Removal of Organic Pollutants from Industrial Wastewater by Treatment with Oxidoreductase Enzymes

  • Edelmira ValeroEmail author
  • María-Isabel González-Sánchez
  • María-Teresa Pérez-Prior
Part of the The Handbook of Environmental Chemistry book series (HEC, volume 32)


Removal of persistent organic pollutants in wastewaters of industrial origin is an increasingly relevant issue in industrialized countries that needs addressing. Remarkable research efforts have been made for the development and implementation of new efficient and eco-friendly treatments capable of reducing, or even eliminating, toxic compounds in effluents prior to their disposal. Enzymatic methods appear to be a promising technology for this task, with a minor impact on ecosystems as compared to physicochemical methods. The applicability of such technology has been explicitly demonstrated in a huge number of publications and patents registered to date.

The present chapter focuses on the application of oxidoreductase enzymes in industrial wastewaters treatment. Numerous redox enzymes, including peroxidases, tyrosinases, and laccases from different sources, and even hemoglobin from animal blood have exhibited their potential for the remediation of a broad spectrum of recalcitrant organic compounds. However the implementation of this technology on an industrial scale still needs further research. Here the most important aspects about the current situation of the subject and future perspectives for the use of redox enzymes in industrial wastewaters treatment are highlighted.


Aromatic pollutants Immobilization Industrial wastewater Oxidation Oxidoreductase enzymes 



