Purıfıcatıon of protease enzyme from çiğdem (crocus bıflorus) tubers and ınvestıgatıon of usabılıty of the purıfıed protease enzyme ın coagulatıon of caseın

It is known that in some diseases (SARS, mad cow, pig and bird flu) that have passed from animals to humans have increased in recent years. In consumers, the use of animal proteases in the production of food products leads to concerns about the spread of these diseases. For these reasons, it has been an increase in demand for the product obtained from plant sources. The fact that only a small proportion of commercially available enzymes are herbaceous, have encouraged researchers to search for new herbal protease sources that can be used commercially. In this study, for the first time protease enzyme was purified from the tubers of a Crocus biflorus and characterized. The protease enzyme was purified 86 fold by ammonium sulfate precipitation and ion exchange chromatography. Optimum pH and temperature was found as 5.5 and 50 °C. The values of Vmax and KM were calculated as 47 mg/mL.min and 0.9 g/L, respectively. The molecular weight of the enzyme was calculated to be 35 kDa by using SDS-PAGE. It was determined that the purified enzyme can be used in the process of casein coagulation, the first step of cheese making.