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Malaysian Journal of Biochemistry and Molecular Biology, vol.25, no.1, pp.104-113, 2022 (Scopus)
© 2022 Malaysian Society for Biochemistry and Molecular Biology. All rights reserved.Azo dyes are widely used in the textile industry due to their bold colors and resistance toward degradation. This has made azo dyes the global burden in wastewater treatment as conventional dye removal methods have their limitations. Laccase has emerged as an alternative due to its ability to decolorise a wide range of dyes, producing water as by-product. In this study, we aim to assess the dye decolorisation capacity of the Basidiomycota, L. rhinocerotis dialysed fraction and produce recombinant laccase expressed in Pichia pastoris. Out of the 3 dyes tested, the dialysed fraction of L. rhinocerotis sclerotia was capable of decolorising around 90% of the Congo Red and Coomassie Brilliant Blue dye at the enzyme unit of 0.018 U/mL. The gDNA of Lac1 [MG210944] was successfully updated in GenBank and the sequence was used to design the pPICZαA/Lac vector, which was successfully cloned and the recombinant Lac1 was successfully expressed using the Pichia pastoris system. The recombinant Lac1 expressed contain the 6x His-tag for purification but the binding towards the Ni-NTA resin was weak, therefore the purification can be optimised through reducing Imidazole concentration to compensate for the weak binding. In conclusion, L. rhinocerotis extract showed promising results in decolorising CR and CBB, while recombinant Lac1 laccase successfully demonstrated its activity and can be further used for dye decolorisation in future studies.