Identification of Protease from Euphorbia amygdaloides Latex and its Use in Cheese Production

Abstract

In this research, protease enzyme was purified and characterized from milk of Euphorbia amygdaloides. (NH4)2SO4 fractionation and CM-cellulose ion exchange chromatography methods were used for purification of the enzyme. The optimum pH value was determined to be 5, and the optimum temperature was determined to be 60 degrees C. The V(max) and K(M) values at optimum pH and 25 degrees C were calculated by means of Linewearver-Burk graphs as 0.27 mg/L min(-1) and 16 mM, respectively. The purification degree was controlled by using SDS-PAGE and molecular weight was found to be 26 kD. The molecular weight of the enzyme was determined as 54 kD by gel filtration chromatography. These results show that the enzyme has two subunits. In the study, it was also researched whether purified and characterized protease can be collapsed to milk. It was determined that protease enzyme can collapse milk and it can be used to produce cheese.

Ginger rhizome as a potential source of milk coagulating cysteine protease

“…Such a high thermal stability shows an excellent scope of the ginger enzyme for use in dairy industry. The optimal temperature was also found to be 60°C for capparin from Capparis spinosa (Demir et al, 2008), protease from Euphorbia amygdaloides (Demir et al, 2005), and Synadenium grantii enzymes (Menon et al, 2002). Procerain from Calotropis procera was optimal in the temperature range of 55-60°C (Dubey and Jagannadham, 2003).…”

“…Plant sources for milk-clotting enzymes have been identified in various studies (Ahmed et al, 2009;Demir et al, 2005Demir et al, , 2008Dubey and Jagannadham, 2003;Lo Piero et al, 2002;Menon et al, 2002;Sousa and Malcata, 2002;Su et al, 2009). Unfortunately, the excessive proteolytic nature of most plant rennet substitutes has limited their use in cheese manufacturing due to lower yields of cheese, bitter flavors and texture defects (Lo Piero et al, 2002).…”

Occurrence and Properties of Proteases in Plant Latices

“…Nevertheless, some of them are rather well characterized (marked with*). The proteolytic enzyme in the latex of Euphorbia amygdaloides L. (Euphorbiaceace) was purified after ammonium sulfate precipitation and ion exchange chromatography on a CM-cellulose column, the molecular mass is 54 kDa and the optimum temperature and pH conditions are 60°C and pH 5, respectively (azocasein as substrate), it was tested for its milk clotting activity and the use in cheese production [78]. Curcain is the protease purified from the latex of Jatropha curcas L. (Euphorbiaceae), a perennial hedge plant grown in India and other tropical countries commonly known as Bagbherenda.…”

The Purification of Protease From Cowslip (Primula Veris) and Its Use in Food Processing

“…1998). The protease from Calotropis procera (family: Asclepiadaceae) was optimal in the range temperature of 55–60C (Dubey and Jagannadham 2003; Demir et al. 2005).…”