Abstract
The mechanisms of industrial dye transformation by versatile peroxidase were elucidated. Purified versatile peroxidase from Bjerkandera adusta was able to decolorize different classes of dyes including azo and phthalocyanines, but unable to transform any of the anthraquinones tested. Kinetic constants for selected dyes were determined and the transformation products were analyzed by EPR spectroscopy and mass spectrometry. The EPR and MS analyses of the enzymatic decolorization products showed the cleavage of the azo bond in azo dyes and the total disruption of the phthalocyaninic ring in phthalocyanine dyes. The EPR analysis on two copper-containing dyes, reactive violet 5 (azo) and reactive blue 72 (phthalocyanine), showed that the transformation can or not break the metal-ion coordination bond according the dye nature. The role of the catalytic Trp172 in the dye transformation by a long-range electron transfer pathway was confirmed and the oxidation mechanisms are proposed and discussed.
Similar content being viewed by others
References
Abadulla E, Tzanov T, Costa S, Robra K-H, Cavaco-Paulo A, Gübitz GM (2000) Decolorization and detoxification of textile dyes with a laccase from Trametes hirsute. Appl Environ Microbiol 66:3357–3362
Ayala M, Baratto MC, Basosi R, Vazquez-Duhalt R, Pogni R (2001) Spectroscopic characterization of manganese-lignin peroxidase hybrid isoenzyme produced by Bjerkandera adusta in the absence of manganese: evidence of a protein centred radical by hydrogen peroxide. J Mol Catal B Enzym 16:159–167
Barr DP, Aust SD (1994) Pollutant degradation by white rot fungi. Environ Contam Toxicol 138:49–72
Blodig W, Smith AT, Winterhalter K, Piontek K (1999) Evidence from spin-trapping for a transient radical on tryptophan residue 171 of lignin peroxidase. Arch Biochem Biophys 370:86–92
Camarero S, Sarkar S, Ruiz-Dueñas 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
Chivukula M, Spadaro JT, Renganathan V (1995) Lignin peroxidase-catalyzed oxidation of sulfonated azo dyes generates novel sulfophenyl hydroperoxides. Biochemistry 34:7765–7772
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 beta of tryptophan 171: a novel radical site formed during the redox cycle. J Mol Biol 286:09–827
Chung K-T, Stevens SE, Cerniglia CE (1992) The reduction of azo dyes y the intestinal microflora. Crit Rev Microbiol 18:175–190
Coen JJF, Smith AT, Candeias LP, Oakes J (2001) New insights into mechanisms of dye degradation by one-electron oxidation processes. J Chem Soc Perkin Trans 2:2125–2129
Conneely A, Smyth WF, Mc Mullan G (1999) Metabolism of the phthalocyanine textile dye remazol turquoise blue by Phanerochaete chrysosporium. FEMS Microbiol Lett 179:333–337
d’Alessandro N, Tonucci L, Bressan M, Dragani LK, Morvillo A (2003) Rapid and selective oxidation of metallosulfophthalocyanines prior to their usefulness as precatalysts in oxidation reactions. Eur J Inorg Chem 2003:1807–1814
Davila-Vazquez G, Tinoco R, Pickard MA, Vazquez-Duhalt R (2005) Transformation of halogenated pesticides by versatile peroxidase from Bjerkandera adusta. Enzym Microb Technol 36:223–231
Doyle WA, Blodig W, Veitch NC, Piontek K, Smith AT (1998) Two substrate interaction sites in lignin peroxidase revealed by site-directed mutagenesis. Biochemistry 37:15097–15105
Forgacs E, Cserháti T, Oros G (2004) Removal of synthetic dyes from wastewaters: a review. Environ Int 30:953–971
Goodwin TW, Morton RA (1946) The spectrophotometric determination of tyrosine and tryptophan in proteins. Biochem J 40:628–632
Goszczynski S, Paszczynski A, Pasti-Grigsby MB, Crawford RL, Crawford DL (1994) New pathway for degradation of sulfonated azo dyes by microbial peroxidases of Phanerochaete chrysosporium and Streptomyces chromofuscus. J Bacteriol 176:1339–1347
Hammel KE, Kalyanaraman B, Kirk TK (1986) Oxidation of polycyclic aromatic hydrocarbons and dibenzo[p]dioxins by Phanerochaete chrysosporium ligninase. J Biol Chem 261:16948–16952
Heinfling A, Bergbauer M, Szewzyk U (1997) Biodegradation of azo and phthalocyanine dyes by Trametes versicolor and Bjerkandera adusta. Appl Microbiol Biotechnol 48:261–266
