Skip to main content
SpringerLink
Log in

Production, Purification and Immobilization of Laccase from Trametes versicolor HBB 7328 for its Role in Decolorization of Textile Dyes

  • ORIGINAL RESEARCH ARTICLE
  • Published:
Indian Journal of Microbiology Aims and scope Submit manuscript

Abstract

Laccase is an extracellular enzyme that is widely used in the decolonization of textile dyes in waste water. The aim of our study was to isolate, purify, characterize and immobilize the laccase enzyme produced by Trametes versicolor HBB 7328. Purified laccase enzyme was immobilized in polyacrylamide gel to explore its ability in decolonization of textile dyes. Laccase purification process was carried out by fractionation using ammonium sulphate (80%) followed by DEAE Sepharose column (30 × 3 cm) chromatography method. Recovery and fold purification in this step were 27.35 and 16.23%. Purified laccase (named as LAC1) revealed its optimum activity at pH 5.0 and 35 °C temperature, and displayed remarkable stability in the range of 30–40 °C and in the pH range (pH 3.0–7.0). The single bands on SDS-PAGE represent the purity of LAC1 with molecular weight of 60 kDa. Both free and immobilized laccase assessed for their ability to decolorize textile dyes. Free laccase decolorized Methyl red to 72.705%, Reactive orange to 57.851%, Reactive blue to 37.231%, Bromophenol blue to 24.412% however Immobilized laccase decolorized Methyl red to 89.823%, Reactive orange to 63.151%, Reactive blue to 59.548%, Bromophenol blue to 49.421% respectively. This study proposes the role of laccase from Trametes versicolor HBB 7328 in decolonization of textile dyes.

Graphical Abstract

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
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Maheshwari K, Agrawal M, Gupta AB (2021) Dye pollution in water and wastewater. In: Muthu S S, Khadir A (eds) Novel materials for dye-containing wastewater treatment. Sustainable textiles: production, processing, manufacturing & chemistry. Springer, Singapore, pp 1–25. https://doi.org/10.1007/978-981-16-2892-4_1

  2. Hemashenpagam N, Selvajeyanthi S (2023) Textile dyes and their effect on human beings. In: Ahmad A, Jawaid M, Mohammad Ibrahim MN, Yaqoob A, Alshammari MB (eds) Nanohybrid materials for treatment of textiles dyes. Smart Nanomaterial Technology. Springer, Singapore, pp 41–60.https://doi.org/10.1007/978-981-99-3901-5_3

  3. Katheresan V, Kansedo J, Lau SY (2018) Efficiency of various recent wastewater dye removal methods. J Environ Chem Eng 6:4676–4697. https://doi.org/10.1016/j.jece.2018.06.060

    Article  CAS  Google Scholar 

  4. Kumar V, Pallavi P, Sen SK, Raut S (2024) Harnessing the potential of white rot fungi and ligninolytic enzymes for efficient textile dye degradation: a comprehensive review. Water Environ Res 96:e10959. https://doi.org/10.1002/wer.10959

    Article  CAS  PubMed  Google Scholar 

  5. Kunamneni A, Ballesteros A, Plou FJ, Alcalde M (2007) Fungal laccase a versatile enzyme for biotechnological applications. Commun Curr Res Educat Topics Trends Appl Microbiol 1:233–245

    Google Scholar 

  6. Zofair SF, Ahmad S, Hashmi MA, Khan SH, Khan MA, Younus H (2022) Catalytic roles, immobilization and management of recalcitrant environmental pollutants by laccases: significance in sustainable green chemistry: J Environ Manage 309:1146–1176. https://doi.org/10.1016/j.jenvman.2022.114676

  7. Bertrand B, Martínez-Morales F, Tinoco-Valencia R, Rojas S, Acosta-Urdapilleta L, Trejo-Hernández MR (2015) Biochemical and molecular characterization of laccase isoforms produced by the white-rot fungus Trametes versicolor under submerged culture conditions. J Mol Catal B Enzym 122:339–347. https://doi.org/10.1016/j.molcatb.2015.10.009

    Article  CAS  Google Scholar 

  8. 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. https://doi.org/10.1007/s00018009-0169-1

    Article  CAS  PubMed  Google Scholar 

  9. Sitarz AK, Mikkelsen JD, Meyer AS (2016) Structure, functionality and tuning up of laccases for lignocellulose and other industrial applications. Crit Rev Biotechnol 36:70–86. https://doi.org/10.3109/07388551.2014.949617

    Article  CAS  PubMed  Google Scholar 

  10. Brijwani K, Rigdon A, Vadlani PV (2010) Fungal laccases: production, function, and applications in food processing. Enzyme Res 2010:10. https://doi.org/10.4061/2010/149748

    Article  CAS  Google Scholar 

  11. Chen L, Zou M, Hong FF (2015) Evaluation of fungal laccase immobilized on natural nanostructured bacterial cellulose. Front Microbiol 6:1245. https://doi.org/10.3389/fmicb.2015.01245

    Article  PubMed  PubMed Central  Google Scholar 

  12. Singh J, Kapoor RK (2019) Immobilization and reusability efficiency of Laccase onto different matrices using different approaches. eIJPPR 9:58–65.

