The production of rennet was studied, using different strains of the fungus Rhizomucor miehei. The selection and preservation of strains, type of growth, media design and operation conditions were evaluated. The experiments were carried out in Erlenmeyer flasks in rotary shaker at 250 rpm and 2.5 eccentricity, and in mechanically stirred fermentors of the New Brunswick type, at 30 degrees C. In the studies concerning strain selection, the best strain was Rhizomucor miehei NRRL 3169. The major titles of enzyme were obtained in batch process at 168 h, with 884 SU/ml, whereas in mechanically stirred fermentors the best value was 1160 SU/ml. These values were far more superior to former ones published by various experts.