Production of Fungal Laccase under Solid State Bioprocessing of Agroindustrial Waste and Its Application in Decolourization of Synthetic Dyes

Main Article Content

Benjamin Vandelun Ado
Abiodun Anthony Onilude
Hyacinth Ocheigwu Apeh Oluma
Daniel Malo Mabitine


Fungal laccases are preferred due to high redox potentials and low substrate specificity to xenobiotics including synthetic dyes. For large-scale applications, low enzyme yield and high cost of production has remained the challenge. Agroindustrial waste such as saw-dust of Terminalia superba abounds locally and was utilized as low-cost alternative substrate for laccase production in Solid State Bioprocessing (SSB) using Trametes sp. isolate G31. The study optimized laccase production using various parameters. Optimal production of laccase was at pH 5.0 - 7.0 with 2356 U/mL - 2369 U/mL and 25°C (2336 U/mL). Among the sources of nitrogen and carbon tested, laccase production in ammonium sulphate and sucrose supplemented media were higher. The effect of activators on laccases production showed that Cu2+ and Ca2+induced high titres of laccase at 4 -5 mM and 2 - 4 mM respectively, while production of laccase by Mn2+ was significantly high at 40 mM. The effect of 2, 2-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS), guaiacol and varatryl alcohol on laccase production was significantly different especially in glycerol. Optimum production for laccase was on day 14 with 2356 U/mL followed by steady declined up to day 34. The purified laccase had a specific activity of 5008 µmol/min/mg, purification factor of 3.85, and a molecular mass of ~40 kDa using N-PAGE. The potential of crude laccase to decolourize diverse dyes was tested. Phenol red achieved 40% decolourization for 1hr, while RBBR (93%), Crystal violet (60%), Methylene blue (53%) and Congo red (51%) for 24 hr, 72 hr, 48 hr and 120 hr respectively. Methyl red and Malachite green attained 42% (72 hr) and 32% (48 hr) decolourization. The enzyme ability to oxidize Phenol red and other synthetic dyes without mediators made it eco-friendly in treating dye wastewaters.

Trametes sp. isolate G31, solid state bioprocessing, sawdust, optimization, laccase, synthetic dyes

Article Details

How to Cite
Vandelun Ado, B., Anthony Onilude, A., Apeh Oluma, H. O., & Mabitine, D. M. (2019). Production of Fungal Laccase under Solid State Bioprocessing of Agroindustrial Waste and Its Application in Decolourization of Synthetic Dyes. Journal of Advances in Biology & Biotechnology, 21(4), 1-17.
Original Research Article

Article Metrics


Iqbal HMN, Asgher M, Bhatti HN. Optimization of physical and nutritional factors for synthesis of lignin degrading enzymes by a novel strain of Trametes versicolor. Bioresources. 2011;6(2):1273-1287.

Kaur S, Dhillon GS, Brar SK. Increasing Trend towards production of high value bioproducts from biomass feedstocks. International Journal of Waste Resources. 2013;3:e105.


Akhator P, Obanor A, Ugege A. Nigerian Wood Waste: A potential resource for economic development. Journal of Applied Sciences and Environmental Management. 2017;21(2):246-251.

Rominiyi OL, Adaramola BA, Ikumapayi OM, Oginni OT, Akinola SA. Potential utilization of sawdust in energy, manufacturing and agricultural industry; waste to wealth. World Journal of Engineering and Technology. 2017;5:526-539.


Anwar Z, Gulfraz M, Irshad M. Agroindustrial lignocellulosic biomass a key to unlock the future bio-energy: A brief review. Journal of Radiation Research and Applied Sciences. 2014;7:163-173.

El-Batal AI, El-kenawy NM, Yassin AS, Amin MA. Laccase production by Pleurotus ostreatus and its application in synthesis of gold nano particles. Biotechnology Report. 2015;5:31-39.

Akpinar M, Urek RO. Induction of fungal laccase production under solid state bioprocessing of new agroindustrial waste and its application on dye decolourization. 3 Biotechnology. 2017;7:98.1-10.

DOI: 10.1007/s13205-017-0742-5

Poojary H, Hoskeri A, Kaur A, Mugeraya G. Comparative production of ligninolytic enzymes from novel isolates of basidiomycetes and their potential to degrade textile dyes. Nature and Science. 2012;10(10):90-96.

Aftab ZH, Ahmad S. Ganoderma lucidum: A case study for laccase biosynthesis. Pakistan Journal of Phytopathology. 2015; 27(01):95-103.

Ado BV, Amande JT, Ebah EE, Mabitine DM. Screening, production and partial characterization of a thermostable laccase from Trametes sp. isolate B7 with biotechnological potentials. Biotechnology Journal International. 2018;22(4):1-16. DOI: 10.9734/BJI/2018/v22i430065.

Elsayed MA, Hassan MM, Elshafei AM, Haroun BM, Othman AM. Optimization of Cultural and Nutritional Parameters for the Production of Laccase by Pleurotus ostreatus ARC280. British Biotechnology Journal. 2012;2(3):115-132.

