The limitations of rennin application in cheese crafting usually urge the discovery of novel proteases. The Withania coagulans fruits are well-known for their caseinolytic activity in cheese production. The study aims to evaluate some of the factors affecting the milk-clotting activity (MCA) of W. coagulans fruit enzymatic extract in even and odd-toed hoofed-mammals milk. The extracts were prepared by distilled water and normal saline and their protein content were evaluated. The time necessary for the appearance of discernible and discrete particles in the mare and cow milk by the two concentrations of saline and hydro W. coagulans extracts (SE and HE, respectively) was assessed at 35 and 40°C while exposing different levels of calcium chloride. The interaction of these factors on MCA was evaluated using mixed-design ANOVA. Three significant interaction patterns considering the maximum number of factors were revealed (p < 0.05). The higher extract concentration and incubation temperature (40°C) was always effective in producing the utmost MCA in these interactions. The SE was faster than HE in milk clot formation. The cow milk was a more suitable substrate than mare milk for the enzyme activity.
Withania coagulans, milk-clotting activity, cow milk, mare milk
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