3 Biotech

, 9:129 | Cite as

Simultaneous laccase production and transformation of bisphenol-A and triclosan using Trametes versicolor

  • Jagdeep Singh
  • Punit Kumar
  • Vicky Saharan
  • Rajeev Kumar KapoorEmail author
Original Article


New age micro-pollutants, bisphenol-A (BPA) and triclosan (TCA), known for their carcinogenic effects in living organisms can effectively be removed from water using laccase from Trametes versicolor. Laccase was produced from T. versicolor JSRK13 in both submerged and solid-state fermentation (SmF and SSF) conditions. In SmF, T. versicolor JSRK13 gave the maximum production of laccase on the 10th day with an activity of 22 U mL− 1, whereas, in SSF 185 U g− 1 of the enzyme was produced on the 17th day. Maximum production of laccase was observed with Parthenium as substrate. Parthenium, with a particle size of 3–5 mm having 60% moisture was found to be a suitable substrate for laccase production and simultaneous transformation (LPST) of BPA in a synergistic manner. A one-step concentration using 85% ammonium sulphate followed by dialysis was sufficient to give 6.7-fold purification of laccase from the crude culture filtrate. Transformation of BPA was achieved in both SmF and SSF conditions along with the production of laccase, whereas TCA was degraded with free enzyme only. Above 90% of BPA (55–5 mg L− 1) was degraded using the LPST strategy with HBT acting as a mediator in the reaction. LPST strategy did not work for TCA as it completely inhibits the growth of T. versicolor JSRK13. TCA was degraded up to 75% (1.5–0.375 mg L− 1) by the free enzyme. Our study of simultaneous laccase production and transformation proved to be efficacious in case of BPA. The results indicate that industrial and sewage wastewater containing BPA can potentially be treated with T. versicolor JSRK13 laccase. The described strategy can further be used to develop a bioprocess which can work both on solid and liquid wastes containing BPA.


Bisphenol-A Triclosan Laccase Laccase production and simultaneous transformation Solid-state fermentation Submerged fermentation Trametes versicolor JSRK13 



The author sincerely acknowledges the University Grants Commission, for sanctioning a Major Research Project on “Optimizing Production of Laccase Enzyme from Selected White-Rot Fungi and Developing a Process for the Degradation of Endocrine Disruptors”. [Grant no. 42-486/2013 (SR)].

Compliance with ethical standards

Conflict of interest

On behalf of all authors, the corresponding author states that there is no conflict of interest.

Supplementary material

13205_2019_1648_MOESM1_ESM.docx (886 kb)
Supplementary material 1 (DOCX 885 KB)