Biochemical oxygen demand


Bisphenol A


Cross-linked enzyme aggregates


Chemical oxygen demand


Compound I of peroxidase


Compound II of peroxidase




Endocrine disrupting compounds




Horseradish peroxidase


Lignin peroxidase




Manganese peroxidase


Polycyclic aromatic hydrocarbons


Polychlorinated biphenyls


Polychlorinated dibenzodioxins


Polychlorinated dibenzofurans


Polyethylene glycol


Persistent organic pollutants


Quantitative structure–activity relationship


Soya bean peroxidase


United States Environmental Protection Agency


White-rot fungi


Wastewater treatment


Wastewater treatment plants


  1. 1.
    ACS (2014) CAS Registry. Accessed 11 Feb 2014
  2. 2.
    European Community (2004) Regulation No 850/2004 of the European Parliament and of the Council of 29 April 2004 on Persistent Organic Pollutants and Amending Directive 79/117/EEC.¼OJ:L:004:158:0007:0049:EN:PDF
  3. 3.
    Busca G, Berardinelli S, Resini C et al (2008) Technologies for the removal of phenol from fluid streams: a short review of recent developments. J Hazard Mater 160:265–288CrossRefGoogle Scholar
  4. 4.
    Husain Q (2006) Potential applications of the oxidoreductive enzymes in the decolorization and detoxification of textile and other synthetic dyes from polluted water: a review. Crit Rev Biotechnol 26:201–221CrossRefGoogle Scholar
  5. 5.
    Klibanov AM, Alberti BN, Morris ED et al (1980) Enzymatic removal of toxic phenols and anilines from waste waters. J Appl Biochem 2:414–421Google Scholar
  6. 6.
    Torres E, Ayala M (2010) Biocatalysis based on heme peroxidases. Springer-Verlag, Berlin/HeidelbergCrossRefGoogle Scholar
  7. 7.
    Cammarota MC, Freire DMG (2006) A review on hydrolytic enzymes in the treatment of wastewater with high oil and grease content. Bioresource Technol 97:2195–2210CrossRefGoogle Scholar
  8. 8.
    EPA (2013) Water: CWA Methods, Priority Pollutants. Accessed 26 Feb 2014
  9. 9.
    Pinheiro HM, Touraud E, Thomas O (2004) Aromatic amines from azo dye reduction: status review with emphasis on direct UV spectrophotometer detection in textile industries wastewater. Dyes Pigments 61:121–139CrossRefGoogle Scholar
  10. 10.
    EPA (2011) Persistent Bioaccumulative and Toxic (PBT) Chemical Program. Dioxins and Furans. Accessed 26 March 2014
  11. 11.
    EPA (2013) Ecosystems & Environment: Wastewater treatment. Accessed 4 March 2014
  12. 12.
    Federal Register, The Daily Journal of the US Government (2013) Endocrine disruptor screening program; final second list of chemicals and substances for tier 1 screening. document 78 FR 35922Google Scholar
  13. 13.
    Vuorinen A, Odermatt A, Schuster D (2013) In silico methods in the discovery of endocrine disrupting chemicals. J Steroid Biochem Mol Biol 137:18–26CrossRefGoogle Scholar
  14. 14.
    Husain Q, Qayyum S (2013) Biological and enzymatic treatment of bisphenol A and other endocrine disrupting compounds: a review. Crit Rev Biotech 33:260–292CrossRefGoogle Scholar
  15. 15.
    Dos Santos AB, Cervantes FJ, van Lier JB (2007) Review paper on current technologies for decolourisation of textile wastewaters: Perspectives for anaerobic biotechnology. Bioresource Technol 98:2369–2385CrossRefGoogle Scholar
  16. 16.
    Husain (2010) Peroxidase mediated decolorization and remediation of wastewater containing industrial dyes: a review. Rev Environ Sci Biotechnol 9:117–140Google Scholar
  17. 17.
    Dunford HB, Jones PA (2010) Peroxidases and catalases: biochemistry, biophysics, biotechnology and physiology. Wiley, New JerseyGoogle Scholar
  18. 18.
    Nicell JA, Bewtra JK, Biswas N et al (1993) Reactor development for peroxidase catalyzed polymerization and precipitation of phenols from wastewater. Water Res 27:1629–1639CrossRefGoogle Scholar
  19. 19.
    Klibanov AM (1982) Enzymatic removal of hazardous pollutants from industrial aqueous effluents. Enzyme Eng 6:319–323CrossRefGoogle Scholar
  20. 20.
    Regalado C, García-Almendárez BE, Duarte-Vázquez MA (2004) Biotechnological applications of peroxidases. Phytochem Rev 3:243–256CrossRefGoogle Scholar
  21. 21.
    González-Sánchez MI, Laurenti M, Rubio-Retama J et al (2011) Fluorescence decrease of conjugated polymers by the catalytic activity of horseradish peroxidase and its application in phenolic compounds detection. Biomacromolecules 12:1332–1338CrossRefGoogle Scholar
  22. 22.
    González-Sánchez MI, Rubio-Retama J, López-Cabarcos E et al (2011) Development of an acetaminophen amperometric biosensor based on peroxidase entrapped in polyacrylamide microgels. Biosen Bioelectron 26:1883–1889CrossRefGoogle Scholar
