Produção Nacional
Revisado por pares
Purification of a lectin from Cratylia mollis crude extract seed by a single step PEG/phosphate aqueous two-phase system
2020; Taylor & Francis; Volume: 50; Issue: 7 Linguagem: Inglês
10.1080/10826068.2020.1725771
ISSN1532-2297
AutoresCynthia O. Nascimento, Romero Marcos Pedrosa Brandão Costa, Paulo Rodinei Soares Lopes, Polyanna Nunes Herculano, Tatiana Souza Porto, Thiago Pajeú Nascimento, Carolina de Albuquerque Lima, Raquel Pedrosa Bezerra, J. A. Teixeira, Luana Cassandra Breitenbach Barroso Coelho, Maria Tereza dos Santos Correia, Maria das Graças Carneiro-da-Cunha, Ana Lúcia Figueiredo Porto,
Tópico(s)Microbial Metabolites in Food Biotechnology
ResumoThe partitioning and purification of lectins from the crude extract of Cratylia mollis seeds (Cramoll 1,4) was investigated in aqueous two-phase systems (ATPS). A factorial design model (24) was used to evaluate the influence of polyethylene glycol (PEG) molar mass (1500–8000 g/mol), PEG concentration (12.5–17.5% w/w), phosphate (10–15% w/w) concentration, and pH (6–8) on the differential partitioning, purification factor, and yield of the lectin. Polymer and salt concentration were the most important variables affecting partition of lectin and used to find optimum purification factor by experimental Box–Behnken design together with the response surface methodology (RSM). ATPS showed best conditions composed by 13.9% PEG1500, 15.3% phosphate buffer at pH 6, which ensured purification factor of 4.70. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis showed a single band of protein with 26.1 kDa. Furthermore, results demonstrated a thermostable lectin presenting activity until 60 °C and lost hemagglutinating activity at 80 °C. According to the obtained data it can be inferred that the ATPS optimization using RSM approach can be applied for recovery and purification of lectins.
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