Empleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosuero

ABSTRACT: The whey is the largest waste subproduct of the dairy industry. Due to its high values of BOD is considered a source of water and soil contamination when discarded without any treatment. An alternative for the disposal of this agroindustrial waste is its application as a culture medium for...

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Autores:
Beltrán Flórez, Laura Julliet
Acosta Cárdenas, Alejandro
Tipo de recurso:
Article of investigation
Fecha de publicación:
2012
Institución:
Universidad de Antioquia
Repositorio:
Repositorio UdeA
Idioma:
spa
OAI Identifier:
oai:bibliotecadigital.udea.edu.co:10495/10193
Acceso en línea:
http://hdl.handle.net/10495/10193
Palabra clave:
Beta-Galactosidasa
Galactosidasa
Hidrólisis enzimática
Kluyveromyces
Lactosuero
Suero de leche
Enzymatic hydrolysis
Whey
Rights
openAccess
License
Atribución-NoComercial-CompartirIgual 2.5 Colombia (CC BY-NC-SA 2.5 CO)
id UDEA2_6f9aef943a9341a5fcfb6cb9ce61f7f3
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network_acronym_str UDEA2
network_name_str Repositorio UdeA
repository_id_str
dc.title.spa.fl_str_mv Empleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosuero
dc.title.alternative.spa.fl_str_mv Usage of a commercial β-galactosidase from Kluyveromyces lactis in the hydrolysis of whey
title Empleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosuero
spellingShingle Empleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosuero
Beta-Galactosidasa
Galactosidasa
Hidrólisis enzimática
Kluyveromyces
Lactosuero
Suero de leche
Enzymatic hydrolysis
Whey
title_short Empleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosuero
title_full Empleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosuero
title_fullStr Empleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosuero
title_full_unstemmed Empleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosuero
title_sort Empleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosuero
dc.creator.fl_str_mv Beltrán Flórez, Laura Julliet
Acosta Cárdenas, Alejandro
dc.contributor.author.none.fl_str_mv Beltrán Flórez, Laura Julliet
Acosta Cárdenas, Alejandro
dc.subject.none.fl_str_mv Beta-Galactosidasa
Galactosidasa
Hidrólisis enzimática
Kluyveromyces
Lactosuero
Suero de leche
Enzymatic hydrolysis
Whey
topic Beta-Galactosidasa
Galactosidasa
Hidrólisis enzimática
Kluyveromyces
Lactosuero
Suero de leche
Enzymatic hydrolysis
Whey
description ABSTRACT: The whey is the largest waste subproduct of the dairy industry. Due to its high values of BOD is considered a source of water and soil contamination when discarded without any treatment. An alternative for the disposal of this agroindustrial waste is its application as a culture medium for various fermentation processes after an enzymatic hydrolysis thus ensuring a greater availability of fermentable sugars for subsequent application in biotechnology, environment and / or food. Objective To evaluate the effects of pH, temperature and enzyme concentration on whey using a commercial β-galactosidase from Kluyveromyces lactis, and determine the kinetic parameters of the enzyme. Materials and methods The hydrolysis of lactose was performed in buffered solutions of lactose and in whey as well, 5 levels were evaluated for pH (4.5-8.5), temperature (25°C-65°C) and enzyme concentration (0.025 g/L to 0.5 g/L), all the experiments were performed in triplicate in Erlenmeyer flask (100 mL) and analyzed using the GraphPad Prism software. Results We found that the best conditions of pH and temperature for the whey hydrolysis were at 6,5°C and 45°C respectively. According to the kinetic results, the enzyme presented competitive inhibition for galactose with a Km equal to 8.01x10-2 M, Ki of 6.85x10-2 M and Vmax of 1.86x10-2 molmg-1min-1. Conclusions The best conditions of pH and temperature were 6.5°C and 45°C respectively, a 100% of hydrolysis was achieved in one hour with 0.1 g/L of enzyme. Furthermore, it was shown that the reaction rate of the enzyme is inhibited by galactose.
