INVESTIGATING THE ENZYMOLOGICAL PROPERTIES OF CASEASE FROM VARIOUS ORGANISMS: KINETICS, INHIBITION, SUBSTRATE SPECIFICITY AND METAL ION DEPENDENT ACTIVITY

Proteolytic enzymes like casease are essential biological catalysts, valued for their ability to break down complex proteins such as casein into smaller, more digestible molecules. Among these, casease a serine protease holds major industrial importance, particularly in the food, pharmaceutical, and environmental sectors, owing to its role in protein hydrolysis and nutrient release. In this study, we isolated, produced, and characterized casease from two bacterial species, Bacillus subtilis and Serratia marcescens, both known for efficient extracellular enzyme secretion. Enzyme activity was confirmed by the formation of clear zones on skim milk agar signifying successful casein degradation. Comparative profiling revealed distinct production dynamics. Bacillus subtilis generated casease early, peaking at 24 hours, while Serratia marcescens showed delayed but more intense activity, reaching its maximum at 48 hours. Characterization studies indicated that Serratia marcescens had higher overall proteolytic activity. The response of enzymes to chemical inhibitors like hydrogen peroxide and hydrochloric acid suggested structural and functional differences between the two. Both enzymes also displayed broad substrate specificity, acting on proteins such as gelatin, glycinin, and zein.Furthermore, enzymatic efficiency was significantly impacted by the presence of metal ions including calcium, copper, iron, magnesium, and zinc which could enhance or inhibit activity. These findings illustrate that both Bacillus subtilis and Serratia marcescens are effective producers of extracellular casease with promising applications in food technology, pharmaceuticals, and environmental biotechnology, particularly for sustainable waste management and protein-rich waste biodegradation.