title: Qualitative and quantitative determination of proteolysis in mastitic milks.
abstract: Proteolytic activity in mastitic skim-milk was often 5-10 fold higher than in normal milk, its level being related to somatic cell count but not precisely correlated with it. In milks with the highest levels of activity plasmin accounted for about one third of the total proteinase. A further third was sedimented with the micellar fraction together with the plasmin, but unlike plasmin, was not inhibited by addition of soyabean trypsin inhibitor (SBTI). The final third remained in the serum phase. Polyacrylamide gel electrophoresis (PAGE) showed that alpha-sl- and beta-caseins were degraded at about the same overall rate. The plasmin produced the usual readily identified fragments from beta-casein, but incubation of mastitic milk also produced changes in patterns in the gamma-casein region differing from plasmin-induced changes, which were also apparent when the micellar fraction was incubated. As they were inhibited by SBTI, a second trypsin-like enzyme in addition to plasmin may also have been present. Other proteinase(s) not inhibited by SBTI was also associated with casein micelles and produced at least 3 characteristic protein fragments seen on PAGE. The serum phase proteinase(s) was likewise not inhibited by SBTI, and did not produce any well-defined electrophoretic bands, suggesting a rather non-specific breakdown of caseins. After separation of mastitic whole milk, a considerable proportion of the proteolytic activity was found in the cream phase. The proportion was enhanced by freezing and thawing, and the enzyme appeared to be identical to the SBTI-resistant micellar proteinase. Because of the considerable proteolysis likely to occur under the time and temperature conditions involved, our results may provide some explanation for the problems encountered in cheesemaking with mastitic milks (e.g. yield losses, poor curd strength and off-flavour development).
mesh_terms: D000818:Animals; D002364:Caseins / Q000378:metabolism*; D002417:Cattle; D010450:Endopeptidases / Q000378:metabolism*; D004731:Endotoxins / Q000633:toxicity; D004926:Escherichia coli; D005260:Female; D005341:Fibrinolysin / Q000378:metabolism; D005615:Freezing; D006868:Hydrolysis; D008414:Mastitis, Bovine / Q000209:etiology / Q000378:metabolism*; D008892:Milk / Q000166:cytology / Q000201:enzymology*; D010958:Plasminogen / Q000378:metabolism; D013290:Streptococcal Infections / Q000662:veterinary; D013292:Streptococcus agalactiae
chemical_list: D002364:Caseins; D004731:Endotoxins; D010958:Plasminogen; D010450:Endopeptidases; D005341:Fibrinolysin