Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

An endo-β-1,6-glucanase involved in Lentinula edodes fruiting body autolysis

Abstract

A β-1,6-glucanase, LePus30A, was purified and cloned from fruiting bodies of the basidiomycete Lentinula edodes. β-1,6-Glucanases degrade β-1,6-glucan polysaccharides, a unique and essential component of fungal cell walls. The complementary DNA of LePus30A includes an open reading frame of 1,575 bp encoding an 18 amino acid signal peptide and the 506 amino acid mature protein. Sequence analysis indicated that LePus30A is a member of glycoside hydrolase family 30, and highly similar genes are broadly conserved among basidiomycetes. The purified LePus30A catalyzed depolymerization of β-1,6-glucan endolytically and was highly specific toward β-1,6-glucan polysaccharide. It is known that the cell walls of fruiting bodies of basidiomycetes are autodegraded after harvesting by means of enzymatic hydrolysis. The transcript level of LePus30A gene (lepus30a) was significantly increased in fruiting bodies after harvesting. Moreover, LePus30A showed hydrolyzing activity against the cell wall components of L. edodes fruiting bodies. These results suggest that LePus30A is responsible for the degradation of the cell wall components during fruiting body autolysis after harvest.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. Adams DJ (2004) Fungal cell wall chitinases and glucanases. Microbiology 150:2029–2035

  2. Adams EL, Rice PJ, Graves B, Ensley HE, Yu H, Brown GD, Gordon S, Monteiro MA, Papp-Szabo E, Lowman DW, Power TD, Wempe MF, Williams DL (2008) Differential high-affinity interaction of dectin-1 with natural or synthetic glucans is dependent upon primary structure and is influenced by polymer chain length and side-chain branching. J Pharmacol Exp 325:115–123

  3. Aimanianda V, Clavaud C, Simenel C, Fontaine T, Delepierre M, Latgé JP (2009) Cell wall β-(1,6)-glucan of Saccharomyces cerevisiae: structural characterization and in situ synthesis. J Biol Chem 284:13401–13012

  4. Bottom CB, Siehr DJ (1979) Structure of an alkali-soluble polysaccharide from the hyphal wall of the basidiomycete Coprinus macrorhizus var. microsporus. Carbohydr Res 77:169–181

  5. Bryant MK, May KJ, Bryan GT, Scott B (2007) Functional analysis of a β-1,6-glucanase gene from the grass endophytic fungus Epichloë festucae. Fungal Genet Biol 44:808–817

  6. Chihara G, Maeda Y, Hamuro J, Sasaki T, Fukuoka F (1969) Inhibition of mouse sarcoma 180 by polysaccharides from Lentinus edodes (Berk.) Sing. Nature 222:687–688

  7. Chizhov AO, Dell A, Morris HR, Reason AJ, Haslam Stuart M, McDowell RA, Chizhov OS, Usov AI (1998) Structural analysis of laminarans by MALDI and FAB mass spectrometry. Carbohydr Res 310:203–210

  8. De la Cruz J, Llobell A (1999) Purification and properties of a basic endo-β-1,6-glucanase (BGN16.1) from the antagonistic fungus Trichoderma harzianum. Eur J Biochem 265:145–151

  9. Djonović S, Pozo MJ, Kenerley CM (2006) Tvbgn3, a β-1,6-glucanase from the biocontrol fungus Trichoderma virens, is involved in mycoparasitism and control of Pythium ultimum. Appl Environ Microbiol 72:7661–7670

  10. Dvir H, Harel M, McCarthy AA, Toker L, Silman I, Futerman AH, Sussman JL (2003) X-ray structure of human acid-β-glucosidase, the defective enzyme in Gaucher disease. EMBO Rep 4:704–709

  11. Enderlin CS, Selitrennikoff CP (1994) Cloning and characterization of a Neurospora crassa gene required for (1,3)β-glucan synthase activity and cell wall formation. Proc Natl Acad Sci USA 91:9500–9504

  12. Fontaine T, Simenel C, Dubreucq G, Adam O, Delepierre M, Lemoine J, Vorgias CE, Diaquin M, Latgé JP (2000) Molecular organization of the alkali-insoluble fraction of Aspergillus fumigatus cell wall. J Biol Chem 275:27594–27607

  13. Fukuda K, Hiraga M, Asakuma S, Arai I, Sekikawa M, Urashima T (2008) Purification and characterization of a novel exo-β-1,3-1,6-glucanase from the fruiting body of the edible mushroom Enoki (Flammulina velutipes). Biosci Biotechnol Biochem 72:3107–3113

  14. Hirano T, Sato T, Okawa K, Kanda K, Yaegashi K, Enei H (1999) Isolation and characterization of the glyceraldehyde-3-phosphate dehydrogenase gene of Lentinus edodes. Biosci Biotechnol Biochem 63:1223–1227

  15. Ishibashi K, Miura NN, Adachi Y, Ohno N, Yadomae T (2001) Relationship between solubility of grifolan, a fungal 1,3-β-d-glucan, and production of tumor necrosis factor by macrophages in vitro. Biosci Biotechnol Biochem 65:1993–2000

  16. Kim YT, Kim EH, Cheong C, Williams DL, Kim CW, Lim ST (2000) Structural characterization of β-d-(1→3, 1→6)-linked glucans using NMR spectroscopy. Carbohydr Res 328:31–41

  17. Kim YG, Kim JH, Kim KJ (2009) Crystal structure of the Salmonella enterica serovar typhimurium virulence factor SrfJ, a glycoside hydrolase family enzyme. J Bacteriol 191:6550–6554

