Proteolytic and lipolytic activity:

Proteolytic and lipolytic activity: Proteolytic activity was assessed using the agar plate assay method 20. Briefly, an overnight culture of each strain (MRS broth, 24 h, 30°C ) was centrifuged at 13,000 g for 5 min and the pellet was resuspended in 20 mM phosphate buffer (pH 7.0). Cell suspension (10 µl) was spotted on the surface of MRS agar supplemented with 10% skimmed milk. After incubation at 30°C for 48 h, the proteolytic activity was determined by measurement of the diameter of the clear zone surrounding the inoculated spots (mm). All tests were carried out in triplicate. Lipolytic activity was assessed by the agar plate on MRS agar supplemented with neutral red and 15% of pork fat 21. Overnight culture (100 µl) was spread on the surface of MRS agar with 15% pork fat as described above. Plates were incubated at 30°C for 48 h, and the red spots, formed by lipolytic strains, were measured. All tests were carried out in triplicate.