Aplicação de enzimas coagulantes de leite de origem fúngica e comercial na obtenção de queijos caprinos minas frescal e avaliação da atividade biológica de seus peptídeos

Abstract

With the increase in public debate on contemporary issues such as population aging and people-related illnesses, the general interest in food and its effects on human health has been increasingly aroused. One of the food products that can act to prevent and reduce some pathologies is the cheese, which presents bioactive peptides capable of modulating a series of reactions in an animal organism. Therefore, it was the objective of this study to obtain and apply Aspergillus tamarii URM6599 milk coagulant enzymes and commercial origin in the preparation of Minas Frescal goat cheeses and to evaluate the biological activities of their peptides. For that, it was necessary to produce and optimize the production of coagulant enzymes by means of the factorial statistical plans 2³ complete and central composite design; to evaluate the production of aflatoxins (B1, G1, B2 and G2) by the microorganism and the toxicity of the enzymatic extract by means of bioassay with Artemia salina and to characterize the obtained enzymatic extract for the optimum pH and temperature, thermal stability and pH, NaCl and CaCl2 and to the effects of ions and inhibitors on coagulant activity. After obtaining and analyzing the enzymatic extract, the cheeses were produced and their peptides extracted for evaluation of antioxidant, antihypertensive and antimicrobial activities. From the statistical planning, the enzymatic production was optimized for an activity of 289.16 U.mL-1; the microorganism did not produce aflatoxins in any of the tests carried out and the LD50 of the A. salina nauplii was 683 μg.mL-1, considered of low toxicity. The enzymes presented optimum performance at 70 ºC and pH at 5.5; activity above 50% at temperatures below 60 °C and activity above 60% at acid and alkaline pH values for 1 hour. The enzymes were inhibited by PMSF and pepstatin A, suggesting the existence of serine and aspartic proteases in the crude extract. The enzymes were also tolerant to metal ions, NaCl and CaCl2. The peptides of both cheeses presented antioxidant, antihypertensive and antimicrobial activities. In view of the above, it is observed that the coagulant enzymes produced in the study demonstrate potential use in the elaboration of Minas coolcal goat cheese. In addition, the results demonstrate that Minas Frescal type cheese produced with goat milk is an excellent source of bioactive peptides.