Bioremoval of toxic malachite green from water through simultaneous decolorization and degradation using laccase immobilized biochar

In this study, decolorization and degradation of malachite green dye was studied using the laccase immobilized pine needle biochar. Successful immobilization of biochar was achieved by adsorption and confirmed through scanning electron microscopy and energy dispersive X-ray analysis (SEM-EDX), Fourier transform infrared spectroscopy (FTIR). High laccase binding of 64.4 U/g and high immobilization yield of 78.1% was achieved using 4U of enzyme at pH3 and temperature 30 °C. The immobilized laccase retained >50% relative activity in the pH range 2-7, >45% relative activity at 65 °C and >55% relative activity at 4 °C for 4 weeks. The re-usability of immobilized enzyme was checked with 2, 2'-azino-bis 3-ethylbenzothiazoline-6-sulphonic acid (ABTS) substrate and enzyme retained 53% of its activity after 6 cycles. Immobilized laccase was used for the degradation and decolorization of azo dye malachite green in aqueous solution. More than 85% removal of malachite green dye (50 mg/L) was observed within 5 h. FTIR and high performance liquid chromatography (HPLC) analysis clearly indicated the breakdown of dye and presence of metabolites (leuco malachite green, methanone, [4-(dimethyl amino)pheny]phenyl and 3-dimethyl-phenyl amine) in gas chromatography-mass spectrometry (GC-MS) analysis confirmed the dye degradation. Phytotoxicity analysis indicated that the enzymatic degradation resulted in lesser toxic metabolites than the original dye. Thus, laccase immobilized biochar can be used as an efficient biocatalytic agent to remove dye from water.