  23. 23.
    Valero E, García-Carmona F (1998) A continuous spectrophotometric method based on enzymatic cycling for determining L-glutamate. Anal Biochem 259:265–271CrossRefGoogle Scholar
  24. 24.
    Hiner A, Hernández-Ruiz J, Williams GA et al (2001) Catalase-like oxygen production by horseradish peroxidase must predominantly be an enzyme-catalyzed reaction. Arch Biochem Biophys 392:295–302CrossRefGoogle Scholar
  25. 25.
    Arnao MB, Acosta M, del Río JA et al (1990) A kinetic study on the suicide inactivation of peroxidase and hydrogen peroxide. Biochim Biophys Acta 1041:43–47CrossRefGoogle Scholar
  26. 26.
    Valderrama B, Ayala M, Vazquez-Duhalt R (2002) Suicide inactivation of peroxidases and the challenge of engineering more robust enzymes. Chem Biol 9:555–565CrossRefGoogle Scholar
  27. 27.
    Karam J, Nicell JA (1997) Potential applications of enzymes in waste treatment. J Chem Techn Biotech 69:141–153CrossRefGoogle Scholar
  28. 28.
    Wagner M, Nicell JA (2001) Peroxidase-catalyzed removal of phenols from a petroleum refinery wastewater. Water Sci Technol 43:253–260Google Scholar
  29. 29.
    Auriol M, Filali-Meknassi Y, Tyagi RD et al (2007) Oxidation of natural and synthetic hormones by the horseradish peroxidase enzyme in wastewater. Chemosphere 68:1830–1837CrossRefGoogle Scholar
  30. 30.
    Zheng W, Colosi LM (2011) Peroxidase-mediated removal of endocrine disrupting compound mixtures from water. Chemosphere 85:553–557CrossRefGoogle Scholar
  31. 31.
    Pramparo L, Stüber F, Font J et al (2010) Immobilisation of horseradish peroxidase on Eupergit C for the enzymatic elimination of phenol. J Hazard Mater 177:990–1000CrossRefGoogle Scholar
  32. 32.
    Alemzadeh I, Nejati S (2009) Phenols removal by immobilized horseradish peroxidase. J Hazard Mater 166:1082–1086CrossRefGoogle Scholar
  33. 33.
    Zhang YP, Liu TH, Wang Q et al (2012) Synthesis of novel poly(N,N-diethylacrylamide-co-acrylic acid) (P(DEA-co-AA)) microgels as carrier of horseradish peroxidase immobilization for pollution treatment. Macromol Res 20:484–489CrossRefGoogle Scholar
  34. 34.
    Niu JF, Xu JJ, Da YR et al (2013) Immobilization of horseradish peroxidase by electrospun fibrous membranes for adsorption and degradation of pentachlorophenol in water. J Hazard Mater 246–247:119–125CrossRefGoogle Scholar
  35. 35.
    Bayramoglu G, Arica MY (2008) Enzymatic removal of phenol and p-chlorophenol in enzyme reactor: Horseradish peroxidase immobilized on magnetic beads. J Hazard Mater 156:148–155CrossRefGoogle Scholar
  36. 36.
    Zhang F, Zheng B, Zhang J et al (2010) Horseradish peroxidase immobilized on graphene oxide: physical properties and applications in phenolic compound removal. J Phys Chem C 114:8469–8473CrossRefGoogle Scholar
  37. 37.
    Zhai R, Zhang B, Wan YZ et al (2013) Chitosan-halloysite hybrid nanotubes: Horseradish peroxidase immobilization and applications in phenol removal. Chem Eng J 214:304–309CrossRefGoogle Scholar
  38. 38.
    Jiang Y, Tang W, Gao J et al (2014) Immobilization of horseradish peroxidase in phospholipid-templated titania and its applications in phenolic compounds and dye removal. Enzyme Microb Technol 55:1–6CrossRefGoogle Scholar
  39. 39.
    McEldoon JP, Pokora AR, Dordick JS (1995) Lignin peroxidase-type activity of soybean peroxidase. Enzyme Microb Technol 17:359–365CrossRefGoogle Scholar
  40. 40.
    Kamal JKA, Behere DV (2002) Thermal and conformational stability of seed coat soybean peroxidase. Biochemistry 41:9034–9042CrossRefGoogle Scholar
  41. 41.
    Hailu G, Weersink A, Cahlík F (2010) Examining the prospects for commercialization of soybean peroxidase. AgBioForum 13:263–273Google Scholar
  42. 42.
    Gómez M, Murcia MD, Ortega S et al (2012) Removal of 4-chlorophenol in a continuous membrane bioreactor using different commercial peroxidases. Desalin Water Treat 37:97–107CrossRefGoogle Scholar
  43. 43.
    Valli K, Wariishi H, Gold MH (1992) Degradation of 2,7-diclorodibenzo-p-dioxin by the lignin-degrading basidiomycete Phanerochaete chrysosporium. J Bacteriol 174:2131–2137Google Scholar
  44. 44.
    Dong SP, Mao L, Luo SQ et al (2014) Comparison of lignin peroxidase and horseradish peroxidase for catalyzing the removal of nonylphenol from water. Environ Sci Pollut Res Int 21:2358–2366CrossRefGoogle Scholar
  45. 45.
    Aitken MD, Irvine RL (1989) Stability testing of ligninase and Mn-peroxidase from Phanerochaete-chrysosporium. Biotechnol Bioeng 34:1251–1260CrossRefGoogle Scholar
