Skip to main content
SpringerLink
Log in

DyP-like peroxidases of the jelly fungus Auricularia auricula-judae oxidize nonphenolic lignin model compounds and high-redox potential dyes

  • Biotechnologically Relevant Enzymes and Proteins
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

The jelly fungus Auricularia auricula-judae produced an enzyme with manganese-independent peroxidase activity during growth on beech wood (∼300 U l−1). The same enzymatic activity was detected and produced at larger scale in agitated cultures comprising of liquid, plant-based media (e.g. tomato juice suspensions) at levels up to 8,000 U l−1. Two pure peroxidase forms (A. auricula-judae peroxidase (AjP I and AjP II) could be obtained from respective culture liquids by three chromatographic steps. Spectroscopic and electrophoretic analyses of the purified proteins revealed their heme and peroxidase nature. The N-terminal amino acid sequence of AjP matched well with sequences of fungal enzymes known as “dye-decolorizing peroxidases”. Homology was found to the N-termini of peroxidases from Marasmius scorodonius (up to 86%), Thanatephorus cucumeris (60%), and Termitomyces albuminosus (60%). Both enzyme forms catalyzed not only the conversion of typical peroxidase substrates such as 2,6-dimethoxyphenol and 2,2′-azino-bis(3-ethylthiazoline-6-sulfonate) but also the decolorization of the high-redox potential dyes Reactive Blue 5 and Reactive Black 5, whereas manganese(II) ions (Mn2+) were not oxidized. Most remarkable, however, is the finding that both AjPs oxidized nonphenolic lignin model compounds (veratryl alcohol; adlerol, a nonphenolic β-O-4 lignin model dimer) at low pH (maximum activity at pH 1.4), which indicates a certain ligninolytic activity of dye-decolorizing peroxidases.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Adler E (1977) Lignin chemistry—past, present and future. Wood Sci Technol 11:169–218

    Article  CAS  Google Scholar 

  • Aitken MD, Irvine RL (1989) Stability testing of ligninase and Mn-peroxidase from Phanerochaete chrysosporium. Biotechnol Bioeng 34:1251–1260

    Article  CAS  Google Scholar 

  • Anh DH, Ullrich R, Benndorf D, Svatos A, Muck A, Hofrichter M (2007) The coprophilous mushroom Coprinus radians secretes a haloperoxidase that catalyzes aromatic peroxygenation. Appl Environ Microbiol 73:5477–5485

    Article  CAS  Google Scholar 

  • Bohlin C, Persson P, Gorton L, Lundquist K, Jönsson LJ (2005) Product profiles in enzymic and non-enzymic oxidations of the lignin model compound erythro-1-(3, 4-dimethoxyphenyl)-2-(2-methoxyphenoxy)-1, 3-propanediol. J Mol Catal B: Enzym 35:100–107

    Article  CAS  Google Scholar 

  • Bollag JM, Sjoblad RD, Liu SY (1979) Characterization of an enzyme from Rhizoctonia praticola which polymerizes phenolic compounds. Can J Microbiol 25:229–233

    Article  CAS  Google Scholar 

  • Camarero S, Sarkar S, Ruiz-Duenas FJ, Martinez MJ, Martinez AT (1999) Description of a versatile peroxidase involved in the natural degradation of lignin that has both manganese peroxidase and lignin peroxidase substrate interaction sites. J Biol Chem 274:10324–10330

    Article  CAS  Google Scholar 

  • Caramelo L, Martinez MJ, Martinez AT (1999) A search for ligninolytic peroxidases in the fungus Pleurotus eryngii involving α-keto-γ-thiomethylbutyric acid and lignin model dimers. Appl Environ Microbiol 65:916–922

    CAS  Google Scholar 

  • Chung N, Aust SD (1995) Inactivation of lignin peroxidase by hydrogen peroxide during the oxidation of phenols. Arch Biochem Biophys 316:851–855

