Separation and characterization of a water soluble protein fraction from Moringa oleifera seeds

Cristóbal Lárez Velásquez


A simple and relatively inexpensive method was developed in order to separate and purify the water soluble fraction of proteins present in dry dehulled Moringa seeds directly collected from local trees. Procedure includes: (a) extraction of the oil fraction from M. oleifera seeds using petroleum ether at room temperature; (b) aqueous extraction of defatted seeds; (c) fractionation of the resulting aqueous phase by cooling at 4 oC and separating the precipitated solid by centrifugation; and (d) freezing and vacuum drying of the wet solid. Protein fraction obtained consists of a mixture of molecular species of different sizes, with those around 26.5, 21.0 and < 14.2 KDa present in greater proportion, which shows interesting acid/base properties. Moreover, two important aspects have emerged from analyses performed to this protein sample: (a) a slightly acidic character was observed by potentiometric and conductimetric titrations which could be justified by the presence of tyrosine phenolic groups in the protein chains forming the mixture, as it has been inferred by FTIR studies, and (b) bathochromic displacements observed during UV-visible studies for the signal associated to π → π* transitions of the peptide bonds can be attributed to conformational changes which modify natural α-helix structure of the protein, especially those caused by rupture of hydrogen bonds probably involving phenolic groups of tyrosine.


Moringa seeds; Protein conformational changes; Conductimetric/Potentiometric titrations; FTIR characterization; Electrophoretic profiles

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