Hostname: page-component-586b7cd67f-t7czq Total loading time: 0 Render date: 2024-11-23T15:41:22.475Z Has data issue: false hasContentIssue false
  • English
  • Français

Isolation and some properties of extracellular heat-stable lipases from Pseudomonas fluorescens strain AFT 36

Published online by Cambridge University Press:  01 June 2009

Patrick F. Fox  and
Leszek Stepaniak
Patrick F. Fox
Affiliation:
Department of Dairy and Food Chemistry, University College, Cork, Irish Republic
Leszek Stepaniak
Affiliation:
Department of Dairy and Food Chemistry, University College, Cork, Irish Republic
Get access Rights & Permissions [Opens in a new window]

Summary

Aeration increased the growth and lipase production in milk by Pseudomonas fluorescens strain AFT 36, isolated from refrigerated bulk milk. A heat-stable lipase was isolated from a shaken milk culture of this microorganism by DEAE-chromatography and gel filtration in Sepharose 6B. The lipase-rich fraction from DEAE cellulose contained 3 lipases that were separated by gel filtration; only the principal lipase, which represented ∼ 71 % of total lipolytic activity, was characterized. The purified enzyme showed maximum activity on tributyrin at pH 8·0 and 35 °C; it had a Km on tributyrin of 3·65 mM. and was inhibited by concentrations of substrate > ∼ 17 mM. The enzyme was very stable over the pH range 6–9; it was relatively heat-labile in phosphate buffer in the temperature range 60–80 °C, where it was stabilized significantly by Ca2+. It was, however, very stable at 100–150 °C: the D values at 150 °C were ∼ 22 s and 28 s in phosphate buffer and synthetic milk serum respectively; the corresponding Z values in the temperature range 100–150 °C were ∼ 40 and ∼ 42 °C and the Ea for inactivation were 7·65 × 104 J mol-1 and 6·97 × 104 J mol-1 respectively.

Type
Original Articles
Information
Journal of Dairy Research , Volume 50 , Issue 1 , February 1983 , pp. 77 - 89
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

Adams, D. M. & Brawley, T. G. 1981 a Heat resistant bacterial lipases and ultra-high temperature sterilization of dairy products. Journal of Dairy Science 64 1951–1957CrossRefGoogle Scholar
Adams, D. M. & Brawley, T. G. 1981 b Factors influencing the heat resistance of a heat-resistant lipase of Pseudomonas. Journal of Food Science 46 673–676CrossRefGoogle Scholar
Adams, D. M. & Brawley, T. G. 1981 c Factors influencing the activity of a heat resistant lipase of Pseudomonas. Journal of Food Science 46 677–680CrossRefGoogle Scholar
Andersson, R. E. 1980 Microbial lipolysis at low temperatures. Applied and Environmental Microbiology 39 36–40CrossRefGoogle ScholarPubMed
Andersson, R. E., Hedlund, C. B. & Jonnson, U. 1979 Thermal inactivation of a heat resistant lipase produced by the psychrotrophic bacterium Pseudomonas fluorescens. Journal of Dairy Science 62 361–367CrossRefGoogle Scholar
Breuil, C. & Kushner, D. J. 1975 Partial purification and characterization of the lipase of a facultatively psychrophilic bacterium (Acinetobacter O16). Canadian Journal of Microbiology 21 434–441CrossRefGoogle ScholarPubMed
Christen, G. L. & Marshall, R. T. 1980 Thermostability of selected lipases produced by psychrotrophic bacteria. Journal of Dairy Science 63 Suppl. 1 46–47Google 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
Dawson, R. M., Elliott, D. C., Elliott, W. H. & Jones, K. M. 1969 Data for Biochemical Research, 2nd edn p. 485Oxford: Clarendon PressGoogle Scholar
Driesskn, 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
Fields, R. 1972 The rapid determination of amino groups with TNBS. Methods in Enzymology 25 464–468CrossRefGoogle ScholarPubMed
Fossum, K. 1970 Proteolytic enzymes and biological inhibitors. I. Comparison between the Kunitz method and the agar gel casein precipitating reaction for determination of the activity of some commercial proteolytic enzymes and inhibitors. Acta Pathologica et Microbiologica Scandinavia B 78 350–362Google ScholarPubMed
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
Jenness, R. & Koops, J. 1902 Preparation and properties of a salt solution which simulates milk ultrafiltrate. Netherlands Milk and Dairy Journal 16 153–164Google Scholar
Khan, I. M., Dill, C. W., Chandan, R. C. & Shahani, K. M. 1907 Production and properties of the extracellular lipase of Achromobacter lipolyticum. Biochimica et Biophysica Acta 132 68–77CrossRefGoogle Scholar
Knaut, T. 1978 Heat resistance of Pseudomonas lipases in milk. 20th International Dairy Congress, Paris E 305–306Google Scholar
Kosugi, Y. & Kamibayashi, A. 1971 Thermostable lipase from Pseudomonas spp. Cultural conditions and properties of the crude enzyme. Journal of Fermentation Technology 49 968–980Google Scholar
Landaas, A. & Solberg, P. 1978 Production and characterization of lipase from a fluorescent pseudomonad. 20th International Dairy Congress, Paris E 304–305Google Scholar
Law, B. A. 1979 Reviews of the progress of Dairy Science: Enzymes of psychrotrophie 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. 1967 Microbial lipases and related esterases. I. Detection, distribution and production of microbial lipases. Dairy Science Abstracts 29 1–8Google 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. E. & Reiter, B. 1967 b Rapid method for the quantitative estimation of microbial lipases. Nature 213 1264–1265CrossRefGoogle Scholar
Lin, Y., Means, G. E. & Feeney, R. E. 1969 The action of proteolytic enzymes on N, N-dimethyl proteins. Journal of Biological Chemistry 244 789–793CrossRefGoogle Scholar
Lu, J. Y. & Liska, B. J. 1969 a Lipase from Pseudomonas fragi. I. Purification of the enzyme. Applied Microbiology 18 104–107CrossRefGoogle ScholarPubMed
Lu, J. Y. & Liska, B. J. 1969 b Lipase from Pseudomonas fragi. II. Properties of the enzyme. Applied Microbiology 18 108–113CrossRefGoogle ScholarPubMed
Mencher, J. R. & Alford, J. A. 1967 Purification and characterization of the lipase of Pseudomonas fragi. Journal of General Microbiology 48 317–328CrossRefGoogle ScholarPubMed
Nashif, S. A. & Nelson, F. E. 1953 The lipase of Pseudomonas fragi. I. Characterization of the enzyme. Journal of Dairy Science 36 459–470CrossRefGoogle Scholar
O'Donnell, E. T. 1978 Heat resistance of lipase enzymes produced by psychrotrophic bacteria. 20th International Dairy Congress, Paris E 307Google Scholar
Stepaniak, L. & Fox, P. F. 1983 Thermal stability of an extracellular proteinase from Pseudomonas fluorescens AFT 36. Journal of Dairy Research 50 in pressCrossRefGoogle ScholarPubMed
Stepaniak, L., Fox, P. F. & Daly, C. 1982 Isolation and general characterization of heat stable proteinase from Pseudomonas fluorescens AFT 36. Biochimica et Biophysica Acta. 717 376–383CrossRefGoogle ScholarPubMed