Overexpression, purification, and biochemical characterization of GumC, an enzyme involved in the biosynthesis of exopolysaccharide by Xylella fastidiosa

dc.creatorde Pieri, C
dc.creatorBeltramini, LM
dc.creatorSelistre-de-Araujo, HS
dc.creatorVettore, AL
dc.creatorda Silva, FR
dc.creatorArruda, P
dc.creatorOliva, G
dc.creatorde Souza, DHF
dc.date2004
dc.dateAPR
dc.date2014-11-14T03:05:47Z
dc.date2015-11-26T17:12:54Z
dc.date2014-11-14T03:05:47Z
dc.date2015-11-26T17:12:54Z
dc.date.accessioned2018-03-29T00:01:18Z
dc.date.available2018-03-29T00:01:18Z
dc.identifierProtein Expression And Purification. Academic Press Inc Elsevier Science, v. 34, n. 2, n. 223, n. 228, 2004.
dc.identifier1046-5928
dc.identifierWOS:000220233400009
dc.identifier10.1016/j.pep.2003.11.003
dc.identifierhttp://www.repositorio.unicamp.br/jspui/handle/REPOSIP/81545
dc.identifierhttp://www.repositorio.unicamp.br/handle/REPOSIP/81545
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/81545
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1281444
dc.descriptionGumC is one of nine enzymes involved in the biosynthesis of fastidian gum, an exopolysaccharide produced by Xylella fastidiosa that may be linked directly to the pathogenicity of the microorganism. GumC may be responsible for gum polymerization or secretion through the membrane of X. fastidiosa. To perform structure and functions studies, we developed an expression system for the production of GumC as a fusion protein with maltose binding protein (MBP) using pMAL-c2x vector. The GumC-MBP fusion protein was expressed as a 94 kDa protein, which strongly reacts with anti-MBP antibodies. GumC-MBP was isolated by affinity chromatography through an amylose column and used to produce antibodies against the fusion protein. After the enzymatic cleavage of MBP, GumC was purified on a Q Sepharose Fast Flow column. GumC showed a molecular weight corresponding to the expected one (52 kDa) and its N-terminal sequence was identical to that deduced from the DNA. The shape of the circular dichroism spectrum was compatible with a folded protein that contains alpha-helical regions in its structure. Therefore, in this study we describe, for the first time, the production of GumC recombinant protein. (C) 2003 Elsevier Inc. All rights reserved.
dc.description34
dc.description2
dc.description223
dc.description228
dc.languageen
dc.publisherAcademic Press Inc Elsevier Science
dc.publisherSan Diego
dc.publisherEUA
dc.relationProtein Expression And Purification
dc.relationProtein Expr. Purif.
dc.rightsfechado
dc.rightshttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.sourceWeb of Science
dc.subjectXylella fastidiosa
dc.subjectcitrus variegated chlorosis
dc.subjectgum operon
dc.subjectfastidian gum
dc.subjectGumC
dc.subjectexopolysaccharide
dc.subjectXanthomonas-campestris
dc.subjectXanthan Gum
dc.subjectBacterial Pathogenicity
dc.subjectPolysaccharide
dc.subjectVirulence
dc.subjectProteins
dc.subjectCleavage
dc.subjectStrains
dc.subjectMutants
dc.subjectPlants
dc.titleOverexpression, purification, and biochemical characterization of GumC, an enzyme involved in the biosynthesis of exopolysaccharide by Xylella fastidiosa
dc.typeArtículos de revistas


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