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Fast decolorization of azo dyes in alkaline solutions by a thermostable metal-tolerant bacterial laccase and proposed degradation pathways

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Abstract

Biocatalytic decolorization of azo dyes is hampered by their recalcitrance and the characteristics of textile effluents. Alkaline pH and heavy metals present in colored wastewaters generally limit the activity of enzymes such as laccases of fungal origin; this has led to an increasing interest in bacterial laccases. In this work, the dye decolorization ability of LAC_2.9, a laccase from the thermophilic bacterial strain Thermus sp. 2.9, was investigated. Its resistance towards different pHs and toxic heavy metals frequently present in wastewaters was also characterized. LAC_2.9 was active and highly stable in the pH range of 5.0 to 9.0. Even at 100 mM Cd+2, As+5 and Ni+2 LAC_2.9 retained 99%, 86% and 75% of its activity, respectively. LAC_2.9 was capable of decolorizing 98% of Xylidine, 54% of RBBR, 40% of Gentian Violet, and 33% of Methyl Orange after 24 h incubation at pH 9, at 60 °C, without the addition of redox mediators. At acidic pH, the presence of the mediator 1-hydroxybenzotriazole generally increased the catalytic effectiveness. We analyzed the degradation products of laccase-treated Xylidine and Methyl Orange by capillary electrophoresis and mass spectrometry, and propose a degradation pathway for these dyes. For its ability to decolorize recalcitrant dyes, at pH 9, and its stability under the tested conditions, LAC_2.9 could be effectively used to decolorize azo dyes in alkaline and heavy metal containing effluents.

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Abbreviations

RBBR:

Remazol Brilliant Blue R

HBT:

1-Hydroxybenzotriazole

ABTS:

2,2′-Azino-di-[3-ethylbenzthiazoline sulfonate]

pHBA:

Para-hydroxybenzoic acid

CE:

Capillary electrophoresis

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Acknowledgements

We thank Irma Fuxan for the technical support in routine laboratory protocols and Dr. Morgan Fetherolf for the critical reading of the manuscript. LN was granted a fellowship from CONICET. RC, LL, and MB are staff members of CONICET. This work was supported by Instituto Nacional de Tecnología Agropecuaria (INTA) (Grant Number PNAIyAV-1130034), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET, Argentina) and University of Buenos Aires.

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LN obtained and purified the enzyme and performed the decolorization experiments. RC performed the CE and MS analysis and the statistical analysis of decolorization experiments. LL and MB conceived the study and MB supervised the project. All authors have contributed to the writing and revising of the manuscript and approved the final version to be submitted.

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Correspondence to Marcelo F. Berretta.

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The authors declare that they have no conflict of interest.

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Communicated by I. Cann.

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Navas, L.E., Carballo, R., Levin, L. et al. Fast decolorization of azo dyes in alkaline solutions by a thermostable metal-tolerant bacterial laccase and proposed degradation pathways. Extremophiles 24, 705–719 (2020). https://doi.org/10.1007/s00792-020-01186-w

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  • DOI: https://doi.org/10.1007/s00792-020-01186-w

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