β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization

Autores
Eberhardt, María Florencia; Irazoqui, Jose Matias; Amadio, Ariel
Año de publicación
2021
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Stabilization ponds are a common treatment technology for wastewater generated by dairy industries. Large proportions of cheese whey are thrown into these ponds, creating an environmental problem because of the large volume produced and the high biological and chemical oxygen demands. Due to its composition, mainly lactose and proteins, it can be considered as a raw material for value-added products, through physicochemical or enzymatic treatments. β-Galactosidases (EC 3.2.1.23) are lactose modifying enzymes that can transform lactose in free monomers, glucose and galactose, or galactooligosacharides. Here, the identification of novel genes encoding β-galactosidases, identified via whole-genome shotgun sequencing of the metagenome of dairy industries stabilization ponds is reported. The genes were selected based on the conservation of catalytic domains, comparing against the CAZy database, and focusing on families with β-galactosidases activity (GH1, GH2 and GH42). A total of 394 candidate genes were found, all belonging to bacterial species. From these candidates, 12 were selected to be cloned and expressed. A total of six enzymes were expressed, and five cleaved efficiently ortho-nitrophenyl-β-galactoside and lactose. The activity levels of one of these novel β-galactosidase was higher than other enzymes reported from functional metagenomics screening and higher than the only enzyme reported from sequence-based metagenomics. A group of novel mesophilic β-galactosidases from diary stabilization ponds’ metagenomes was successfully identified, cloned and expressed. These novel enzymes provide alternatives for the production of value-added products from dairy industries’ by-products.
EEA Rafaela
Fil: Eberhardt, María Florencia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina
Fil: Eberhardt, María Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.
Fil: Irazoqui, Jose Matias. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina
Fil: Irazoqui, Jose Matias. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.
Fil: Amadio, Ariel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina
Fil: Amadio, Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.
Fuente
Microorganisms 9 (1) : 55 (2021)
Materia
Galactosidasas
Beta Galactosidasa
Industria Lechera
Genómica
Expresión Génica
Galactosidases
Beta Galactosidase
Dairy Industry
Genomics
Gene Expression
β-Galactosidases
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
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oai_identifier_str oai:localhost:20.500.12123/9010
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spelling β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and CharacterizationEberhardt, María FlorenciaIrazoqui, Jose MatiasAmadio, ArielGalactosidasasBeta GalactosidasaIndustria LecheraGenómicaExpresión GénicaGalactosidasesBeta GalactosidaseDairy IndustryGenomicsGene Expressionβ-GalactosidasesStabilization ponds are a common treatment technology for wastewater generated by dairy industries. Large proportions of cheese whey are thrown into these ponds, creating an environmental problem because of the large volume produced and the high biological and chemical oxygen demands. Due to its composition, mainly lactose and proteins, it can be considered as a raw material for value-added products, through physicochemical or enzymatic treatments. β-Galactosidases (EC 3.2.1.23) are lactose modifying enzymes that can transform lactose in free monomers, glucose and galactose, or galactooligosacharides. Here, the identification of novel genes encoding β-galactosidases, identified via whole-genome shotgun sequencing of the metagenome of dairy industries stabilization ponds is reported. The genes were selected based on the conservation of catalytic domains, comparing against the CAZy database, and focusing on families with β-galactosidases activity (GH1, GH2 and GH42). A total of 394 candidate genes were found, all belonging to bacterial species. From these candidates, 12 were selected to be cloned and expressed. A total of six enzymes were expressed, and five cleaved efficiently ortho-nitrophenyl-β-galactoside and lactose. The activity levels of one of these novel β-galactosidase was higher than other enzymes reported from functional metagenomics screening and higher than the only enzyme reported from sequence-based metagenomics. A group of novel mesophilic β-galactosidases from diary stabilization ponds’ metagenomes was successfully identified, cloned and expressed. These novel enzymes provide alternatives for the production of value-added products from dairy industries’ by-products.EEA RafaelaFil: Eberhardt, María Florencia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Eberhardt, María Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.Fil: Irazoqui, Jose Matias. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Irazoqui, Jose Matias. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.Fil: Amadio, Ariel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Amadio, Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.MDPI2021-03-31T13:55:08Z2021-03-31T13:55:08Z2021info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/9010https://www.mdpi.com/2076-2607/9/1/552076-2607https://doi.org/10.3390/microorganisms9010055Microorganisms 9 (1) : 55 (2021)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repograntAgreement/INTA/PNBIO-1131033/AR./Genómica y biotecnología aplicada a la cría animal.info:eu-repograntAgreement/INTA/PNAIyAV-1130033/AR./Tecnologías de preservación de alimentos y de aprovechamiento de subproductos.info: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)2025-09-04T09:48:49Zoai:localhost:20.500.12123/9010instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-04 09:48:49.921INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization
title β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization
