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. 1982 Feb;69(2):368–376. doi: 10.1172/JCI110460

Intestinal Phospholipase, a Novel Enzyme

Charles M Mansbach II 1,2,3, Gerard Pieroni 1,2,3, Robert Verger 1,2,3
PMCID: PMC370986  PMID: 7056853

Abstract

We evaluated phospholipase activity in the intestine of rats and other species. Phospholipase activity was assayed by a surface barostat technique or an egg yolk titration system. Mucosal activity was found only by the surface barostat technique with phosphatidylglycerol as substrate; it was not found with phosphatidylcholine as substrate in assays by either technique. In gut luminal fluid activity was found when both phosphatidylcholine and phosphatidylglycerol were used as substrate in assays by the surface barostat technique, and phosphatidylcholine as substrate yielded activity in egg yolk titration. In rats in which pancreatic juice had been diverted, mucosal and gut luminal phospholipase activity was greater than in controls, thus demonstrating that enzyme activity was not due to pancreatic phospholipase. Bacterial origin of phospholipase activity was excluded in that phospholipase activity was found in germ-free rats; gastric and salivary gland origins were excluded in that continued phospholipase activity was found in rats with gastric fistula. The physiological importance of the enzyme was established by the finding that rats with pancreatic fistula absorbed 111 μmol of phosphatidylcholine and that controls absorbed 119 μmol of a 135-μmol load. Activity was found to be three times greater in the distal than in the proximal intestine; in cryptal cells it was 10 times greater than in villus tip cells. 65% of the activity in the gut lumen was tightly bound to particulate matter. We propose that intestinal phospholipase may be important in gut bacterial control, in the digestion of vegetable matter (phosphatidylglycerol is a major phospholipid in both plants and bacteria), and in the digestion of phospholipids in the gut lumen.

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Selected References

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  1. Alpers D. H., Isselbacher K. J. Protein synthesis by rat intestinal mucosa. The role of ribonuclease. J Biol Chem. 1967 Dec 10;242(23):5617–5622. [PubMed] [Google Scholar]
  2. Alpers D. H., Solin M. The characterization of rat intestinal amylase. Gastroenterology. 1970 Jun;58(6):833–842. [PubMed] [Google Scholar]
  3. Amic J., Lairon D., Hauton J. Technique de dosage automatique de l'orthophosphate de grande fiabilité. Clin Chim Acta. 1972 Aug;40(1):107–114. doi: 10.1016/0009-8981(72)90256-2. [DOI] [PubMed] [Google Scholar]
  4. Arnesjö B., Nilsson A., Barrowman J., Borgström B. Intestinal digestion and absorption of cholesterol and lecithin in the human. Intubation studies with a fat-soluble reference substance. Scand J Gastroenterol. 1969;4(8):653–665. doi: 10.3109/00365526909180651. [DOI] [PubMed] [Google Scholar]
  5. Belleville J., Clément J. Comparaison de l'activité phospholipasique A de préparations de pancréas et de suc pancréatique sur les phospholipides liés aux lipoprotéines du jaune d'oeuf et sur les phospholipides extraits du jaune d'oeuf. C R Acad Sci Hebd Seances Acad Sci D. 1968 Feb 26;266(9):959–962. [PubMed] [Google Scholar]
