1887

Abstract

SUMMARY: Lipase from has been purified 4800-fold from crude culture supernatant. The purified enzyme preparation had no assayable protease, hyaluronate lyase or acid phosphatase activities. The molecular weight of the lipase was 46 770 as determined by gel filtration. Sodium dodecyl sulphatepolyacrylamide gel electrophoresis revealed a major protein component (mol. wt 41 190) together with two minor protein components (mol. wt 67 000 and 125 900). The lipase had a pH optimum of 68, was most stable in the pH range 5·0 to 6·0 and was completely inactivated after 30 min at 60 °C. The lipase hydrolysed trilaurin, triolein, trimyristin and tripalmitin at decreasing rates and did not exhibit phospholipase activity. Analysis of the reaction products from the hydrolysis of triolein by lipase did not demonstrate an accumulation of 2-monoolein which suggested that the enzyme did not exhibit a positional specificity for the 1-position of the triacylglycerol.

Crude lipase preparations contained an aggregated high molecular weight form of the enzyme which was eluted with the void volume from Sephadex G-200. This aggregated form was dissociated to produce the lower molecular weight lipase species by subsequent dialysis and elution from Sephadex G-200 using buffer with a higher ionic strength.

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1981-06-01
2021-05-18
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References

  1. Breuil C., Kushner D.J. 1974; Partial purification and characterization of the lipase of a facultatively psychrophilic bacterium (Acinetobacter O16). Canadian Journal of Microbiology 21:434–441
    [Google Scholar]
  2. Desnuelle P. 1972; The lipases. In The Enzymes 8 pp. 575–616 Boyer P.D. Edited by New York & London:: Academic Press.;
    [Google Scholar]
  3. Dole V.P., Meinertz H. 1960; Microdetermination of long chain fatty acids in plasma and tissues. Journal of Biological Chemistry 235:2595–2599
    [Google Scholar]
  4. Ferguson D.A., Cummins C.S. 1978; Nutritional requirements of anaerobic coryneforms. Journal of Bacteriology 135:858–867
    [Google Scholar]
  5. Freinkel R.K., Shen Y. 1969; The origin of free fatty acids in sebum. II. Assay of the lipases of the cutaneous bacteria and effects of pH. Journal of Investigative Dermatology 53:422–427
    [Google Scholar]
  6. Fulton J.E., Noble N.L., Bradley S., Awad W.M. 1974; The glycerol ester hydrolase (EC 3·1·1·3) from Corynebacterium acnes: a serine lipase. Biochemistry 13:2320–2326
    [Google Scholar]
  7. Greten H., Walter B. 1973; Purification of rat adipose tissue lipoprotein lipase. FEBS Letters 35:36–40
    [Google Scholar]
  8. Hagen J.H., Hagen P.B. 1962; An enzymic method for the estimation of glycerol in blood and its use to determine the effect of noradrenaline on the concentration of glycerol in blood. Canadian Journal of Biochemistry and Physiology 40:1129–1139
    [Google Scholar]
  9. Hassing G.S. 1971; Partial purification and some properties of a lipase from Corynebacterium acnes . Biochimica et biophysica acta 242:381–394
    [Google Scholar]
  10. Henderson C., Hodgkiss W. 1973; An electron microscopic study of Anaerovibrio lipolytica (strain 5 S) and its lipolytic enzyme. Journal of General Microbiology 76:389–393
    [Google Scholar]
  11. Holland K.T., Cunliffe W.J., Roberts C.D. 1978; The role of bacteria in acne-a new approach. Clinical and Experimental Dermatology 3:253–257
    [Google Scholar]
  12. Holland K.T., Greenman J., Cunliffe W.J. 1979; Growth of cutaneous propionibacteria on synthetic medium: growth yields and exoenzyme production. Journal of Applied Bacteriology 47:383–394
    [Google Scholar]
  13. Ingham E., Holland K.T., Gowland G., Cunliffe W.J. 1979; Purification and partial characterization of hyaluronate lyase (EC 4·2·2·1) from Propionibacterium acnes . Journal of General Microbiology 115:411–418
    [Google Scholar]
