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A new method for the detection of microbial proteolytic enzymes in milk

SUMMARY Optimum conditions have been determined for the use of Hide Powder Azure (HPA) to estimate extracellular proteinases produced by psychrotrophic Gram-negative bacteria. This assay is quicker, more precise and 3·2 times more sensitive than assays of soluble N released from casein. It provides a simple alternative method for monitoring proteinase purification schemes. With the modification described it is now possible to determine proteinase activities directly in milk. In its simplest form the HPA assay is sufficiently sensitive to detect proteinases produced by as few as 1·5 x 106 c.f.u. ml-1 psychrobtrophs without the necessity for any sample processing.