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dc.contributor.author
Torres, Pedro  
dc.contributor.author
Manzo, Ricardo Martín  
dc.contributor.author
Rubiolo, Amelia Catalina  
dc.contributor.author
Batista Viera, Francisco  
dc.contributor.author
Mammarella, Enrique José  
dc.date.available
2016-12-12T21:11:14Z  
dc.date.issued
2014-03  
dc.identifier.citation
Torres, Pedro; Manzo, Ricardo Martín; Rubiolo, Amelia Catalina; Batista Viera, Francisco; Mammarella, Enrique José; Purification of an l-arabinose isomerase from Enterococcus faecium DBFIQ E36 employing a biospecific affinity strategy; Elsevier Science; Journal Of Molecular Catalysis B: Enzymatic; 102; 3-2014; 99-105  
dc.identifier.issn
1381-1177  
dc.identifier.uri
http://hdl.handle.net/11336/9194  
dc.description.abstract
l-Arabinose isomerase is an intracellular enzyme that can convert l-arabinose to l-ribulose and d-galactose to d-tagatose, a promising but rare nutraceutical. Most of l-arabinose isomerases purified upto date employed the combination between DNA recombinant technology and affinity chromatographybased on poly-histidine tail recognition, but few of the enzymes were obtained and purified in a non-recombinant way. For these reasons, a specific affinity bioadsorbent containing l-arabitol as ligand, acompetitive inhibitor of the enzyme, was designed and synthesized for achieving pure preparations ofthe enzyme l-arabinose isomerase from wild-type Enterococcus faecium DBFIQ E36 strain, isolated fromraw cow milk.The two-step purification procedure consisted in fractionation by ammonium sulphate precipitationfollowed by affinity chromatography with obtained bioadsorbent, allowing the purification, to elec-trophoretical homogeneity, of target enzyme. Characterization studies were performed with purifiedl-arabinose isomerase in order to increase knowledge of their physicochemical properties. In this sense,enzyme exhibited an optimum temperature of 50?C and optimum pH of 7.0, maintaining good sta-bility in the ranges 20–45?C and pH 6.5–8. Kiwere calculated, employing d-galactose as substrate, forl-arabitol and l-ribitol, achieving values of 7.9 mM and 183 mM, respectively. Kmand Vmaxvalues obtainedwere 35 mM and 81 U mg-1at 50?C, respectively. Mass spectrometry assay revealed a 48 kDa monomerwhereas gel permeation chromatography achieved a 187 kDa molecular weight for native enzyme. Finally,2D-electrophoresis and isoelectrofocusing analysis revealed an isoelectric point value of 3.80. Resultshave unveiled both an acidic nature and promising properties for l-arabinose isomerase isolated from E.faecium DBFIQ E36.  
dc.format
application/pdf  
dc.language.iso
eng  
dc.publisher
Elsevier Science  
dc.rights
info:eu-repo/semantics/openAccess  
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/  
dc.subject
L-Arabinose Isomerase  
dc.subject
D-Tagatose D-Galactos  
dc.subject
Affinity Chromatography  
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Enterococcus Faecium Strain  
dc.subject.classification
Bioprocesamiento Tecnológico, Biocatálisis, Fermentación  
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Biotecnología Industrial  
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INGENIERÍAS Y TECNOLOGÍAS  
dc.title
Purification of an l-arabinose isomerase from Enterococcus faecium DBFIQ E36 employing a biospecific affinity strategy  
dc.type
info:eu-repo/semantics/article  
dc.type
info:ar-repo/semantics/artículo  
dc.type
info:eu-repo/semantics/publishedVersion  
dc.date.updated
2016-12-12T13:47:56Z  
dc.journal.volume
102  
dc.journal.pagination
99-105  
dc.journal.pais
Países Bajos  
dc.journal.ciudad
Amsterdam  
dc.description.fil
Fil: Torres, Pedro. Universidad de la Republica; Uruguay  
dc.description.fil
Fil: Manzo, Ricardo Martín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina  
dc.description.fil
Fil: Rubiolo, Amelia Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina  
dc.description.fil
Fil: Batista Viera, Francisco. Universidad de la Republica; Uruguay  
dc.description.fil
Fil: Mammarella, Enrique José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Santa Fe. Instituto de Desarrollo Tecnológico para la Industria Química (i); Argentina  
dc.journal.title
Journal Of Molecular Catalysis B: Enzymatic  
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1016/j.molcatb.2014.01.023  
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1381117714000344