Mate DM, Alcalde M. Laccase: A multipurpose biocatalyst at the forefront of biotechnology. Microbial Biotechnology. 2017;10(6):1457-1467.

Zhang J, Sun L, Zhang H, Wang S, Zhang X, Geng A. A novel homodimer laccase from Cerrena unicolor BBP6: Purification, characterization, and potential in dye decolourization and denim bleaching. Plos One. 2018;13(8):e0202440.

Asgher M, Kamal S, Iqbal HMN. Improvement of catalytic efficiency, thermo-stability and dye decolourization capability of Pleurotus ostreatus IBL-02 laccase by hydrophobic sol gel entrapment. Chemistry Central Journal. 2012;6(1):110.

DOI: 10.1186/1752-153X-6-110.

Pang S, Wu Y, Zhang X, Li B, Ouyang J, Ding M. Immobilization of laccase via adsorption onto bimodal mesoporous Zr-MOF. Process Biochemistry 2016;51(2): 229-239.

Zemin F, Xiaoman L, Liyuan C, Ya S. Identification of a Laccase GLac 15 from Ganoderma lycidum 77002 and it’s application in Biotechnology for Biofuels. Biotechnology for Biofuels. 2015;8(1):54.

Zhu C, Bao G, Huang S. Optimization of laccase production in the white-rot fungus Pleurotus ostreatus (ACCC 52857) induced through yeast extract and copper. Biotechnology and Biotechnological Equipment. 2016;30(2):270-276.

DOI: 10.1080/13102818.2015.1135081.

Ado BV, Onilude AA, Amande T. Production and optimization of laccase by Trametes sp. isolate B7 and its’ dye decolourization potential. Journal of Advances in Microbiology. 2018;13(1): 1-14.

DOI: 10.9734/JAMB/2018/44218.

Songserm P, Sihanonth P, Sangvanich P, Karnchanatat A. Decolourization of textile dyes by Polyporus pseudobetulinus and extracellular laccase. African Journal of Microbiology Research. 2012;6(4):779-792.

DOI: 10.5897/AJMR11.988.

Wakil SM, Eyiolawi SA, Salawu KO, Onilude AA. Decolourization of synthetic dyes by laccase enzyme produced by Kluyveromyces dobzhanskii DW1 and Pichia manshurica DW2. African Journal of Biotechnology. 2019;18(1):1-11.

DOI: 10.5897/AJB2018.16674.

Pramanik S, Chaudhuri S. Laccase activity and azo dye decolourization potential of Podoscypha elegans. Mycobiology. 2018; 46(1):79-83.

Villegas LGC, Mashhadi N, Chen M, Mukherjee D, Taylor KE, Biswas N. A short review of techniques for phenol removal from wastewater. Current Pollution Reports. 2016;2(3): 157-167.

Shervedani R, Amini A. Direct electrochemistry of dopamine on gold-laccase enzyme electrode: Characterization and quantitative detection. Bioelectrochemistry. 2012;84: 25-31.

Dunn M, Domsch K, Gams W, Anderson T. Compendium of soil fungi. Taxon. 1982; 31(3):600.

Patrick F, Mtui G, Mshandete AM, Kivaisi A. Optimization of laccase and manganese peroxidase production in submerged culture of Pleurotus sajorcaju. African Journal of Biotechnology. 2011;10(50): 10166-10177.

Pointing SB. Qualitative methods for the determination of lignocellulolytic enzyme production by tropical fungi. Fungi Diversity. 1999;2:17-33.

Dhouib A, Hamza M, Zouari H, Mechichi T, H’midi R, Labat M, et al. Autochthonous fungal strains with high ligninolytic activities from Tunisian biotopes. African Journal of Biotechnology. 2005;4(5):431- 436.

Markson AA, Madunagu BE, Enyiko ED. Growth performance of Pleurotus ostreatus (Jacq. et. Fr.) Kummer on different substrates treated with used automobile engine oil. International Journal of Recent Scientific Research. 2012;3(5): 384 -388.

Osibe DA, Chiejina NV. Assessment of palm press fibre and sawdust based substrate formulas for efficient carpophore production of Lentinus squarrosulus (Mont.) singer. Mycobiology. 2015;43(4): s467-474. Available:

Gomes E, Aguiar AP, Carvalho CC, Bonfa MRB, Silva R, Boscolo M. Ligninase production by basidiomycetes strains on lignocellulosic agricultural residues and their application in the decolourization of synthetics dyes. Brazilian Journal of Microbiology. 2009;40:31-39.

Urairuj C, Khanongnuch C, Lumyoung S. Ligninolytic enzymes from tropical endophytic Xylariaceae. Fungal Diversity. 2003;13:209-219.

Sivakumar R, Rajendran R, Balakumar C, Tamilvendan M. Isolation, screening and optimization of production medium for thermostable laccase production from Ganoderma sp. International Journal of Engineering Science and Technology. 2010;2(12):7133-7141.

Singh N, Abraham J. Isolation of laccase producing fungus from compost soil and partial characterization of laccase. Advances in Applied Science Research. 2013;4(5):91-98.

