Lignin-Degrading Enzyme Activities

Over the past three decades, the activities of four kinds of enzyme have been purported to furnish the mechanistic foundations for macromolecular lignin depolymerization in decaying plant cell walls. The pertinent fungal enzymes comprise lignin peroxidase (with a relatively high redox potential), manganese peroxidase, an alkyl aryl etherase, and laccase. The peroxidases and laccase, but not the etherase, are expressed extracellularly by white-rot fungi. A number of these microorganisms exhibit a marked preference toward lignin in their degradation of lignocellulose. Interestingly, some white-rot fungi secrete both kinds of peroxidase but no laccase, while others that are equally effective express extracellular laccase activity but no peroxidases. Actually, none of these enzymes has been reported to possess significant depolymerase activity toward macromolecular lignin substrates that are derived with little chemical modification from the native biopolymer. Here, the assays commonly employed for monitoring the traditional fungal peroxidases, alkyl aryl etherase, and laccase are described in their respective contexts. A soluble native polymeric substrate that can be isolated directly from a conventional milled-wood lignin preparation is characterized in relation to its utility in next-generation lignin-depolymerase assays.

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

Access this chapter

Subscribe and save

Springer+ Basic €32.70 /Month

Buy Now

Price includes VAT (France)

eBook EUR 106.99 Price includes VAT (France)

Softcover Book EUR 137.14 Price includes VAT (France)

Hardcover Book EUR 147.69 Price includes VAT (France)

Tax calculation will be finalised at checkout

Purchases are for personal use only

Similar content being viewed by others

Ligninolytic Fungi from the Indian Subcontinent and Their Contribution to Enzyme Biotechnology

