The objective of the investigations was to compare the hydrolytic activity of enzymatic preparations obtained from post-culture fluids of two yeast strains: Yarrowia lipolytica PII6a and A-101, and to test the possibility of using them in the hydrolysis of pigskins. The enzymatic preparations were obtained from the yeast cultured in media with two different sources of carbon (oil or glucose). It was stated that the collagenolytic activity of all enzymatic preparations investigated was low. Their proteolytic and lipolytic activity was determined, and, on this basis, the preparations obtained from the PII6a strain were selected for the hydrolysis of pigskins. The efficiency of enzymatic hydrolysis of pigskins, expressed as an amount of: released hydroxyproline, free amine groups, and protein, was the highest in the case of applying a mixture of enzymatic preparations obtained from the cultures developed in the two media as indicated above. Moreover, it was stated that the appropriate selection of the fitting process conditions (temperature, pH, and raw material comminution) might significantly enhance the process efficiency. The results of the investigations performed confirm that this inexpensive source of enzymes could be widely used for the degradation of proteins in pigskins.