Heterologous expression of bacterial CotA-laccase, characterization and its application for biodegradation of malachite green

Laccase-mediated Malachite green bioremediation of aquaculture pond wastewater. [Display omitted] •CotA laccase gene of B. subtilis was cloned and overexpressed in E. coli.•CotA-laccase concentration was increased for MG bioremediation from aquatic ponds.•MG decolorization efficiency recorded 90 and...

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Bibliographic Details
Published in:Bioresource technology 2021-11, Vol.340, p.125708-125708, Article 125708
Main Authors: Cheng, Chiu-Min, Patel, Anil Kumar, Singhania, Reeta Rani, Tsai, Cheng-Hsian, Chen, Shen-Yi, Chen, Chiu-Wen, Dong, Cheng Di
Format: Article
Language:English
Subjects:
ABTS
CotA-laccase
Malachite green
Overexpression
Recombinant-CotA
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Summary:Laccase-mediated Malachite green bioremediation of aquaculture pond wastewater. [Display omitted] •CotA laccase gene of B. subtilis was cloned and overexpressed in E. coli.•CotA-laccase concentration was increased for MG bioremediation from aquatic ponds.•MG decolorization efficiency recorded 90 and 93% from fresh and saline wastewaters.•Cytotoxicity analysis confirmed non-toxic byproducts of MG biodegradation.•N-methylation was the key process for laccase mediated MG degradation. Malachite green (MG) is used as fungicide/parasiticide in aquaculture, its persistence is detrimental as it exhibits carcinogenic effects to aquatic organisms. Bacterial laccase evaluated as the best enzyme at extreme condition for aquatic MG removal. Study aims to increase laccase concentration, CotA-laccase from Bacillus subtilis was cloned and overexpressed in Escherichia coli. Optimal catalysis for purified CotA-laccase were at pH 5.0, 60 °C, and 1 mM of (2,2-azino-di-[3-ethylbenzothiazoline-sulphonate-(6)]) with Km and Kcat 0.087 mM and 37.64 S-1 respectively. MG biodegradation by CotA-laccase in clam and tilapia pond wastewaters and cytotoxic effect of biodegraded products in grouper fin-1 cells were determined. MG degradation by CotA-laccase was equally efficient, exhibiting upto 90–94% decolorization at freshwater and saline conditions and treated solution was non-toxic to GF-1 cells. Thus, recombinant-CotA-laccase could be an environmentally-friendly enzyme for aquaculture to remove MG, thereby effective to reduce its accumulation in aquatic organisms and ensuring safe aquaculture products.
ISSN:0960-8524
1873-2976
DOI:10.1016/j.biortech.2021.125708