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dc.contributor.author
Salinas, Silvina Rosa
dc.contributor.author
Bianco, Maria Isabel
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Barreras, Maximo
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Ielpi, Luis
dc.date.available
2017-02-09T18:35:56Z
dc.date.issued
2011-07
dc.identifier.citation
Salinas, Silvina Rosa; Bianco, Maria Isabel; Barreras, Maximo; Ielpi, Luis; Expression, purification and biochemical characterization of GumI, a monotopic membrane GDP-mannose:glycolipid 4-{beta}-D-mannosyltransferase from Xanthomonas campestris pv. campestris; Oxford Univ Press Inc; Glycobiology; 21; 7; 7-2011; 903-913
dc.identifier.issn
0959-6658
dc.identifier.uri
http://hdl.handle.net/11336/12803
dc.description.abstract
We describe the first biochemical characterization of the gumI gene product, an essential protein for xanthan polysaccharide synthesis. Cellular fractionation experiments reveal the presence of a protein associated with the membrane fraction, even in the absence of the other proteins responsible for the synthesis of glycolipid intermediates and the proteins involved in the polymerization and transport of the xanthan chains. By alkaline buffer extraction and detergent phase partitioning, GumI was categorized as a monotopic membrane protein. GumI was overexpressed in Escherichia coli, solubilized and purified in an active and stable form using a simple and reproducible two-step procedure. The purified recombinant GumI is a nonprocessive β-mannosyltransferase that uses GDP-Man as a donor substrate and glucuronic acid-β-1,2-mannose-α-1,3-glucose-β-1,4-glucose-PP-polyisoprenyl as an acceptor. We also established the optimal biochemical conditions for GumI enzymatic activity. Sequence analysis revealed the presence of a conserved domain for glycosyltransferases (GTs) of the GT-B superfamily and homologous proteins in several prokaryote organisms. On the basis of this biochemical characterization, GumI may represent the founding member of a new GT family in the Carbohydrate-Active EnZymes classification.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Oxford Univ Press Inc
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Glycosyltransferase
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Mannosyltransferase
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Membrane-Associated Proteins
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Xanthan
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Xanthomonas Campestris
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Bioquímica y Biología Molecular
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Ciencias Biológicas
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CIENCIAS NATURALES Y EXACTAS
dc.title
Expression, purification and biochemical characterization of GumI, a monotopic membrane GDP-mannose:glycolipid 4-{beta}-D-mannosyltransferase from Xanthomonas campestris pv. campestris
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2017-02-07T13:52:18Z
dc.journal.volume
21
dc.journal.number
7
dc.journal.pagination
903-913
dc.journal.pais
Reino Unido
dc.journal.ciudad
Oxford
dc.description.fil
Fil: Salinas, Silvina Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; Argentina
dc.description.fil
Fil: Bianco, Maria Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; Argentina
dc.description.fil
Fil: Barreras, Maximo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; Argentina
dc.description.fil
Fil: Ielpi, Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; Argentina
dc.journal.title
Glycobiology
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://academic.oup.com/glycob/article-lookup/doi/10.1093/glycob/cwr022
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1093/glycob/cwr022
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