Produção e caracterização físico-química e biológica da cadeia alfa da lectina recombinante de Canavalia brasiliensis

• Autor(a) principal: Oliveira, Antônia Simoni de
• Data de Publicação: 2017
• Tipo de documento: Tese
• Idioma: por
• Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
• Texto Completo: http://www.repositorio.ufc.br/handle/riufc/27407
• topic: Lectina; Recombinante; Canavalia brasiliensis

The ability to express and purify desired recombinant proteins in large quantities allows its biochemical characterization and use in industrial processes. Numerous plant lectins have been produced in heterologous systems, especially in Escherichia coli. By recognizing and specifically binding to carbohydrates, these molecules can perform various biological functions, such as cell-cell recognition, cell adhesion, fertilization, and defense mechanisms. Among plant lectins, leguminous lectins have been extensively investigated. The lectin of Canavalia brasiliensis, isolated by the first time in 1984 by Moreira and Cavada is a widely studied protein and has shown an immunostimulatory, andidepressive, antinociceptive, neroptotective, and antiproliferative effect in human leucemia cells, among onther biological activities tested. The biological properties, however, of many lectins have been attributed to isolectin blends, since it is difficult to separate isoforms using standard techniques. In this study the recombinant lectin (α chain) of Canavalia brasiliensis was produced from the synthetic gene inserted in the vector pET28a e. The strains DH5α and BL21 (DE3) were used for cloning and expression, respectively. The rConBr was expressed under all conditions tested and the condition of 16oC, 16 hours and 1,0 mM IPTG (isopropil-β-D-galatosídeo) was chosen for protein production. The protein was purified, being obtained in its active form. Lectin was active when tested against rabbit erytrocytes and was inhibited by mannose and α-methyl-mannopyranoside. The activity of rConBr is optimum at pH 4.0 to 7.0, remains stable up to 60 ° C and is not dependent on divalent cations. In addition, rConBr was not toxic against Artemia sp. and showed no cytotoxic effect on glioma cells C6 lineage.