PARTIAL PURIFICATION OF MILK-CLOTTING ENZYME FROM THE SEEDS OF MORINGA OLEIFERA

Abstract

The aim of the present study was to search for milk clotting substitute from different parts (flowers, seeds, stem, leaves, ripe and unripe fruits) of Moringa oleifera. The samples were blended and extracted using different types of extracting solutions. The most reliable, quick and efficient enzyme extracting solution was found to be 5% NaCl in 100 mM sodium acetate buffer, pH 5.0, which was used throughout the study. The milk clotting activity was only observed in the seeds extract while the other parts were either deficient or has very low milk clotting activity. Thus, the moringa seeds were used as source of milk clotting enzyme. The extracted proteins were fractionated with ammonium sulfate at concentration of 20, 30, 40, 50 and 60 %. Highest milk clotting activity was observed in the 20 % fraction. This fraction was assumed to contain the clotting enzymes and characterized for its heating stability (30 – 90°C) and optimum temperature (30 – 90°C). The results demonstrated that moringa seeds milk clotting enzyme is stable up to 50°C with an optimum milk clotting activity of 70°C. The high ratio of milk-clotting to proteolytic activity of the partially purified enzyme indicates the potential of this enzyme as suitable rennet substitute in dairy industry. However, further study is needed to completely purify and characterize this promising milk clotting enzyme from moringa seeds.