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Measurement of proteolysis in milk and cheese using trinitrobenzene sulphonic acid and a new dissolving reagent

Published online by Cambridge University Press:  01 June 2009

Gérard Humbert
Affiliation:
Laboratory of Applied Biochemistry University of Nancy 1, BP 239, 54506- Vandoeuvre-Les-Nancy, France
Marie-France Guingamp
Affiliation:
Laboratory of Applied Biochemistry University of Nancy 1, BP 239, 54506- Vandoeuvre-Les-Nancy, France
Rosalie Kouomegne
Affiliation:
Laboratory of Applied Biochemistry University of Nancy 1, BP 239, 54506- Vandoeuvre-Les-Nancy, France
Guy Linden
Affiliation:
Laboratory of Applied Biochemistry University of Nancy 1, BP 239, 54506- Vandoeuvre-Les-Nancy, France

Extract

Milk contains indigenous plasmin-like and thrombin-like proteinases and also exogenous proteinases, mainly extracellular enzymes of psychrotrophic bacteria. Their activities lead to protein hydrolysis which may have important effects on milk quality and cheese yield. It is also useful to estimate proteolysis and its development during cheese ripening.

Milk opacity or turbidity due to calcium phosphocaseinate micelles and fat globules necessitates preliminary sample treatment, i.e. precipitation and filtration before spectrophotometric measurement. It is possible to eliminate these firs steps of sample treatment by dispersing the colloidal constituents. Some authors have already used different dissolving mixtures for sample analysis. Nakai & Anh Chi Le (1970) were the first to determine protein contents of milk and dairy products by absorbance measurement at 280 nm after clarification. Bosset & Blanc (1977) used the Biuret method. Owen & Andrews (1984) measured free amino groups with 2,4,6-trinitrobenzene sulphonic acid (TNBS). Humbert et al. (1982) and Stead (1987) used synthetic or specific substrates for proteinase determination.

In this work, we report the application of a new dissolving reagent to measure free amino groups in milk and cheese.

Type
Short Communication
Copyright
Copyright © Proprietors of Journal of Dairy Research 1990

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References

REFERENCES

Barlow, I. E., Lloyd, G. T. & Ramshaw, E. H. 1986 The measurement of proteolysis in Cheddar cheese: a comparison of trinitrobenzene sulphonic acid procedures. Australian Journal of Dairy Technology 41 7981Google Scholar
Bosset, J. O. & Blanc, B. H. 1977 [New, simple and economical method for routine determination of protein content of milk and some milk products.] Le Laitier Romand 53 697699.Schweizerische Milchzeitung 103 325–326Google Scholar
Grieve, P. A. & Kitchen, B. J. 1985 Proteolysis in milk: the significance of proteinases originating from milk leucocytes and a comparison of the action of leucocyte, bacterial and natural milk proteinases on casein. Journal of Dairy Research 52 101112CrossRefGoogle Scholar
Humbert, G., Desnouveaux, R. & Linden, G. 1982 [Application of complete solubilization of the colloidal components of milk to determination of proteolytic activity.] Lait 62 427434CrossRefGoogle Scholar
Linden, G., Humbert, G. & Guingamp, M. F. 1987 Reagent for rendering biological fluid transparent and its analytical applications. European Patent Application EP 0 246 978 A1, 10 ppGoogle Scholar
Nakai, S. & Anh Chi, Le 1970 Spectrophotometric determination of protein and fat in milk simultaneously. Journal of Dairy Science 53 276278CrossRefGoogle Scholar
Owen, A. J. & Andrews, A. T. 1984 A procedure for the complete clarification of milk of various species and its suitability for use with colorimetric measurements. Journal of Dairy Research 51 307315CrossRefGoogle Scholar
Polychroniadou, A. 1988 A simple procedure using trinitrobenzenesulphonic acid for monitoring proteolysis in cheese. Journal of Dairy Research 55 585596CrossRefGoogle 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 6063CrossRefGoogle Scholar
Stead, D. 1987 Assay of proteinases of psychrotrophic bacteria in milk using Hide Powder Azure and a simple procedure for clarification of milk. Journal of Dairy Research 54 295302CrossRefGoogle Scholar