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
Heinfling A, Ruiz-Dueñas J, 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
Heinfling A, Martinez MJ, Martinez AT, Bergbauer M, Szewzyk U (1998c) Transformation of industrial dyes by manganese peroxidases from Bjerkandera adusta and Pleurotus eryngii in a manganese-independent reaction. Appl Environ Microbiol 64:2788–2793
Heinfling-Weidtmann A, Reemtsma T, Storm T, Szewzyk U (2001) Sulfophthalimide as major metabolite formed from sulfonated phthalocyanine dyes by the white-rot fungus Bjerkandera adusta. FEMS Microbiol Lett 203:179–183
Holcapek M, Jandera P, Prikryl J (1999) Analysis of sulphonated dyes and intermediates by electrospray mass spectrometry. Dye Pigment 43:127–137
Janusz G, Kucharzyk KH, Pawlik A, Staszczak M, Paszczynski AJ (2013) Fungal laccase, manganese peroxidase and lignin peroxidase: gene expression and regulation. Enzym Microb Technol 52:1–12
Jarosz-Wilkolazka A, Kochmanska-Rdest J, Malarczyk E, Wardas W, Leonowicz A (2002) Fungi and their ability to decolourize azo and anthraquinonic dyes. Enzym Microb Technol 30:66–557
Johjima T, Itoh N, Kabuto M, Tokimura F, Nakagawa T, Wartishi H, Tanaka H (1999) Direct interaction of lignin and lignin peroxidase from Phanerochaete chrysosporium. Proc Natl Acad Sci U S A 96:1989–1994
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
Lopez C, Moreira MT, Feijoo G, Lema JM (2004) Dye decolorization by manganese peroxidase in an enzymatic membrane bioreactor. Biotechnol Prog 20:74–81
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
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
Moawad H, El-Rahim WM, Khalafallah M (2003) Evaluation of biotoxicity of textile dyes using two bioassays. J Basic Microbiol 43:218–229
Morales M, Mate MJ, Romero A, Martínez MJ, Martínez AT, Ruiz-Dueñas FJ (2012) Two oxidation sites for low redox potential substrates: a directed mutagenesis, kinetic, and crystallographic study on Pleurotus eryngii versatile peroxidase. J Biol Chem 287:41053–41067
Moreira MT, Mielgo I, Feijoo G, Lema JM (2000) Evaluation of different fungal strains in the decolourisation of sythetic dyes. Biotechnol Lett 22:1499–1503
Moreira PR, Bouillenne F, Almeida-Vara E, Malcata FX, Frère JM, Cardoso Duarte J (2006) Purification, kinetics and spectral characterisation of a new versatile peroxidase from a Bjerkandera sp. isolate. Enzym Microb Technol 38:28–33
Paszczynski A, Pasti-Grigsby MB, Goszczynski S, Crawford RL, Crawford DL (1992) Mineralization of sulfonated azo dyes and sulfanilic acid by Phanerochaete chrysosporium and Streptomyces chromofuscus. Appl Environ Microbiol 58:3598–3604
Pearce CI, Lloyd JR, Guthrie JT (2003) The removal of colour from textile wastewater using whole bacterial cells: a review. Dye Pigment 58:179–196
Piontek K, Smith AT, Blodig W (2001) Lignin peroxidase structure and function. Biochem Soc Trans 29:111–116
Pogni R, Baratto MC, Giansanti S, Teutloff C, Verdin J, Valderrama B, Lendzian F, Lubitz W, Vazquez-Duhalt R, Basosi R (2005) Tryptophan-based radical in the catalytic mechanism of versatile peroxidase from Bjerkandera adusta. Biochemistry 44:4267–4274
Pogni R, Baratto MC, Teutloff C, Giansanti S, Ruiz-Dueñas FJ, Choinowski T, Piontek K, Martínez AT, Lendzian F, Basosi R (2006) A tryptophan neutral radical in the oxidized state of versatile peroxidase from Pleurotus eryngii: a combined multifrequency EPR and density functional theory study. J Biol Chem 281:9517–9526
Pointing SB (2001) Feasibility of bioremediation by white-rot fungi. Appl Microbiol Biotechnol 57:20–33
Rakhit G, Antholine WE, Froncisz W, Hyde JS, Pilbrow JR, Sinclair JR, Sarkar B (1985) Direct evidence of nitrogen coupling in the copper(II) complex of bovine serum albumin by S-band electron spin resonance technique. J Inorg Biochem 25:217–224
Reddy CA (1995) The potential for white-rot fungi in the treatment of pollutants. Curr Opin Biotechnol 6:320–328
Robinson T, McMullan G, Marchant R, Nigam P (2001) Remediation of dyes in textile effluent: a critical review on current treatment technologies with a proposed alternative. Bioresour Technol 77:247–255