  46. 46.
    Aitken MD, Massey IJ, Chen TP et al (1994) Characterization of reaction-products from the enzyme-catalyzed oxidation of phenolic pollutants. Water Res 28:1879–1889CrossRefGoogle Scholar
  47. 47.
    Carmichael R, Fedoruk PM, Pickar MA (1985) Oxidation of phenols by chloroperoxidase. Biotechnol Lett 7:289–294CrossRefGoogle Scholar
  48. 48.
    Dr D, Corbett MD (1991) Peroxygenation mechanism for chloroperoxidase-catalyzed N-oxidation of arylamines. Chem Res Toxicol 4:556–560CrossRefGoogle Scholar
  49. 49.
    Zhang J, Feng MY, Jiang YC et al (2012) Eficient decolorization/degradation of aqueous azo dyes using buffered H2O2 oxidation catalyzed by a dosage below ppm level of chloroperoxidase. Chem Eng J 191:236–242CrossRefGoogle Scholar
  50. 50.
    Ortiz-Leon M, Velasco L, Vazquez-Duhalt R (1995) Biocatalytic oxidation of polycyclic aromatic hydrocarbons by hemoglobin and hydrogen peroxide. Biochem Biophys Res Commun 215:968–973CrossRefGoogle Scholar
  51. 51.
    González-Sánchez MI, Manjabacas MC, García-Carmona F et al (2009) Mechanism of acetaminophen oxidation by the peroxidase-like activity of methemoglobin. Chem Res Toxicol 22:1841–1850CrossRefGoogle Scholar
  52. 52.
    González-Sánchez MI, García-Carmona F, Macià H et al (2011) Catalase-like activity of human methemoglobin: a kinetic and mechanistic study. Arch Biochem Biophys 516:10–20CrossRefGoogle Scholar
  53. 53.
    Woodward J, Allen BF, Scott MA (1984) Measurement of phenol concentrations using hemoglobin. Biotechnol Bioeng Symp 14:435–438Google Scholar
  54. 54.
    Chapsal JM, Bourbigot MM, Thomas D (1986) Oxidation of aromatic compounds by hemoglobin. Water Res 20:709–713CrossRefGoogle Scholar
  55. 55.
    Pérez-Prior MT, Gómez-Bombarelli R, González-Sánchez MI et al (2012) Biocatalytic oxidation of phenolic compounds by bovine methemoglobin in the presence of H2O2. Quantitative structure-activity relationships. J Hazard Mater 241–242:207–215CrossRefGoogle Scholar
  56. 56.
    Liu J, Guan J, Lu M et al (2012) Hemoglobin immobilized with modified “fish-in-net” approach for the catalytic removal of aniline. J Hazard Mater 217–218:156–163CrossRefGoogle Scholar
  57. 57.
    Ortiz de Montellano PR, Catalano CE (1985) Epoxidation of styrene by hemoglobin and myoglobin. Transfer of oxidizing equivalents to the protein surface. J Biol Chem 260:9265–9271Google Scholar
  58. 58.
    Torres E, Vazquez-Duhalt R (2000) Chemical modification of hemoglobin improves biocatalytic oxidation of PAHs. Biochem Biophys Res Commun 273:820–823CrossRefGoogle Scholar
  59. 59.
    Laveille P, Falcimaigne A, Chamouleau F et al (2010) Hemoglobin immobilized on mesoporous silica as effective material for the removal of polycyclic aromatic hydrocarbons pollutants from water. New J Chem 34:2153–2165CrossRefGoogle Scholar
  60. 60.
    Liu Q, Yu J, Xu Y et al (2013) Bioelectrocatalytic dechlorination of trichloroacetic acid at gel-immobilized hemoglobin on multiwalled carbon nanotubes modified graphite electrode: kinetic modeling and reaction pathways. Electrochim Acta 92:153–160CrossRefGoogle Scholar
  61. 61.
    Valero E, Varón R, García-Carmona F (2002) Tyrosinase-mediated oxidation of acetaminophen to 4-acetamido-o-benzoquinone. Biol Chem 383:1931–1939CrossRefGoogle Scholar
  62. 62.
    Valero E, Escribano J, García-Carmona F (1988) Reactions of 4-methyl-o-benzoquinone, generated chemically or enzymatically, in the presence of l-proline. Phytochemistry 27:2055–2061CrossRefGoogle Scholar
  63. 63.
    Valero E, Carrión P, Varón R et al (2003) Quantification of acetaminophen by oxidation with tyrosinase in the presence of Besthorn’s hydrazone. Anal Biochem 318:187–195CrossRefGoogle Scholar
  64. 64.
    Mukherjee S, Basak B, Bhunia B et al (2013) Potential use of polyphenol oxidases (PPO) in the bioremediation of phenolic contaminants containing industrial wastewater. Rev Environ Sci Biotechnol 12:61–73CrossRefGoogle Scholar
  65. 65.
    Atlow SC, Bonadonna-Aparo L, Klibanov AM (1984) Dephenolization of industrial wastewaters catalyzed by polyphenol oxidase. Biotechnol Bioeng 26:599–603CrossRefGoogle Scholar
  66. 66.
    Wada S, Ichikawa H, Tatsumi K (1995) Removal of phenols and aromatic amines from wastewater by a combination treatment with tyrosinase and a coagulant. Biotechnol Bioeng 45:304–309CrossRefGoogle Scholar
  67. 67.
    Amjad K, Qayyum H (2007) Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes. J Environ Sci 19:396–402CrossRefGoogle Scholar
