A Simplified Method for the Purification of an Intrinsically Disordered Coagulant Protein from Defatted Moringa oleifera Seeds
Document Type
Article
Publication Date
2016
Digital Object Identifier (DOI)
https://doi.org/10.1016/j.procbio.2016.04.021
Abstract
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.
Was this content written or created while at USF?
Yes
Citation / Publisher Attribution
Process Biochemistry, v. 51, issue 8, p. 1085-1091
Scholar Commons Citation
Dezfooli, Seyedehsara M.; Uversky, Vladimir N.; Saleem, Mussarat; Baharudin, Farah Salma; Hitam, Sharifah M.; and Bachmann, Robert T., "A Simplified Method for the Purification of an Intrinsically Disordered Coagulant Protein from Defatted Moringa oleifera Seeds" (2016). Molecular Medicine Faculty Publications. 364.
https://digitalcommons.usf.edu/mme_facpub/364
