Hostname: page-component-78c5997874-xbtfd Total loading time: 0 Render date: 2024-11-05T19:49:28.517Z Has data issue: false hasContentIssue false
  • English
  • Français

A new method for the detection of microbial proteolytic enzymes in milk

Published online by Cambridge University Press:  01 June 2009

Allan J. Cliffe  and
Barry A. Law
Allan J. Cliffe
Affiliation:
National Institute for Research in Dairying, Shinfield, Reading RG2 9AT, UK
Barry A. Law
Affiliation:
National Institute for Research in Dairying, Shinfield, Reading RG2 9AT, UK
Get access Rights & Permissions [Opens in a new window]

Summary

Optimum conditions have been determined for the use of Hide Powder Azure (HPA) to estimate extracellular proteinases produced by psychrotrophic Gram-negative bacteria. This assay is quicker, more precise and 3·2 times more sensitive than assays of soluble N released from casein. It provides a simple alternative method for monitoring proteinase purification schemes. With the modification described it is now possible to determine proteinase activities directly in milk. In its simplest form the HPA assay is sufficiently sensitive to detect proteinases produced by as few as 1·5 x 106 c.f.u. ml-1 psychrobtrophs without the necessity for any sample processing.

Type
Original Articles
Information
Journal of Dairy Research , Volume 49 , Issue 2 , May 1982 , pp. 209 - 219
Copyright
Copyright © Proprietors of Journal of Dairy Research 1982

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

Alichanidis, E. & Andrews, A. T. 1977 Some properties of the extracellular protease produced by the psychrotrophic bacterium Pseudomonas fluorescens strain AR-11. Biochimica et Biophysica Acta 485 424–433CrossRefGoogle Scholar
Adams, D. M., Barach, J. T. & Speck, M. L. 1975 Heat resistant proteases produced in milk by psychrotrophie bacteria of dairy origin Journal of Dairy Science 58 828–834Google Scholar
Cousins, C. M., Sharpe, M. E. & Law, B. A. 1977 The bacteriological quality of milk for Cheddar cheesemaking. Dairy Industries International 42(7) 12–17Google Scholar
Dawson, R. M. C., Elliott, D. C., Elliott, W. H. & Jones, K. M. 1969 pH, buffers, and physiological media. In Data for Biochemical Research 2nd edn pp. 484–485. London: Oxford University PressGoogle Scholar
Feuillat, M., Le Guennec, S. & Olsson, A. 1976 [Proteolysis of refrigerated milks and effects on yield in the manufacture of soft cheese] Lait 56 521–536CrossRefGoogle Scholar
Griffiths, M. W., Phillips, J. D. & Muir, D. D. 1981 Thermostability of proteases and lipases from a number of species of psychrotrophic bacteria of dairy origin Journal of Applied Bacteriology 50 289–303CrossRefGoogle ScholarPubMed
Harrigan, W. F. & McCance, M. E. 1976 Biochemical tests for bacteria, in Laboratory Methods in Food and Dairy Microbiology 2nd edn pp. 67–68London: Academic PressGoogle Scholar
Hendrie, M. S. & Shewan, J. M. 1966 The identification of certain Pseudomonas species. In Technical Series, Society for Applied Bacteriology 1 1–7Google Scholar
Hull, M. E. 1947 Studies on milk proteins. II. Colorimetric determination of the partial hydrolysis of the proteins in milk Journal of Dairy Science 30 881–884CrossRefGoogle Scholar
Juffs, H. S. 1973 Proteolysis detection in milk. I. Interpretation of tyrosine value data for raw milk supplies in relation to natural variation, bacterial counts and other factors Journal of Dairy Research 40 371–381CrossRefGoogle Scholar
Law, B. A. 1979 Reviews of the progress of Dairy Science: Enzymes of psychrotrophic bacteria and their effects on milk and milk products. Journal of Dairy Research 46 573–588CrossRefGoogle Scholar
Law, B. A., Andrews, A. T., Cliffe, A. J., Sharpe, M. E. & Chapman, H. R. 1979 a Effect of proteolytic raw milk psychrotrophs on Cheddar cheese-making with stored milk. Journal of Dairy Research 46 497–509CrossRefGoogle Scholar
Law, B. A., Andrews, A. T. & Sharpe, M. E. 1977 Gelation of ultra-high-temperature-sterilized milk by proteases from a strain of Pseudomonas fluorescens isolated from raw milk. Journal of Dairy Research 44 145–148CrossRefGoogle Scholar
Law, B. A., Cousins, C. M., Sharpe, M. E. & Da Vies, F. L. 1979 b Psychrotrophs and their effects on milk and dairy products. In Technical Series, Society for Applied Bacteriology 13 137–152Google Scholar
Law, B. A., Sharpe, M. E. & Chapman, H. R. 1976 The effect of lipolytic Gram-negative psychrotrophs in stored milk on the development of rancidity in Cheddar cheese. Journal of Dairy Research 43 459–468CrossRefGoogle Scholar
Law, B. A., Sharpe, M. E., Mabbitt, L. A. & Cole, C. B. 1973 Microflora of Cheddar cheese and some of the metabolic products. In Technical Series, Society for Applied Bacteriology 7 1–9Google Scholar
Moore, S. & Stein, W. H. 1954 A modified ninhydrin reagent for the photometric determination of amino acids and related compounds. Journal of Biological Chemistry 211 907–913CrossRefGoogle ScholarPubMed
Reimerdes, E. H. & Klostermeyer, H. 1976 Determination of proteolytic activities on casein substrates. Methods in Enzymology 45 26–28CrossRefGoogle ScholarPubMed
Rinderknecht, H., Geokas, M. C., Silverman, P. & Haverback, B. J. 1968 A new ultrasensitive method for the determination of proteolytic activity. Clinica Chimica Acta 21 197–203CrossRefGoogle ScholarPubMed
Shewan, J. M., Hobbs, G. & Hodgkiss, W. 1960 A determinative scheme for the identification of certain genera of Gram-negative bacteria, with special reference to the Pseudomonadaceae. Journal of Applied Bacteriology 23 379–390CrossRefGoogle Scholar
Stanier, R. Y., Palleroni, N. J. & Doudoroff, M. 1966 The aerobic Pseudomonads: a taxonomic study. Journal of General Microbiology 43 159–271CrossRefGoogle ScholarPubMed
Sunada, H. & Nagai, Y. 1980 A rapid micro-assay method for gelatinolytic activity using tritium-labeled heat-denatured polymeric collagen as a substrate and its application to the detection of enzymes involved in collagen metabolism. Journal of Biochemistry 87 1765–1771CrossRefGoogle Scholar