dc.creator | Campos, Eleonora | |
dc.creator | Negro Alvarez, María José | |
dc.creator | Sabarís Di Lorenzo, Gonzalo Julián | |
dc.creator | González, Sergio Alberto | |
dc.creator | Rorig, Marcela | |
dc.creator | Talia, Paola | |
dc.creator | Grasso, Daniel Hector | |
dc.creator | Sáez, Felicia | |
dc.creator | Manzanares Secades, Paloma | |
dc.creator | Ballesteros Perdices, Mercedes | |
dc.creator | Cataldi, Ángel Adrián | |
dc.date.accessioned | 2019-10-08T21:07:11Z | |
dc.date.accessioned | 2022-10-15T08:39:15Z | |
dc.date.available | 2019-10-08T21:07:11Z | |
dc.date.available | 2022-10-15T08:39:15Z | |
dc.date.created | 2019-10-08T21:07:11Z | |
dc.date.issued | 2013-05 | |
dc.identifier | Campos, Eleonora; Negro Alvarez, María José; Sabarís Di Lorenzo, Gonzalo Julián; González, Sergio Alberto; Rorig, Marcela; et al.; Purification and characterization of a GH43 β-xylosidase from Enterobacter sp. identified and cloned from forest soil bacteria; Elsevier Gmbh; Microbiological Research; 169; 2-3; 5-2013; 213-220 | |
dc.identifier | 0944-5013 | |
dc.identifier | http://hdl.handle.net/11336/85402 | |
dc.identifier | CONICET Digital | |
dc.identifier | CONICET | |
dc.identifier.uri | https://repositorioslatinoamericanos.uchile.cl/handle/2250/4366013 | |
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 2.92mM and 1.32seg-1, respectively. Activity was also demonstrated on xylobiose (X2), with Km 17.8mM and Kcat 380s-1. These results demonstrated that Xyl43 is a functional beta-xylosidase and it is the first evidence of this activity for Enterobacter sp. | |
dc.language | eng | |
dc.publisher | Elsevier Gmbh | |
dc.relation | info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0944501313000906 | |
dc.relation | info:eu-repo/semantics/altIdentifier/doi/https://doi.org/10.1016/j.micres.2013.06.004 | |
dc.rights | https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ | |
dc.rights | info:eu-repo/semantics/openAccess | |
dc.subject | BETA-XYLOSIDASE | |
dc.subject | ENTEROBACTER | |
dc.subject | GH43 | |
dc.subject | LIGNOCELLULOSIC BIOFUELS | |
dc.title | Purification and characterization of a GH43 β-xylosidase from Enterobacter sp. identified and cloned from forest soil bacteria | |
dc.type | info:eu-repo/semantics/article | |
dc.type | info:ar-repo/semantics/artículo | |
dc.type | info:eu-repo/semantics/publishedVersion | |