Chapter © 2021

Fungal Diversity and Enzymes Involved in Lignin Degradation

Chapter © 2019

Lignin Degrading Fungal Enzymes

Chapter © 2016

References

  1. Wong DWS (2009) Structure and action mechanism of ligninolytic enzymes. Appl Biochem Biotechnol 157:174–209 ArticleCASGoogle Scholar
  2. Davin LB, Patten AM, Jourdes M, Lewis NG (2008) Lignins: a twenty-first century challenge. In: Himmel ME (ed) Biomass recalcitrance—deconstructing the plant cell wall for bioenergy. Blackwell, Oxford, pp 213–305 Google Scholar
  3. Rencoret J, Gutiérrez A, Nieto L, Jiménez-Barbero J, Faulds CB, Kim H, Ralph J, Martínez ÁT, del Río JC (2011) Lignin composition and structure in young versus adult Eucalyptus globulus plants. Plant Physiol 155:667–682 ArticleCASGoogle Scholar
  4. Lawoko M, Henriksson G, Gellerstedt G (2005) Structural differences between the lignin–carbohydrate complexes present in wood and chemical pulps. Biomacromolecules 6:3467–3473 ArticleCASGoogle Scholar
  5. Balakshin MY, Capanema EA, Chang H-M (2007) MWL fraction with a high concentration of lignin–carbohydrate linkages: isolation and 2D NMR spectroscopic analysis. Holzforschung 61:1–7 ArticleCASGoogle Scholar
  6. Gellerstedt G (2007) Lignin complexity: fundamental and applied issues. http://rfparois.free.fr/LIG2G/Seminaire%20LIG2G-WEB-vs-tout-public.htm.
  7. Tien M, Kirk TK (1983) Lignin-degrading enzyme from the hymenomycete Phanerochaete chrysosporium Burds. Science 221:661–663 ArticleCASGoogle Scholar
  8. Kern HW, Haider K, Pool W, de Leeuw JW, Ernst L (1989) Comparison of the action of Phanerochaete chrysosporium and its extracellular enzymes (lignin peroxidases) on lignin preparations. Holzforschung 43:375–384 ArticleCASGoogle Scholar
  9. Haemmerli SD, Leisola MSA, Fiechter A (1986) Polymerization of lignins by ligninases from Phanerochaete chrysosporium. FEMS Microbiol Lett 35:33–36 ArticleCASGoogle Scholar
  10. Lundell T, Schoemaker H, Hatakka A, Brunow G (1993) New mechanism of the Cα–Cβ cleavage in non-phenolic arylglycerol β-aryl ether lignin substructures catalyzed by lignin peroxidase. Holzforschung 47:219–224 ArticleCASGoogle Scholar
  11. Hammel KE, Jensen KA Jr, Mozuch MD, Landucci LL, Tien M, Pease EA (1993) Ligninolysis by a purified lignin peroxidase. J Biol Chem 268:12274–12281 CASGoogle Scholar
  12. Hall PL (1980) Enzymatic transformations of lignin: 2. Enzyme Microb Technol 2:170–176 ArticleCASGoogle Scholar
  13. Forney LJ, Reddy CA, Tien M, Aust SD (1982) The involvement of hydroxyl radical derived from hydrogen peroxide in lignin degradation by the white rot fungus Phanerochaete chrysosporium. J Biol Chem 257:11455–11462 CASGoogle Scholar
  14. Gold MH, Kutsuki H, Morgan MA (1983) Oxidative degradation of lignin by photochemical and chemical radical generating systems. Photochem Photobiol 38:647–651 ArticleCASGoogle Scholar
  15. Glenn JK, Gold MH (1985) Purification and characterization of an extracellular Mn(II)-dependent peroxidase from the lignin-degrading basidiomycete, Phanerochaete chrysosporium. Arch Biochem Biophys 242:329–341 ArticleCASGoogle Scholar
  16. 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 ArticleCASGoogle Scholar
  17. Bourbonnais R, Paice MG (1990) Oxidation of non-phenolic substrates—an expanded role for laccase in biodegradation. FEBS Lett 267:99–102 ArticleCASGoogle Scholar
  18. Nutsubidze NN, Sarkanen S, Schmidt EL, Shashikanth S (1998) Consecutive polymerization and depolymerization of kraft lignin by Trametes cingulata. Phytochemistry 49:1203–1212 ArticleCASGoogle Scholar
  19. Li K, Horanyi PS, Collins R, Phillips RS, Eriksson K-E (2001) Investigation of the role of 3-hydroxyanthranilic acid in the degradation of lignin by white-rot fungus Pycnoporus cinnabarinus. Enzyme Microb Technol 28:301–307 ArticleCASGoogle Scholar
  20. Hammel KE, Kapich AN, Jensen KA Jr, Ryan ZC (2002) Reactive oxygen species as agents of wood decay by fungi. Enzyme Microb Technol 30:445–453 ArticleCASGoogle Scholar
  21. Blanchette RA, Krueger EW, Haight JE, Akhtar M, Akin DE (1996) Cell wall alterations in loblolly pine wood decayed by the white-rot fungus Ceriporiopsis subvermispora. J Biotechnol 53:203–213 ArticleGoogle Scholar
  22. Srebotnik E, Messner K, Foisner R (1988) Penetrability of white-rot-degraded pine wood by the lignin peroxidase of Phanerochaete chrysosporium. Appl Environ Microbiol 54:2608–2614 CASGoogle Scholar
  23. Wei D, Houtman CJ, Kapich AN, Hunt CG, Cullen D, Hammel KE (2010) Laccase and its role in production of extracellular reactive oxygen species during wood decay by the brown rot basidiomycete Postia placenta. Appl Environ Microbiol 76:2091–2097 ArticleCASGoogle Scholar
  24. Chen Y-R, Sarkanen S (2010) Macromolecular replication during lignin biosynthesis. Phytochemistry 71:453–462 ArticleCASGoogle Scholar
  25. Choinowski T, Blodig W, Winterhalter KH, Piontek K (1999) The crystal structure of lignin peroxidase at 1.70 Å resolution reveals a hydroxy group on the C β of tryptophan 171: a novel radical site formed during the redox cycle. J Mol Biol 286:809–827 ArticleCASGoogle Scholar
  26. Sundaramoorthy M, Kishi K, Gold MH, Poulos TL (1997) Crystal structures of substrate binding site mutants of manganese peroxidase. J Biol Chem 272:17574–17580 ArticleCASGoogle Scholar
  27. Ruiz-Dueñas FJ, Morales M, Pérez-Boada M, Choinowski T, Martínez MJ, Piontek K, Martínez ÁT (2007) Manganese oxidation site in Pleurotus eryngii versatile peroxidase: a site-directed mutagenesis, kinetic, and crystallographic study. Biochemistry 46:66–77 ArticleGoogle Scholar
