A New Affinity Matrixe Synthesized from Aminobenzohydrazide Derivatives for Purification of Lactoperoxidase Enzyme

In this study, a new affinity process was developed for the purification of Lactoperoxidase with synthesized sixteen aminobenzohydrazide derivatives. For this purpose, ligands were covalently bound to CNBr‐activated Sepharose‐4B−L‐tyrosine matrix and affinity columns were prepared, and LPO was purified in one step with high yield and purity. Among all synthesized molecules, the 4‐amino‐3‐bromo‐2‐methylbenzohydrazide molecule had a high usable potential in the purification of Lactoperoxidase from mammalian milk. Lactoperoxidase was purified 411.8 times with a yield of 17.38 % from goat milk, 187.25 times with a yield of 9.72 % buffalo milk, 2772.4 times with a yield of 18.98 % from bovine milk, and 1246.65 times with a yield of 4.43 % from sheep milk. It was demonstrated for the first time that aminobenzohydrazide molecules could be used as ligands in the purification of Lactoperoxidase enzyme. The lactoperoxidase system has a very wide range of application areas, such as in the dairy industry, the preservation of milk during transport, the collection of raw milk and delivery to processing centers. In this context, a new affinity process was developed for the purification of Lactoperoxidase with synthesized aminobenzohydrazide derivatives.