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The use of lignocellulosic biomass for second generation biofuels requires optimization of enzymatic breakdown of plant cell walls. In this work, cellulolytic bacteria were isolated from a native and two cultivated forest soil samples. Amplification of glycosyl hydrolases was attempted by using a low stringency-degenerate primer PCR strategy, using total soil DNA and bulk DNA pooled from positive colonies as template. A set of primers was designed based
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dc.contributor.author | Campos, Eleonora | |
dc.contributor.author | Negro Alvarez, María José | |
dc.contributor.author | Sabaris Di Lorenzo, Gonzalo Julián | |
dc.contributor.author | Gonzalez, Sergio Alberto | |
dc.contributor.author | Rorig, Marcela Laura | |
dc.contributor.author | Talia, Paola Mónica | |
dc.contributor.author | Grasso, Daniel Horacio | |
dc.contributor.author | Sáez, Felicia | |
dc.contributor.author | Manzanares Secades, Paloma | |
dc.contributor.author | Ballesteros Perdices, Mercedes | |
dc.contributor.author | Cataldi, Angel Adrian | |
dc.date.accessioned | 2019-01-23T11:42:11Z | |
dc.date.available | 2019-01-23T11:42:11Z | |
dc.date.issued | 2014-03 | |
dc.identifier.issn | 0944-5013 | |
dc.identifier.issn | 1618-0623 | |
dc.identifier.other | https://doi.org/10.1016/j.micres.2013.06.004 | |
dc.identifier.uri | https://www.sciencedirect.com/science/article/pii/S0944501313000906 | |
dc.identifier.uri | http://hdl.handle.net/20.500.12123/4313 | |
dc.description.abstract | The use of lignocellulosic biomass for second generation biofuels requires optimization of enzymatic breakdown of plant cell walls. In this work, cellulolytic bacteria were isolated from a native and two cultivated forest soil samples. Amplification of glycosyl hydrolases was attempted by using a low stringency-degenerate primer PCR strategy, using total soil DNA and bulk DNA pooled from positive colonies as template. A set of primers was designed based on Acidothermus cellulolyticus genome, by search of conserved domains of glycosyl hydrolases (GH) families of interest. Using this approach, a fragment containing an open reading frame (ORF) with 98% identity to a putative GH43 beta-xylosidase coding gene from Enterobacter cloacae was amplified and cloned. The full protein was expressed in Escherichia coli as N-terminal or C-terminal His-tagged fusions and purified under native conditions. Only N-terminal fusion protein, His-Xyl43, presented beta-xylosidase activity. On pNPX, optimal activity was achieved at pH 6 and 40 °C and Km and Kcat values were 2.92 mM and 1.32 seg−1, respectively. Activity was also demonstrated on xylobiose (X2), with Km 17.8 mM and Kcat 380 s−1. These results demonstrated that Xyl43 is a functional beta-xylosidase and it is the first evidence of this activity for Enterobacter sp. | eng |
dc.format | application/pdf | es_AR |
dc.language.iso | eng | es_AR |
dc.publisher | Elsevier | es_AR |
dc.rights | info:eu-repo/semantics/openAccess | es_AR |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-sa/4.0/ | |
dc.source | Microbiological Research 169 (2–3) : 213-220 (February–March 2014) | es_AR |
dc.subject | Biocarburante | es_AR |
dc.subject | Biofuels | eng |
dc.subject | Lignocelulosa | es_AR |
dc.subject | Lignocellulose | eng |
dc.subject | Enterobacter | es_AR |
dc.subject | Bacterias del Suelo | es_AR |
dc.subject | Soil Bacteria | eng |
dc.subject.other | Biocombustibles | es_AR |
dc.subject.other | GH43 | es_AR |
dc.title | Purification and characterization of a GH43 β-xylosidase from Enterobacter sp. identified and cloned from forest soil bacteria | es_AR |
dc.type | info:ar-repo/semantics/artículo | es_AR |
dc.type | info:eu-repo/semantics/article | es_AR |
dc.type | info:eu-repo/semantics/publishedVersion | es_AR |
dc.rights.license | Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) | |
dc.description.origen | Instituto de Biotecnología | es_AR |
dc.description.fil | Fil: Campos, Eleonora. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina | es_AR |
dc.description.fil | Fil: Negro Alvarez, María José. Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT). Unidad Biocarburantes; España | es_AR |
dc.description.fil | Fil: Sabaris Di Lorenzo, Gonzalo Julián. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina | es_AR |
dc.description.fil | Fil: Gonzalez, Sergio Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina | es_AR |
dc.description.fil | Fil: Rorig, Marcela Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Suelos; Argentina | es_AR |
dc.description.fil | Fil: Talia, Paola Mónica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina | es_AR |
dc.description.fil | Fil: Grasso, Daniel Horacio. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Suelos; Argentina | es_AR |
dc.description.fil | Fil: Sáez, Felicia. Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT). Unidad Biocarburantes; España | es_AR |
dc.description.fil | Fil: Manzanares Secades, Paloma. Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT). Unidad Biocarburantes; España | es_AR |
dc.description.fil | Fil: Ballesteros Perdices, Mercedes. Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT). Unidad Biocarburantes; España | es_AR |
dc.description.fil | Fil: Cataldi, Angel Adrian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina | es_AR |
dc.subtype | cientifico |
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