  13. Asgher M, Shahid M, Kamal S, Iqbal HM (2014) Recent trends and valorization of immobilization strategies and ligninolytic enzymes by industrial biotechnology. J Mol Catal B Enzym 101:56–66. https://doi.org/10.1016/j.molcatb.2013.12.016

    Article  CAS  Google Scholar 

  14. Liu EK, He WQ, Yan CR (2014) ‘White revolution ‘to ‘white pollution’ agricultural plastic film mulch in China. Environ Res Lett 9:091001. https://doi.org/10.1088/17489326/9/9/091001

    Article  Google Scholar 

  15. Gong T, Zhang X, Liu W (2018) tracing the sources of iodine species in a non saline wastewater. Chemosphere 205:8. https://doi.org/10.1016/j.chemosphere.2018.04.147

    Article  CAS  Google Scholar 

  16. Mani S, Chowdhary P, Bharagava R N (2019) Textile wastewater dyes: toxicity profile and treatment approaches. In: Bharagava R, Chowdhary P (eds) Emerging and eco-friendly approaches for waste management. Springer, Singapore, pp 219–244. https://doi.org/10.1007/978-981-10-8669-4_11

  17. Sarkar P, Roy A, Pal S et al (2017) Enrichment and characterization of hydrocarbon degrading bacteria from petroleum refinery waste as potent bio augmentation agent for in situ bioremediation. Bio Resour Technol 242:2. https://doi.org/10.1016/j.biortech.2017.05.010

    Article  CAS  Google Scholar 

  18. Kaushik P, Malik A (2009) Fungal dye decolorization: recent advances and future potential. Environ Int 35:127–141. https://doi.org/10.1016/j.envint.2008.05.010

    Article  CAS  PubMed  Google Scholar 

  19. Jeyabalan J, Veluchamy A, Kumar A, Chandrasekar R, Narayanasamy S (2023) A review on the laccase assisted decolorization of dyes: recent trends and research progress. J Taiwan Inst Chem Eng 151:105081. https://doi.org/10.1016/j.jtice.2023.105081

    Article  CAS  Google Scholar 

  20. Vasdev K, Kuhad RC (1994) Induction of laccase production in Cyathus bulleri under shaking and static culture conditions. Folia Microbiol 39:30. https://doi.org/10.1007/BF02814322

    Article  Google Scholar 

  21. Kumar VV, Kirupha SD, Periyaraman P, Sivanesan S (2011) Screening and induction of laccase activity in fungal species and its application in dye decolorization. African J Microbiol Res 5:1261–1267. https://doi.org/10.5897/AJMR10.894

    Article  Google Scholar 

  22. Kumar VP, Naik C, Sridhar M (2015) Production, purification and characterization of novel laccase produced by Schizophyllum commune NI-07 with potential for delignification of crop residues. Appl Biochem Microbiol 51:432–441. https://doi.org/10.1134/S0003683815040080

    Article  CAS  Google Scholar 

  23. Pointing SB, Jones EB, Vrijmoed LL (2000) Optimization of laccase production by Pycnoporus sanguineus in submerged liquid culture. Mycologia 92:139–144. https://doi.org/10.2307/3761458

    Article  CAS  Google Scholar 

  24. 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. https://doi.org/10.1128/aem.64.5.1601-1606.1998

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Brazkova M, Mercati A, Hristova I, Lante A, Krastanov A (2016) Isolation, purification and characterization of laccase from the white-rot fungus Trametes Versicolor. Food Sci Eng Technol 62:155–162

    Google Scholar 

  26. Umar A, Ahmed S (2022) Optimization, purification and characterization of laccase from Ganoderma leucocontextum along with its phylogenetic relationship. Sci Rep 12:24–16. https://doi.org/10.1038/s41598-022-06111-z

    Article  CAS  Google Scholar 

  27. Umar A, Abid I, Elshikh MS, Dufossé L, Abdel-Azeem AM, Ali I (2023) Agitation role (Dissolved Oxygen) in production of laccase from newly identified Ganoderma multistipitatum sp. nov. and its effect on mycelium morphology. BMC Microbiol 23:280. https://doi.org/10.1186/s12866-023-03009-2

  28. Dong JL, Zhang YZ (2004) Purification and characterization of two laccase isoenzymes from a ligninolytic fungus Trametes gallica. Prep Biochem Biotechnol 34:179–194. https://doi.org/10.1081/PB-120030876

