Bitter taste of enzymic hydrolysates of casein. I. Isolation, structural and sensorial analysis of peptides from tryptic hydrolysates of beta-casein.

beta-Casein A2 was isolated from milk of a homozygous cow and hydrolysed with trypsin. The hydrolysate was separated by RP-HPLC into 18 peptides, all but one of which could be attributed to the sequence of beta-casein on the basis of the amino acid composition. Some peptides overlapped. In total, they represented about 97% of the protein sequence. Only three peptides had a bitter taste, namely I49-N68 (recognition threshold 1.0 mg/ml, 0.45 mmol/l), I49-K97 (1.5 mg/ml, 0.28 mmol/l) and G203-V209 (0.175 mg/ml, 0.23 mmol/l). The contribution of the three peptides to the overall bitterness of the beta-casein hydrolysate (2.67 mg/ml) was about 11, 21, and 60%, respectively. Peptide I49-K97 was present in the hydrolysate together with its fragments I49-N68 and S69-K97. Remarkably, the smaller and more hydrophobic fragment I49-N68 was less bitter than I49-K97 on a molar basis, whereas the larger and more hydrophilic fragment S69-K97 had a neutral taste. These results show that in the case of larger peptides neither hydrophobicity nor size are responsible alone for bitter potency, but that conformational parameters must be of great importance. Furthermore, it can be concluded that only a part of the structure is responsible for the contact with the receptor. The bitterness of G203-V209 is discussed in connection with related synthetic peptides in the literature.