  1. Annamalai J, Namasivayam V (2015) Endocrine disrupting chemicals in the atmosphere: Their effects on humans and wildlife. Environ Int 76:78–97. CrossRefPubMedGoogle Scholar
  2. Anwar Z, Gulfraz M, Irshad M (2014) Agro-industrial lignocellulosic biomass a key to unlock the future bio-energy: a brief review. J Radiat Res Appl Sci 7:163–173. CrossRefGoogle Scholar
  3. Aparicio I, Martín J, Abril C et al (2018) Determination of household and industrial chemicals, personal care products and hormones in leafy and root vegetables by liquid chromatography-tandem mass spectrometry. J Chromatogr A 1533:49–56. CrossRefPubMedGoogle Scholar
  4. Aristiawan Y, Aryana N, Putri D, Styarini (2015) Analytical method development for Bisphenol-A in Tuna by using High-performance liquid chromatography—UV. Proc Chem 16:202–208CrossRefGoogle Scholar
  5. Ashe B, Nguyen LN, Hai FI et al (2016) Impacts of redox-mediator type on trace organic contaminants degradation by laccase: degradation efficiency, laccase stability and effluent toxicity. Int Biodeterior Biodegrad 113:169–176. CrossRefGoogle Scholar
  6. Asif MB, Hai FI, Kang J, Price WE, Nghiem LD (2018) Biocatalytic degradation of pharmaceuticals, personal care products, industrial chemicals, steroid hormones and pesticides in a membrane distillation-enzymatic bioreactor. Biores Technol 247:528–536CrossRefGoogle Scholar
  7. Aurand C (2012) Comprehensive SPE sample preparation and fast HPLC method for bisphenol-A in drinking water. Supelco Analytical. Report US Rapid Determ Packag Contam Food Beverages 30.1:8–9Google Scholar
  8. Baweja M, Nain L, Kawarabayasi Y, Shukla P (2016) Current technological improvements in enzymes toward their biotechnological applications. Front Microbiol 7:1–13. CrossRefGoogle Scholar
  9. Becker D, Rodriguez-Mozaz S, Insa S et al (2017) Removal of endocrine disrupting chemicals in wastewater by enzymatic treatment with fungal laccases. Org Process Res Dev 21:480–491. CrossRefGoogle Scholar
  10. Bernier MR, Vandenberg LN (2017) Handling of thermal paper: Implications for dermal exposure to bisphenol-A and its alternatives. PLoS One 12:e0178449. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Cao X-L, Corriveau J, Popovic S (2009) Levels of Bisphenol-A in canned soft drink products in canadian markets. CrossRefGoogle Scholar
  12. Chandra R, Sharma P, Yadav S, Tripathi S (2018) Biodegradation of endocrine-disrupting chemicals and residual organic pollutants of pulp and paper mill effluent by biostimulation. Front Microbiol. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Chattu D, Supervisor C, Donkor K (2016) Determination of triclosan in personal care products and swimming pool samples by liquid chromatography-mass spectrometry. In: Proceedings of the annual thompson rivers university undergraduate research and innovation conference, vol 10(1), Article 3.
  14. Chhaya R, Modi HA (2013) Comparative Study of laccase production by streptomyces chartreusis in solid state and submerged. Indian J Fundam Appl Life Sci 3:73–84Google Scholar
  15. Chiang C, Mahalingam S, Flaws J (2017) Environmental contaminants affecting fertility and somatic health. Semin Reprod Med 35:241–249. CrossRefPubMedPubMedCentralGoogle Scholar
  16. Dhillon GS, Kaur S, Pulicharla R et al (2015) Triclosan: current status, occurrence, environmental risks and bioaccumulation potential. Int J Environ Res Public Health 12:5657–5684. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Dionex Corporation (2012) Determination of inorganic anions in environmental waters using a hydroxide-selective column. Application Note 154; Sunnyvale, pp 42–50Google Scholar
  18. Falade AO, Mabinya LV, Okoh AI, Nwodo UU (2018) Ligninolytic enzymes: versatile biocatalysts for the elimination of endocrine-disrupting chemicals in wastewater. MicrobiologyOpen 7:1–17. CrossRefGoogle Scholar
  19. Garcia-Morales R, Rodríguez-Delgado M, Gomez-Mariscal K et al (2015) Biotransformation of endocrine-disrupting compounds in groundwater: bisphenol-A, nonylphenol, ethynylestradiol and triclosan by a laccase cocktail from Pycnoporus sanguineus CS43. Water Air Soil Pollut. CrossRefPubMedPubMedCentralGoogle Scholar