spellingShingle β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization
Eberhardt, María Florencia
Galactosidasas
Beta Galactosidasa
Industria Lechera
Genómica
Expresión Génica
Galactosidases
Beta Galactosidase
Dairy Industry
Genomics
Gene Expression
β-Galactosidases
title_short β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization
title_full β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization
title_fullStr β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization
title_full_unstemmed β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization
title_sort β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization
dc.creator.none.fl_str_mv Eberhardt, María Florencia
Irazoqui, Jose Matias
Amadio, Ariel
author Eberhardt, María Florencia
author_facet Eberhardt, María Florencia
Irazoqui, Jose Matias
Amadio, Ariel
author_role author
author2 Irazoqui, Jose Matias
Amadio, Ariel
author2_role author
author
dc.subject.none.fl_str_mv Galactosidasas
Beta Galactosidasa
Industria Lechera
Genómica
Expresión Génica
Galactosidases
Beta Galactosidase
Dairy Industry
Genomics
Gene Expression
β-Galactosidases
topic Galactosidasas
Beta Galactosidasa
Industria Lechera
Genómica
Expresión Génica
Galactosidases
Beta Galactosidase
Dairy Industry
Genomics
Gene Expression
β-Galactosidases
dc.description.none.fl_txt_mv Stabilization ponds are a common treatment technology for wastewater generated by dairy industries. Large proportions of cheese whey are thrown into these ponds, creating an environmental problem because of the large volume produced and the high biological and chemical oxygen demands. Due to its composition, mainly lactose and proteins, it can be considered as a raw material for value-added products, through physicochemical or enzymatic treatments. β-Galactosidases (EC 3.2.1.23) are lactose modifying enzymes that can transform lactose in free monomers, glucose and galactose, or galactooligosacharides. Here, the identification of novel genes encoding β-galactosidases, identified via whole-genome shotgun sequencing of the metagenome of dairy industries stabilization ponds is reported. The genes were selected based on the conservation of catalytic domains, comparing against the CAZy database, and focusing on families with β-galactosidases activity (GH1, GH2 and GH42). A total of 394 candidate genes were found, all belonging to bacterial species. From these candidates, 12 were selected to be cloned and expressed. A total of six enzymes were expressed, and five cleaved efficiently ortho-nitrophenyl-β-galactoside and lactose. The activity levels of one of these novel β-galactosidase was higher than other enzymes reported from functional metagenomics screening and higher than the only enzyme reported from sequence-based metagenomics. A group of novel mesophilic β-galactosidases from diary stabilization ponds’ metagenomes was successfully identified, cloned and expressed. These novel enzymes provide alternatives for the production of value-added products from dairy industries’ by-products.
EEA Rafaela
Fil: Eberhardt, María Florencia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina
Fil: Eberhardt, María Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.
Fil: Irazoqui, Jose Matias. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina
Fil: Irazoqui, Jose Matias. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.
Fil: Amadio, Ariel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina
Fil: Amadio, Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina.
description Stabilization ponds are a common treatment technology for wastewater generated by dairy industries. Large proportions of cheese whey are thrown into these ponds, creating an environmental problem because of the large volume produced and the high biological and chemical oxygen demands. Due to its composition, mainly lactose and proteins, it can be considered as a raw material for value-added products, through physicochemical or enzymatic treatments. β-Galactosidases (EC 3.2.1.23) are lactose modifying enzymes that can transform lactose in free monomers, glucose and galactose, or galactooligosacharides. Here, the identification of novel genes encoding β-galactosidases, identified via whole-genome shotgun sequencing of the metagenome of dairy industries stabilization ponds is reported. The genes were selected based on the conservation of catalytic domains, comparing against the CAZy database, and focusing on families with β-galactosidases activity (GH1, GH2 and GH42). A total of 394 candidate genes were found, all belonging to bacterial species. From these candidates, 12 were selected to be cloned and expressed. A total of six enzymes were expressed, and five cleaved efficiently ortho-nitrophenyl-β-galactoside and lactose. The activity levels of one of these novel β-galactosidase was higher than other enzymes reported from functional metagenomics screening and higher than the only enzyme reported from sequence-based metagenomics. A group of novel mesophilic β-galactosidases from diary stabilization ponds’ metagenomes was successfully identified, cloned and expressed. These novel enzymes provide alternatives for the production of value-added products from dairy industries’ by-products.
publishDate 2021
dc.date.none.fl_str_mv 2021-03-31T13:55:08Z
2021-03-31T13:55:08Z
2021
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/20.500.12123/9010
https://www.mdpi.com/2076-2607/9/1/55
2076-2607
https://doi.org/10.3390/microorganisms9010055
url http://hdl.handle.net/20.500.12123/9010
https://www.mdpi.com/2076-2607/9/1/55
https://doi.org/10.3390/microorganisms9010055
identifier_str_mv 2076-2607
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repograntAgreement/INTA/PNBIO-1131033/AR./Genómica y biotecnología aplicada a la cría animal.
info:eu-repograntAgreement/INTA/PNAIyAV-1130033/AR./Tecnologías de preservación de alimentos y de aprovechamiento de subproductos.
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
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv Microorganisms 9 (1) : 55 (2021)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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