The catalytic and kinetic characterization of Bacillus subtilis MK775302 milk clotting enzyme: comparison with calf rennet as a coagulant in white soft cheese manufacture
Background
Calfrennetis considered the traditional source of milk clotting enzyme (MCE). However, increasing cheese consumption with decreasing the calfrennetsupply had encouraged the quest for new rennet alternatives. The purpose of this study is to acquire more information about the catalytic and kinetic properties of partially purifiedBacillus subtilisMK775302 MCE and to assess the role of enzyme incheese manufacture.
Results
B. subtilisMK775302 MCE was partially purified by 50% acetone precipitation with 5.6-foldpurification. The optimum temperature and pH of the partially purified MCE were 70 °C and 5.0, respectively. The activation energy was calculated as 47.7 kJ/mol. The calculatedKmandVmaxvalues were 36 mg/ml and 833 U/ml, respectively. The enzyme retained full activity at NaCl concentration of 2%. Compared to the commercial calf rennet, the ultra-filtrated whitesoft cheeseproduced from the partially purifiedB. subtilisMK775302 MCE exhibited higher total acidity, highervolatile fatty acids, and improved sensorial properties.
Conclusions
The partially purified MCE obtained in this study is a promising milk coagulant that can replace calf rennet at a commercial scale to produce better-quality cheese with improved texture and flavor.