Biophysical and biochemical studies of a major endoglucanase secreted by Xanthomonas campestris pv. campestris.

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dc.contributor.authorRosseto, Flavio Rodolfo
dc.contributor.authorManzine, Livia Regina
dc.contributor.authorNeto, Mario de Oliveira [UNESP]
dc.contributor.authorPolikarpov, Igor
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2018-11-26T17:55:53Z
dc.date.available2018-11-26T17:55:53Z
dc.date.issued2016-09-01
dc.description.abstractEndoglucanases are the main cellulolytic enzymes secreted by the bacterium Xanthomonas campestris pv. campestris (Xcc). The major endoglucanase exported by this bacterium into an external milieu is an enzyme XccCel5A, which belongs to GH5 family subfamily 1 and is encoded by the gene engXCA. We purified XccCel5A using ammonium sulfate precipitation followed by size exclusion chromatography and identified it by zymogram analysis. Circular dichroism and fluorescence spectroscopy studies showed that XccCel5A is stable in a wide pH range and up to about 55 degrees C and denatures at the higher temperatures. The optimal conditions for enzyme activity were identified as T = 45 degrees C and pH = 7.0. Under the optimum conditions the catalytic efficiency (k(cat)/K-M) of the enzyme was determined as 5.16 x 10(4)s(-1) M-1 using carboxymethylcellulose (CMC) as a substrate. Our SAXS studies revealed extended tadpole-shape molecular assembly, typical for cellulases, and allowed to determine an overall shape of the enzyme and a relative position of the catalytic and cellulose binding domains. (C) 2016 Elsevier Inc. All rights reserved.en
dc.description.affiliationUniv Sao Paulo, Inst Fis Sao Carlos, Ave Trabalhador Sao 400 Ctr, BR-13560970 Sao Carlos, SP, Brazil
dc.description.affiliationUniv Estadual Paulista, Inst Biociencias, Distrito Rubiao Jr S-N, BR-18618970 Botucatu, SP, Brazil
dc.description.affiliationUnespUniv Estadual Paulista, Inst Biociencias, Distrito Rubiao Jr S-N, BR-18618970 Botucatu, SP, Brazil
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.description.sponsorshipIdFAPESP: 2008/56255-9
dc.description.sponsorshipIdFAPESP: 2009/54035-4
dc.description.sponsorshipIdFAPESP: 2010/08370-3
dc.description.sponsorshipIdFAPESP: 2010/16542-9
dc.description.sponsorshipIdCNPq: 482166/2010-0
dc.format.extent1-7
dc.identifierhttp://dx.doi.org/10.1016/j.enzmictec.2016.05.007
dc.identifier.citationEnzyme And Microbial Technology. New York: Elsevier Science Inc, v. 91, p. 1-7, 2016.
dc.identifier.doi10.1016/j.enzmictec.2016.05.007
dc.identifier.fileWOS000381321700001.pdf
dc.identifier.issn0141-0229
dc.identifier.lattes8213371495151651
dc.identifier.urihttp://hdl.handle.net/11449/164735
dc.identifier.wosWOS:000381321700001
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofEnzyme And Microbial Technology
dc.relation.ispartofsjr0,754
dc.rights.accessRightsAcesso aberto
dc.sourceWeb of Science
dc.subjectXanthomonas campestris
dc.subjectEndoglucanases
dc.subjectHydrolysis
dc.subjectSAXS
dc.titleBiophysical and biochemical studies of a major endoglucanase secreted by Xanthomonas campestris pv. campestris.en
dc.typeArtigo
dcterms.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dcterms.rightsHolderElsevier B.V.
unesp.advisor.lattes8213371495151651
unesp.author.orcid0000-0001-9496-4174[4]

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