publishDate 2012
dc.date.issued.none.fl_str_mv 2012
dc.date.accessioned.none.fl_str_mv 2018-10-04T21:49:57Z
dc.date.available.none.fl_str_mv 2018-10-04T21:49:57Z
dc.type.spa.fl_str_mv info:eu-repo/semantics/article
dc.type.coarversion.fl_str_mv http://purl.org/coar/version/c_970fb48d4fbd8a85
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dc.type.local.spa.fl_str_mv Artículo de investigación
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dc.identifier.citation.spa.fl_str_mv Beltrán Flórez, LJ, Acosta Cárdenas, A. . Empleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosuero. Hechos Microbiol. 2012; 3(2);25-35.
dc.identifier.issn.none.fl_str_mv 2145-8898
dc.identifier.uri.none.fl_str_mv http://hdl.handle.net/10495/10193
dc.identifier.eissn.none.fl_str_mv 2805-6485
identifier_str_mv Beltrán Flórez, LJ, Acosta Cárdenas, A. . Empleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosuero. Hechos Microbiol. 2012; 3(2);25-35.
2145-8898
2805-6485
url http://hdl.handle.net/10495/10193
dc.language.iso.spa.fl_str_mv spa
language spa
dc.relation.ispartofjournalabbrev.spa.fl_str_mv Hechos Microbiol.
dc.rights.*.fl_str_mv Atribución-NoComercial-CompartirIgual 2.5 Colombia (CC BY-NC-SA 2.5 CO)
dc.rights.spa.fl_str_mv info:eu-repo/semantics/openAccess
dc.rights.uri.*.fl_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/co/
dc.rights.accessrights.spa.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.rights.creativecommons.spa.fl_str_mv https://creativecommons.org/licenses/by-nc-sa/4.0/
rights_invalid_str_mv Atribución-NoComercial-CompartirIgual 2.5 Colombia (CC BY-NC-SA 2.5 CO)
https://creativecommons.org/licenses/by-nc-sa/2.5/co/
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eu_rights_str_mv openAccess
dc.format.mimetype.spa.fl_str_mv application/pdf
dc.publisher.spa.fl_str_mv Universidad de Antioquia, Escuela de Microbiología
dc.publisher.group.spa.fl_str_mv Grupo de Biotransformación
dc.publisher.place.spa.fl_str_mv Medellín, Colombia
institution Universidad de Antioquia
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spelling Beltrán Flórez, Laura JullietAcosta Cárdenas, Alejandro2018-10-04T21:49:57Z2018-10-04T21:49:57Z2012Beltrán Flórez, LJ, Acosta Cárdenas, A. . Empleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosuero. Hechos Microbiol. 2012; 3(2);25-35.2145-8898http://hdl.handle.net/10495/101932805-6485ABSTRACT: The whey is the largest waste subproduct of the dairy industry. Due to its high values of BOD is considered a source of water and soil contamination when discarded without any treatment. An alternative for the disposal of this agroindustrial waste is its application as a culture medium for various fermentation processes after an enzymatic hydrolysis thus ensuring a greater availability of fermentable sugars for subsequent application in biotechnology, environment and / or food. Objective To evaluate the effects of pH, temperature and enzyme concentration on whey using a commercial β-galactosidase from Kluyveromyces lactis, and determine the kinetic parameters of the enzyme. Materials and methods The hydrolysis of lactose was performed in buffered solutions of lactose and in whey as well, 5 levels were evaluated for pH (4.5-8.5), temperature (25°C-65°C) and enzyme concentration (0.025 g/L to 0.5 g/L), all the experiments were performed in triplicate in Erlenmeyer flask (100 mL) and analyzed using the GraphPad Prism software. Results We found that the best conditions of pH and temperature for the whey hydrolysis were at 6,5°C and 45°C respectively. According to the kinetic results, the enzyme presented competitive inhibition for galactose with a Km equal to 8.01x10-2 M, Ki of 6.85x10-2 M and Vmax of 1.86x10-2 molmg-1min-1. Conclusions The best conditions of pH and temperature were 6.5°C and 45°C respectively, a 100% of hydrolysis was achieved