Cloning, expression and characterization of Bauhinia variegata trypsin inhibitor BvTI

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dc.contributor.authorSouza, A. F. de
dc.contributor.authorTorquato, RJS
dc.contributor.authorTanaka, A. S.
dc.contributor.authorSampaio, CAM
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.date.accessioned2016-01-24T12:38:09Z
dc.date.available2016-01-24T12:38:09Z
dc.date.issued2005-11-01
dc.description.abstractA Bauhinia variegata trypsin inhibitor (BvTI) cDNA fragment was cloned into the pCANTAB5E phagemid. the clone pAS 1.1.3 presented a cDNA fragment of 733 bp, including the coding region for a mature BvTI protein comprising 175 amino acid residues. the deduced amino acid sequence for BvTI confirmed it as a member of the Kunitz-type plant serine proteinase inhibitor family. the BvTI cDNA fragment encoding the mature form was cloned into the expression vector, pET-14b, and expressed in E coli BL2I (DE3) pLysS in an active form. in addition, a BvTI mutant form, r(mut)BvTI, with a Pro residue as the fifth amino acid in place of Leu, was produced. the recombinant proteins, rBvTI and r(mut)BvTI, were purified on a trypsin-Sepharose column, yielding 29 and 1.44 mg/I of active protein, respectively, and showed protein bands of approximately 21.5 kDa by SDS-PAGE. Trypsin inhibition activity was comparable for rBvTI (K-I=4 nm) and r(mut)BvTI (K-i=6 nm). Our data suggest that the Leu to Pro substitution at the fifth amino-terminal residue was not crucial for proteinase inhibition.en
dc.description.affiliationUNIFESP, EPM, Dept Bioquim, BR-04044020 São Paulo, Brazil
dc.description.affiliationUnifespUNIFESP, EPM, Dept Bioquim, BR-04044020 São Paulo, Brazil
dc.description.sourceWeb of Science
dc.format.extent1185-1189
dc.identifierhttp://dx.doi.org/10.1515/BC.2005.135
dc.identifier.citationBiological Chemistry. Berlin: Walter de Gruyter & Co, v. 386, n. 11, p. 1185-1189, 2005.
dc.identifier.doi10.1515/BC.2005.135
dc.identifier.issn1431-6730
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/28546
dc.identifier.wosWOS:000233703800012
dc.language.isoeng
dc.publisherWalter de Gruyter & Co
dc.relation.ispartofBiological Chemistry
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.subjectBauhinia variegataen
dc.subjectcloningen
dc.subjectexpressionen
dc.subjectKunitz-type serine proteinase inhibitoren
dc.subjectLeguminosaeen
dc.subjecttrypsin inhibitoren
dc.titleCloning, expression and characterization of Bauhinia variegata trypsin inhibitor BvTIen
dc.typeinfo:eu-repo/semantics/article
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