Estimation of Residual Pepsin and Chymosin Activity in Curd,

Pasteurized milk (225 g) adjusted to pH 6.2 was set with 3.5 milk clotting units of chymosin (EC 3.4.23.4). The same amount of milk at pH 5.8 was set with 3.5 milk clotting units of porcine pepsin (EC 3.4.23.1). Fifteen minutes after clotting, the curd was broken, and curd and whey were separated by centrifugation at 3500 × g for 20 min. The curd (30 g) was extracted at pH 6.8 in 450 ml water or at pH 6.2 (chymosin) or 5.8 (pepsin) in 450 ml 1 M sodium chloride.Chymosin was completely released from the curd and accounted for by both methods of extraction. Pepsin was completely released and accounted for after extraction in 1 M sodium chloride at pH 5.8 but was partly inactivated during extraction at pH 6.8.Assay of curd extracts and whey by a linear agar diffusion test accounted for 102 ± 6% of the pepsin activity added to milk when the curd was extracted in 1 M sodium chloride. Extraction at pH 6.8 allowed recovery of only 63% of the activity. Chymosin recovery was 100 ± 5% by both methods of curd extraction.