Biodegradation of Malachite Green by Extracellular Laccase Producing Bacillus thuringiensis RUN1


Laccase, malachite green, decolorization, biodegradation, Bacillus thuringiensis.

How to Cite

Olumide D. Olukanni, Aliu Adenopo, Ayodeji O. Awotula, & Akinniyi A. Osuntoki. (2021). Biodegradation of Malachite Green by Extracellular Laccase Producing Bacillus thuringiensis RUN1. Journal of Basic & Applied Sciences, 9, 543–549.


A bacteria strain Bacillus thuringiensis RUN1identifiedby 16s RNA gene phylogenetic analysiswas used to decolorize malachite green, a triphenylmethane dye in a simulated wastewater. The ability of the organism to produce extracellular laccase and degrade the dye were also investigated. Results showed that the organism decolorized (84.67 ± 1.19 %) malachite green at 40 mgl-1 within 6 h; and the decolorization was associated to laccase production by the organism experimentally. Laccase activity increased as the decolorization process progressed, with the highest activity value of 0.1043 ± 0.02 U/min/ mg protein recorded after 24 h of incubation using ABTS as substrate. In addition, FTIR analysis showed that the strain actually degraded the dye. It was therefore concluded that this strain of Bacillus thuringiensis will be relevant in the biotreatment of industrial effluent containing malachite green and in the production of laccase, an industrially important enzyme.


Boer CG, Obici L, Souza CG, Peralta RM. Decolorization of synthetic dyes by solid state cultures of Lentinula (Lentinus) edodes producing manganese peroxidase as the main lignolytic enzyme. Bioresour Technol 2004; 94: 107-112.

Rajamohan N, Karthikeyan C. Kinetic studies of dye effluent degradation by Pseudomonas stutzeri. ECO services International 2006. 061103.html

Lal N, Srivastava AK. Decolorization of malachite green by newly isolated Bacillus Strain MTCC -3330 Arch Environ Sci 2011; 5: 71-76. LalN2011.pdf

Mass R, Chaudhari S. Adsorption and biological decolorization of azo dye reactive red 2 in semicontinous anaerobic reactors. Process Biochem 2005; 40: 699-705.

Moturi B, Singara-Charya MA. Decolorization of crystal violet and malachite green by Fungi. Sci World J. 2009; 4(4): 28-33. 42151

Hai FI, Fukushi K, Yamamoto K. Treatment of textile wastewater: Membrane bioreactor with special dye-degrading microorganism. Proceeding of the Asian waterqual, Bangkok 2003. Article # 2Q3F16.

Culp SJ, Beland FA. Malachite green: A toxicological review. J Am Coll Toxicology 1996; 15: 219-238.

Alderman DJ. Malachite green: A review. J Fish Dis 1984; 8: 289-298.

Cha C, Doerge DR, Cerniglia CE. Biotransformation of malachite green by the Fungus Cunninghamella elegans. Appl Environ Microbiol 2001; 67(9): 4358-4360.

Parshetti G, Kalme S, Saratale G, Govindwar S. Biodegradation of malachite green by Kocuria rosea MTCC 1532. Acta Chim Slov 2006; 53: 492–498.

Olukanni OD, Osuntoki AA, Gbenle GO. Textile effluent biodegradation potentials of textile effluent-adapted and non adapted bacteria. Afr J Biotech 2006; 5 (20): 1980-1984.

Jayasinghe C, Imtiaj A, Lee GW, Im KH, Hur H, Lee MW, Yang H, Lee T. Degradation of three aromatic dyes by white rot Fungi and the production of ligninolytic enzymes. Mycobiology 2008; 36(2): 114-120.

Fu Y, Viraraghavan T. Fungal decolorization of dye wastewaters: A review. Bioresour Technol 2001; 79: 251-262.

Olukanni OD, Osuntoki AA, Keyalni DC, Gbenle GO., Govindwar S. Decolorization and biodegradation of reactive blue 13 by Proteus mirabilis LAG. J Hazard Mater 2010; 184: 290-298.

Reiss R, Ihssen J, Thony-meyer L. Bacillus punilus laccase; a heat stable enzyme with a wide substrate spectrum. BMC Biotechnol 2011; 11: 9-14.

Telke AA, Kalyani DC, Jadhav UU, Parshetti GK, Govindwar SP. Purification and characterization of an extracellular laccase from a Pseudomonas sp. LBC1 and its application for the removal of bisphenol A. J Mol Catal B: Enzymatic 2009; 61: 252-60.

Chen KC, Wu JY, Liou DJ, Hwang SCJ. Decolorization of textile dyes by newly isolated bacterial strains. J Biotechnol 2003; 101(1): 57-68.

Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 2007; 24: 1596-1599.

Wolfenden B, Willson R. Radical-cations as reference chromogens in kinetic studies of one-electron transfer reactions. J Chem Soc Perkin Trans 1982; 2: 805-812.

D’Souza DT, Tiwari R, Shah AK, Chandralata R. Enhanced production of laccase by a marine fungus during the treatment of colored effluents and synthetic dyes. Enzyme Microb Technol 2006; 38: 504-511.

Youssef AS, El-Sherif MF, El-assar SA. Studies on the decolorization of malachite green by local isolate Acremonium kiliense. Biotechnology 2008; 7(2): 213-223.

Broderick NA, Raffa KF, Handelsman J. Midgut bacteria required for Bacillus thuringiensis insecticidal activity. Proc Natl Acad Sci USA 2006; 103(41): 15196-9.

Kunjadia PD, Patel FD, Nagee A, Mukhopadhyaya NP, Dave GS. Biodegradation of crystal violet. Bioresources 2012; 7(1): 1189-1199.

Zouari-Mechichi H, Mechichi T, Dhouib A, Sayadi S, Mart´ınez AT, Mart´ınez MJ. Laccase purification and characterization from Trametes trogii isolated in Tunisia: decolorization of textile dyes by the purified enzyme. Enzyme Microbial Tech 2006; 39: 141-148.

Tom-Sinoy, ES, Mohan DA, Shaikh H. Biodegradation of textile dyes by Pseudomonas Species and E.Coli. VSRD-TNTJ 2011; 2 (5): 238-248. vsrdindex.html

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.