Author(s):
Nascimento, Cynthia Oliveira ; Costa, Romero Marcos Pedrosa Brandão ; Soares, Paulo Antônio ; Herculano, Polyanna Nunes ; Porto, Tatiana Souza ; Nascimento, Thiago Pajeú ; Lima, Carolina de Albuquerque ; Bezerra, Raquel Pedrosa ; Teixeira, J. A. ; Coelho, Luana Cassandra Breitenbach Barroso ; Correia, Maria Tereza dos Santos ; Carneiro-da-Cunha, Maria das Graças ; Porto, Ana Lúcia Figueiredo
Date: 2020
Persistent ID: https://hdl.handle.net/1822/66919
Origin: RepositóriUM - Universidade do Minho
Subject(s): ATPS; Cratylia mollis seeds; Lectins; Partitioning; Purification factor
Description
The 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 (15008000g/mol), PEG concentration (12.517.5% w/w), phosphate (1015% w/w) concentration, and pH (68) 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 BoxBehnken 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 sulfatepolyacrylamide gel electrophoresis showed a single band of protein with 26.1kDa. 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.