    Article  CAS  PubMed  Google Scholar 

  29. Patel H, Gupte S, Gahlout M, Gupte A (2014) Purification and characterization of an extracellular laccase from solid state culture of Pleurotus ostreatus HP 1.3Biotech 4: 77–84. https://doi.org/10.1007/s13205-013-0129-1

  30. Zheng F, An Q, Meng G, Wu XJ, Dai YC, Si J, Cui BK (2017) A novel laccase from white rot fungus Trametes orientalis: purification, characterization, and application. Int J Biol Macromol 102:758–770. https://doi.org/10.1016/j.ijbiomac.2017.04.089

    Article  CAS  PubMed  Google Scholar 

  31. Guo M, Lu F, Liu M, Li T, Pu J, Wang N, Liang P, Zhang C (2008) Purification of recombinant laccase from Trametes versicolor in Pichia methanolica and its use for the decolorization of anthraquinone dye. Biotech Lett 30:2091–2096. https://doi.org/10.1007/s10529-008-9817-z

    Article  CAS  Google Scholar 

  32. Moon-Jeong H, Hong-Gyu S (2005) Purification and characterization of laccase from the white rot fungus Trametes versicolor. J Microbiol 43:555–560

    Google Scholar 

  33. Galhaup C, Wagner H, Hinterstoisser B, Haltrich D (2002) Increased production of laccase by the wood degrading basidiomycete Trametes pubescens. Enzyme Microb Technol 30:529–536. https://doi.org/10.1016/S0141-0229(01)00522-1

    Article  CAS  Google Scholar 

  34. Ezike TC, Ezugwu AL, Udeh JO, Eze SO, Chilaka FC (2020) Purification and characterization of new laccase from Trametes polyzona WRF03. Biotechnol Reports 28:00566. https://doi.org/10.1016/j.btre.2020.e00566

    Article  Google Scholar 

  35. Devasia S, Nair A (2022) Purification and characterization of laccase from Arthrographis ksf. Asian J Microbiol Biotechnol Environ Sci 24:528–538. https://doi.org/10.53550/AJMBES.2022.v24i03.0016

  36. More SS, Renuka PS, Malini S (2011) Isolation, purification, and characterization of fungal laccase from Pleurotus sp. Enzyme Res 2011:7. https://doi.org/10.4061/2011/248735

    Article  CAS  Google Scholar 

  37. Gökgöz M, Altinok H (2012) Immobilization of laccase on polyacrylamide and polyacrylamide κ carragennan based semi interpenetrating polymer networks. Artif Cells Blood Subst Biotechnol 40:326–330. https://doi.org/10.3109/10731199.2012.658469

    Article  CAS  Google Scholar 

  38. Huber D, Pellis A, Daxbacher A, Nyanhongo GS, Guebitz GM (2016) Polymerization of various lignins via immobilized Myceliophthora thermophila Laccase (MtL). Polymers 8:280. https://doi.org/10.3390/polym8080280

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Nyanhongo GS, Gomes J, Gübitz GM, Zvauya R, Read J, Steiner W (2002) Decolorization of textile dyes by laccases from a newly isolated strain of Trametes modesta. Water Res 36:1449–1456. https://doi.org/10.1016/S0043-1354(01)00365-7

    Article  CAS  PubMed  Google Scholar 

  40. Ben Younes S, Mechichi T, Sayadi S (2007) Purification and characterization of the laccase secreted by the white rot fungus Perenniporia tephropora and its role in the decolourization of synthetic dyes. J Appl Microbiol 102:1033–1042. https://doi.org/10.1111/j.1365-2672.2006.03152.x

    Article  CAS  PubMed  Google Scholar 

  41. Shraddha SR, Sehgal S, Kamthania M, Kumar A (2011) Laccase: microbial sources, production, purification and potential biotechnological applications. Enzyme Res 2011:217861. https://doi.org/10.4061/2011/217861

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Elisashvili V, Kachlishvili E, Penninckx M (2008) Effect of growth substrate, method of fermentation, and nitrogen source on lignocellulose-degrading enzymes production by white-rot basidiomycetes. J Ind Microbiol Biotechnol 35:531–538. https://doi.org/10.1007/s10295-008-0454-2

    Article  CAS  Google Scholar 

  43. Wasak A, Drozd R, Grygorcewicz B, Jankowiak D, Rakoczy R (2018) Purification and recovery of laccase produced by submerged cultures of Trametes versicolor by three-phase partitioning as simple and highly efficient technique. Pol J Chem Technol 20:88–95. https://doi.org/10.2478/pjct-2018-0059

    Article  CAS  Google Scholar 

  44. Zapata-Castillo P, Villalonga-Santana M (2012) Purification and characterization of laccase from Trametes hirsutaBm2 and its contribution to dye and effluent decolorization. Afr J Biotech 11:3603–3611. https://doi.org/10.5897/AJB11.2050