in one hour with 0.1 g/L of enzyme. Furthermore, it was shown that the reaction rate of the enzyme is inhibited by galactose.RESUMEN: El lactosuero es el mayor subproducto de desecho de la industria láctea. Por sus elevados valores de DBO se considera un foco de contaminación de aguas y suelo cuando se descarta sin ningún tratamiento. Una alternativa para la disposición final de este residuo agroindustrial, es su utilización como medio de cultivo para varios procesos fermentativos, después de una hidrólisis enzimática, de esta manera garantizar una mayor disponibilidad de azucares fermentables, para su posterior aplicación en biotecnología, medio ambiente y/o alimentos. Objetivo Evaluar los efectos del pH, temperatura y concentración de la enzima sobre lactosuero empleando una β-galactosidasa comercial de Kluyveromyces lactis y determinar los parámetros cinéticos de la enzima. Materiales y métodos La hidrólisis de la lactosa se desarrolló tanto en soluciones tamponadas de lactosa como en lactosuero, se evaluaron 5 niveles para el pH (4,5 - 8,5), temperatura (25°C-65°C) y concentración de enzima (0,025 g/L - 0,5 g/L), todos los experimentos se realizaron por triplicado en matraz de 100 mL y los datos fueron analizados empleando el software GraphPad Prism. Resultados A un pH de 6,5 y una temperatura de 45°C, se alcanzaron las mejores condiciones de hidrolisis del lactosuero. De acuerdo a los resultados cinéticos, la enzima presentó inhibición competitiva por galactosa con un Km igual a 8,01x10-2M, Ki de 6,85x10-2M y Vmax de 1,86x10-2molmg-1min-1. Conclusión Las mejores condiciones de temperatura y pH fueron de 6,5°C y 45°C respectivamente, se alcanzó un 100% de hidrólisis en una hora con 0,1 g/L de enzima. Además, se evidenció que la velocidad de reacción de la enzima fue inhibida por galactosa.COL0066991application/pdfspaUniversidad de Antioquia, Escuela de MicrobiologíaGrupo de BiotransformaciónMedellín, Colombiainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_2df8fbb1https://purl.org/redcol/resource_type/ARTArtículo de investigaciónhttp://purl.org/coar/version/c_970fb48d4fbd8a85Atribución-NoComercial-CompartirIgual 2.5 Colombia (CC BY-NC-SA 2.5 CO)info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/co/http://purl.org/coar/access_right/c_abf2https://creativecommons.org/licenses/by-nc-sa/4.0/Beta-GalactosidasaGalactosidasaHidrólisis enzimáticaKluyveromycesLactosueroSuero de lecheEnzymatic hydrolysisWheyEmpleo de una ß-galactosidasa comercial de Kluyveromyces lactis en la hidrólisis de lactosueroUsage of a commercial β-galactosidase from Kluyveromyces lactis in the hydrolysis of wheyHechos Microbiol.Hechos Microbiológicos253532ORIGINALBeltranLaura_2012_GalactosidasacomercialHidrolisis.pdfBeltranLaura_2012_GalactosidasacomercialHidrolisis.pdfArtículo de investigaciónapplication/pdf412744http://bibliotecadigital.udea.edu.co/bitstream/10495/10193/1/BeltranLaura_2012_GalactosidasacomercialHidrolisis.pdfad9614ba0a7ee70353aa6f6699292484MD51CC-LICENSElicense_urllicense_urltext/plain; charset=utf-849http://bibliotecadigital.udea.edu.co/bitstream/10495/10193/2/license_url4afdbb8c545fd630ea7db775da747b2fMD52license_textlicense_texttext/html; charset=utf-80http://bibliotecadigital.udea.edu.co/bitstream/10495/10193/3/license_textd41d8cd98f00b204e9800998ecf8427eMD53license_rdflicense_rdfLicenciaapplication/rdf+xml; charset=utf-80http://bibliotecadigital.udea.edu.co/bitstream/10495/10193/4/license_rdfd41d8cd98f00b204e9800998ecf8427eMD54LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://bibliotecadigital.udea.edu.co/bitstream/10495/10193/5/license.txt8a4605be74aa9ea9d79846c1fba20a33MD5510495/10193oai:bibliotecadigital.udea.edu.co:10495/101932022-04-22 10:16:02.444Repositorio Institucional Universidad de Antioquiaandres.perez@udea.edu.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