A systematic study was made of the ability of aminopeptidase N from Lactococcus lactis subsp, cremoris Wg2 to hydrolyse different peptide substrates. The enzyme showed a marked preference for substrates containing arginine as the N-terminal residue but, to a lesser extent, was also capable of cleaving other residues such as lysine and leucine. There was a tendency for the activity to increase with the hydrophobicity index of the C-terminal residue of dipeptide substrates. It was also observed that the enzyme tended to have higher affinities but lower V-max values for tripeptides with hydrophobic C-terminal residues. The values determined for K-m and V-max increased with chain length for oligopeptides of the general formula Lys-Phe-(Gly)(n), the optimum, as determined from V-max/K-m, being when n = 3. Typical K-m values for the most effective substrates were in the range 0.2-0.6 mM.