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Purification of endo polygalacturonases from Sclerotinia sclerotiorum: Multiplicity of the complex enzyme system

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Abstract

Fourteen forms of endopolygalacturonases were purified from the culture medium of Sclerotinia sclerotiorum with ion exchange chromatographies. Individual forms differ in their isoelectric point but exhibit similar molecular masses. On the basis of the cross-reactions to antibodies raised against a purified acidic endopolygalacturonase, these forms could be divided into two related groups. Polygalacturonase activities constitute a complex enzyme system in which the extensive multiplicity is due to the expression of a large gene family.

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Literature Cited

  1. Alghisi P, Favaron F (1995) Pectin-degrading enzymes and plant parasite interactions. Eur J Plant Pathol 101: 365–375

    Article  CAS  Google Scholar 

  2. Bateman D, Basham H (1976) Degradation of plant cell walls and membranes by microbial enzymes. In: Heitefuss R, Williams P (eds) Physiological plant pathology. Berlin: Springer-Verlag, pp 316–355

    Google Scholar 

  3. Bussink H, Buxton F, Fraaye B, de Graaff L, Visser J (1992) The polygalacturonases of Aspergillus niger are encoded by a family of diverged genes. Eur J Biochem 208: 83–90

    Article  PubMed  CAS  Google Scholar 

  4. Byrde RJW (1979) Role of polysaccharide-degrading enzymes in microbial pathogenicity. In: Berkeley RCW, Gooday GW, Ellwood DC (eds) Microbial polysaccharides and polysaccharases. London: Academic Press, pp 417–436

    Google Scholar 

  5. Caprari C, Bergmann C, Micheli Q, Salvi C, Alberheim P, Darvill A, Cervone F, De Lorenzo G (1993) Fusarium moniliforme secretes four endopolygalacturonases derived from a single gene product. Physiol Mol Plant Pathol 43: 453–462

    Article  CAS  Google Scholar 

  6. Favaron F, Alghisi P, Marciano P (1992) Characterization of two Sclerotinia sclerotiorum polygalacturonases with different abilities to elicit glyceollin in soybean. Plant Sci 83: 7–13

    Article  CAS  Google Scholar 

  7. Fraissinet-Tachet L, Reymond-Cotton P, Fèvre M (1995) Characterization of a multigene family encoding an endopolygalacturonase in Sclerotinia sclerotiorum. Curr Genet 29: 96–100

    Article  PubMed  CAS  Google Scholar 

  8. Honda S, Nishimura Y, Takahashi M, Chiba H, Kakehi K (1982) A manual method for the spectrophotometric determination of reducing carbohydrates with 2-cyanoacetamide. Anal Biochem 119: 194–199

    Article  PubMed  CAS  Google Scholar 

  9. Keon JPR, Waksman G (1990) Common amino acid domain among endo polygalacturonases of Ascomycetes fungi. Appl Environ Microbiol 56: 2522–2528

    PubMed  CAS  Google Scholar 

  10. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 222: 680–685

    Article  Google Scholar 

  11. Marciano P, Di Lenna P, Magro P (1982) Polygalacturonase isoenzymes produced by Sclerotinia sclerotiorum in vivo and in vitro. Physiol Plant Pathol 20: 201–212

    Article  CAS  Google Scholar 

  12. Reymond P, Deléage G, Rascle C, Fèvre M (1994) Cloning and sequence analysis of a polygalacturonase-encoding gene from the phytopathogenic fungus S. sclerotiorum. Gene 146: 233–237

    Article  PubMed  CAS  Google Scholar 

  13. Riou C, Freyssinet G, Fèvre M (1991a) Production of cell wall-degrading enzymes by the phytopathogenic fungus Sclerotinia sclerotiorum. Appl Environ Microbiol 57: 1478–1484

    PubMed  CAS  Google Scholar 

  14. Riou C, Fraissinet-Tachet L, Freyssinet G, Fèvre M (1991b) Purification and characterization of extracellular pectinolytic enzymes produced by Sclerotinia sclerotiorum. Environ Microbiol 58: 578–583

    Google Scholar 

  15. Riou C, Fraissinet-Tachet L, Freyssinet G, Fèvre M (1992) Secretion of pectic isoenzymes by Sclerotinia sclerotiorum. FEMS Microbiol Lett 91: 231–238

    Article  CAS  Google Scholar 

  16. Smith P, Krohn R, Hermanson G, Mallia A, Gartner F, Provenzano M, Fujimoto E, Goeke N, Olson B, Klenk D (1985) Measurement of protein using bincinchoninic acid. Anal Biochem 150: 76–85

    Article  PubMed  CAS  Google Scholar 

  17. Stratilova E, Markovic O, Strovinova D, Rexova-Benkova L, Jornvall H (1993) Aspergillus sp. polygalacturonase: multiplicity, divergence and structural patterns linking fungal, bacterial and plant polygalacturonases. J Prot Chem 12: 15–22

    Article  CAS  Google Scholar 

  18. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354

    Article  PubMed  CAS  Google Scholar 

  19. Waksman G, Keon JP, Turner G (1991) Purification and characterization of two endopolygalacturonases from Sclerotinia sclerotiorum. Biochim Biophys Acta 1073: 43–48

    PubMed  CAS  Google Scholar 

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  1. Laboratoire de Biologie Cellulaire Fongique, Centre de Génétique Moléculaire et Cellulaire, UMR-CNRS 106, Université Lyon I (Bât. 405), 43 Bd du, 69622, Villeurbanne Cedex, France

    Marie-Bénédicte Martel, Robert Létoublon & Michel Fěvre

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  1. Marie-Bénédicte Martel
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  2. Robert Létoublon
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  3. Michel Fěvre
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Martel, MB., Létoublon, R. & Fěvre, M. Purification of endo polygalacturonases from Sclerotinia sclerotiorum: Multiplicity of the complex enzyme system. Current Microbiology 33, 243–248 (1996). https://doi.org/10.1007/s002849900107

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  • DOI: https://doi.org/10.1007/s002849900107

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