression of recombinant S. cerevisiae which expresses an acidic PG from Aspergillus kawachii was studied in batch and fed batch cultures. Kinetics and stoichiometric parameters of the recombinant yeast were determined in batch cultures in a synthetic medium. In these cultures, the total biomass concentration, protein concentration, and enzyme activity achieved were 2.2 g/l, 10 mg/l, and 3 U/ml, respectively, to give a productivity of 0.06 U/ml.h. In fed batch cultures, various strategies for galactose feeding were used: a) after a glucose growth phase, the addition of a single pulse of galactose which gave a productivity of 0.19 U/ml.h.; b) after a glucose growth phase, a double pulse of galactose at the same final concentration, resulting in a productivity of 0.21 U/ml.h.; c) a simultaneous feeding of glucose and galactose, yielding a productivity of 1.32 U/ml.h. Based on these results, the simultaneous feeding of glucose and galactose was by far the most suitable strategy for the production of this enzyme. Moreover, some biochemical characteristics of the recombinant enzyme such as a MW of ~60 kDa, an isoelectric point of 3.7 and its ability to hydrolyze polygalacturonic acid at pH 2.5 were determined.
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: Ortiz, Gastón Ezequiel. 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: Chesini, Mariana. 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: Baruque, Diego Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Cavalitto, Sebastian Fernando. 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 Polygalacturonases (PG; EC 3.2.1.15) catalyze the hydrolysis of pectin and/or pectic acid and are useful for industrial applications such as juice clarification and pectin extraction. Growth and heterologous expression of recombinant S. cerevisiae which expresses an acidic PG from Aspergillus kawachii was studied in batch and fed batch cultures. Kinetics and stoichiometric parameters of the recombinant yeast were determined in batch cultures in a synthetic medium. In these cultures, the total biomass concentration, protein concentration, and enzyme activity achieved were 2.2 g/l, 10 mg/l, and 3 U/ml, respectively, to give a productivity of 0.06 U/ml.h. In fed batch cultures, various strategies for galactose feeding were used: a) after a glucose growth phase, the addition of a single pulse of galactose which gave a productivity of 0.19 U/ml.h.; b) after a glucose growth phase, a double pulse of galactose at the same final concentration, resulting in a productivity of 0.21 U/ml.h.; c) a simultaneous feeding of glucose and galactose, yielding a productivity of 1.32 U/ml.h. Based on these results, the simultaneous feeding of glucose and galactose was by far the most suitable strategy for the production of this enzyme. Moreover, some biochemical characteristics of the recombinant enzyme such as a MW of ~60 kDa, an isoelectric point of 3.7 and its ability to hydrolyze polygalacturonic acid at pH 2.5 were determined.
publishDate 2011
dc.date.none.fl_str_mv 2011-02
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/11336/184061
Rojas, Natalia Lorena; Ortiz, Gastón Ezequiel; Chesini, Mariana; Baruque, Diego Jorge; Cavalitto, Sebastian Fernando; Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures; University of Zagreb. Faculty of Food Technology and Biotechnology; Food Technology and Biotechnology; 49; 2-2011; 316-321
1330-9862
1334-2606
CONICET Digital
CONICET
url http://hdl.handle.net/11336/184061
identifier_str_mv Rojas, Natalia Lorena; Ortiz, Gastón Ezequiel; Chesini, Mariana; Baruque, Diego Jorge; Cavalitto, Sebastian Fernando; Optimization of the Production of a Polygalacturonase from Aspergillus kawachii cloned in Saccharomyces cerevisiae in batch and fed-batch cultures; University of Zagreb. Faculty of Food Technology and Biotechnology; Food Technology and Biotechnology; 49; 2-2011; 316-321
1330-9862
1334-2606
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.ftb.com.hr/index.php/component/content/article/56-alias/70-optimization-of-the-production-of-polygalacturonase-from-aspergillus-kawachii-cloned-in-saccharomyces-cerevisiae-in-batch-and-fed-batch-cultures
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
dc.publisher.none.fl_str_mv University of Zagreb. Faculty of Food Technology and Biotechnology
publisher.none.fl_str_mv University of Zagreb. Faculty of Food Technology and Biotechnology
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
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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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