COLLATION OF CASEINOLYTIC ENZYMES YIELD FROM BACILLUS CEREUS 13BN USING ABUNDANT TYPES OF MEDIA POTENTIALLY AS BLOOD ANTICOAGULANTS

Abstract

Previously, a proteolytic enzyme producer from one of Malaysian fermented food - belacan was successfully isolated and named Bacillus cereus 13BN. This caseinolytic enzyme is a systemic enzyme, operationally needed to break down the extra amount of fibrin of the blood clots that belongs to the family of serine protease. Considering the potential of this enzyme as an alternative to the commercially available anticoagulant, the best condition for the yield of the enzyme of “B. Cereus 13BN” was optimised by slightly modifying the arrangement of broth of nutrient, using five types of carbon source, which are lactose, galactose, maltose, glucose, and glycerol. Five types of nitrogen sources which include casein, gelatine, and peptone of proteose, ammonium nitrate, and ammonium were also used in modifying the composition of nutrient broth, as well as the influence of different incubation periods. It showed that maltose and casein had constantly increased caseinolytic activity during incubation. Both sources had the highest caseinolytic and activity at 36 hours of incubation, which was 5.824 ± 0.065 U/mL for maltose, and during 72 hours of incubation, which was 3.948 ± 0.772 U/mL for casein, respectively. As for the control, the highest caseinolytic activity was at 36 hours of incubation, which was 1.390 ± 0.074 U/mL respectively. After combining these two sources, the analysis using statistical software showed that the highest caseinolytic enzymes can be optimised during 72 hours of incubation with desirability of 0.540.