Effect of the activity levels of the added proteolytic enzyme mixture on free amino acids in ripening Ossau-Iraty cheese

A proteolytic enzymatic preparation (using one of three enzyme concentrations and, hence, one of three different enzymatic activity levels) was added (before clotting) to the milk used to manufacture Ossau-Iraty ewes'-milk cheese. The free amino acids were analysed by reversed-phase high-performance liquid chromatography and the sulphosalicylic acid-soluble N fraction was quantified by the trinitrobenzenesulphonic acid method for use as an index of proteolysis during ripening. Sensory analysis of the cheeses began after two months of ripening. Use of the enzymatic preparation increased the rate of release of amino acids in an amount proportional to the enzyme concentration employed. The effect of the preparation was more pronounced in the early months of ripening, with the differences in the free amino acid contents of the various batches decreasing as ripening progressed. Levels of certain free amino acids, such as taurine, tyrosine and valine, were virtually unaffected by the addition of the enzymatic preparation, whereas levels of such amino acids as serine, glycine, arginine and proline were reduced. Texture defects in the cheeses were observed, namely, reduced elasticity and creaminess and increased brittleness. Similarly, enzymatic treatment also gave rise to bitter flavours that were not characteristic of the normal taste and aftertaste of Ossau-Iraty cheese and these changes were proportional to the quantity of enzyme added.