Comparative detoxification of Remazol Rrilliant Blue R by free and immobilized laccase of Oudemansiella canarii

Abstract

A laccase from Oudemansiella canarii was immobilized using the crosslinked enzyme aggregate (CLEA) methodology and applied in the degradation of the anthraquinonic dye Remazol Brilliant Blue R (RBBR). The immobilized laccase was superior to the free laccase in both thermal and storage stabilities. Both immobilized and free laccase decolourized 100 mg/L RBBR within 24 h at 30 °C and pH 5.0, but the former was still efficient in degrading the dye after at least 6 cycles. The relationship between the decolorization rate and the RBBR concentration obeyed Michaelis–Menten kinetics, with K_M of 0.126 ± 0.044 mmol/L and V_max of 1.412 ± 0.295 µmol/min for free laccase and K_M of 0.159 ± 0.050 mmol/L and V_max of 1.214 ± 0.242 µmol/min for immobilized laccase. Fourier transform infra-red spectroscopy (FTIR) and mass spectrometry allowed to conclude that the O. canarii laccase acts not only on the chromophore group of the dye, but that it also cleaves other covalent bonds, causing an effective fragmentation of the molecule. The Microtox assay detected a significant diminution in toxicity, a finding corroborated by the phytotoxicity test performed with lettuce seeds. Our results indicate that the immobilized laccase from O. canarii could be useful in biological strategies aiming at degrading unwanted dyes in the environment.

Keywords:

• Biodegradation
• detoxification
• enzyme immobilization
• laccase
• Remazol Brilliant Blue R
• white-rot fungi

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

Author T.M. Uber thanks CAPES Coordenação de Aperfeiçoamento do Pessoal do Ensino Superior) for the financial support provided for her graduate studies in Universidade Estadual de Maringá. A. Bracht, R.A. Peralta, R.F. Peralta-Muniz-Moreira, and R.M. Peralta are research grant recipients of CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico). The authors thank the CNPq (Proc. 304406/2019-8 and Proc. 404898/2016-5) for funding this study.