Purification of microbial Rennet from Mucor Miehei

In order to make cheese an enzyme preparation ,called rennet, is added to milk. Rennet hydrolysis the phe(105)-met(106) bound in kappa casein. Result of this cleavage is curd production. Three common experimental techniques were used in purification process of Rennet in the present study. Clear culture supernatant was separated from mycelia of Mucor Miehei by centrifugation at 5000 rev/min for 15 min and was concentrated in two stages. In the first stage, an ultrafiltration system with membranes of 20000 dal. por sizes , was used. In this stage total activity of enzyme was recovered. In the second stage ammonium sulfate salt, in six different saturation percents(30,40,50,60,70 and 80) were studied. Protein samples which were precipitated by 80% (w/v) ammonium sulfate show the best result and sixty percent of total enzyme activity were recovered. These samples were loaded on DEAE-sepharose® ion exchange chromatography in batch Results showed that using these steps, with 55 percent recovery, the enzyme was purified 15.33 times. mode

کلیدواژه ها:

Cheese ، Rennet ، Mucor Miehei ، ion exchange chromatography