Skip to main content

Official Journal of the Japan Wood Research Society

Journal of Wood Science Cover Image

Extracellular peroxidase activity at the hyphal tips of the white-rot fungus Phanerochaete crassa WD1694

Abstract

The white-rot fungus Phanerochaete crassa WD1694 was cultivated and peroxidase activity staining was performed to determine the sites at which the extracellular peroxidase reaction actually occurs in vivo. Although the ligninolytic peroxidases were found in the culture filtrates, the culture medium did not show a color reaction. However, a particularly strong color reaction was observed on the hyphal tips. Visible spectra and absorbance of the staining were analyzed by microspectrophotometry, and the catalytic rates of the peroxidase reaction at the hyphal tips were calculated. The estimated catalytic rate of the peroxidase reaction at the hyphal tips peaked at 794 μM/min, expressed as the consumption rate of H2O2, on day 3 of the cultivation. Analysis of the extracellular enzyme eluted with 0.1% Tween 80 from the mycelium revealed that manganese peroxidase accounted for 89% of all the peroxidase activity measured. The results clearly showed the existence of the concentrated manganese peroxidase reaction around the hyphal tips of the organism.

References

  1. 1.

    Nakano J (1979) Lignin in plants (in Japanese). In: Nakano J (ed) Chemistry of lignin, basic science and application. Uni, Tokyo, pp 20–36

    Google Scholar 

  2. 2.

    Higuchi T (1985) Biosynthesis of lignin. In: Higuchi T (ed) Biosynthesis and biodegradation of wood components. Academic, London, pp 141–161

    Google Scholar 

  3. 3.

    Chen CL, Chang HM (1985) Chemistry of lignin biodegradation. In: Higuchi T (ed) Biosynthesis and biodegradation of wood components. Academic, London, pp 535–556

    Google Scholar 

  4. 4.

    Ander P (1990) Biodegradation of lignin. In: Eriksson KEL, Blanchette RA, Ander P (eds) Microbial and enzymatic degradation of wood and wood components. Springer, Berlin Heidelberg New York, pp 225–333

    Google Scholar 

  5. 5.

    Glenn JK, Morgan MA, Mayfield MB, Kuwahara M, Gold MH (1983) An extracellular H2O2-requiring enzyme preparation involved in lignin biodegradation by the white-rot basidiomycete Phanerochaete chrysosporium. Biochem Biophys Res Commun 114:1077–1083

    CAS  Article  PubMed  Google Scholar 

  6. 6.

    Kuwahara M, Glenn JK, Morgan MA, Gold MH (1984) Separation and characterization of two extracellular H2O2-dependent oxidases from ligninolytic cultures of Phanerochaete chrysosporium. FEBS Lett 169:247–250

    CAS  Article  Google Scholar 

  7. 7.

    Tien M, Kirk TK (1983) Lignin-degrading enzyme from the hymenomycete Phanerochaete chrysosporium Burds. Science 221:661–663

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    Gold MH, Alic M (1993) Molecular biology of the lignin-degrading basidiomycete Phanerochaete chrysosporium. Microbiol Rev 57: 605–622

    CAS  PubMed  PubMed Central  Google Scholar 

  9. 9.

    Kirk TK (1987) Enzymatic “combustion”: the microbial degradation of lignin. Ann Rev Microbiol 41:465–505

    CAS  Article  Google Scholar 

  10. 10.

    Gold MH, Wariishi H, Valli K (1989) Extracellular peroxidases involved in lignin degradation by the white rot basidiomycete Phanerochaete chrysosporium. In: Biocatalysis in agricultural biotechnology, ACS Symposium Series 389:127–140

    CAS  Article  Google Scholar 

  11. 11.

    Higuchi T (1985) Degradative pathways of lignin model compounds. In: Higuchi T (ed) Biosynthesis and biodegradation of wood components. Academic, London, pp 557–578

    Google Scholar 

  12. 12.

    Umezawa T (1988) Mechanisms for chemical reactions involved in lignin biodegradation by Phanerochaete chrysosporium. Wood Res 75:21–79

    CAS  Google Scholar 

  13. 13.

    Daniel G (1994) Use of electron microscopy for aiding our understanding of wood biodegradation. FEMS Microbial Rev 13:199–233

    CAS  Article  Google Scholar 

  14. 14.

    Daniel G, Nilsson T, Pettersson B (1989) Intra-and extracellular localization of lignin peroxidase during the degradation of solid wood and wood fragments by Phanerochaete chrysosporium by using transmission electron microscopy and immuno-gold labeling. Appl Environ Microbiol 55:871–881

    CAS  PubMed  PubMed Central  Google Scholar 

  15. 15.

    Joseleau JP, Ruel K (1992) Ultrastructual examination of lignin and polysaccharide degradation in wood by white-rot fungi. In: Kuwahara M, Shimada M (eds) Biotechnology in pulp and paper industry. Uni, Tokyo, pp 195–201

    Google Scholar 

  16. 16.

    Daniel G, Pettersson B, Volc J, Nilsson T (1990) Spatial distribution of lignin-and manganese peroxidase(s) during degradation of wood and wood fragments by Phanerochaete chrysosporium as revealed by T.E.M. immunogold labeling. In: Kirk TK, Chang H-M (eds) Biotechnology in pulp and paper manufacture, applications and fundamental investigations. Butterworth-Heinemann, Boston, pp 99–110

    Google Scholar 

  17. 17.