  20. Ghazarian AA, Trabert B, Robien K et al (2018) Maternal use of personal care products during pregnancy and risk of testicular germ cell tumours in sons. Environ Res 164:109–113. CrossRefPubMedGoogle Scholar
  21. Jasim Hashim A (2011) Determination of optimal conditions for laccase production by Pleurotus ostreatus using sawdust as solid medium and its use in phenol degradation. Baghdad Sci J 9(3):491–498Google Scholar
  22. Kapoor RK, Rajan K, Carrier DJ (2015) Applications of Trametes versicolor crude culture filtrates in detoxification of biomass pretreatment hydrolyzates. Bioresour Technol 189:99–106CrossRefGoogle Scholar
  23. Kaur M, Aggarwal NK, Kumar V, Dhiman R (2014) Effects and management of Parthenium hysterophorus: a weed of global significance. Int Sch Res Not 2014:1–12. CrossRefGoogle Scholar
  24. Kim M, Day DF (2011) Composition of sugar cane, energy cane, and sweet sorghum suitable for ethanol production at Louisiana sugar mills. J Ind Microbiol Biotechnol 38:803–807. CrossRefPubMedGoogle Scholar
  25. Koumaki E, Mamais D, Noutsopoulos C (2018) Assessment of the environmental fate of endocrine disrupting chemicals in rivers. Sci Total Environ 628–629:947–958. CrossRefPubMedGoogle Scholar
  26. Kumar V, Baweja M, Liu H, Shukla P (2017) Microbial enzyme engineering: applications and perspectives. Recent advances in applied microbiology. Springer, Singapore, pp 259–273CrossRefGoogle Scholar
  27. Lassouane F, Aït-Amar H, Amrani S, Rodriguez-Couto S (2019) A promising laccase immobilization approach for Bisphenol-A removal from aqueous solutions. Bioresour Technol 271:360–367. CrossRefPubMedGoogle Scholar
  28. Lecomte S, Habauzit D, Charlier DC, Pakdel F (2017) Emerging estrogenic pollutants in the aquatic environment and breast cancer. Genes (Basel) 8:2–21. CrossRefGoogle Scholar
  29. Lind PM, Lind L (2018) Endocrine-disrupting chemicals and risk of diabetes: an evidence-based review. Diabetologia 61:1495–1502. CrossRefPubMedGoogle Scholar
  30. Liu T, Wu D (2012) High-performance liquid chromatographic determination of triclosan and triclocarban in cosmetic products. Int J Cosmet Sci 34:489–494. CrossRefPubMedGoogle Scholar
  31. Llorca M, Badia-Fabregat M, Rodríguez-Mozaz S et al (2017) Fungal treatment for the removal of endocrine disrupting compounds from reverse osmosis concentrate: identification and monitoring of transformation products of benzotriazoles. Chemosphere 184:1054–1070. CrossRefPubMedGoogle Scholar
  32. McConnachie AJ, Strathie LW et al (2011) Current and potential geographical distribution of the invasive plant Parthenium hysterophorus (Asteraceae) in eastern and southern Africa. Weed Res 51:71–84. CrossRefGoogle Scholar
  33. Meehnian H, Jana AK, Jana MM (2016) Effect of particle size, moisture content, and supplements on selective pretreatment of cotton stalks by Daedalea flavida and enzymatic saccharification. 3 Biotech 6:235. CrossRefPubMedPubMedCentralGoogle Scholar
  34. Mohapatra DP, Brar SK, Tyagi RD, Surampalli RY (2011) Occurrence of bisphenol-A in wastewater and wastewater sludge of CUQ treatment plant. J Xenobiot 1:9–16. CrossRefGoogle Scholar
  35. Murthy K (2010) Chemical and biochemical properties of Parthenium and Chormolaena compost. Int J Sci Nat 1(2):166–171Google Scholar
  36. Nag SK, Das Sarkar S, Manna SK (2018) Triclosan—an antibacterial compound in water, sediment and fish of River Gomti, India. Int J Environ Health Res. CrossRefPubMedGoogle Scholar
  37. Nagy KZ, László B, Fleit E, Czihat MK (2018) Behavior of organic micropollutants during river bank filtration in Budapest, Hungary. Water 10:2–13Google Scholar
  38. Nguyen LN, Hai FI, Yang S et al (2013) Removal of trace organic contaminants by an MBR comprising a mixed culture of bacteria and white-rot fungi. Bioresour Technol 148:234–241. CrossRefPubMedGoogle Scholar
  39. Niladevi KN, Sukumaran RK, Prema P (2007) Utilization of rice straw for laccase production by Streptomyces psammoticus in solid-state fermentation. J Ind Microbiol Biotechnol 34:665–674. CrossRefPubMedGoogle Scholar