  14. Ingham E., Holland K.T., Gowland G., Cunliffe W.J. 1980; Purification and partial characterization of an acid phosphatase (EC 3·1·3·2) produced by Propionibacterium acnes . Journal of General Microbiology 118:59–65
    [Google Scholar]
  15. Ishihara H., Okuyama H., Ikezawa H., Tejima S. 1975; Studies on lipase from Mucor javanicus. I. Purification and properties. Biochimica et biophysica acta 388:413–422
    [Google Scholar]
  16. Kates M. 1972; Techniques of lipidology, isolation, analysis and identification of lipids. Laboratory Techniques in Biochemistry and Molecular Biology 3:428–436
    [Google Scholar]
  17. Kimura H., Kitamura T., Tsuji M. 1972; Studies on human pancreatic lipase. I. Interconversion between low and high molecular weight forms of human pancreatic lipase. Biochimica et biophysica acta 270:307–316
    [Google Scholar]
  18. Kligman A.M. 1974; An overview of acne. Journal of Investigative Dermatology 62:268–287
    [Google Scholar]
  19. Lawrence R.C., Fryer T.F., Reiter B. 1967; The production and characterization of lipases from a micrococcus and a pseudomonad. Journal of General Microbiology 48:401–418
    [Google Scholar]
  20. Marples R.R., McGinley K.J. 1974; Coryne-bacterium acnes and other anaerobic diphtheroids from human skin. Journal of Medical Microbiology 7:349–357
    [Google Scholar]
  21. Marples R.R., Downing D.T., Kligman A.M. 1971; Control of free fatty acids in human surface lipids by Corynebacterium acnes . Journal of Investigative Dermatology 56:127–131
    [Google Scholar]
  22. Moore W.E.C., Holdeman L.V. 1974; Genus I.Propionibacterium . In Bergey’s Manual of Determinative Bacteriology, 8th. pp. 633–641 Buchanan R.E., Gibbons N.E. Edited by Baltimore:: Williams & Wilkins.;
    [Google Scholar]
  23. Nieman C. 1954; Influence of trace amounts of fatty acids on the growth of microorganisms. Bacteriological Reviews 18:147–163
    [Google Scholar]
  24. Noble W.C. 1968; Observations on the surface flora of the skin and on the skin pH. British Journal of Dermatology 80:279–281
    [Google Scholar]
  25. Pablo G., Hammons A., Bradley S., Fulton J.E. 1974; Characteristics of the extracellular lipases from Corynebacterium acnes and Staphylococcus epidermidis . Journal of Investigative Dermatology 63:231–238
    [Google Scholar]
  26. Puhvel S.M., Reisner R.M. 1970; Effect of fatty acids on the growth of Corynebacterium acnes in vitro . Journal of Investigative Dermatology 54:48–52
    [Google Scholar]
  27. Puhvel S.M., Sakamoto M. 1977; A reevaluation of fatty acids as inflammatory agents in acne. Journal of Investigative Dermatology 68:93–97
    [Google Scholar]
  28. Rebello T., Hawk J.L.M. 1978; Skin surface glycerol levels in acne vulgaris. Journal of Investigative Dermatology 70:352–354
    [Google Scholar]
  29. Rietsch J., Pattus F., Desnuelle P., Verger R. 1977; Further studies of mode of action of lipolytic enzymes. Journal of Biological Chemistry 252:4313–4318
    [Google Scholar]
  30. Tomizuka N., Ota Y., Yamada K. 1966; Studies on the lipase from Candida cylindracea. Part I. Purification and properties. Agricultural and Biological Chemistry 30:576–584
    [Google Scholar]
  31. Vadehra D.V. 1974; Staphylococcal lipases. Lipids 9:158–165
    [Google Scholar]
  32. Voss J.G. 1970; Differentiation of two groups of Corynebacterium acnes . Journal of Bacteriology 101:392–397
    [Google Scholar]
  33. Weber K., Pringle J.R., Osborn M. 1972; Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods in En-zymology 26:3–27
    [Google Scholar]
  34. Weeks J.G., McCarty L., Black T., Fulton J.E. 1977; The inability of a bacterial lipase inhibitor to control acne vulgaris. Journal of Investigative Dermatology 69:236–243
    [Google Scholar]
  35. Whiteside J.A., Voss J.G. 1973; Incidence and lipolytic activity of Propionibacterium acnes (Corynebacterium acnes group I) and P. granulosum(C.acnes group II) in acne and in normal skin. Journal of Investigative Dermatology 60:94–97
    [Google Scholar]
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