  18. Kollár R, Reinhold BB, Petráková E, Yeh HJ, Ashwell G, Drgonová J, Kapteyn JC, Klis FM, Cabib E (1997) Architecture of the yeast cell wall: β(1→6)-glucan interconnects mannoprotein, β(1→3)-glucan, and chitin. J Biol Chem 272:17762–17775

  19. Kües U (2000) Life history and developmental processes in the basidiomycete Coprinus cinereus. Microbiol Mol Biol Rev 64:316–353

  20. Mahadevan PR, Mahadkar UR (1970) Role of enzymes in growth and morphology of Neurospora crassa: cell-wall-bound enzymes and their possible role in branching. J Bacteriol 101:941–947

  21. Minato K, Kawakami S, Nomura K, Tsuchida H, Mizuno M (2004) An exo β-1,3 glucanase synthesized de novo dgrades lentinan during storage of Lentinula edodes and diminishes immunomodulationg activity of the mushroom. Carbohydr Polym 56:279–286

  22. Montero M, Sanz L, Rey M, Monte E, Llobell A (2005) BGN16.3, a novel acidic β-1,6-glucanase from mycoparasitic fungus Trichoderma harzianum CECT 2413. FEBS J 272:3441–3448

  23. Moy M, Li HM, Sullivan R, White JF Jr, Belanger FC (2002) Endophytic fungal β-1,6-glucanase expression in the infected host grass. Plant Physiol 13:1298–1308

  24. Nagai M, Kawata M, Watanabe H, Ogawa M, Saito K, Takesawa T, Kanda K, Sato T (2003) Important role of fungal intracellular laccase for melanin synthesis: purification and characterization of an intracellular laccase from Lentinula edodes fruit bodies. Microbiology 149:2455–2462

  25. Nishikawa Y, Tanaka M, Shibata S, Fukuoka F (1970) Polysaccharides of lichens and fungi. IV. Antitumour active O-acetylated pustulan-type glucans from the lichens of Umbilicaria species. Chem Pharm Bull 18:1431–1434

  26. Ohno N, Furukawa M, Miura NN, Adachi Y, Motoi M, Yadomae T (2001) Antitumor β-glucan from the cultured fruit body of Agaricus blazei. Biol Pharm Bull 24:820–828

  27. Ooi VEC, Liu F (2000) Immunomodulation and anti-cancer activity of polysaccharide–protein complexes. Curr Med Chem 7:715–729

  28. Oyama S, Yamagata Y, Abe K, Nakajima T (2002) Cloning and expression of an endo-1,6-β-d-glucanase gene (neg1) from Neurospora crassa. Biosci Biotechnol Biochem 66:1378–1381

  29. Pitson SM, Seviour RJ, McDougall BM, Stone BA, Sadek M (1996) Purification and characterization of an extracellular (1→6)-β-glucanase from the filamentous fungus Acremonium persicinum. Biochem J 316:841–846

  30. Sakamoto Y, Irie T, Sato T (2005a) Isolation and characterization of a fruiting body-specific exo-β-1,3-glucanase-encoding gene, exg1, from Lentinula edodes. Curr Genet 47:244–252

  31. Sakamoto Y, Minato K, Nagai M, Kawakami S, Mizuno M, Sato T (2005b) Characterization of the Lentinula edodes exg2 gene encoding a lentinan-degrading exo-β-1,3-glucanase. Curr Genet 48:195–203

  32. Sakamoto Y, Watanabe H, Nagai M, Nakade K, Takahashi M, Sato T (2006) Lentinula edodes tlg1 encodes a thaumatin-like protein that is involved in lentinan degradation and fruiting body senescence. Plant Physiol 141:793–801

  33. Sakamoto Y, Nakade K, Sato T (2009) Characterization of the post-harvest changes in gene transcription in the gill of the Lentinula edodes fruiting body. Curr Genet 55:409–423

  34. Seiler S, Plamann M (2003) The genetic basis of cellular morphogenesis in the filamentous fungus Neurospora crassa. Mol Biol Cell 14:4352–4364

  35. Shida M, Ushioda Y, Nakajima T, Matsuda K (1981) Structure of the alkali-insoluble skeletal glucan of Lentinus edodes. J Biochem 90:1093–1100

  36. Somogyi M (1952) Notes on sugar determination. J Biol Chem 195:19–23

  37. Suzuki T, Ohno N, Saito K, Yadomae T (1992) Activation of the complement system by (1→3)-β-d-glucans having different degrees of branching and different ultrastructures. J Pharmacobiodyn 15:277–285

  38. Tabata K, Itoh W, Hirata A, Sugawara I, Mori S (1990) Preparation of polyclonal antibodies to an anti-tumor (1→3)-β-d-glucan, schizophyllan. Agric Biol Chem 54:1953–1959

  39. Wessels JG, Niederpruem DJ (1967) Role of a cell-wall glucan-degrading enzyme in mating of Schizophyllum commune. J Bacteriol 94:1594–1602

  40. Zhang Y, Xu X, Xu J, Zhang L (2007) Dynamic viscoelastic behavior of triple helical lentinan in water: effects of concentration and molecular weight. Polymer 48:6681–6690

Download references

Acknowledgments

We thank Miss A. Kimura for her help with the experiments. This research was supported by a Grant-in-Aid for Scientific Research to N. K. (no. 21880050) from the Japan Society for the Promotion of Science (JSPS).

Author information

Correspondence to Naotake Konno.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Konno, N., Sakamoto, Y. An endo-β-1,6-glucanase involved in Lentinula edodes fruiting body autolysis. Appl Microbiol Biotechnol 91, 1365–1373 (2011). https://doi.org/10.1007/s00253-011-3295-2

Download citation

Keywords

  • β-1,6-glucanase
  • Fungal cell wall
  • Fruiting body autolysis
  • Basidiomycete