  6. Bereziat G., Wolf C., Colard O., Polonovski J. Phospholipases of plasmic membranes of adipose tissue. Possible intermediaries for insulin action. Adv Exp Med Biol. 1978;101:191–199. doi: 10.1007/978-1-4615-9071-2_18. [DOI] [PubMed] [Google Scholar]
  7. Bonnefis M. J., Lloveras J., Douste-Blazy L. Spécificité d'action des phospholipases de la muqueuse intestinale du rat. C R Acad Sci Hebd Seances Acad Sci D. 1975 Feb 3;280(5):661–664. [PubMed] [Google Scholar]
  8. Bonnefis M. J., Marmouyet J., Rey G., Thouvenot J. P., Douste-Blazy L. Influence des lipides de la muqueuse intestinale de rat sur son activité phospholipasique. Biochimie. 1978 Sep 4;60(5):521–527. doi: 10.1016/s0300-9084(78)80868-2. [DOI] [PubMed] [Google Scholar]
  9. Bonnefis M. J., Rey G., Thouvenot J. P., Douste-Blazy L. Phospholipases de l'intestin de rat: mode d'action. Biochimie. 1977;59(4):355–361. doi: 10.1016/s0300-9084(77)80311-8. [DOI] [PubMed] [Google Scholar]
  10. DAVIDSON F. M., LONG C. The structure of the naturally occurring phosphoglycerides. 4. Action of cabbage-leaf phospholipase D on ovolecithin and related substances. Biochem J. 1958 Jul;69(3):458–466. doi: 10.1042/bj0690458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dobbins W. O., 3rd, Ruffin J. M. A light- and electron-microscopic study of bacterial invasion in Whipple's disease. Am J Pathol. 1967 Aug;51(2):225–242. [PMC free article] [PubMed] [Google Scholar]
  12. EPSTEIN B., SHAPIRO B. Intestinal lecithinase. Nature. 1957 Aug 24;180(4582):387–387. doi: 10.1038/180387a0. [DOI] [PubMed] [Google Scholar]
  13. EPSTEIN B., SHAPIRO B. Lecithinase and lysolecithinase of intestinal mucosa. Biochem J. 1959 Apr;71(4):615–619. doi: 10.1042/bj0710615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
  15. Franson R., Dobrow R., Weiss J., Elsbach P., Weglicki W. B. Isolation and characterization of a phospholipase A2 from an inflammatory exudate. J Lipid Res. 1978 Jan;19(1):18–23. [PubMed] [Google Scholar]
  16. GALLAI-HATCHARD J. J., THOMPSON R. H. PHOSPHOLIPASE-A ACTIVITY OF MAMMALIAN TISSUES. Biochim Biophys Acta. 1965 Feb 1;98:128–136. doi: 10.1016/0005-2760(65)90017-2. [DOI] [PubMed] [Google Scholar]
  17. Garcia A., Newkirk J. D., Mavis R. D. Lung surfactant synthesis: a Ca++-dependent microsomal phospholipase A2 in the lung. Biochem Biophys Res Commun. 1975 May 5;64(1):128–135. doi: 10.1016/0006-291x(75)90228-4. [DOI] [PubMed] [Google Scholar]
  18. Goracci G., Porcellati G., Woelk H. Subcellular localization and distribution of phospholipases A in liver and brain tissue. Adv Prostaglandin Thromboxane Res. 1978;3:55–67. [PubMed] [Google Scholar]
  19. Gratecos D., Knibiehler M., Benoit V., Sémériva M. Plasma membranes from rat intestinal epithelial cells at different stages of maturation. I. Preparation and characterization of plasma membrane subfractions originating from crypt cells and from villous cells. Biochim Biophys Acta. 1978 Oct 4;512(3):508–524. doi: 10.1016/0005-2736(78)90161-x. [DOI] [PubMed] [Google Scholar]
  20. Harrison D. D., Webster H. L. Proximal to distal variations in enzymes of the rat intestine. Biochim Biophys Acta. 1971 Aug 19;244(2):432–436. doi: 10.1016/0304-4165(71)90246-7. [DOI] [PubMed] [Google Scholar]
