Proteolysis of milk during storage by two strains of Pseudomonas NCIMB 702085 (414) and NCIMB 701274 (416) was investigated using the Urea PAGE method. Pseudomonas fluorescens enzymes were also extracted and purified by dialysis before inoculation into UHT skim milk in an attempt to partially purify the enzyme. Results showed that dialysis removed some peptides and amino acids which would interfere with the assay procedure. The method also confirmed that Pseudomonas fluorescens NCIMB 701274 (416) was more proteolytic than Pseudomonas NCIMB 702085 (414). Thus, Urea PAGE is a useful method for monitoring proteolysis in milk by Pseudomonas fluorescens.