Rodriguez E, Pickard MA, Vazquez-Duhalt R (1999) Industrial dyes decolorization by laccases from ligninolytic fungi. Curr Microbiol 39:27–32
Ruiz-Dueñas FJ, Martinez MJ, Martinez AT (1999a) Molecular characterization of a novel peroxidase isolated from the ligninolytic fungus Pleurotus eryngii. Mol Microbiol 31:223–235
Ruiz-Dueñas FJ, Martinez MJ, Martinez AT (1999b) Heterologous expression of Pleurotus eryngii peroxidase confirms its ability to oxidize Mn(2+) and different aromatic substrates. Appl Environ Microbiol 65:4705–4707
Ruiz-Dueñas FJ, Morales M, Gracía E, Miki Y, Martinez MJ, Martinez AT (2009) Substrate oxidation sites in versatile peroxidase and other basidiomycete peroxidases. J Exp Bot 60:441–452
Sarkar S, Martinez AT, Martinez MJ (1997) Biochemical and molecular characterization of a manganese peroxidase isoenzyme from Pleurotus ostreatus. Biochim Biophys Acta 1339:23–30
Shin K, Oh I, Kim C (1997) Production and purification of remazol brilliant blue R decolorizing peroxidase from the culture filtrate of Pleurotus ostreatus. Appl Environ Microbiol 63:1744–1748
Spadaro JT, Renganathan V (1994) Peroxidase-catalyzed oxidation of azo dyes: mechanism of disperse yellow 3 degradation. Arch Biochem Biophys 312:301–307
Spande TF, Witkop B (1967) Determination of the tryptophan content of proteins with N-bromosuccinimide. Methods Enzymol 11:498–506
Taboada-Puig R, Lú-Chau T, Eibes G, Moreira MT, Feijoo G, Lema JM (2011) Biocatalytic generation of Mn(III)-chelate as a chemical oxidant of different environmental contaminants. Biotechnol Prog 27:668–676
ten Have R, Teunissen PJ (2001) Oxidative mechanisms involved in lignin degradation by white-rot fungi. Chem Rev 101:2297–3413
Tien M, Kirk TK (1988) Lignin peroxidase of Phanerochaete chrysosporium. Methods Enzymol 161:238–248
Tsukihara T, Honda Y, Sakai R, Watanabe T, Watanabe T (2008) Mechanism for oxidation of high-molecular-weight substrates by a fungal versatile peroxidase, MnP2. Appl Environ Microbiol 74:2873–2881
Vazquez-Duhalt R, Westlake DWS, Fedorak PM (1994) Lignin peroxidase oxidation of aromatic compounds in systems containing organic solvents. Appl Environ Microbiol 60:459–466
Wang Y, Vazquez-Duhalt R, Pickard MA (2001) Effect of growth conditions on the production of manganese peroxidase by three strains of Bjerkandera adusta. Can J Microbiol 47:277–282
Wang Y, Vazquez-Duhalt R, Pickard MA (2002) Purification, characterization, and chemical modification of manganese peroxidase from Bjerkandera adusta UAMH 8258. Curr Microbiol 43:77–87
Wang Y, Vazquez-Duhalt R, Pickard MA (2003) Manganese-lignin peroxidase hybrid from Bjerkandera adusta oxidizes polyaromatic hydrocarbons more actively in the absence of manganese. Can J Microbiol 49:675–682
Wariishi H, Valli K, Gold MH (1992) Manganese(II) oxidation by manganese peroxidase from the basidiomycete Phanerochaete chrysosporium. J Biol Chem 267:23688–23695
Wesenberg D, Kyriakidos I, Agathos SN (2003) White-rot fungi and their enzymes from the treatment of industrial dye effluent. Biotechnol Adv 22:161–187
Whitwam RE, Brown KR, Musick M, Natan MJ, Tien M (1997) Mutagenesis of the Mn2+-binding site of manganese peroxidase affects oxidation of Mn2+ by both compound I and compound II. Biochemistry 36:9766–9773
Wong DW (2009) Structure and action mechanism of ligninolytic enzymes. Appl Biochem Biotechnol 157:174–209
Acknowledgments
We thank Rosa Roman for her technical assistance. This project was funded by the National Council of Science and Technology of Mexico (CONACYT) and the Italian MIUR PRIN 2009 STNWX3 and the Italian CSGI Consortium.
Author information
Authors and Affiliations
Corresponding author
Additional information
Responsible editor: Philippe Garrigues
Rights and permissions
About this article
Cite this article
Baratto, M.C., Juarez-Moreno, K., Pogni, R. et al. EPR and LC-MS studies on the mechanism of industrial dye decolorization by versatile peroxidase from Bjerkandera adusta . Environ Sci Pollut Res 22, 8683–8692 (2015). https://doi.org/10.1007/s11356-014-4051-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11356-014-4051-9