  21. Joubert F. J., Van der Walt S. J. Naja melanoleuca (forest cobra) venom. Purification and some properties of phospholipases A. Biochim Biophys Acta. 1975 Feb 27;379(2):317–328. doi: 10.1016/0005-2795(75)90140-3. [DOI] [PubMed] [Google Scholar]
  22. King E. J. The enzymic hydrolysis of lecithin. Biochem J. 1931;25(3):799–811. doi: 10.1042/bj0250799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kunze H., Bohn E. Phospholipase A2 and prostaglandins in human seminal plasma. Adv Prostaglandin Thromboxane Res. 1978;3:159–165. [PubMed] [Google Scholar]
  24. Kwong W. K., Seetharam B., Alpers D. H. Effect of exchange exocrine pancreatic insufficiency on small intestine in the mouse. Gastroenterology. 1978 Jun;74(6):1277–1282. [PubMed] [Google Scholar]
  25. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  26. Lloveras J., Record M., Ribbes G., Douste-Blazy L. Phospholipases A of Krebs II ascites cells: specificity, release and subcellular localization. Adv Exp Med Biol. 1978;101:185–190. doi: 10.1007/978-1-4615-9071-2_17. [DOI] [PubMed] [Google Scholar]
  27. Louvard D., Maroux S., Baratti J., Desnuelle P., Mutaftschiev S. On the preparation and some properties of closed membrane vesicles from hog duodenal and jejunal brush border. Biochim Biophys Acta. 1973 Feb 16;291(3):747–763. doi: 10.1016/0005-2736(73)90478-1. [DOI] [PubMed] [Google Scholar]
  28. Louvard M. N., Puigserver A. On bovine and porcine anionic trypsinogens. Biochim Biophys Acta. 1974 Nov 5;371(1):177–185. doi: 10.1016/0005-2795(74)90167-6. [DOI] [PubMed] [Google Scholar]
  29. MAGEE W. L., GALLAI-HATCHARD J., SANDERS H., THOMPSON R. H. The purification and properties of phospholipase A from human pancreas. Biochem J. 1962 Apr;83:17–25. doi: 10.1042/bj0830017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mansbach C. M., 2nd Complex lipid synthesis in hamster intestine. Biochim Biophys Acta. 1973 Feb 14;296(2):386–402. doi: 10.1016/0005-2760(73)90097-0. [DOI] [PubMed] [Google Scholar]
  31. Mansbach C. M., 2nd, Parthasarathy S. Regulation of de novo phosphatidylcholine synthesis in rat intestine. J Biol Chem. 1979 Oct 10;254(19):9688–9694. [PubMed] [Google Scholar]
  32. Mansbach C. M., 2nd The origin of chylomicron phosphatidylcholine in the rat. J Clin Invest. 1977 Aug;60(2):411–420. doi: 10.1172/JCI108790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Meyer H., Puijk W. C., Dijkman R., Foda-van der Hoorn M. M., Pattus F., Slotboom A. J., de Haas G. H. Comparative studies of tyrosine modification in pancreatic phospholipases. 2. Properties of the nitrotyrosyl, aminotyrosyl, and dansylaminotyrosyl derivatives of pig, horse, and ox phospholipases A2 and their zymogens. Biochemistry. 1979 Aug 7;18(16):3589–3597. doi: 10.1021/bi00583a024. [DOI] [PubMed] [Google Scholar]
  34. Nachbaur J., Colbeau A., Vignais P. M. Distribution of membrane-confined phospholipases A in the rat hepatocyte. Biochim Biophys Acta. 1972 Aug 9;274(2):426–446. doi: 10.1016/0005-2736(72)90189-7. [DOI] [PubMed] [Google Scholar]
  35. Newkirk J. D., Waite M. Phospholipid hydrolysis by phospholipase A 1 and A 2 in plasma membranes and microsomes of rat liver. Biochim Biophys Acta. 1973 Mar 29;298(3):562–576. doi: 10.1016/0005-2736(73)90074-6. [DOI] [PubMed] [Google Scholar]
