Hostname: page-component-77c89778f8-5wvtr Total loading time: 0 Render date: 2024-07-16T11:32:54.133Z Has data issue: false hasContentIssue false

A fluorimetric method for the determination of Pseudomonas fluorescens AR11 lipase in milk

Published online by Cambridge University Press:  01 June 2009

Donald Stead
Affiliation:
National Institute for Research in Dairying, Shinfield, Reading RG2 9 AT, UK

Summary

A method has been developed, using the fluorogenic substrate 4-methylumbelliferyl oleate, to measure the activity of the extracellular lipase of the psychrotroph Pseudomonas fluorescens AR11 in milk. Inhibition of lipolysis by constituents of skim-milk was prevented by including both sodium taurocholate and cetyltrimethylammonium bromide in the reaction mixture. Cream also interfered with the reaction and skim-milk was used for high sensitivity. Natural milk lipase was readily inactivated by heat treatment to which AR11 lipase was relatively resistant.

Type
Original Articles
Copyright
Copyright © Proprietors of Journal of Dairy Research 1983

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

Brockerhoff, H. & Jensen, R. G. 1974 Lipolytic enzymes. London: Academic PressGoogle Scholar
Cogan, T. M. 1980 Heat resistant lipases and proteinases and the quality of dairy products. Annual Bulletin, International Dairy Federation Document 118 2632Google Scholar
Cousin, M. A. 1981 Proteolytic activity of psychrotrophic microorganisms on milk and dairy products. In Psychrotrophic Micro-organisms in Spoilage and Pathogenicity 6372 (Eds Roberts, T. A., Hobbs, G., Christian, J. H. B. and Skovgaard, N.) (InternationalSymposium on Food Microbiology no. 11, 1980) London: Academic Press.Google Scholar
Dooijewaard-Kloosterziel, A. M. P. & Wouters, J. T. M. 1976 Some properties of the lipase of Geotrichum candidum evaluated by a fluorimetric assay technique. Journal of Applied Bacteriology 40 293299Google Scholar
Downey, W. K. 1980 Review of the progress of Dairy Science: flavour impairment from pre- and post-manufacture lipolysis in milk and dairy products. Journal of Dairy Research 47 237252CrossRefGoogle Scholar
Downey, W. K. & Andrews, P. 1965 a Milk lipases and the association of lipases with casein micelles studied by gel-filtration. Biochemical Journal 94 33PGoogle Scholar
Downey, W. K. & Andrews, P. 1965 b Gel filtration applied to the study of lipases and other esterases. Biochemical Journal 94 642650Google Scholar
Driessen, F. M. & Stadhouders, J. 1975 Lipolysis in hard cheese made from pasteurized milk. Annual Bulletin, International Dairy Federation Document 86 101107Google Scholar
Dumont, J. P., Delespaul, G., Miguot, B. & Adda, J. 1977 [Effect of psychrotrophic bacteria on the organoleptic qualities of soft cheeses] Lait 57 619630CrossRefGoogle Scholar
Feuillat, M., Le Guennec, S. & Olsson, A. 1976 [Proteolysis in refrigerated milks and its effect on the yield of manufactured soft cheeses] Lait 56 521536Google Scholar
Fraser, G. P. & Nicol, A. D. 1966 Studies on human pancreatic lipase. Clinica Chimica Acta 13 552562Google Scholar
Gaffney, P. J., Harper, W. J. & Gould, I. A. 1966 Distribution of lipase among components of a water extract of rennet casein. Journal of Dairy Science 49, 921924CrossRefGoogle ScholarPubMed
Jacks, T. J. & Kircher, H. W. 1967 Fluorometric assay for the hydrolytie activity of lipase using fatty acyl esters of 4-methylumbelliferone. Analytical Biochemistry 21 279285Google Scholar
Kishonti, E. 1975 Influence of heat resistant lipases and proteases in psychrotrophic bacteria on product quality. Annual Bulletin, International Dairy Federation Document 86 121124Google Scholar
Kishonti, E. & Sjöström, G. 1970 Influence of heat resistant lipases and proteases in psychrotrophic bacteria on product quality. International Dairy Congress 18, Sydney 1E 501Google 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 573588CrossRefGoogle 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 145148CrossRefGoogle Scholar
Law, B. A., Cousins, C. M., Sharpe, M. E. & Davies, F. L. 1979 Psychrotrophs and their effects on milk and dairy products. In Cold Tolerant Microbes in Spoilage and the Environment 137152 (Eds Russell, A. D. and Fuller, R.) London: Academic Press (Society for Applied Bacteriology Technical Series 13)Google Scholar
Law, B. A., Sharpe, M. E. & Chapman, H. R. 1976 The effect of lipolytic Gram-negative psyehrotrophs in stored milk on the development of rancidity in Cheddar cheese. Journal of Dairy Research 43 459468Google Scholar
Leaback, D. H. 1975 An Introduction to the Fluorimetric Estimation of Enzyme Activities 2nd edn.Colnbrook, Berks: Koch-Light Laboratories LtdGoogle Scholar
Muir, D. D., Kelly, M. E. & Phillips, J. D. 1978 The effect of storage temperature on bacterial growth and lipolysis in raw milk. Journal of the Society of Dairy Technology 31 203208CrossRefGoogle Scholar
Olivecrona, T. 1980 Biochemical aspects of lipolysis in bovine milk. Annual Bulletin, International Dairy Federation Document 118 1925Google Scholar
Pinheiro, A. J. R., Liska, B. J. & Parmelee, C. E. 1965 Heat stability of lipases of selected psychrophilic bacteria in milk and Purdue Swiss-type cheese. Journal of Dairy Science 48 983984CrossRefGoogle ScholarPubMed
Roy, R. N. 1980 Fluorimetric assay of the activity of extracellular lipases of Pseudomonas fluorescens and Serratia marcescens. Journal of Applied Bacteriology 49 265271Google Scholar
Skean, J. D. & Overcast, W. W. 1961 Apparent location of lipase in cream. Journal of Dairy Science 44 823832.Google Scholar
Stewart, D. B., Murray, J. G. & Neill, S. D. 1975 Lipolytic activity of organisms isolated from refrigerated bulk milk. Annual Bulletin, International Dairy Federation Document 86 3850Google Scholar
Wills, E. D. 1965 Lipases. Advances in Lipid Research 3 197240Google Scholar