Heterologous expression and characterisation of a laccase from Colletotrichum lagenarium and decolourisation of different synthetic dyes

Abstract

Laccases have received considerable attention in recent decades because of their ability to oxidise a large spectrum of phenolic and non-phenolic organic substrates and highly recalcitrant environmental pollutants. In this research, a laccase gene from Colletotrichum lagenarium was chemically synthesised using yeast bias codons and expressed in Pichia pastoris. The molecular mass of the recombinant laccase was estimated to be 64.6 kDa by SDS-PAGE, and the enzyme exhibited maximum activity at pH 3.6-4.0 but more stability in buffer with higher pH (>pH 3.6). The optimal reaction temperature of the enzyme was 40 °C, beyond which stability significantly decreased. By using 2,2'-azino-bis-(3-ethylbenzothiazoline)-6-sulphonate (ABTS) as a substrate, K m and V max values of 0.34 mM and 7.11 mM min(-1) mg(-1), respectively, were obtained. Using ABTS as a mediator, the laccase could oxidise hydroquinone to p-benzoquinone and decolourise the synthetic dyes malachite green, crystal violet and orange G. These results indicated that the laccase could be used to treat industrial effluents containing artificial dyes.

Purification and characterization of a fungal laccase from the ascomycete Thielavia sp. and its role in the decolorization of a recalcitrant dye

“…In presence of 10 mM Zn 2+ and Mn 2+ , TaLac1 showed higher activity compared to that obtained by Cerrena unicolor GSM-01 laccase [2]. Moreover, there was inactivation of TaLac1 by more than 10 mM Cu 2+ , which is consistent with many reports showing the inhibitory effect associated with the increase of Cu 2+ in the reactions [26,41]. These results also suggest that TaLac1 could be suitable for catalyzing reactions of industrial effluents treatment containing high amount of diverse metal ions, such as textile dye wastewater.…”

Heterologous expression and characterization of a laccase from Laccaria bicolor in Pichia pastoris and Arabidopsis thaliana

“…ABTS is necessary for the decolorization of crystal violet by the two laccases. A similar result could be found in a heterologous laccase from Colletotrichum lagenarium [29]. In the presence of ABTS, the decolorization rates of crystal violet by laccases in P. pastoris or A. thaliana reached 90.7％ and 83.6%, respectively (Fig.…”

A halotolerant laccase from Chaetomium strain isolated from desert soil and its ability for dye decolourization

“…The decolourization based on the use of enzymatic methods is shown as a potential alternative to the chemical methods, and it is investigated in many studies using fungal laccase systems as a promising tool (Chen and Yien Ting 2015;Manai et al 2016). Multiple studies have shown the treatment of industrial synthetic dyes by using purified laccases (Wong and Yu 1999;Wesenberg et al 2003;Zouari-Mechichi et al 2006;Koyani et al 2013;Xu et al 2016;Wang et al 2016). In this work, we demonstrated the ability of the purified Chaetomium sp.…”