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1 CoRFiLaC, Regione Siciliana, 97100 Ragusa, Italy
2 Department of Food and Nutritional Sciences, University College, Cork, Ireland
3 Dipartimento di Scienze Agronomiche, Agrochimiche e delle Produzioni Animali, Catania University, 95100 Catania, Italy
Corresponding author: V. Fallico; e-mail: enzofallico{at}corfilac.it.
The appearance of undesirable bitter taste in Ragusano cheese was investigated by comparing the composition of 9 bitter cheeses with that of 9 reference cheeses of good quality by means of chemical, electrophoretic, and chromatographic analyses. Rates of proteolysis were significantly affected in cheeses of different quality. Primary proteolysis, as measured by pH 4.6-soluble N, was significantly greater in bitter cheeses compared with reference samples. Urea-PAGE profiles showed an almost complete breakdown of caseins in bitter cheeses and the further degradation of primary peptides into smaller compounds not detectable by this technique. Cheeses with defects had significantly lower levels of secondary proteolysis as reflected by the percentage of pH 4.6-soluble N soluble in 12% trichloroacetic acid and the amounts of total free amino acids. Peptides separated by reversed phase-HPLC revealed that the large and significant differences in peptide profiles of the soluble fractions between bitter and reference cheeses were mainly due to a much higher proportion of hydrophobic peptides in the former. The occurrence of bitterness in Ragusano cheese was therefore attributable to unbalanced levels of proteolysis and peptidolysis. Extensive degradation of caseins and primary peptides by activities of proteases produced large amounts of small- and medium-sized hydrophobic peptides that were not adequately removed by peptidases of microflora and therefore accumulated in cheese potentially contributing to its bitter taste. The presence of these compounds in bitter cheeses was related to high salt-in-moisture and low moisture contents that limited the enzymatic activities of microflora important in secondary proteolysis. Combining salt-in-moisture and the ratio of hydrophobic-to-hydrophilic soluble peptides resulted in the best logistic partial least squares regression model predicting cheese quality. Although bitterness is known to be rarely encountered in cheese at salt-in-moisture levels >5.0, all of the bitter cheeses analyzed in this study had salt-in-moisture levels much greater than this value. According to the logistic model, a risk of bitterness development may exist for cheeses with a midrange (5 to 10%) salt-in-moisture content but with an inadequate level of secondary proteolysis.
Key Words: Ragusano cheese • bitterness • hydrophobic peptide • secondary proteolysis
Abbreviation key: EIN = 70% ethanol-insoluble nitrogen, ESN = 70% ethanol-soluble nitrogen, FAA = free AA, HHIP = ratio of hydrophobic-to-hydrophilic insoluble peptides, HHSP = ratio of hydrophobic-to-hydrophilic soluble peptides, PDO = Protected Denomination of Origin, pH4.6SN = pH 4.6 acetate buffer-soluble N, RP-HPLC = reversed-phase HPLC, S/M = salt-in-moisture, TCASN = 12% TCA-soluble N
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