A Simplified Method for the Purification of an Intrinsically Disordered Coagulant Protein from Defatted Moringa oleifera Seeds

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Coagulant proteins from Moringa oleifera (MO) seeds were successfully purified by removing seed oil followed by a protein salting out method at 40% (NH4)2SO4 combined with subsequent dialysis and heat treatment. A microtiter plate-based coagulation activity assay was then performed using colloidal bentonite solution and alum as positive control. The results show an oil recovery of 40 ± 2% while the water-soluble protein fraction in the defatted MO seed was 12.5 ± 0.5 mg/g or 4.4% of the total seed protein content. Heating the desalted protein fraction to 121 °C helped to obtain a 94% pure protein of approximately 7 kDa. Computational analyses support the hypothesis that the coagulating MO protein is an intrinsically disordered protein, providing reason for its stability at extreme temperatures (121 °C). The purified, thermo-stable coagulating protein retained its coagulating activity paving the way to economically produce a sterile natural coagulant from MO seeds.

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Process Biochemistry, v. 51, issue 8, p. 1085-1091