β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization
- Autores
- Eberhardt, Maria Florencia; Irazoqui, José Matías; Amadio, Ariel Fernando
- Año de publicación
- 2020
- 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.
Fil: Eberhardt, Maria Florencia. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; Argentina
Fil: Irazoqui, José Matías. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; Argentina
Fil: Amadio, Ariel Fernando. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; Argentina - Materia
-
SEQUENCE-BASED METAGENOMICS
STABILIZATION PONDS
VALUE-ADDED PRODUCTS
Β-GALACTOSIDASE - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/184847
Ver los metadatos del registro completo
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β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and CharacterizationEberhardt, Maria FlorenciaIrazoqui, José MatíasAmadio, Ariel FernandoSEQUENCE-BASED METAGENOMICSSTABILIZATION PONDSVALUE-ADDED PRODUCTSΒ-GALACTOSIDASEhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Stabilization 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.Fil: Eberhardt, Maria Florencia. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; ArgentinaFil: Irazoqui, José Matías. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; ArgentinaFil: Amadio, Ariel Fernando. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; ArgentinaMultidisciplinary Digital Publishing Institute2020-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/184847Eberhardt, Maria Florencia; Irazoqui, José Matías; Amadio, Ariel Fernando; β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization; Multidisciplinary Digital Publishing Institute; Microorganisms; 9; 1; 12-2020; 1-142076-2607CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/2076-2607/9/1/55info:eu-repo/semantics/altIdentifier/doi/10.3390/microorganisms9010055info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:10:43Zoai:ri.conicet.gov.ar:11336/184847instacron: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:34982025-09-29 10:10:44.208CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
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, Maria Florencia SEQUENCE-BASED METAGENOMICS STABILIZATION PONDS VALUE-ADDED PRODUCTS Β-GALACTOSIDASE |
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, Maria Florencia Irazoqui, José Matías Amadio, Ariel Fernando |
author |
Eberhardt, Maria Florencia |
author_facet |
Eberhardt, Maria Florencia Irazoqui, José Matías Amadio, Ariel Fernando |
author_role |
author |
author2 |
Irazoqui, José Matías Amadio, Ariel Fernando |
author2_role |
author author |
dc.subject.none.fl_str_mv |
SEQUENCE-BASED METAGENOMICS STABILIZATION PONDS VALUE-ADDED PRODUCTS Β-GALACTOSIDASE |
topic |
SEQUENCE-BASED METAGENOMICS STABILIZATION PONDS VALUE-ADDED PRODUCTS Β-GALACTOSIDASE |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
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. Fil: Eberhardt, Maria Florencia. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; Argentina Fil: Irazoqui, José Matías. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; Argentina Fil: Amadio, Ariel Fernando. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Santa Fe. Instituto de Investigacion de la Cadena Lactea. - Instituto Nacional de Tecnologia Agropecuaria. Centro Regional Santa Fe. Estacion Experimental Agropecuaria Rafaela. Instituto de Investigacion de la Cadena Lactea.; 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 |
2020 |
dc.date.none.fl_str_mv |
2020-12 |
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/11336/184847 Eberhardt, Maria Florencia; Irazoqui, José Matías; Amadio, Ariel Fernando; β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization; Multidisciplinary Digital Publishing Institute; Microorganisms; 9; 1; 12-2020; 1-14 2076-2607 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/184847 |
identifier_str_mv |
Eberhardt, Maria Florencia; Irazoqui, José Matías; Amadio, Ariel Fernando; β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization; Multidisciplinary Digital Publishing Institute; Microorganisms; 9; 1; 12-2020; 1-14 2076-2607 CONICET Digital CONICET |
dc.language.none.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/2076-2607/9/1/55 info:eu-repo/semantics/altIdentifier/doi/10.3390/microorganisms9010055 |
dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by/2.5/ar/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by/2.5/ar/ |
dc.format.none.fl_str_mv |
application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Multidisciplinary Digital Publishing Institute |
publisher.none.fl_str_mv |
Multidisciplinary Digital Publishing Institute |
dc.source.none.fl_str_mv |
reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
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CONICET Digital (CONICET) |
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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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1844613999549743104 |
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13.070432 |