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Evaluation of a fluorimetric assay on the lipases from strains of milk psychrotrophic bacteria

Published online by Cambridge University Press:  01 June 2009

Donald Stead
Donald Stead
Affiliation:
*National Institute for Research in Dairying, (University of Reading), ShinfieldReadingRG2 9ATUK
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Summary

A fluorimetric assay, developed for the assay in milk of the lipase of the raw milk psychrotroph Pseudomonas fluorescens AR11 (Stead, 1983), was evaluated in both buffer and in milk using cell-free lipase preparations from 11 strains of lipolytic psychrotrophic bacteria isolated from milk. Lipases of all strains were more active against 4-methylumbelliferyl nonanoate (4-MUN) than against 4-methylumbelliferyl oleate (4-MUO) in the buffer system but most were less active against 4-MUN than against 4-MUO in the milk assay system. In both systems, 4-MUO had a much lower rate of non-enzymic hydrolysis than did 4-MUN. Lipase activities measured by radial diffusion in tributyrin agar and trioctanoin agar reflected activities against 4-MUO and 4-MUN, except that lipase from a strain of coliform was relatively more reactive against the 4-MU esters. When compared with assay in the buffer system the most effectively activated lipase in the milk system was that from AR11; of those from other strains, 6 were 50–69% as effectively activated, 2 were about 17% and 2 were 7–8% activated. This difference in behaviour was related to the direct inhibition of lipases of some of the strains by the mixture of sodium taurocholate (NaTC) and cetyltrimethylammonium bromide (CTAB) needed in the milk assay system to dissociate lipase from milk protein. Increasing the proportion of NaTC to CTAB increased the activities of the more weakly activated lipases but decreased those of others. The assay in milk may therefore underestimate the lipase from certain strains of psychrotrophic bacteria.

Type
Original Articles
Information
Journal of Dairy Research , Volume 51 , Issue 1 , February 1984 , pp. 123 - 130
Copyright
Copyright © Proprietors of Journal of Dairy Research 1984

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References

REFERENCES

Breuil, C. & Kushner, D. J. 1975 Partial purification and characterization of the lipase of a facultatively psychrophilic bacterium (Acinetobacter 016). Canadian Journal of Microbiology 21 434–441CrossRefGoogle Scholar
Cliffe, A. J. & Law, B. A. 1982 A new method for the detection of microbial proteolytic enzymes in milk. Journal of Dairy Research 49 209–219CrossRefGoogle Scholar
Cogan, T. M. 1977 A review of heat resistant lipases and proteinases and the quality of dairy products. Irish Journal of Food Science and Technology 1 95–105Google Scholar
Cousins, C. M. & Bramley, A. J. 1981 The microbiology of raw milk. In Dairy Microbiology 1 119–163 (Ed. Robinson, R. K.) Barking Essex: Applied Science PublishersGoogle 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 40 293–299CrossRefGoogle Scholar
Driessen, F. M. & Stadhouders, J. 1974 Thermal activation and inactivation of exocellular lipases of some Gram-negative bacteria common in milk. Netherlands Milk and Dairy Journal 28 10–22Google Scholar
Dumont, J. P., Delespaul, G., Migout, B. & Adda, J. 1977 Influence of psychrotrophic bacteria on the organoleptic qualities of soft curd cheese. Lait 57 619–630CrossRefGoogle 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
Henderson, C. 1968 A study of the lipase of Anaerovibrio lipolytica, a rumen bacterium. Ph.D. Thesis. University of Aberdeen.Google 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., 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
Lawrence, R. C., Fryer, T. F. & Reiter, B. 1967 a The production and characterization of lipases from a micrococcus and a pseudomonad. Journal of General Microbiology 48 401–418CrossRefGoogle Scholar
Lawrence, R. C., Fryer, T. F. & Reiter, B. 1967 b Rapid method for the quantitative estimation of microbial lipases. Nature 213 1264–1265CrossRefGoogle Scholar
Marks, T. A., Quinn, J. G., Sampugna, J. & Jensen, R. G. 1968 Studies on the specificity of a lipase system from Geotrichum candidum. Lipids 3 143–146CrossRefGoogle Scholar
Nantel, G. & Proulx, P. 1973 Lipase activity in E. coli. Biochimica et Biophysica Acta 316 156–161CrossRefGoogle Scholar
Patel, V., Goldberg, H. S. & Blenden, D. 1964 Characterization of Leptospiral lipase. Journal of Bacteriology 88 877–884CrossRefGoogle ScholarPubMed
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 48 983–984CrossRefGoogle ScholarPubMed
Roberts, T. L. & Rosenkrantz, H. 1966 Lipase and acetyl esterase activities in intact Bacillus coagulans spores. Canadian Journal of Biochemistry 44 677–685CrossRefGoogle ScholarPubMed
Roy, R. N. 1980 Fluorimetric assay of the activity of extracellular lipases of Pseudomonas fluorescens and Serratia marcescens. Journal of Applied Biochemistry 44 677–685Google Scholar
Sémériva, M. & Dufour, C. 1972 Further studies on the exocellular lipase of Rhizopus arrhizus. Biochimica et Biophysica Acta 260 393–400CrossRefGoogle ScholarPubMed
Shah, D. B. & Wilson, J. B. 1965 Egg yolk factor of Staphylococcus aureus. II. Characterization of the lipase activity. Journal of Bacteriology 89 949–953CrossRefGoogle ScholarPubMed
Stead, D. 1983 A fluorimetric method for the determination of Pseudomonas fluorescens AR11 lipase in milk. Journal of Dairy Research 50 491–502CrossRefGoogle Scholar
Weaber, K., Freedman, R. & Eudy, W. W. 1971 Tetracycline inhibition of a lipase from Corynebacterium acnes. Applied Microbiology 21 639–642CrossRefGoogle ScholarPubMed
Witter, L. D. 1961 Psychrophilic bacteria – a review. Journal of Dairy Science 44 983–1015CrossRefGoogle Scholar