Milk Enzymes

There are several enzymes in bovine milk. They occur in various states including free in solution, as associated or integral parts of membrane fractions of the fat globule membrane or membrane vesicles (in skim milk), and associated with casein micelles or microsomal particles. Approximately 60 indigenous enzymes are excreted from secretory cells in the mammary gland including lipoprotein lipase, plasmin, phosphatases, lactate dehydrogenase, catalase, lactoperoxidase, and xanthine oxidase. Several have important functions in dairy processing and quality control including lipoprotein lipase, plasmin, and alkaline phosphatase. Esterases also occur in milk. Three types have been determined: the A - type carboxylic ester that hydrolyzes aromatic residues, the B - type that hydrolyzes alphaphilic esters rapidly and aromatic esters slowly, and the C - type that hydrolyzes alkaline esters. Several enzymes in milk have antibacterial functions including lysozyme, lactoferrin, and lactoperoxidase. The enzymes with major functions in milk and milk processing are discussed below.

 Lysozyme Lysozyme consists of 148 amino acids and it is a relatively small, single polypeptide protein. It is found in low concentrations in bovine milk (0 to 2 mg/L). This enzyme s level rises significantly during infection with mastitis. Lactoperoxidase The lactoperoxidase enzyme consists of 712 amino acids and has antimicrobial activity. Its optimum activity is at pH 6.0, but it is stable over pH 5.0 to 10.0. Higher concentrations of this enzyme are present in bovine milk compared to human milk. It chelates to metals including iron.

 Oxidoreductases Milk contains oxidoreductases such as xanthine oxidase, sulfhydryl oxidase, and superoxide dismutase. Xanthine oxidase is a dimeric metallo - avoprotein that associates with the fat globule membrane and is approximately 30 kDa. It is a nonspeci c oxidoreductase that catalyzes the oxidation of a variety of substances, including nitrate to nitrite in cheese, which inhibits the growth of butyric acid bacteria. Higher concentrations are present in bovine milk than human milk. Superoxidase dismutase is located in the milk serum and catalyzes the dismutation of the superoxide anion to hydrogen peroxide and triplet oxygen. It may inhibit the oxidation of milk constituents including the autoxidation of lipids. Sulfhydryl oxidase is associated with lipoproteins in the fat globule membrane. It catalyzes the oxidation of sulfhydryl groups to disulfides and is inactivated partially by pasteurization.

 Plasminogn and Plasmin Plasminogen is the principal proteolytic enzyme in milk. It remains inactive until converted to plasmin via plasminogen activators. There are three plasmin types. Plasmin is responsible for the production of - casein and proteose peptone by hydrolysis of - casein. It also hydrolyzes many other proteins. Its optimal activity is at 37 C and pH 8.0; however, approximately 82% of the enzyme s activity is lost after pasteurization and is reduced by UV light. The concentration of plasminogen and plasmin is affected by lactation stage, disease status, diet, age, breed, and hormone use.

Lipoprotein Lipase Lipoprotein lipase is the principal lipase in milk bound mostly to casein micelles. It is a glycoprotein that is approximately 10 kDa, which hydrolyzes triglycerides and diglycerides into fatty acids. Its optimum activity is at 37 C in the pH range of 8.4 to 9.2. It is inhibited by its products (i.e., fatty acids), UV light, heat, acid, calcium chloride, manganese chloride, and oxidizing agents. It is stimulated by the presence of bovine serum albumin (BSA), sodium chloride, magnesium chloride, and calcium. The fat globule membrane protects most lipids from enzyme activity. However, damage to the membrane results in lipase activity and the production of short - chain fatty acids that increase rancidity and produce off flavors, which render milk unacceptable for consumption. Lipolysis can occur from vigorous agitation and homogenization.

Phosphatases
Two main phosphatases occur in bovine milk: alkaline phosphatase and acid phospha-tase. Both hydrolyze phosphoric esters and have roles in the processing of milk and milk products. Alkaline phosphatase is an enzyme situ-ated in the fat globule membrane that hydro-lyzes phosphoric esters. Its optimum activity is at pH 8.5 at 37 C. It is activated by divalent metal ions and inhibited by metal chela-tors, orthophosphates, and pasteurization. Its concentration in bovine milk depends on the individuality of the cow and the lactation stage. Acid phosphatase is an enzyme that hydrolyzes tyrosine phosphorylated proteins and aryl and acyl phosphates; it is approximately 42 kDa. It is predominantly located in the milk serum and its optimum activity is at 37 C and pH 7.9. Its concentration in bovine milk depends on the lactation state and disease status of the cow. Its activity is lower than alkaline phosphatise; however, its heat stability is not affected by pasteurization. Its activity is inactivated by UV light, and inhibited by heavy metals, oxidizing agents, orthophosphosphates, and polyphosphates. It may in uence heat stability in dairy products.

Enzymes in Bovine milk