    Article  CAS  Google Scholar 

  45. Zouari-Mechichi H, Mechichi T, Dhouib A, Sayadi S, Martínez AT, Martinez MJ (2006) Laccase purification and characterization from Trametes trogii isolated in Tunisia: decolorization of textile dyes by the purified enzyme. Enzyme Microb Technol 39:141–148. https://doi.org/10.1016/j.enzmictec.2005.11.027

    Article  CAS  Google Scholar 

  46. Vaithanomsat P, Sangnam A, Boonpratuang T, Choeyklin R, Promkiam on P, Chuntranuluck S, Kreetachat T (2013) Wood degradation and optimized laccase production by resupinate white rot fungi in northern Thailand. Bio Resources 8:6342–6360. https://doi.org/10.15376/biores.8.4.6342-6360

  47. Minovska V, Winkel hausen E, Kuzmanova S (2005) Lipase immobilized by different techniques on various support materials applied in oil hydrolysis. J Serb Chem Soc 70:609–624. https://doi.org/10.2298/JSC0504609M

    Article  Google Scholar 

  48. Yinghui D, Qiuling W, Shiyu F (2002) Laccase stabilization by covalent binding immobilization on activated polyvinyl alcohol carrier. Lett Appl Microbiol 35:4516. https://doi.org/10.1046/j.1472-765X.2002.01196.x

    Article  Google Scholar 

  49. Al Adhami AJ, Bryjak J, Greb Markiewicz B, Peczyńska zoch W, (2002) Immobilization of wood rotting fungi laccases on modified cellulose and acrylic carriers. Process Biochem 37:1387–1394. https://doi.org/10.1016/S0032-9592(02)00023-7

    Article  CAS  Google Scholar 

  50. Forootanfar H, Moezzi A, Aghaie-Khozani M, Mahmoudjanlou Y, Ameri A, Niknejad F, Faramarzi MA (2012) Synthetic dye decolorization by three sources of fungal laccase. Iranian Journal of Environmental Health Science & Engineering 27:1–10. https://doi.org/10.1186/1735-2746-9-27

    Article  CAS  Google Scholar 

  51. Kuhar F, Castiglia V, Levin L (2015) Enhancement of laccase production and malachite green decolorization by co culturing Ganoderma lucidum and Trametes versicolor in solid state fermentation. Int Biodeterior Biodegradation 104:238–243. https://doi.org/10.1016/j.ibiod.2015.06.017

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the Department of Microbiology, Maharshi Dayanand University Rohtak, Haryana, 124001 India and Department of Microbiology, CCS Haryana Agricultural University Hisar, 125004 India.

Funding

Not applicable.

Author information

Authors and Affiliations

  1. Department of Microbiology, CCS Haryana Agricultural University, Hisar, 125004, India

    Nikita Goyat & Baljeet Singh Saharan

  2. Department of Microbiology, Maharshi Dayanand University, Rohtak, Haryana, 124001, India

    Rajeev Kumar Kapoor

  3. Department of Molecular Biology, Biotechnology and Bioinformatics, CCS Haryana Agricultural University, Hisar, 125004, India

    Prexha Kapoor

  4. Department of Biochemistry, CCS Haryana Agricultural University, Hisar, 125004, India

    Kajal Kumari

  5. Department of Bio and Nano Technology, Guru Jambheshwar University of Science and Technology Hisar, Haryana, 124001, India

    Namita Singh

Authors
  1. Nikita Goyat
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rajeev Kumar Kapoor
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Baljeet Singh Saharan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Prexha Kapoor
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kajal Kumari
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Namita Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Nikita Goyat: carried out the experiments; data curation; formal analysis; investigation; methodology; software; validation; visualization; writing original draft. Rajeev Kumar Kapoor: designed the experiment: conceptualization; formal analysis; investigation; resources; super vision; validation; visualization; review. Baljeet Singh Saharan: formal analysis; investigation; super vision; validation; visualization; writing-review and editing. Prexha Kapoor: writing-review and editing; statistical analysis. Kajal Kumari: writing review and editing. Namita Singh: supervision; writing-review and editing.

Corresponding author

Correspondence to Baljeet Singh Saharan.

Ethics declarations

Conflict of interests

The authors declare that they have no known competing financial interests or personal relationship that could have appeared to influence the work reported in this paper.

Consent to Participate

Not applicable.

Consent for Publication

Not applicable.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Goyat, N., Kapoor, R.K., Saharan, B.S. et al. Production, Purification and Immobilization of Laccase from Trametes versicolor HBB 7328 for its Role in Decolorization of Textile Dyes. Indian J Microbiol (2024). https://doi.org/10.1007/s12088-024-01260-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s12088-024-01260-3

Keywords

Search

Navigation