Hostname: page-component-78c5997874-dh8gc Total loading time: 0 Render date: 2024-11-12T14:28:18.287Z Has data issue: false hasContentIssue false
  • English
  • Français

A simple procedure using trinitrobenzenesulphonic acid for monitoring proteolysis in cheese

Published online by Cambridge University Press:  01 June 2009

Anna Polychroniadou
Anna Polychroniadou
Affiliation:
Laboratory of Dairy Technology, Faculty of Agriculture, University of Thessaloniki, 540 06 Thessaloniki, Greece
Get access Rights & Permissions [Opens in a new window]

Summary

A simple, rapid and sensitive spectrophotometric assay was developed and evaluated for monitoring proteolysis during cheese ripening, based on the fact that α-amino groups released by hydrolysis of cheese proteins react with trinitrobenzenesulphonic acid to form products that absorb strongly at 420 nm. A linear relationship was shown to exist between A420 and concentration of free α amino groups up to 0·5 HIM (r = 0·999, 38 df, P < 0·001). Repeatability of the method was satisfactory. The coefficient of variance was 0·53% for amino acid solutions and 1·19% for cheese extracts. Average recovery of glycine added to the cheese was 104 ± 2·9%. A comparison of the above method with that of determination of water-soluble N to total N ratio showed that there was good agreement between these two methods of assessment of proteolysis in cheese (r = 0·857, 32 df, P < 0·001). Mainly Feta and Teleme cheese were examined, but a similar correlation was obtained with hard Greek cheeses. Analytical conditions of the procedure are discussed.

Type
Original articles
Information
Journal of Dairy Research , Volume 55 , Issue 4 , November 1988 , pp. 585 - 596
Copyright
Copyright © Proprietors of Journal of Dairy Research 1988

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Adler-Nissen, J. 1979 Determination of the degree of hydrolysis of food protein hydrolysates by trinitrobenzenesulfonic acid. Journal of Agricultural and Food Chemistry 27 1256–1262CrossRefGoogle ScholarPubMed
Alichanidis, E., Anifantakis, E. M., Polychroniadou, A. & Nanou, M. 1984 Suitability of some microbial coagulants for Feta cheese manufacture. Journal of Dairy Research 51 141–147Google Scholar
Alichanidis, E., Polychroniadou, A., Tzanetakis, N. & Vafopoulou, A. 1981 Teleme cheese from deep-frozen curd. Journal of Dairy Science 64 732–739Google Scholar
Arnott, D. R., Morris, H. A. & Combs, W. B. 1957 Effect of certain chemical factors on the melting quality of process cheese. Journal of Dairy Science 40 957–963CrossRefGoogle Scholar
Association Of Official Analytical Chemists 1980 Official Methods of Analysis of the AOAC. 13th edn p. 265 (Ed. Horwitz, W.). Washington, DC: Association of Official Analytical ChemistsGoogle Scholar
Beeby, R. 1980 The use of fluorescamine at pH 6·0 to follow the action of chymosin on κ-casein and to estimate this protein in milk. New Zealand Journal of Dairy Science and Technology 15 99–108Google Scholar
Church, F. C., Catignani, G. L. & Swaisgood, H. E. 1981 Hydrolysis of milk proteins by immobilized Streptomyces griseus pronase. Journal of Dairy Science 64 724–731Google Scholar
Church, F. C., Swaisgood, H. E., Porter, D. H. & Catignani, G. L. 1983 Spectrophotometric assay using o−phthaldialdehyde for determination of proteolysis in milk and isolated milk proteins. Journal of Dairy Science 66 1219–1227CrossRefGoogle Scholar
Davis, J. G. 1976 Cheese. III. Manufacturing Methods. London: Churchill LivingstoneGoogle Scholar
Fields, R. 1971 The measurement of amino groups in proteins and peptides. Biochemical Journal 124 581–590Google Scholar
Freedman, R. B. & Radda, G. K. 1968 The reaction of 2,4,6-trinitrobenzenesulphonic acid with amino acids, peptides and proteins. Biochemical Journal 108 383–391Google Scholar
Habeeb, A. F. S. A. 1966 Determination of free amino groups in proteins by trinitrobenzenesulfonic acid. Analytical Biochemistry 14 328–336Google Scholar
Kuchroo, C. N. & Fox, P. F. 1982 Soluble nitrogen in Cheddar cheese: comparison of extraction procedures. Milchwissenschaft 37 331–335Google Scholar
Kuchroo, C. N., Rahilly, J. & Fox, P. F. 1983 Assessment of proteolysis in cheese by reaction with trinitrobenzene sulphonic acid. Irish Journal of Food Science and Technology 7 129–133Google Scholar
Kwan, K. K. H., Nakai, S. & Skura, B. J. 1983 Comparison of four methods for determining protease activity in milk. Journal of Food Science 48 1418–1421Google Scholar
McKellar, R. C. 1981 Development of off-flavors in ultra-high temperature and pasteurized milk as a function of proteolysis. Journal of Dairy Science 64 2138–2145CrossRefGoogle Scholar
Polychroniadou, A. & Vlachos, J. 1979 [Amino acids of Teleme cheese]. Lait 59 234–243Google Scholar
Reimerdes, E. H. & Klostermeyer, H. 1976 Determination of proteolytic activities on casein substrates. Methods in Enzymology. 45 26–28Google Scholar
Samples, D. R., Richter, R. L. & Dill, C. W. 1984 Measuring proteolysis in Cheddar cheese slurries: Comparison of Hull and trinitrobenzene sulfonic acid procedures. Journal of Dairy Science 67 60–63Google Scholar
Spadaro, A. C. C., Draghetta, W., Nassif Del Lama, S., Camargo, A. C. M. & Greene, L. J. 1979 A convenient manual trinitrobenzenesulfonic acid method for monitoring amino acids and peptides in Chromatographie column effluents. Analytical Biochemistry 96 317–321Google Scholar
Vakaleris, D. G. & Price, W. V. 1959 A rapid Spectrophotometric method for measuring cheese ripening. Journal of Dairy Science 42 264–276Google Scholar
Wakkers, P. J. M., Hellendoorn, H. B. A., Op De Weegh, G. J. & Heerspink, W. 1975 Applications of statistics in clinical chemistry. A critical evaluation of regression lines. Clinica Chimica Acta 64 173–184Google Scholar
Yamasaki, R. B., Shimer, D. A. & Feeney, R. E. 1981 Colorimetrie determination of arginine residues in proteins by p−nitrophenylglyoxal. Analytical Biochemistry 111 220–226CrossRefGoogle Scholar
Youden, W. J. & Steiner, E. H. 1975 Statistical Manual of the AOAC. Washington, DC: Association of Official Analytical ChemistsGoogle Scholar