Design of an innovative technique for application of the immobilized Rhizomucor miehei (CBS: 370.65) rennin-like enzyme on paraffin wax in cheese-making process and the kinetic properties of the immobilized enzyme.

ABSTRACT

This research aimed to invent a new method for cheese making using Rennin-like enzyme from fungus with high efficiency and reusability. Accordingly, Rhizomucor miehei (CBS 370.65) showed a promising milk clotting (MCF) activity and the mycotoxin test was negative. The partially purified enzyme was immobilized by entrapment in paraffin wax using different techniques. Wax-enzyme tablets preparation exhibited complete immobilization yield (100%). Ca2+ had a marked stimulating effect on the activities of both the free and immobilized enzyme forms. The immobilized enzyme (MCI) exhibited more than sixteen effective reuses to produce cheese in a batch reactor. The free and the immobilized forms recorded their optimum activities at pH 5.6 and 55 °C, respectively. The immobilization process reduced the consumed activation energy (Ea) to 39%. The immobilized enzyme was more stable than the free form. Among all the used substrates, buffalo milk and full cream milk showed the highest immobilized enzyme activity (7142.9 U). km value was unaffected by the immobilization process and was 600 mg reaction-1, for both. Schematic setup was used as semi-pilot example for a repeated batch of MCI wax tablets. This design solved the clotting problem completely by the refine bundle nominated its agreeability in the cheese-making process.