    Takano M, Nishida A, Nakamura M (2001) Screening of wood-rotting fungi for kraft pulp bleaching by the Poly R decoloration test and biobleaching of hardwood kraft pulp by Phanerochaete crassa WD1694. J Wood Sci 47:63–68

    CAS  Article  Google Scholar 

  18. 18.

    Takano M, Nakamura M, Nishida A, Ishihara M (2004) Manganese peroxidase from Phanerochaete crassa WD1694. Bull For For Prod Res Inst 3:7–13

    CAS  Google Scholar 

  19. 19.

    Wariishi H, Valli K, Gold MH (1992) Manganese (II) oxidation by manganese peroxidase from the basidiomycete Phanerochaete chrysosporium. J Biol Chem 267:23688–23695

    CAS  PubMed  Google Scholar 

  20. 20.

    Mester T, Field JA (1998) Characterization of a novel manganese peroxidase-lignin peroxidase hybrid isozyme produced by Bjerkandera species strain BOS55 in the absence of manganese. J Biol Chem 273:15412–15417

    CAS  Article  PubMed  Google Scholar 

  21. 21.

    Sprey B (1988) Cellular and extracellular localization of endocellulase in Trichoderma reesei. FEMS Microbiol Lett 55:283–294

    CAS  Article  Google Scholar 

  22. 22.

    Pugh D, Cawson RA (1977) The cytochemical localization of acid hydrolases in four common fungi. Cell Mol Biol 22:125–132

    CAS  Google Scholar 

  23. 23.

    Chung PLY, Trevithick JR (1970) Biochemical and histochemical localization of invertase in Neurospora crassa during conidial germination and hyphal growth. J Bacteriol 102:423–429

    CAS  PubMed  PubMed Central  Google Scholar 

  24. 24.

    Cai YJ, Chapman SJ, Buswell JA, Chang S (1999) Production and distribution of endoglucanase, cellobiohydrolase, and β-glucosidase components of the cellulolytic system of Volvariella volvacea, the edible straw mushroom. Appl Environ Microbiol 65:553–559

    CAS  PubMed  PubMed Central  Google Scholar 

  25. 25.

    Wösten HAB, Moukha SM, Siestsma JH, Wessels JGH (1991) Localization of growth and secretion of proteins in Aspergillus niger. J Microbiol 137:2017–2023

    Google Scholar 

  26. 26.

    Wessels JGH (1993) Wall growth, protein secretion and morphogenesis in fungi. New Phytol 123:397–413

    CAS  Article  Google Scholar 

  27. 27.

    Wariishi H, Valli K, Gold MH (1991) In vitro depolymerization of lignin by manganese peroxidase of Phanerochaete chrysosporium. Biochem Biophys Res Commun 176:269–275

    CAS  Article  PubMed  Google Scholar 

  28. 28.

    Hammel KE, Moen MA (1991) Depolymerization of a synthetic lignin in vitro by lignin peroxidase. Enzyme Microb Technol 13:15–18

    CAS  Article  Google Scholar 

  29. 29.

    Haemmerli SD, Leisola MSA, Fiechter A (1986) Polymerization of lignins by ligninases from Phanerochaete chrysosporium. FEMS Microbiol lett 35:33–36

    CAS  Article  Google Scholar 

  30. 30.

    Evans RC, Stempen H, Stewart SJ (1981) Development of hyphal sheaths in Bipolaris maydis race T. Canad J Bot 59:453–459

    Article  Google Scholar 

  31. 31.

    Abu AR, Murphy RJ, Dickinson DJ (1999) Investigation of the extracellular mucilaginous materials produced by some wood decay fungi. Mycol Res 103:1453–1461

    Article  Google Scholar 

  32. 32.

    Palmer JG, Muranis L, Highly TL (1983) Visualization of hyphal sheath in wood-decay hymenomycetes. I. Brown-rotters. Mycologia 75:995–1004

    Article  Google Scholar 

  33. 33.

    Palmer JG, Muranis L, Highly TL (1983) Visualization of hyphal sheath in wood-decay hymenomycetes. II. White-rotters. Mycologia 75:1005–1010

    Article  Google Scholar 

  34. 34.

    Ruel K, Joseleau JP (1991) Involvement of an extracellular glucan sheath during degradation of Populus wood by Phanerochaete chrysosporium. Appl Environ Microbiol 57:374–384

    CAS  PubMed  PubMed Central  Google Scholar 

  35. 35.

    Barrasa JM, Gutiérrez A, Escaso V, Guillén F, Martínez MJ, Martínez AT (1998) Electron and fluorescence microscopy of extracellular glucan and aryl-alcohol oxidase during wheat-straw degradation by Pleurotus eryngii. Appl Environ Microbiol 64:325–332

    CAS  PubMed  PubMed Central  Google Scholar 

  36. 36.

    Bes B, Pettersson B, Lentholm H, Iversen T, Eriksson KE (1987) Synthesis, structure, and enzymatic degradation of an extracellular glucan produced in nitrogen-starved cultures of the white rot fungus Phanerochaete chrysosporium. Biotechnol Appl Biochem 9:310–318

    CAS  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Mariko Takano.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Takano, M., Abe, H. & Hayashi, N. Extracellular peroxidase activity at the hyphal tips of the white-rot fungus Phanerochaete crassa WD1694. J Wood Sci 52, 429–435 (2006). https://doi.org/10.1007/s10086-005-0781-0

Download citation

Key words

  • White-rot fungi
  • Phanerochaete crassa
  • Microspectrophotometry
  • Manganese peroxidase
  • Hyphal tips