  36. Nieuwenhuizen W., Kunze H., de Haas G. H. Phospholipase A2 (phosphatide acylhydrolase, EC 3.1.1.4) from porcine pancreas. Methods Enzymol. 1974;32:147–154. doi: 10.1016/0076-6879(74)32018-6. [DOI] [PubMed] [Google Scholar]
  37. OTTOLENGHI A. ESTIMATION AND SUBCELLULAR DISTRIBUTION OF LECITHINASE ACTIVITY IN RAT INTESTINAL MUCOSA. J Lipid Res. 1964 Oct;5:532–537. [PubMed] [Google Scholar]
  38. Ottolenghi A. Phospholipase activity of rat tissues and its modification by trypsin(1,2). Lipids. 1967 Jul;2(4):303–307. doi: 10.1007/BF02532116. [DOI] [PubMed] [Google Scholar]
  39. Rahman Y. E., Verhagen J. Evidence of a membrane-bound phospholipase A in rat liver lysosomes. Biochem Biophys Res Commun. 1970 Feb 20;38(4):670–677. doi: 10.1016/0006-291x(70)90633-9. [DOI] [PubMed] [Google Scholar]
  40. Record M., Lloveras J., Ribes G., Douste-Blazy D. Phospholipases A1 and A2 in subcellular fractions and plasma membranes of Krebs II ascites cells. Cancer Res. 1977 Dec;37(12):4372–4377. [PubMed] [Google Scholar]
  41. Rock C. O., Snyder F. Rapid purification of phospholipase A2 from Crotalus adamanteus venom by affinity chromatography. J Biol Chem. 1975 Aug 25;250(16):6564–6566. [PubMed] [Google Scholar]
  42. SCHMIDT G., BESSMAN M. J., THANNHAUSER S. J. Enzymic hydrolysis of cephalin in rat intestinal mucosa. Biochim Biophys Acta. 1957 Jan;23(1):127–138. doi: 10.1016/0006-3002(57)90294-9. [DOI] [PubMed] [Google Scholar]
  43. SMITH H. W. OBSERVATIONS ON THE FLORA OF THE ALIMENTARY TRACT OF ANIMALS AND FACTORS AFFECTING ITS COMPOSITION. J Pathol Bacteriol. 1965 Jan;89:95–122. [PubMed] [Google Scholar]
  44. Sahu S., Lynn W. S. Characterization of phospholipase A from pulmonary secretions of patients with alveolar proteinosis. Biochim Biophys Acta. 1977 Nov 24;489(2):307–317. doi: 10.1016/0005-2760(77)90150-3. [DOI] [PubMed] [Google Scholar]
  45. Sarzala M. G. Endogenous lipolysis in mitochondria and microsomes from intestinal mucosa and liver. Bull Acad Pol Sci Biol. 1969;17(5):285–290. [PubMed] [Google Scholar]
  46. Scow R. O., Stein Y., Stein O. Incorporation of dietary lecithin and lysolecithin into lymph chylomicrons in the rat. J Biol Chem. 1967 Nov 10;242(21):4919–4924. [PubMed] [Google Scholar]
  47. Skipski V. P., Barclay M., Reichman E. S., Good J. J. Separation of acidic phospholipids by one-dimensional thin-layer chromatography. Biochim Biophys Acta. 1967 Feb 14;137(1):80–89. doi: 10.1016/0005-2760(67)90010-0. [DOI] [PubMed] [Google Scholar]
  48. Subbaiah P. V., Ganguly J. Studies on the phospholipases of rat intestinal mucosa. Biochem J. 1970 Jun;118(2):233–239. doi: 10.1042/bj1180233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Takagi R., Sasaki T. Phospholipid-deacylating enzymes of rat small intestinal mucosa. J Biochem. 1979 Jan;85(1):29–39. doi: 10.1093/oxfordjournals.jbchem.a132323. [DOI] [PubMed] [Google Scholar]
  50. Tocanne J. F., Ververgaert P. H., Verkleij A. J., van Deenen L. L. A monolayer and freeze-etching study of charged phospholipids. I. Effects of ions and pH on the ionic properties of phosphatidylglycerol and lysylphosphatidylglycerol. Chem Phys Lipids. 1974 May;12(3):201–219. doi: 10.1016/0009-3084(74)90075-9. [DOI] [PubMed] [Google Scholar]
