Antonie van Leeuwenhoek

, Volume 31, Issue 1, pp 323–340 | Cite as

Thetrans-eliminative breakdown of Na-polygalacturonate byPseudomonas fluorescens

  • A. Fuchs
Article
Received:

Abstract

Three strains ofPseudomonas fluorescens were isolated, which produced an inducible enzyme capable of converting Na-polygalacturonate into several reaction products which strongly absorbed ultraviolet light with a maximum at 232 mµ. Paper and column chromatography revealed that these products constituted a homologous series of oligo-uronides. The fastest moving compound appeared to be an unsaturated mono-uronic acid, 4-deoxy-l-threo-5-hexoseulose uronic acid, whereas indirect evidence showed the others to consist of the unsaturated mono-uronic acid linked by glycosidic bonding to 1, 2, 3, or 4 galacturonic acid moieties. Therefore, the enzyme concerned appears to be an endo-polygalacturonic acidtrans-eliminase. Enzyme extracts prepared from the isolates did not show a hydrolytic activity.

The effects of some factors (pH, substrate and enzyme concentrations, inhibitory substances) on enzyme kinetics were investigated.

Study of the literature on bacterial pectolytic enzymes suggests that many of these have been wrongly described as depolymerases, polygalacturonases, macerating enzymes, etc., and are better classified astrans-eliminases.

