A comparative study on different culture media for the production of a rennin-like enzyme from Rhizomucor pusillus 1653 was carried out for preparation of an economic medium to be used for production of rennin like enzyme from Rhizomucor pusillus 1653. The investigated media were prepared mainly from whey permeate supplemented with oilseed meals as sources of nitrogen. The oilseed meals included: sesame meal, sunflower meal, peanut meal, cottonseed meal, as well as two degossypolized cottonseed meals. Results, revealed that the ratio of milk clotting activity (MCA) to proteolytic activity (PA) was taken as an index for the selection between different media. On this basis results showed that the milk clotting enzyme from Rhizomucor pusillus 1653 was produced by all the media supplemented with oilseed meals in the following order: Sesame meal > sunflower meal > cottonseed meal (A:H:W)>cottonseed meal (acetic: ether)>cottonseed meal > peanut meal. The activity of milk clotting enzyme resulting from whey permeate supplemented with sesame meal decreased rapidly as heat treatment increased over 55°C and pH 6.0. Furthermore, the enzyme activity decreased gradually as Nacl concentration increased but was improved greatly by increasing the  calcium ion concentration. Utilization of the Rhizomucor pusillus rennet proved to be compatible with calf rennet for the preparation of Ras cheese slurry.