    Article  CAS  Google Scholar 

  • Cullen D, Kersten PJ (2004) Enzymology and molecular biology of lignin degradation. In: Brambl R, Marzluf GA (eds) The Mycota, vol 3. Biochemistry and molecular biology, 2nd edn. Springer, Berlin, pp 249–273

    Google Scholar 

  • Eggert C, Temp U, Dean JF, Eriksson KE (1996) A fungal metabolite mediates degradation of non-phenolic lignin structures and synthetic lignin by laccase. FEBS Lett 391:144–148

    Article  CAS  Google Scholar 

  • Faraco V, Piscitelli A, Sannia G, Giardina P (2007) Identification of a new member of the dye-decolorizing peroxidase family from Pleurotus ostreatus. World J Microbiol Biotechnol 23:889–893

    Article  CAS  Google Scholar 

  • Gamauf C, Metz B, Seiboth B (2007) Degradation of plant cell wall polymers by fungi. In: Kubicek CP, Druzhinina IS (eds) The Mycota, vol 4. Environmental and microbial relationships, 2nd edn. Springer, Berlin, pp 325–339

    Google Scholar 

  • Gazarian IG, Lagrimini LM, George SJ, Thorneley RN (1996) Anionic tobacco peroxidase is active at extremely low pH: veratryl alcohol oxidation with a pH optimum of 1.8. Biochem J 320:369–372

    CAS  Google Scholar 

  • Gelpke MDS, Lee J, Gold MH (2002) Lignin peroxidase oxidation of veratryl alcohol: effects of the mutants H82A, Q222A, W171A, and F267L. Biochemistry 41:3498–3506

    Article  CAS  Google Scholar 

  • Gold MH, Kuwahara M, Chiu AA, Glenn JK (1984) Purification and characterization of an extracellular H2O2-requiring diarylpropane oxygenase from the white rot basidiomycete, Phanerochaete chrysosporium. Arch Biochem Biophys 234:353–362

    Article  CAS  Google Scholar 

  • Hatakka A (2001) Biodegradation of lignin. In: Hofrichter M, Steinbüchel A (eds) Biopolymers. Lignin, humic substances and coal, vol 1. Wiley-VCH, Weinheim, pp 129–180

    Google Scholar 

  • Hatakka A, Lundell TK, Tervilä-Wilo ALM, Brunow G (1991) Metabolism of non-phenolic β-O-4 lignin model compounds by the white-rot fungus Phlebia radiata. Appl Microbiol Biotechnol 36:270–277

    Article  CAS  Google Scholar 

  • Heinfling A, Martinez MJ, Martinez AT, Bergbauer M, Szewzyk U (1998a) Purification and characterization of peroxidases from the dye-decolorizing fungus Bjerkandera adusta. FEMS Microbiol Lett 165:43–50

    Article  CAS  Google Scholar 

  • Heinfling A, Ruiz-Duenas FJ, Martinez MJ, Bergbauer M, Szewzyk U, Martinez AT (1998b) A study on reducing substrates of manganese-oxidizing peroxidases from Pleurotus eryngii and Bjerkandera adusta. FEBS Lett 428:141–146

    Article  CAS  Google Scholar 

  • Heinzkill M, Bech L, Halkier T, Schneider P, Anke T (1998) Characterization of laccases and peroxidases from wood-rotting fungi (family Coprinaceae). Appl Environ Microbiol 64:1601–1606

    CAS  Google Scholar 

  • Hibbett DS (2006) A phylogenetic overview of the Agaricomycotina. Mycologia 98:917–925

    Article  Google Scholar 

  • Hofrichter M, Ziegenhagen D, Vares T, Friedrich M, Jager MG, Fritsche W, Hatakka A (1998) Oxidative decomposition of malonic acid as basis for the action of manganese peroxidase in the absence of hydrogen peroxide. FEBS Lett 434:362–366

    Article  CAS  Google Scholar 

  • Hofrichter M, Lundell T, Hatakka A (2001) Conversion of milled pine wood by manganese peroxidase from Phlebia radiata. Appl Environ Microbiol 67:4588–4593

