Heat-Stable Proteases from Psychrotrophs in Milk

Twelve gram-negative psychrotrophic bacteria producing heat-resistant proteases that hydrolyzed casein were isolated from refrigerated raw milk. All were pseudomonads and the enzymes of the six most proteolytic cultures were examined further. The proteases were partially purified, and gel electrophoresis indicated that only a single enzyme was present in the preparation. The molecular weight of most of the proteases was approximately 45,000. All six enzymes retained some activity after being heated at 121 C for 10 min and casein was hydrolyzed at pH levels found in normal milk and many cultured dairy products. Although proteolysis was highest at about 40 C, considerable activity was evident at refrigeration temperatures.