  40. Noonan GO, Ackerman LK, Begley TH (2011) Concentration of Bisphenol-A in highly consumed canned foods on the U.S. market. J Agric Food Chem 59:7178–7185. CrossRefPubMedGoogle Scholar
  41. Oates RP, Longley G, Hamlett P, Klein D (2017) Pharmaceutical and endocrine disruptor compounds in surface and wastewater in San Marcos, Texas. Water Environ Res 89:2021–2030. CrossRefPubMedGoogle Scholar
  42. Ozcirak Ergun S, Ozturk Urek R (2017) Production of ligninolytic enzymes by solid state fermentation using Pleurotus ostreatus. Ann Agrar Sci 15:273–277. CrossRefGoogle Scholar
  43. Pandiyan K, Tiwari R, Singh S et al (2014) Optimization of enzymatic saccharification of alkali pretreated Parthenium sp. using response surface methodology. Enzyme Res 2014:1–8. CrossRefGoogle Scholar
  44. Parenti CC, Ghilardi A, Torre DC, Giacco DL, Binelli A (2019) Environmental concentrations of triclosan activate cellular defence mechanism and generate cytotoxicity on zebrafish (Danio rerio) embryos A. Sci Total Environ 650:1752–1758CrossRefGoogle Scholar
  45. Patel S (2011) Harmful and beneficial aspects of Parthenium hysterophorus: an update. 3 Biotech 1:1–9. CrossRefPubMedPubMedCentralGoogle Scholar
  46. Puig DRT (2012) Removal of endocrine disrupting chemicals by the ligninolytic enzyme versatile peroxidase. Group of Environmental Biotechnology Doctoral thesisGoogle Scholar
  47. Raisibe F, Adegbenro LP, DasoJonathan OO (2017) Occurrence and environmental levels of triclosan and triclocarban in selected wastewater treatment plants in Gauteng Province, South Africa. Emerg Contam 3:107–114. CrossRefGoogle Scholar
  48. Rodríguez-Fernández DE, Rodríguez-León JA, de Carvalho JC et al (2012) Influence of airflow intensity on phytase production by solid-state fermentation. Bioresour Technol 118:603–606. CrossRefPubMedGoogle Scholar
  49. Saini A, Aggarwal NK, Sharma A et al (2014) Utility potential of Parthenium hysterophorus for its strategic management. Adv Agric 2014:1–16. CrossRefGoogle Scholar
  50. Sarnthima R, Khammuang S (2009) Laccase activity from fresh fruiting bodies of Ganoderma sp. MK05: purification and remazol brilliant blue R decolorization. J Biol Sci 9:83–87. CrossRefGoogle Scholar
  51. Sharma B, Dangi AK, Shukla P (2018) Contemporary enzyme based technologies for bioremediation: a review. J Environ Manag 210:10–22. CrossRefGoogle Scholar
  52. Shimelis O, Halpenny M, Ken E, Schultz K (2014) Determination of triclosan in environmental waters using polymeric SPE cleanup and HPLC with mass spectrometric detection. Supelco Anal Report 32(2):22–23Google Scholar
  53. Sifakis S, Androutsopoulos VP, Tsatsakis AM, Spandidos DA (2017) Human exposure to endocrine disrupting chemicals: effects on the male and female reproductive systems. Environ Toxicol Pharmacol 51:56–70. CrossRefPubMedGoogle Scholar
  54. Sin J-C, Lam S-M, Mohamed AR, Lee K-T (2012) Degrading endocrine disrupting chemicals from wastewater by photocatalysis: a review. Int J Photoenergy 2012:1–23. CrossRefGoogle Scholar
  55. Singh JP, Kapoor RK (2014) Microbial laccases: a mini-review on their production, purification and applications. Int J Pharm Arch 3:528–536Google Scholar
  56. Singh S, Moholkar VS, Goyal A (2014) Bioethanol production by pretreatment, hydrolysis and fermentation of Parthenium Hysterophorus. Int J Appl Eng Res 9:1149–1150Google Scholar
  57. Singh J, Saharan V, Kumar S, Gulati P, Kapoor RK (2017) Laccase grafted membranes for advanced water filtration systems: a green approach to water purification technology. Crit Rev Biotechnol. CrossRefPubMedGoogle Scholar
  58. Siracusa JS, Yin L, Measel E et al (2018) Effects of bisphenol-A and its analogs on reproductive health: a mini review. Reprod Toxicol 79:96–123. CrossRefPubMedGoogle Scholar
  59. Sivakumar R, Rajendran R, Balakumar C et al (2010) Isolation, screening and optimization of production medium for thermostable laccase production from Ganoderma sp. Int J Eng Sci 2:7133–7141Google Scholar
