On the mechanism of milk clotting by rennin

Abstract

SummaryTwo possible hypotheses for the mechanism of milk clotting were tested. The results obtained constituted strong evidence against one and suggested that the second is improbable. Milk was separated into a 5-fold-concentrated casein micelle suspension and milk serum. Pre-renneting of the serum did not reduce the rate of clotting on subsequent addition to the micelle suspension whether or not the conditions were such that the para-κ-casein became extensively aggregated. Washing of casein micelles up to 3 times with milk dialysate at 23°C extracted very little casein from the micelles and did not increase the clotting time of micelles resuspended to about the same concentration as in milk. The results appear to constitute decisive evidence against the hypothesis of milk clotting proposed by Parry & Carroll (1969). S-carboxymethyl-κ-casein, S-carboxymethyl-κ-casein containing 2·5 dimethylaminonaphthalene sulphonyl residues per mole, and rennin-treated dimethylaminonaphthalene sulphonated-S-carboxymethyl-κ-casein all bound Ca to the same extent at 30°C and pH 6·5, over the range 0·5–15·3 mM-CaCl2. This adds support to existing evidence that milk clotting does not involve formation of Ca bridges between casein micelles.