In the present study, biochemical, chemical and texture changes in Tulum cheeses made using calf rennet and microbial rennets (Aspergillus niger protease and Rhizomucor miehei protease) were compared during ripening for up to 90 days. A total of 15 free fatty acids (FFAs) were detected in the cheese samples. The peroxide values (PV) of the cheeses increased significantly (P < 0.05) during ripening and the cheese made with calf rennet had the highest PV. Proteolysis in the cheeses increased as the ripening time increased. αs1-casein and β-casein degradation was higher in cheeses manufactured with R. miehei protease. Cheeses made with calf rennet were significantly (P < 0.05) harder, more adhesive, more cohesive and more resilient than those made with microbial rennet.