  60. Skledar DG, Carino A, Trontelj J, Troberg J, Distrutti E, Marchianò S, Tomašič T et al (2019) Endocrine activities and adipogenic effects of bisphenol-AF and its main metabolite. Chemosphere 2015:870–880. (Epub 2018 Oct 18) CrossRefGoogle Scholar
  61. Tsioulpas A, Dimou D, Iconomou D, Aggelis G (2002) Phenolic removal in olive oil mill wastewater by strains of Pleurotus spp. in respect to their phenol oxidase (laccase) activity. Bioresour Technol 84:251–257CrossRefGoogle Scholar
  62. Vasdev K, Kuhad RC (1994) Induction of laccase production in Cyathus bulleri under shaking and static culture conditions. Folia Microbiol 39:326–330. CrossRefGoogle Scholar
  63. Vastrad BM, Neelagund SE (2012) Optimization of process parameters for rifamycin b production under solid state fermentation from amycolatopsis mediterranean MTCC 14. Int J Curr Pharm Res 4(2):101–106Google Scholar
  64. Williams M, Kookana RS, Mehta A, Yadav SK, Tailor BL, Maheshwari B (2019) Emerging contaminants in a river receiving untreated wastewater from an Indian urban centre. Sci Total Environ 647:1256–1265. CrossRefPubMedGoogle Scholar
  65. Xie H (2013) Production of a recombinant laccase from Pichia pastoris and biodegradation of chlorpyrifos in a laccase/vanillin system. J Microbiol Biotechnol 23:864–871. CrossRefPubMedGoogle Scholar
  66. Xu ZX, Wu Q, Duan Y et al (2016) Development of a novel spectrophotometric method based on diazotization-coupling reaction for determination of bisphenol-A. J Braz Chem Soc 28:1475–1482. CrossRefGoogle Scholar
  67. Yalçin MS, Geçgel C, Battal D (2016) Determination of bisphenol-A in thermal paper receipts. J Turk Chem Soc Sect A Chem 3:167–174. CrossRefGoogle Scholar
  68. Yang S, Hai FI, Nghiem LD et al (2013) Understanding the factors controlling the removal of trace organic contaminants by white-rot fungi and their lignin modifying enzymes: a critical review. Bioresour Technol 141:97–108. CrossRefPubMedGoogle Scholar
  69. Yang Y, Yang Y, Zhang J, Shao B, Yin J (2019) Assessment of bisphenol A alternatives in paper products from the Chinese market and their dermal exposure in the general population. Environ Pollut 244:238–246. CrossRefPubMedGoogle Scholar
  70. Yao L, Lv Y-Z, Zhang L-J et al (2018) Determination of 24 personal care products in fish bile using hybrid solvent precipitation and dispersive solid phase extraction cleanup with ultrahigh performance liquid chromatography-tandem mass spectrometry and gas chromatography-mass spectrometry. J Chromatogr A 1551:29–40. CrossRefPubMedGoogle Scholar
  71. Yu H, Caldwell DJ, Suri RP (2019) In vitro estrogenic activity of representative endocrine disrupting chemicals mixtures at environmentally relevant concentrations. Chemosphere 215:396–403. (Epub 2018 Oct 11) CrossRefPubMedGoogle Scholar
  72. Zeng S, Zhao J, Xia L (2017) Simultaneous production of laccase and degradation of bisphenol-A with Trametes versicolor cultivated on agricultural wastes. Bioprocess Biosyst Eng 40:1237–1245. CrossRefPubMedPubMedCentralGoogle Scholar
  73. Zhou Y, Cheng G, Chen K, Lu J, Lei J, Pu S (2019a) Adsorptive removal of bisphenol-A, chloroxylenol, and carbamazepine from waterusing a novel β-cyclodextrin polymer. Ecotoxicol Environ Saf 170:278–285. CrossRefPubMedGoogle Scholar
  74. Zhou X, Yang Z, Luo Z, Li H, Chen G (2019b) Endocrine disrupting chemicals in wild freshwater fishes: species, tissues, sizes and human health risks. Environ Pollut 244:462–468. CrossRefPubMedGoogle Scholar
  75. Zhu C, Bao G, Huang S (2016) Optimization of laccase production in the white-rot fungus Pleurotus ostreatus (ACCC 52857) induced through yeast extract and copper. Biotechnol Biotechnol Equip 30:270–276. CrossRefGoogle Scholar

Copyright information

© King Abdulaziz City for Science and Technology 2019

Authors and Affiliations

  • Jagdeep Singh
    • 1
  • Punit Kumar
    • 2
  • Vicky Saharan
    • 1
  • Rajeev Kumar Kapoor
    • 1
    Email author
  1. 1.Enzyme Biotechnology Laboratory, Department of MicrobiologyMaharshi Dayanand UniversityRohtakIndia
  2. 2.Department of BiotechnologyUniversity Institute of Engineering and Technology, Maharshi Dayanand UniversityRohtakIndia

Personalised recommendations