    Article  CAS  Google Scholar 

  • Hushpulian DM, Poloznikov AA, Savitski PA, Rozhkova AM, Chubar TA, Fechina VA, Orlova MA, Tishkov VI, Gazaryan IG, Lagrimini LM (2007) Glutamic acid-141: a heme ‘bodyguard’ in anionic tobacco peroxidase. Biol Chem 388:373–380

    Article  CAS  Google Scholar 

  • Kim SJ, Shoda M (1999) Purification and characterization of a novel peroxidase from Geotrichum candidum Dec 1 involved in decolorization of dyes. Appl Environ Microbiol 65:1029–1035

    CAS  Google Scholar 

  • Kirk TK, Schulz E, Connors WJ, Lorenz LF, Zeikus JG (1978) Influence of culture parameters on lignin metabolism by Phanerochaete chrysosporium. Arch Microbiol 177:277–285

    Article  Google Scholar 

  • Kirk TK, Tien M, Kersten PJ, Mozuch MD, Kalyanaraman B (1986) Ligninase of Phanerochaete chrysosporium. Mechanism of its degradation of the non-phenolic arylglycerol-β-aryl ether substructure of lignin. Biochem J 236:279–287

    CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  • Liers C, Ullrich R, Steffen KT, Hatakka A, Hofrichter M (2006) Mineralization of 14C-labelled synthetic lignin and extracellular enzyme activities of the wood-colonizing ascomycetes Xylaria hypoxylon and Xylaria polymorpha. Appl Microbiol Biotechnol 69:573–579

    Article  CAS  Google Scholar 

  • Lundell T, Hatakka A (1994) Participation of Mn(II) in the catalysis of laccase, manganese peroxidase and lignin peroxidase from Phlebia radiata. FEBS Lett 348:291–296

    Article  CAS  Google Scholar 

  • Lundell TK, Kileläinen IA, Brunow G, Schoemaker HE, Hatakka A (1992) Degradation of macromolecular lignin models by lignin peroxidase from Phlebia radiata and catalytic properties of the enzyme. Paper presented at the 5th International Conference on Biotechnology in the Pulp and Paper Industry. Kyoto, Japan

  • Lundell T, Wever R, Floris R, Harvey P, Hatakka A, Brunow G, Schoemaker H (1993) Lignin peroxidase L3 from Phlebia radiata. Pre-steady-state and steady-state studies with veratryl alcohol and a non-phenolic lignin model compound 1-(3, 4-dimethoxyphenyl)-2-(2-methoxyphenoxy)propane-1, 3-diol. Eur J Biochem 211:391–402

    Article  CAS  Google Scholar 

  • Martinez MJ, Ruiz-Duenas FJ, Guillen F, Martinez AT (1996) Purification and catalytic properties of two manganese peroxidase isoenzymes from Pleurotus eryngii. Eur J Biochem 237:424–432

    Article  CAS  Google Scholar 

  • McEldoon JP, Pokora AR, Dordick JS (1995) Lignin peroxidase-type activity of soybean peroxidase. Enzyme Microb Technol 17:359–365

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • Morgenstern I, Klopman S, Hibbett DS (2008) Molecular evolution and diversity of lignin degrading heme peroxidases in the Agaricomycetes. J Mol Evol 66:243–257

    Article  CAS  Google Scholar 

  • Pecyna M, Ullrich R, Bittner B, Clemens A, Schubert R, Scheibner K, Hofrichter M (2009) Molecular characterization of aromatic peroxygenase from Agrocybe aegerita. Appl Microbiol Biotechnol. doi:10.1007/s00253-009-2000-1:

    Google Scholar 

  • Poulos TL, Edwards SL, Wariishi H, Gold MH (1993) Crystallographic refinement of lignin peroxidase at 2 A. J Biol Chem 268:4429–4440

    CAS  Google Scholar 

  • Robene-Soustrade I, Lung-Escarmant B, Bono JJ, Taris B (1992) Identification and partial characterization of an extracellular manganese-dependent peroxidase in Armillaria ostoyae and Armillaria mellea. Eur J For Pathol 22:227–236

    Article  Google Scholar 

  • Scheibner M, Hulsdau B, Zelena K, Nimtz M, de Boer L, Berger RG, Zorn H (2008) Novel peroxidases of Marasmius scorodonius degrade β-carotene. Appl Microbiol Biotechnol 77:1241–1250

    Article  CAS  Google Scholar 

  • Schmidt O (2006) Wood and tree fungi. Biology, damage, protection, and use. Springer, Berlin

    Google Scholar 

  • Schoemaker HE, Lundell TK, Hatakka A, Piontek K (1994) The oxidation of veratryl alcohol, dimeric lignin models and lignin by lignin peroxidase: redox cycle revisited. FEMS Microbiol Rev 13:321–332

    Article  CAS  Google Scholar 

  • Shimokawa T, Hirai M, Shoda M, Sugano Y (2008) Efficient dye decolorization and production of dye decolorizing enzymes by the basidiomycete Thanatephorus cucumeris Dec 1 in a liquid and solid hybrid culture. J Biosci Bioeng 106:481–487

    Article  CAS  Google Scholar 

  • Shimokawa T, Shoda M, Sugano Y (2009) Purification and characterization of two DyP isozymes from Thanatephorus cucumeris Dec 1 specifically expressed in an air-membrane surface bioreactor. J Biosci Bioeng 107:113–115

    Article  CAS  Google Scholar 

  • Sugano Y (2009) DyP-type peroxidases comprise a novel heme peroxidase family. Cell Mol Life Sci 66:1387–1403

    Article  CAS  Google Scholar 

  • Sugano Y, Sasaki K, Shoda M (1999) cDNA cloning and genetic analysis of a novel decolorizing enzyme, peroxidase gene dyp from Geotrichum candidum Dec 1. J Biosci Bioeng 87:411–417

    Article  CAS  Google Scholar 

  • Sugano Y, Nakano R, Sasaki K, Shoda M (2000) Efficient heterologous expression in Aspergillus oryzae of a unique dye-decolorizing peroxidase, DyP, of Geotrichum candidum Dec 1. Appl Environ Microbiol 66:1754–1758

    Article  CAS  Google Scholar 

  • Sugano Y, Muramatsu R, Ichiyanagi A, Sato T, Shoda M (2007) DyP, a unique dye-decolorizing peroxidase, represents a novel heme peroxidase family: ASP171 replaces the distal histidine of classical peroxidases. J Biol Chem 282:36652–36658

    Article  CAS  Google Scholar 

  • Sugano Y, Matsushima Y, Tsuchiya K, Aoki H, Hirai M, Shoda M (2009) Degradation pathway of an anthraquinone dye catalyzed by a unique peroxidase DyP from Thanatephorus cucumeris Dec 1. Biodegradation 20:433–440

    Article  CAS  Google Scholar 

  • Ten Have R, Hartmans S, Teunissen PJ, Field JA (1998) Purification and characterization of two lignin peroxidase isozymes produced by Bjerkandera sp. strain BOS55. FEBS Lett 422:391–394

    Article  Google Scholar 

  • Tien M, Kirk TK (1988) Lignin peroxidase of Phanerochaete chrysosporium. Methods Enzymol 161:238–249

    Article  CAS  Google Scholar 

  • Tuisel H, Sinclair R, Bumpus JA, Ashbaugh W, Brock BJ, Aust SD (1990) Lignin peroxidase H2 from Phanerochaete chrysosporium: purification, characterization and stability to temperature and pH. Arch Biochem Biophys 279:158–166

    Article  CAS  Google Scholar 

  • Ullrich R, Nuske J, Scheibner K, Spantzel J, Hofrichter M (2004) Novel haloperoxidase from the agaric basidiomycete Agrocybe aegerita oxidizes aryl alcohols and aldehydes. Appl Environ Microbiol 70:4575–4581

    Article  CAS  Google Scholar 

  • Ullrich R, Liers C, Schimpke S, Hofrichter M (2009) Purification of homogeneous forms of fungal peroxygenase. J Biotechnol. doi:10.1002/biot.200900076:

    Google Scholar 

  • Ward G, Hadar Y, Bilkis I, Dosoretz CG (2003) Mechanistic features of lignin peroxidase-catalyzed oxidation of substituted phenols and 1, 2-dimethoxyarenes. J Biol Chem 278:39726–39734

    Article  CAS  Google Scholar 

  • Weiß M, Oberwinkler F (2001) Phylogenetic relationships in Auriculariales and related groups—hypotheses derived from nuclear ribosomal DNA sequences. Mycol Res 105:403–415

    Article  Google Scholar 

  • Welinder KG (1992) Superfamily of plant, fungal and bacterial peroxidases. Curr Opin Struct Biol 2:388–393

    Article  CAS  Google Scholar 

  • Zubieta C, Joseph R, Krishna SS, McMullan D, Kapoor M, Axelrod HL, Miller MD, Abdubek P, Acosta C, Astakhova T, Carlton D, Chiu HJ, Clayton T, Deller MC, Duan L, Elias Y, Elsliger MA, Feuerhelm J, Grzechnik SK, Hale J, Han GW, Jaroszewski L, Jin KK, Klock HE, Knuth MW, Kozbial P, Kumar A, Marciano D, Morse AT, Murphy KD, Nigoghossian E, Okach L, Oommachen S, Reyes R, Rife CL, Schimmel P, Trout CV, van den Bedem H, Weekes D, White A, Xu Q, Hodgson KO, Wooley J, Deacon AM, Godzik A, Lesley SA, Wilson IA (2007a) Identification and structural characterization of heme binding in a novel dye-decolorizing peroxidase, TyrA. Proteins 69:234–243

    Article  CAS  Google Scholar 

  • Zubieta C, Krishna SS, Kapoor M, Kozbial P, McMullan D, Axelrod HL, Miller MD, Abdubek P, Ambing E, Astakhova T, Carlton D, Chiu HJ, Clayton T, Deller MC, Duan L, Elsliger MA, Feuerhelm J, Grzechnik SK, Hale J, Hampton E, Han GW, Jaroszewski L, Jin KK, Klock HE, Knuth MW, Kumar A, Marciano D, Morse AT, Nigoghossian E, Okach L, Oommachen S, Reyes R, Rife CL, Schimmel P, van den Bedem H, Weekes D, White A, Xu Q, Hodgson KO, Wooley J, Deacon AM, Godzik A, Lesley SA, Wilson IA (2007b) Crystal structures of two novel dye-decolorizing peroxidases reveal a β-barrel fold with a conserved heme-binding motif. Proteins 69:223–233

    Article  CAS  Google Scholar 

Download references

Acknowledgment

We thank Martin Kluge (Inge), Matthias Kinne (Konrad), and Elisabet Aranda for their useful comments and discussions as well as Ulrike Schneider and Monika Brandt for their technical assistance. The work was supported by the European Union (integrated project “Biorenew”) and the German Environmental Foundation (DBU, project 13225-32).

Author information

Authors and Affiliations

  1. Unit of Environmental Biotechnology, International Graduate School of Zittau, Markt 23, 02763, Zittau, Germany

    Christiane Liers, Caroline Bobeth, Marek Pecyna, René Ullrich & Martin Hofrichter

Authors
  1. Christiane Liers
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Caroline Bobeth
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Marek Pecyna
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. René Ullrich
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Martin Hofrichter
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Christiane Liers.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liers, C., Bobeth, C., Pecyna, M. et al. DyP-like peroxidases of the jelly fungus Auricularia auricula-judae oxidize nonphenolic lignin model compounds and high-redox potential dyes. Appl Microbiol Biotechnol 85, 1869–1879 (2010). https://doi.org/10.1007/s00253-009-2173-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-009-2173-7

Keywords

Search

Navigation