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Brazilian Journal of Chemical Engineering

Print version ISSN 0104-6632On-line version ISSN 1678-4383


FALCO, A.L.P.; DURRANT, L.R.  and  FRANCO, T.T.. Purification of a-galactosidase from seeds of Sesbania marginata. Braz. J. Chem. Eng. [online]. 2000, vol.17, n.4-7, pp.819-826. ISSN 0104-6632.

Alpha-galactosidase taken from a raw extract of Sesbania marginata legume seeds was purified by partitioning in aqueous two-phase systems (ATPS). Initially, galactomannan/dextran 2,000,000 systems were used for the purification, and the partition coefficients of a -galactosidase varied from 1.5 to 4.0. However, mass transport in these systems was poor due to the high viscosity of the employed polymers. Therefore, partitioning in polyethyleneglycol (PEG)/ sodium phosphate systems and the effect of sodium chloride upon the enzyme purification and the yield of a -galactosidase were also investigated. The purification achieved in a single-step was 5.7 with a recovery of 144% of a -galactosidase, possibly due to the removal of materials which inhibited a -galactosidase activity before the purification. The removal of the main protein contaminants and the highest yields were achieved in PEG 4,000/ sodium phosphate + 6% NaCl system at pH 5.0. Further purification by preparative on-exchange chromatography was also developed.

Keywords : alpha-galactosidase; legume seeds; enzyme purification; aqueous two phase systems.

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