1887

Abstract

SUMMARY: The lipase of ( -25) was purified 75-to 100-fold with an overall recovery of . 20%. The enzyme was found to exist in heavy and light forms exhibiting the same position specificity for triglycerides, Michaelis constants (), and apparent pH and temperature optima. The light form of the enzyme appeared to be present in the heavy one, probably in a complexed state. The purified lipase was found to hydrolyze only glycerol esters of fatty acids; it required a water-fat interface and exhibited a 1,3-position specificity for triglycerides. The optimum pH for the purified enzyme with purified tributyrin as substrate was calculated to be 8.6 to 8.7 from initial velocity measurements with a pH-stat at 25°. The for the purified lipase with tributyrin was found to be 0.9 x 10 at 25°and pH 7.2. Exposure of the purified enzyme to 40°for 10 min. caused a complete loss of activity. A 50% loss of activity-occurred after 10 min. exposure to about 35°. When exposed to pH values ranging from 5.3 to 9.5 for 1 hr at 2°and then assayed at pH 7.0 and 35°, the purified enzyme was found to be stable in the pH range 6.6-7.8.

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1967-09-01
2021-08-04
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References

  1. Alford J. A., Pierce D. A. 1963; Production of lipase by Pseudomonas fragi in a synthetic medium. J. Bad 86:24
    [Google Scholar]
  2. Alford J. A., Pierce D. A., Suggs F. G. 1964; Activity of microbial lipases on natural fats and synthetic triglycerides. J. Lipid Res 5:390
    [Google Scholar]
  3. Alford J. A., Pierce D. A., Sulzbacher W. L. 1963; Microbial lipases and their potential importance to the meat industry. Proc. 15th Res. Conf. Am. Meat Inst. Foundation p. 11
    [Google Scholar]
  4. Benzonana G., Desnuelle P. 1965; Etude cinetique de l’action de la lipase pancreatique sur des trigylcerides en emulsion. Essai d’une enzymologie en milieu heterogene. Biochim. biophys. Acta 105:121
    [Google Scholar]
  5. Clement G., Clement J., Bezard J. 1962; Action de la lipase pancreatique humaine sur des triglycerides mixtes, symetriques de synthese comportant des acides gras long et de l’acide butyrique. Archs Sci. physiol 16:213
    [Google Scholar]
  6. Crenshaw E. C., San Clemente C. L. 1964; Studies on a lipase prepared from Staphylococcus aureus. Bact. Proc p. 61
    [Google Scholar]
  7. Davis B. J. 1964; Disc electrophoresis. II. Method and application to human serum proteins. Am. N.Y. Acad. Sci 121:404
    [Google Scholar]
  8. DeHaas G. H., Sarda L., Roger J. 1965; Positional specific hydrolysis of phospholipids by pancreatic lipase. Biochim. biophys. Acta 106:638
    [Google Scholar]
  9. Desnuelle P. 1961; Pancreatic lipase. Adv. Enzymol 23:129
    [Google Scholar]
  10. Desnuelle P., Savary P. 1963; Specificities of lipases. J. Lipid Res. 4:369
    [Google Scholar]
  11. Desnuelle P., Constantin M. J., Baldy J. 1955; Technique potentiometrique pour la mesure de l’activite de la lipase pancreatique. Bull. Soc. Chim. biol 37:285
    [Google Scholar]
  12. Fukumoto J., Iwai M., Tsujisaka Y. 1964; Studies on lipase. IV. Purification and properties of a lipase secreted by Rhizopus delemar. J. gen. appl. Microbiol 10:257
    [Google Scholar]
  13. Gelotte B. 1964; Separation of pancreatic enzymes by gel filtration. Acta chem. scand 18:1283
    [Google Scholar]
  14. Enzyme Nomenclature; 1965 Standing Committee on Enzymes International Union of Biochemistry p. 128 New York: Elsevier Publishing Co.;
    [Google Scholar]
  15. Iwai M., Tsujisaka Y., Fukumoto J. 1964; Studies of lipase. II. Hydrolytic and esterifying actions of crystalline lipase of Aspergillus niger. J. gen. appl. Microbiol. 10:13
    [Google Scholar]
  16. Kates M. 1964; Simplified procedure for hydrolysis or methanolysis of lipids. J. Lipid Res 5:132
    [Google Scholar]
  17. Koshland D. E. 1963; Correlation of structure and function in enzyme action. Science 142:1533
    [Google Scholar]
  18. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. J. biol. Chem 193:265
    [Google Scholar]
  19. McDonald I. J. 1964; Characteristics of proteinases of three strains of Staphylococcus lactis isolated from Cheddar cheese. J. Dairy Res 31:147
    [Google Scholar]
  20. Mencher J. R., Ng H., Alford J. A. 1965; The extracellular nature of the lipase of Pseudomonas fragi. Biochim. biophys. Acta 106:628
    [Google Scholar]
  21. Nashif S. A., Nelson F. E. 1953; The lipase of Pseudomonas fragi. I. Characterization of the enzyme. J. Dairy Sci 36:459
    [Google Scholar]
  22. Ornstein L. 1964; Disc electrophoresis. I. Background and theory. Ann. N.Y. Acad. Sci 121:321
    [Google Scholar]
  23. Peterson E. A., Sober H. A. 1962; Column chromatography of proteins: Substituted celluloses. Meth. Enzymol 5:25
    [Google Scholar]
  24. Pollock M. R. 1965; Purification and properties of penicillinases from two strains of Bacillus licheniformis: A chemical physico-chemical and physiological comparison. Biochem. J 94:666
    [Google Scholar]
  25. Reithel F. J. 1963; The dissociation and association of protein structures. Adv. Protein Chem 18:123
    [Google Scholar]
  26. Rottem S., Razin S. 1964; Lipase activity of mycoplasma. J. gen. Microbiol 37:123
    [Google Scholar]
  27. Sarda L., Maylie M. F., Roger J., Desnuelle P. 1964; Comportement de la lipase pancreatique sur Sephadex. Application a la determination du poids moleculaire de cet enzyme. Biochim. biophys. Acta 89:183
    [Google Scholar]
  28. Shahani K. M., Chandan R. C. 1965; Activity of purified milk lipase in the presence of milk constituents. Arch. Biochem. Biophys257
    [Google Scholar]
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