β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization
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 com...
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| Format: | Artículo |
| Language: | Inglés |
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MDPI
2021
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| Online Access: | http://hdl.handle.net/20.500.12123/9010 https://www.mdpi.com/2076-2607/9/1/55 https://doi.org/10.3390/microorganisms9010055 |
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| author | Eberhardt, María Florencia Irazoqui, Jose Matias Amadio, Ariel |
| author_browse | Amadio, Ariel Eberhardt, María Florencia Irazoqui, Jose Matias |
| author_facet | Eberhardt, María Florencia Irazoqui, Jose Matias Amadio, Ariel |
| author_sort | Eberhardt, María Florencia |
| collection | INTA Digital |
| 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. |
| format | Artículo |
| id | INTA9010 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | MDPI |
| publisherStr | MDPI |
| record_format | dspace |
| spelling | INTA90102021-03-31T14:06:16Z β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization Eberhardt, María Florencia Irazoqui, Jose Matias Amadio, Ariel Galactosidasas Beta Galactosidasa Industria Lechera Genómica Expresión Génica Galactosidases Beta Galactosidase Dairy Industry Genomics Gene Expression β-Galactosidases 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. 2021-03-31T13:55:08Z 2021-03-31T13:55:08Z 2021 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng 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. 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) application/pdf MDPI Microorganisms 9 (1) : 55 (2021) |
| spellingShingle | Galactosidasas Beta Galactosidasa Industria Lechera Genómica Expresión Génica Galactosidases Beta Galactosidase Dairy Industry Genomics Gene Expression β-Galactosidases Eberhardt, María Florencia Irazoqui, Jose Matias Amadio, Ariel 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 |
| 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_short | 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 |
| topic | Galactosidasas Beta Galactosidasa Industria Lechera Genómica Expresión Génica Galactosidases Beta Galactosidase Dairy Industry Genomics Gene Expression β-Galactosidases |
| url | http://hdl.handle.net/20.500.12123/9010 https://www.mdpi.com/2076-2607/9/1/55 https://doi.org/10.3390/microorganisms9010055 |
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