Investigation of fresh sheep and goat milk cheese coagulated with aspartic protease produced from artichoke (Cynara scolymus L.) pistil cell cultures with melatonin and salicylic acid treatment

In this study, traditional fresh sheep and goat cheeses were produced by adding artichoke flower extract, artichoke suspension cell culture extracts treated with salicylic acid (50 µM) and melatonin (5 µM), culture extract without salicylic acid or melatonin application (control) and rennin enzyme. In this study, along with the determination of milk clotting and proteolytic activity properties of callus cell culture suspensions developed from artichoke (Cynara scolymus L.) flowers; it was aimed to examine the effectiveness of melatonin (5µM and 10µM) and salicylic acid (50 µM and 100 µM) applications in obtaining aspartic protease in artichoke flower pistil suspension cultures. Milk coagulation activity values of the enzymes were measured in the range of 0.52 - 0.74. The enzymes with the highest proteolytic activity were determined as melatonin supplemented (5µM) and Cynara scolymus L. callus with salicylic acid (50 µM) enzymes, respectively. The effect of enzymes developed by suspended cell culture method was found to be significantly effective on titratable acidity, water soluble nitrogen, ripening index values of goat and sheep cheeses (p<0.05). When the urea-Page results were analysed, it was found that the enzymes obtained from liquid cell culture had a statistically significant (p<0.05) effect on αs-casein and β-casein in goat and sheep cheeses and the difference between other degradation products. The highest degradation level for γ - casein and other peptides (in both types of cheese) was observed in cheese produced with melatonin added enzyme, followed by cheese produced with salicylic acid added enzyme. Hardness values of goat cheeses were determined in the range of 18.88 - 97.68 N. In sheep cheeses, this value was determined as 33.46 - 111.95 N. It was determined that all of the values obtained as a result of texture profile analysis in goat and sheep cheeses were statistically different from the cheeses produced with rennin enzyme. The enzymes prepared with suspended cell cultures caused a significant effect on L*, a* and b* values in both cheese types (p<0.05). However, total mesophilic aerobic bacteria, lactobacilli and lactococci counts in both cheese groups were significantly lower than the other cheese groups in the samples in which salicylic acid and melatonin added enzymes were used. In the study, it was determined that melatonin and salicylic acid applied during the suspended cell culture process had a slight positive effect on both the properties of the enzyme and the cheese qualities, although statistical differences were determined in the analyses. It was determined that the enzymes obtained by artichoke-derived suspended cell culture method could not produce cheese as successful as animal-derived enzymes in terms of chemical, textural and biochemical properties, and the cheeses produced with goat and sheep milk were softer, less acidic, and less mature.