Wuyep PA, Khan AU, Nok AJ. Production and regulation of lignin degrading enzymes from Lentinus squarrosulus (Mont.) Singer and Psathyrella atroumbonata Pegler. African Journal of Biotechnology. 2003; 2(11):444-447.

Kumar VV, Kirupha SD, Periyaraman P, Sivanesan S. Screening and induction of laccase activity in fungal species and its application in dye decolourization. African Journal of Microbiology Research. 2011; 5(11):1261-1267.

Aslam S, Asgher M. Partial purification and characterization of ligninolytic enzymes produced by Pleurotus ostreatus during solid state fermentation. African Journal of Biotechnology. 2011;10(77):17875-17883.

Masalu RJ. Ligninolytic enzymes of the fungus isolated from soil contaminated with cow dung. Tanzanian Journal of Science. 2016;42:85-93.

Moturi B, Singara-Charya MA. Decolourization of crystal violet and malachite green by fungi. Science World Journal. 2009;4(4):1597-6343.

Stoilova I, Krastanov A, Stanchev V. Properties of crude laccase from Trametes versicolor produced by solid-state fermentation. Advances in Bioscience and Technology. 2010;1:208-215.

Ducan DB. Multiple range and multiple F tests. Biometrics. 1955;11(1):1-42.

Ding Z, Peng L, Chen Y, Zhang L, Gu Z, Shi V, et al. Production and characterization of thermostable laccase from the mushroom, Ganoderma lucidum, using submerged fermentation. African Journal of Microbiology Research. 2012; 6(6):1147-1157.

Elisashvili V, Kachlishvili E. Physiological regulation of laccase and manganese peroxidase production by white-rot basidiomycetes. Journal of Biotechnology. 2009;144:37-42.

Bhattacharya S, Bhardwaj S, Das A, Anand S. Utilization of sugar cane bassage for solid state fermentation and characterization of α-amylase from Aspergillus flavus isolated from Muthupettai mangrove, Tamil Nadu. Australian Journal of Basic and Applied Sciences. 2011;5(12):1012-1022.

Schulte PM. The effects of temperature on aerobic metabolism: Towards a mechanistic understanding of the responses of ectotherms to a changing environment. The Journal of Experimental Biology. 2015;218.1856-1866.


Stajia M, Vukojevia J, Lausevia SD. Influence of the cultivation conditions on ligninolytic enzyme production in Pleurotus pulmonarius. Proceedings Nature and Science. 2007;113:303-312.

Kachlishvili E, Metreveli E, Elisashvili V. Modulation of Cerrena unicolor laccase and manganese peroxidase production. Springer Plus. 2014;3:463.

Fonseca MI, Shimizu E, Zapata PD, Villalba LL. Copper inducing effect on laccase production of white rot fungi native from Misiones (Argentina). Enzyme Microbial Technology; 2010.

DOI: 10.1016/j.enzmictec.2009.12.017

Irshad M, Asgher M. Production and optimization of ligninolytic enzyme by white rot fungus Schizophyllum commune IBL-06 in solid state fermentation medium banana stalk. African Journal of Biotechnology. 2011;10:18234-18242.

Ire FS, Ahuekwe EF. Production of fungal laccase using orange peelings as substrate by submerged static fermentation. British Microbiology Research Journal. 2016;15(5):1-19.

Zheng ZM, Obbard JP. Effect of nonionic surfactants on elimination of polycyclic aromatic hydrocarbons (PAHs) in soil slurry by Phanerochaete chrysosporium. Journal of Chemical Technology Biotechnology. 2001;76:423-429.

Zhu XD, Gibbons J, Garcia-Rivera J, Casadevall A, Williamson PR. Laccase of Cryptococcus neoformans is a cell wall associated virulence factor. Infection and Immunity. 2001;69:5589-5596.

Saparrat MCN, Guillen F, Arambarri AM, Martinez AT, Martinez MJ. Induction, isolation, and characterization of two laccases from the white rot basidiomycete Coriolopsis rigida. Applied and Environmental Microbiology. 2002;68(4): 1534-1540.

Majolagbe ON, Oloke JK, Deka-Boruah HP, Bordoloi AK, Borah M. Extraction of extracellular laccase from wild, mutants and hybrid strains of two white-rot fungus and its applications in decolourization and ligninolysis. Journal of Science and Technology. 2012;2:301-317.

Suwannawong P, Khammuang S, Sarnthima R. Decolourization of rhodamine B and congo red by partial purified laccase from Lentinus polychrous Lev. Journal of Biochemistry and Technology. 2010;2(3): 182-186.

Zille A, Gornacka B, Rehorek A, Cavaco-Paulo A. Degradation of azo dyes by Trametes villosa laccase over long periods of oxidative conditions. Applied and Environmental Microbiology. 2005;71(11): 6711-6718.

Almeida PH, De Oliveira ACC, De Souza GPN, Friedrich JC, Linde GA, Colauto NB, Valle JSD. Decolourization of remazol brilliant blue R with laccase from Lentinus crinitus grown in agro-industrial by-products. Anais da Academia Brasileira de Ciências.2018;90(4):3463-3473. Available: