Attempts were made to eliminate the bacterial factor in Cheddar cheeseripening, through the agency of chloroform, and an assessment of the role of rennet was thus rendered possible.

After investigating the effect of chloroform and toluene upon the proteolytic action of rennet and trypsin in milk, the results obtained were applied in studies upon the ripening of cheese in the presence of chloroform. Cheese of normal manufacture was chloroformed at the salting stage or one week or more after manufacture (when the curd had lost its rubbery nature and the germicide was more easily incorporated). Owing to the weakened activity of enzymes in the presence of germicides, it was necessary to employ larger proportions than usual of rennet, measured additions generally being made at the time when the chloroform was added. Under these conditions the general course of protein degradation, as measured by determinations of soluble nitrogen and non-protein nitrogen, was found to be identical with that occurring in normal control cheeses. Strong evidence was thus afforded that rennet is the only important agent attacking cheese protein during the ripening process.

In the partition between the non-protein constituents of the normal and chloroformed cheeses, respectively, there were observed definite differences which tended to remain constant over the greater part of the ripening period. The relatively higher proportion of subpeptone nitrogen in the normal cheese was shown to be due, at least in some measure, to bacterial action.

Cheese ripened in the presence of chloroform developed no volatile acid, and no cheese flavour could be detected.