Behavior of Aspartic Proteases From Artichoke Flower ( <i>Cynara cardunculus L. var Scolymus</i> ) in the Hydrolysis of Buffalo Milk Casein

The objective of this study was to assess the activity of proteases from artichoke flower extract in the enzymatic hydrolysis of casein from buffalo milk and to investigate its potential application in cheese production. Artichoke flower extract and microbial chymosin were assessed for their proteolytic activity (PA) and milk clotting activity (MCA) under varying conditions of pH, temperature, CaCl₂, and NaCl concentration. The peptides obtained from the PA were analyzed using reverse phase high performance liquid chromatography, while the clots obtained from the MCA were analyzed through electrophoresis and mid-infrared spectroscopy. Mini cheeses were produced under optimal conditions established by the specificity ratio (SR) value to evaluate the technological application of artichoke flower extract. The artichoke flower extract showed optimal PA conditions at pH 6.15 and temperature 45.2°C, and for MCA it showed maximum activity at pH 5.8 and temperature 56.9°C, values close to those found for chymosin. In the RP-HPLC, SDS-PAGE, and MIR, it was observed that the extract hydrolyzed casein in peptide bonds other than chymosin; however, its yield in cheese production remained unchanged. The artichoke flower has demonstrated potential as coagulant for buffalo cheese production, offering an alternative means to add economic value to these products.