  28. Pérez-Boada M, Ruiz-Dueñas FJ, Pogni R, Basosi R, Choinowski T, Martínez MJ, Piontek K, Martínez AT (2005) Versatile peroxidase oxidation of high redox potential aromatic compounds: site-directed mutagenesis, spectroscopic and crystallographic investigation of three long-range electron transfer pathways. J Mol Biol 354:385–402 ArticleGoogle Scholar
  29. Mester T, Ambert-Balay K, Ciofi-Baffoni S, Banci L, Jones AD, Tien M (2001) Oxidation of a tetrameric nonphenolic lignin model compound by lignin peroxidase. J Biol Chem 276:22985–22990 ArticleCASGoogle Scholar
  30. Hofrichter M, Ullrich R, Pecyna MJ, Liers C, Lundell T (2010) New and classic families of secreted fungal heme peroxidases. Appl Microbiol Biotechnol 87:871–897, and references therein ArticleCASGoogle Scholar
  31. Baciocchi E, Fabbri C, Lanzalunga O (2003) Lignin peroxidase-catalyzed oxidation of nonphenolic trimeric lignin model compounds: fragmentation reactions in the intermediate radical cations. J Org Chem 68:9061–9069 ArticleCASGoogle Scholar
  32. Yokota S, Umezawa T, Higuchi T (1991) Degradation of phenolic β–O–4 lignin model dimers by lignin peroxidase of Phanerochaete chrysosporium. Mokuzai Gakkaishi 37:535–541 CASGoogle Scholar
  33. Archibald FS (1992) A new assay for lignin-type peroxidases employing the dye Azure B. Appl Environ Microbiol 58:3110–3116 CASGoogle Scholar
  34. Bourbonnais R, Paice MG (1988) Veratryl alcohol oxidases from the lignin-degrading basidiomycete Pleurotus sajor-caju. Biochem J 255:445–450 CASGoogle Scholar
  35. de la Rubia T, Linares A, Pérez J, Muñoz-Dorado J, Romera J, Martínez J (2002) Characterization of manganese-dependent peroxidase isoenzymes from the ligninolytic fungus Phanerochaete flavido-alba. Res Microbiol 153:547–554 ArticleGoogle Scholar
  36. 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 ArticleCASGoogle Scholar
  37. Hammel KE, Cullen D (2008) Role of fungal peroxidases in biological ligninolysis. Curr Opin Plant Biol 11:349–355 ArticleCASGoogle Scholar
  38. Tuor U, Wariishi H, Schoemaker HE, Gold MH (1992) Oxidation of phenolic arylglycerol β-aryl ether lignin model compounds by manganese peroxidase from Phanerochaete chrysosporium: oxidative cleavage of an α-carbonyl model compound. Biochemistry 31:4986–4995 ArticleCASGoogle Scholar
  39. 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 ArticleCASGoogle Scholar
  40. Tien M, Kirk TK, Bull C, Fee JA (1986) Steady-state and transient-state kinetic studies on the oxidation of 3,4-dimethoxybenzyl alcohol catalyzed by the ligninase of Phanerochaete chrysosporium Burds. J Biol Chem 261:1687–1693 Google Scholar
  41. Caramelo L, Martínez MJ, Martínez ÁT (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 CASGoogle Scholar
  42. Otsuka Y, Sonoki T, Ikeda S, Kajita S, Nakamura M, Katayama Y (2003) Detection and characterization of a novel extracellular fungal enzyme that catalyzes the specific and hydrolytic cleavage of lignin guaiacylglycerol β-aryl ether linkages. Eur J Biochem 270:2353–2362 ArticleCASGoogle Scholar
  43. Masai E, Katayama Y, Nishikawa S, Yamasaki M, Morohoshi N, Haraguchi T (1989) Detection and localization of a new enzyme catalyzing the β-aryl ether cleavage in the soil bacterium (Pseudomonas paucimobilis SYK-6). FEBS Lett 249:348–352 ArticleCASGoogle Scholar
  44. Giardina P, Faraco V, Pezzella C, Piscitelli A, Vanhulle S, Sannia G (2010) Laccases: a never-ending story. Cell Mol Life Sci 67:369–385, and references therein ArticleCASGoogle Scholar
  45. Bourbonnais R, Leech D, Paice MG (1998) Electrochemical analysis of the interactions of laccase mediators with lignin model compounds. Biochim Biophys Acta 1379:381–390 ArticleCASGoogle Scholar
  46. Leontievsky A, Myasoedova N, Pozdnyakova N, Golovleva L (1997) ‘Yellow’ laccase of Panus tigrinus oxidizes non-phenolic substrates without electron-transfer mediators. FEBS Lett 413:446–448 ArticleCASGoogle Scholar
  47. Leontievsky AA, Vares T, Lankinen P, Shergill JK, Pozdnyakova NN, Myasoedova NM, Kalkkinen N, Golovleva LA, Cammack R, Thurston CF, Hatakka A (1997) Blue and yellow laccases of ligninolytic fungi. FEMS Microbiol Lett 156:9–14 ArticleCASGoogle Scholar
  48. Soden DM, O’Callaghan J, Dobson ADW (2002) Molecular cloning of a laccase isozyme gene from Pleurotus sajor-caju and expression in the heterologous Pichia pastoris host. Microbiology 148:4003–4014 CASGoogle Scholar
  49. Guan S-Y, Mlynár J, Sarkanen S (1997) Dehydrogenative polymerization of coniferyl alcohol on macromolecular lignin templates. Phytochemistry 45:911–918 ArticleCASGoogle Scholar
  50. Lundquist K, Ohlsson B, Simonson R (1977) Isolation of lignin by means of liquid-liquid extraction. Svensk Papperstidn 80(5):143–144 CASGoogle Scholar
  51. Contreras S, Gaspar AR, Guerra A, Lucia LA, Argyropoulos DS (2008) Propensity of lignin to associate: light scattering photometry study with native lignins. Biomacromolecules 9:3362–3369 ArticleCASGoogle Scholar
  52. Wyatt PJ (1993) Light scattering and the absolute characterization of macromolecules. Anal Chim Acta 272:1–40 ArticleCASGoogle Scholar
  53. Sarkanen S (1991) Enzymatic lignin degradation—an extracurricular view. ACS Symp Ser 460:247–269 ArticleCASGoogle Scholar

Acknowledgments

The authors wish to acknowledge a subcontract from the BioEnergy Science Center, which is a US Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science.

Author information

Authors and Affiliations

  1. Department of Bioproducts and Biosystems Engineering, University of Minnesota, St. Paul, MN, USA Yi-ru Chen, Simo Sarkanen & Yun-Yan Wang
  1. Yi-ru Chen