Direct heat treatment of two milk types, skimmed and nonstandardised full-fat, was performed by instant steam infusion and compared with indirect heating. Infusion conditions were temperatures of 72–120°C combined with holding times of 100–700 ms, and indirect heat conditions were 72°C/15 s and 85°C/30 s. The activity of indigenous enzymes such as alkaline phosphatase, lactoperoxidase, xanthine oxidase and γ-glutamyl transpeptidase was evaluated. Infusion temperature was the main determinant of inactivation. Whey protein denaturation represented by β-lactoglobulin increased significantly with infusion temperature. The nonstandardised milk had a higher denaturation rate than skimmed milk. The effect of instant infusion on pH and milk fat globule size in relation to whey protein denaturation and association is discussed.