  68. 68.
    Toscano G, Colarieti ML, Greco G Jr (2003) Oxidative polymerisation of phenols by a phenol oxidase from green olives. Enzyme Microb Technol 33:47–54CrossRefGoogle Scholar
  69. 69.
    Saitoh T, Asano K, Hiraide M (2011) Removal of phenols in water using chitosan-conjugated thermo-responsive polymers. J Hazard Mater 185:1369–1373CrossRefGoogle Scholar
  70. 70.
    Xu DY, Yang Z (2013) Cross-linked tyrosinase aggregates for elimination of phenolic compounds from wastewater. Chemosphere 92:391–398CrossRefGoogle Scholar
  71. 71.
    Sirim D, Wagenr F, Wang L et al (2011) The laccase engineering database: a classification and analysis system for laccases and related multicopper oxidases. Database (Oxford). doi: 10.1093/database/bar006 Google Scholar
  72. 72.
    Thurston CF (1994) The structure and function of fungal laccases. Microbiology 140:19–26CrossRefGoogle Scholar
  73. 73.
    Reiss R, Ihssen J, Richter M et al (2013) Laccase versus Laccase-like multi-copper oxidase: a comparative study of similar enzymes with diverse substrate spectra. PLoS One 8:e65633CrossRefGoogle Scholar
  74. 74.
    Bourbonnais R, Paice MG (1990) Oxidation of non-phenolic substrates: an expanded role of laccase in lignin biodegradation. FEBS Lett 267:99–102CrossRefGoogle Scholar
  75. 75.
    Husain M, Husain Q (2008) Applications of redox mediators in the treatment of organic pollutants by using oxidoreductive enzymes: a review. Crit Rev Environ Sci Technol 38:1–42CrossRefGoogle Scholar
  76. 76.
    Baldrian P (2006) Fungal laccases-occurrence and properties. FEMS Microbiol Lett 30:215–242CrossRefGoogle Scholar
  77. 77.
    Ba S, Haroune L, Cruz-Morató C et al (2014) Synthesis and characterization of combined cross-linked laccase and tyrosinase aggregates transforming acetaminophen as a model phenolic compound in wastewaters. Sci Total Environ 487:748–755CrossRefGoogle Scholar
  78. 78.
    Ba S, Arsenault A, Hassani T et al (2013) Laccase immobilization and insolubilization: from fundamentals to applications for the elimination of emerging contaminants in wastewater treatment. Crit Rev Biotechnol 33:404–418CrossRefGoogle Scholar
  79. 79.
    Durán N, Rosa MA, D’Annibale A et al (2002) Applications of laccases and tyrosinases (phenoloxidases) immobilized on different supports: a review. Enzyme Microb Technol 31:907–931CrossRefGoogle Scholar
  80. 80.
    Majeau JA, Brar SK, Tyagi RD (2010) Laccases for removal of recalcitrant and emerging pollutants. Bioresource Technol 101:2331–2350CrossRefGoogle Scholar
  81. 81.
    Ikehata K, Buchanan ID, Smith DW (2004) Recent developments in the production of extracellular fungal peroxidases and laccases for waste treatment. Environ Eng Sci 3:1–19CrossRefGoogle Scholar
  82. 82.
    Songulashvili G, Elisashvili V, Wasser SP et al (2007) Basidiomycetes laccase and manganese peroxidase activity in submerged fermentation of food industry wastes. Enzyme Microb Technol 41:57–61CrossRefGoogle Scholar
  83. 83.
    Elisashvili V, Penninckx M, Kachlishvili E et al (2008) Lentinus edodes and Pleurotus species lignocellulolytic enzymes activity in submerged and solid-state fermentation of lignocellulosic wastes of different composition. Bioresource Technol 99:457–462CrossRefGoogle Scholar
  84. 84.
    Qiu W, Zhang W, Chen H (2014) Flavonoid-rich plants used as sole substrate to induce the solid-state fermentation of laccase. Appl Biochem Biotechnol 172:3583–3592CrossRefGoogle Scholar
  85. 85.
    Margot J, Bennati-Granier C, Maillard J et al (2013) Bacterial versus fungal laccase: potential for micropollutant degradation. AMB Express 3:63CrossRefGoogle Scholar
  86. 86.
    Camarero S, Cañas AI, Martínez A et al (2010) Laccases having high redox potential obtained through directed evolution. Patent WO2010058057 A1Google Scholar
  87. 87.
    Debaste F, Songulashvili G, Penninckx MJ (2014) The potential of Cerrena unicolor laccase immobilized on mesoporous silica beads for removal of organic micropollutants in wastewaters. Desalin Water Treat 52:2344–2347CrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2014

Authors and Affiliations

  • Edelmira Valero
    • 1
    Email author
  • María-Isabel González-Sánchez
    • 1
  • María-Teresa Pérez-Prior
    • 2
  1. 1.Department of Physical Chemistry, School of Industrial EngineeringUniversity of Castilla-La ManchaAlbaceteSpain
  2. 2.Department of Materials Science and Chemical EngineeringUniversity Carlos III of MadridMadridSpain

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