Keywords

Column Chromatography Indirect Evidence Enzyme Extract Ultraviolet Light Enzyme Concentration 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Albersheim, P. 1963. Auxin induced product inhibition of pectin transeliminase as shown by ozonolysis, Plant Physiol.38: 426–429.CrossRefPubMedGoogle Scholar
  2. Albersheim, P. andKillias, U. 1962. Studies relating to the purification and properties of pectin transeliminase. Arch. Biochem. Biophys.97: 107–115.CrossRefPubMedGoogle Scholar
  3. Albersheim, P., Neukom, H. andDeuel, H. 1960a. Splitting of pectin chain molecules in neutral solutions. Arch. Biochem. Biophys.90: 46–51.CrossRefPubMedGoogle Scholar
  4. Albersheim, P., Neukom, H. undDeuel, H. 1960b. Über die Bildung von ungesättigten Abbauprodukten durch ein pektinabbauendes Enzym. Helv. Chim. Acta43: 1422–1426.CrossRefGoogle Scholar
  5. Ceponis, M. J. andFriedman, B. A. 1959. Pectolytic enzymes ofPseudomonas marginalis and their effects on lettuce. Phytopathology49: 141–144.Google Scholar
  6. Collins, R. P. andSledjeski, W. F. 1960. Pectolytic enzymes produced byBotryosphaeria ribis andPenicillium italicum. Mycologia52: 455–459.Google Scholar
  7. Dorey, M. J. 1959. Some properties of a pectolytic soilFlavobacterium. J. Gen. Microbiol.20: 91–104.PubMedGoogle Scholar
  8. Echandi, E., Van Gundy, S. D. andWalker, J. C. 1957. Pectolytic enzymes secreted by soft-rot bacteria. Phytopathology47: 549–552.Google Scholar
  9. Fernando, M. 1937. Studies of the physiology of parasitism. XV. Effect of the nutrient medium upon the secretion and properties of pectinase. Ann. Botany (London) N.S.1: 727–745.Google Scholar
  10. French, D. andWild, G. M. 1953. Correlation of carbohydrate structure with papergram mobility. J. Am. Chem. Soc.75: 2612–2616.CrossRefGoogle Scholar
  11. Fuchs, A. 1959. On the synthesis and breakdown of levan by bacteria. Thesis, University of Leiden.Google Scholar
  12. Fuchs, A. 1965. On the gelation of pectin by plant extracts and its inhibition. Some considerations in relation to host-parasite interactions. Acta Bot. Neerl.14 (In press).Google Scholar
  13. Gregg, M. 1952. Studies in the physiology of parasitism. XVII. Enzyme secretion by strains ofBacterium carotovorum and other pathogens in relation to parasitic vigour. Ann. Botany (London) N. S.16: 235–250.Google Scholar
  14. Grossmann, F. 1962. Untersuchungen über die Hemmung pektolytischer Enzyme vonFusarium oxysporum f.lycopersici. I. Hemmung durch definierte Substanzen in vitro. Phytopathol. Z.44: 361–380.Google Scholar
  15. Hasegawa, S. andNagel, C. W. 1962. The characterization of an α,β-unsaturated digalacturonic acid. J. Biol. Chem.237: 619–621.PubMedGoogle Scholar
  16. Kraght, A. J. andStarr, M. P. 1953. Pectic enzymes ofErwinia carotovora. Arch. Biochem. Biophys.42: 271–277.CrossRefPubMedGoogle Scholar
  17. Lanning, M. C. andCohen, S. S. 1951. The detection and estimation of 2-ketohexonic acids. J. Biol. Chem.189: 109–114.PubMedGoogle Scholar
  18. Lapwood, D. H. 1957. Studies in the physiology of parasitism. XXIII. On the parasitic vigour of certain bacteria in relation to their capacity to secrete pectolytic enzymes. Ann. Botany (London) N.S.21: 167–184.Google Scholar
  19. Lineweaver, H. andBurk, D. 1934. The determination of enzyme dissociation constants. J. Am. Chem. Soc.56: 658–666.CrossRefGoogle Scholar
  20. Macmillan, J. D., Phaff, H. J. andVaughn, R. H. 1964. The pattern of action of an exopolygalacturonic acid-trans-eliminase fromClostridium multifermentans. Biochemistry3: 572–578.CrossRefPubMedGoogle Scholar
  21. Macmillan, J. D. andVaughn, R. H. 1964. Purification and properties of a polygalacturonic acid-trans-eliminase produced byClostridium multifermentans. Biochemistry3: 564–572.CrossRefPubMedGoogle Scholar
  22. Nagel, C. W. andVaughn, R. H. 1961. The degradation of oligogalacturonides by the polygalacturonase ofBacillus polymyxa. Arch. Biochem. Biophys.94: 328–332.CrossRefPubMedGoogle Scholar
  23. Nagel, C. W. andVaughn, R. H. 1962. Comparison of growth and pectolytic enzyme production byBacillus polymyxa. J. Bacteriol.83: 1–5.CrossRefPubMedGoogle Scholar
  24. Nortje, B. K. andVaughn, R. H. 1953. The pectolytic activity of species of the genusBacillus: Qualitative studies withBacillus subtilis andBacillus pumilis in relation to the softening of olives and pickles. Food Res.18: 57–69.Google Scholar
  25. Oxford, A. E. 1944. Production of a soluble pectinase in a simple medium by certain plant-pathogenic bacteria belonging to the genusPseudomonas. Nature154: 271–272.Google Scholar
  26. Preiss, J. andAshwell, G. 1963. Polygalacturonic acid metabolism in bacteria. I. Enzymatic formation of 4-deoxy-l-threo-5-hexoseulose uronic acid. J. Biol. Chem.238: 1571–1576.PubMedGoogle Scholar
  27. Schwane, R. A. andKowkabany, G. N. 1963. The brown front in paper chromatography. Anal. Chem.35: 1660–1662.CrossRefGoogle Scholar
  28. Starr, M. P. andMoran, F. 1962. Eliminative split of pectic substances by phytopathogenic soft-rot bacteria. Science135: 920–921.PubMedGoogle Scholar
  29. Stolp, H. 1961. Neue Erkenntnisse über phytopathogene Bakterien und die von ihnen verursachten Krankheiten. I. Verwandtschaftsbeziehungen zwischen phytopathogenenPseudomonas-„Arten“ und saprophytischen Fluoreszenten auf der Grundlage von Phagenreaktionen, Phytopathol. Z.42: 197–262.Google Scholar
  30. Tribe, H. T. 1955. Studies in the physiology of parasitism. XIX. On the killing of plant cells by enzymes fromBotrytis cinerea andBacterium aroideae. Ann. Botany (London) N.S.19: 351–368.Google Scholar
  31. Tuttobello, R. andMill, P. J. 1961. The pectic enzymes ofAspergillus niger. 1. The production of active mixtures of pectic enzymes. Biochem. J.79: 51–57.PubMedGoogle Scholar
  32. Umbarger, H. E. andMagasanik, B. 1952. Isoleucine and valine metabolism inEscherichia coli. III. A method for the quantitative determination of α-keto acid analogs of isoleucine and valine, J. Am. Chem. Soc.74: 4253–4255.CrossRefGoogle Scholar
  33. Warren, L. 1960. Thiobarbituric acid spray reagent for deoxy sugars and sialic acids. Nature186: 237.PubMedGoogle Scholar
  34. Wieringa, G. W. 1963. The influence of the pH on the pectinolytic activity of flax-retting bacteria. Antonie van Leeuwenhoek29: 84–88.CrossRefPubMedGoogle Scholar
  35. Wood, R. K. S. 1951. Pectic enzymes produced byBacterium aroideae. Nature167: 771.PubMedGoogle Scholar
  36. Wood, R. K. S. 1955. Studies in the physiology of parasitism. XVIII. Pectic enzymes secreted byBacterium aroideae. Ann. Botany (London) N.S.19: 1–27.Google Scholar

Copyright information

© Swets & Zeitlinger, Publishers 1965

Authors and Affiliations

  • A. Fuchs
    • 1
  1. 1.Laboratory of PhytopathologyAgricultural UniversityWageningenThe Netherlands

Personalised recommendations