Characterization of two GH5 endoglucanases from termite microbiome using synthetic metagenomics
Here, we characterize two novel GH5 endoglucanases (GH5CelA and GH5CelB) from an uncultured bacterium identified in termite gut microbiomes. Both genes were codon-optimized, synthetized, cloned, and expressed as recombinant proteins in Escherichia coli for subsequent purification. Both enzymes showe...
| Autores principales: | , , , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
| Publicado: |
Springer
2020
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/7767 https://link.springer.com/article/10.1007/s00253-020-10831-5 https://doi.org/10.1007/s00253-020-10831-5 |
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| author | Ben Guerrero, Emiliano Marrero Diaz De Villegas, Ruben Soria, Marcelo Abel Santangelo, María De La Paz Campos, Eleonora Talia, Paola Mónica |
| author_browse | Ben Guerrero, Emiliano Campos, Eleonora Marrero Diaz De Villegas, Ruben Santangelo, María De La Paz Soria, Marcelo Abel Talia, Paola Mónica |
| author_facet | Ben Guerrero, Emiliano Marrero Diaz De Villegas, Ruben Soria, Marcelo Abel Santangelo, María De La Paz Campos, Eleonora Talia, Paola Mónica |
| author_sort | Ben Guerrero, Emiliano |
| collection | INTA Digital |
| description | Here, we characterize two novel GH5 endoglucanases (GH5CelA and GH5CelB) from an uncultured bacterium identified in termite gut microbiomes. Both genes were codon-optimized, synthetized, cloned, and expressed as recombinant proteins in Escherichia coli for subsequent purification. Both enzymes showed activity on the pNPC and barley β-glucan substrates, whereas GH5CelB also showed low activity on carboxymethyl cellulose. The optimum conditions for both enzymes were an acid pH (5) and moderate temperature (35 to 50 °C). The enzymes differed in the kinetic profiles and patterns of the generated hydrolysis products. A structural-based modeling analysis indicated that both enzymes possess a typical (β/α)8-barrel fold characteristic of GH5 family, with some differential features in the active site cleft. Also, GH5CelB presents a putative secondary binding site. Furthermore, adjacent to the active site of GH5CelA and GH5CelB, a whole subdomain rarely found in GH5 family may participate in substrate binding and thermal stability.
Therefore, GH5CelA may be a good candidate for the production of cello-oligosaccharides of different degrees of polymerization applicable for feed and food industries, including prebiotics. On the other hand, GH5CelB could be useful in an enzymatic cocktail for the production of lignocellulosic bioethanol, because of the production of glucose as a hydrolysis product. |
| format | info:ar-repo/semantics/artículo |
| id | INTA7767 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2020 |
| publishDateRange | 2020 |
| publishDateSort | 2020 |
| publisher | Springer |
| publisherStr | Springer |
| record_format | dspace |
| spelling | INTA77672020-08-25T13:02:31Z Characterization of two GH5 endoglucanases from termite microbiome using synthetic metagenomics Ben Guerrero, Emiliano Marrero Diaz De Villegas, Ruben Soria, Marcelo Abel Santangelo, María De La Paz Campos, Eleonora Talia, Paola Mónica Genomas Glucanos Isoptera Identificación Genomes Glucans Identification Endoglucanases Termites Here, we characterize two novel GH5 endoglucanases (GH5CelA and GH5CelB) from an uncultured bacterium identified in termite gut microbiomes. Both genes were codon-optimized, synthetized, cloned, and expressed as recombinant proteins in Escherichia coli for subsequent purification. Both enzymes showed activity on the pNPC and barley β-glucan substrates, whereas GH5CelB also showed low activity on carboxymethyl cellulose. The optimum conditions for both enzymes were an acid pH (5) and moderate temperature (35 to 50 °C). The enzymes differed in the kinetic profiles and patterns of the generated hydrolysis products. A structural-based modeling analysis indicated that both enzymes possess a typical (β/α)8-barrel fold characteristic of GH5 family, with some differential features in the active site cleft. Also, GH5CelB presents a putative secondary binding site. Furthermore, adjacent to the active site of GH5CelA and GH5CelB, a whole subdomain rarely found in GH5 family may participate in substrate binding and thermal stability. Therefore, GH5CelA may be a good candidate for the production of cello-oligosaccharides of different degrees of polymerization applicable for feed and food industries, including prebiotics. On the other hand, GH5CelB could be useful in an enzymatic cocktail for the production of lignocellulosic bioethanol, because of the production of glucose as a hydrolysis product. Instituto de Biotecnología Fil: Ben Guerrero, Emiliano. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Marrero Diaz De Villegas, Ruben. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Agrobiotecnología y Biología Molecular; Argentin Fil: Soria, Marcelo Abel. Universidad de Buenos Aires. Facultad de Agronomía. Cátedra de Microbiología Agrícola; Argentina Fil: Santangelo, María De La Paz. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Campos, Eleonora. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Talia, Paola Mónica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología. Instituto de Agrobiotecnología y Biología Molecular. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Agrobiotecnología y Biología Molecular; Argentina 2020-08-25T12:56:46Z 2020-08-25T12:56:46Z 2020-08 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/7767 https://link.springer.com/article/10.1007/s00253-020-10831-5 0175-7598 1432-0614 https://doi.org/10.1007/s00253-020-10831-5 eng info:eu-repograntAgreement/INTA/PNAIyAV/1130034/AR./Desarrollo de procesos para la transformación de biomasa en bioenergía. info:eu-repo/semantics/restrictedAccess application/pdf Springer Applied Microbiology and Biotechnology (2020) |
| spellingShingle | Genomas Glucanos Isoptera Identificación Genomes Glucans Identification Endoglucanases Termites Ben Guerrero, Emiliano Marrero Diaz De Villegas, Ruben Soria, Marcelo Abel Santangelo, María De La Paz Campos, Eleonora Talia, Paola Mónica Characterization of two GH5 endoglucanases from termite microbiome using synthetic metagenomics |
| title | Characterization of two GH5 endoglucanases from termite microbiome using synthetic metagenomics |
| title_full | Characterization of two GH5 endoglucanases from termite microbiome using synthetic metagenomics |
| title_fullStr | Characterization of two GH5 endoglucanases from termite microbiome using synthetic metagenomics |
| title_full_unstemmed | Characterization of two GH5 endoglucanases from termite microbiome using synthetic metagenomics |
| title_short | Characterization of two GH5 endoglucanases from termite microbiome using synthetic metagenomics |
| title_sort | characterization of two gh5 endoglucanases from termite microbiome using synthetic metagenomics |
| topic | Genomas Glucanos Isoptera Identificación Genomes Glucans Identification Endoglucanases Termites |
| url | http://hdl.handle.net/20.500.12123/7767 https://link.springer.com/article/10.1007/s00253-020-10831-5 https://doi.org/10.1007/s00253-020-10831-5 |
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