  51. Van Golde L. M., Fleischer B., Fleischer S. Some studies on the metabolism of phospholipids in Golgi complex from bovine and rat liver in comparison to other subcellular fractions. Biochim Biophys Acta. 1971 Oct 12;249(1):318–330. doi: 10.1016/0005-2736(71)90109-x. [DOI] [PubMed] [Google Scholar]
  52. Verger R., De Haas G. H. Enzyme reactions in a membrane model. 1. A new technique to study enzyme reactions in monolayers. Chem Phys Lipids. 1973 Feb;10(2):127–136. doi: 10.1016/0009-3084(73)90009-1. [DOI] [PubMed] [Google Scholar]
  53. Verger R., Rietsch J., Van Dam-Mieras M. C., de Haas G. H. Comparative studies of lipase and phospholipase A2 acting on substrate monolayers. J Biol Chem. 1976 May 25;251(10):3128–3133. [PubMed] [Google Scholar]
  54. Waite M., Sisson P. Partial purification and characterization of the phospholipase A 2 from rat liver mitochondria. Biochemistry. 1971 Jun 8;10(12):2377–2383. doi: 10.1021/bi00788a031. [DOI] [PubMed] [Google Scholar]
  55. Wassef M. K., Lin Y. N., Horowitz M. I. Phospholipid-deacylating enzymes of rat stomach mucosa. Biochim Biophys Acta. 1978 Mar 30;528(3):318–330. doi: 10.1016/0005-2760(78)90021-8. [DOI] [PubMed] [Google Scholar]
  56. Weiser M. M. Intestinal epithelial cell surface membrane glycoprotein synthesis. I. An indicator of cellular differentiation. J Biol Chem. 1973 Apr 10;248(7):2536–2541. [PubMed] [Google Scholar]
  57. Weiss J., Franson R. C., Beckerdite S., Schmeidler K., Elsbach P. Partial characterization and purification of a rabbit granulocyte factor that increases permeability of Escherichia coli. J Clin Invest. 1975 Jan;55(1):33–42. doi: 10.1172/JCI107915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Wells M. A., Hanahan D. J. Studies on phospholipase A. I. Isolation and characterization of two enzymes from Crotalus adamanteus venom. Biochemistry. 1969 Jan;8(1):414–424. doi: 10.1021/bi00829a057. [DOI] [PubMed] [Google Scholar]
  59. Wu T. W., Tinker D. O. PhospholipA2se A from Crotalus atrox venom. I. Purification and some properties. Biochemistry. 1969 Apr;8(4):1558–1568. doi: 10.1021/bi00832a035. [DOI] [PubMed] [Google Scholar]
  60. Yang S. F., Freer S., Benson A. A. Transphosphatidylation by phospholipase D. J Biol Chem. 1967 Feb 10;242(3):477–484. [PubMed] [Google Scholar]
  61. de Haas G. H., Postema N. M., Nieuwenhuizen W., van Deenen L. L. Purification and properties of phospholipase A from porcine pancreas. Biochim Biophys Acta. 1968 Apr 24;159(1):103–117. doi: 10.1016/0005-2744(68)90248-9. [DOI] [PubMed] [Google Scholar]
  62. de Haas G. H., Slotboom A. J., Verheij H. M., Jansen E. H., de Araujo P. S., Vidal J. C. Interaction of phospholipase A2 with lipid-water interfaces. Adv Prostaglandin Thromboxane Res. 1978;3:11–21. [PubMed] [Google Scholar]
  63. van den Bosch H., Aarsman A. J., van Deenen L. L. Isolation and properties of a phospholipase A1 activity from beef pancreas. Biochim Biophys Acta. 1974 May 29;348(2):197–209. doi: 10.1016/0005-2760(74)90231-8. [DOI] [PubMed] [Google Scholar]
  64. van den Bosch H. Intracellular phospholipases A. Biochim Biophys Acta. 1980 Sep 30;604(2):191–246. doi: 10.1016/0005-2736(80)90574-x. [DOI] [PubMed] [Google Scholar]

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