Efficient secretion of three fungal laccases from Saccharomyces cerevisiae and their potential for decolorization of textile industry effluent—A comparative study

Abstract

Laccases are enzymes with a broad range of biotechnological applications and have, for example, the ability to oxidize many xenobiotics including synthetic dyes. In order to obtain an efficient laccase for the decolorization of dyes which spoil wastewater from the textile industry, genes encoding three various laccase enzymes were expressed in Saccharomyces cerevisiae. The expression of laccases from ascomycete Myceliophthora thermophila (MtL), and two basidiomycetes Trametes versicolor (TvL) and Trametes trogii (TtL) was optimized via selection of plasmids, promoters, media composition, and cultivation conditions. For the first time, the activity of the three secreted laccases was directly compared with the use of various substrates, including different dyes and a wastewater sample. A strong constitutive ADH1 promoter, minimal growth medium, optimized combination of copper and organic nitrogen source, and low cultivation temperature were shown to significantly increase the yields and relative activities of secreted laccases. Heterologous expression of three fungal laccases was successfully achieved in S. cerevisiae being the highest for MtL and the lowest for TvL. MtL, and particularly TtL, showed the decolorization capacity. This is the first report which compared decolorization of synthetic dyes and wastewater by several recombinant laccases and suggested MtL and TtL to be applicable in the ecofriendly enzymatic treatment of colored industry effluent. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:69-80, 2018.

Biodecolorization of Anthraquinone and Azo Dyes by Newly Isolated Indonesian White-Rot Fungi

“…Synthetic dyes in wastewater discharged by textile industry becomes one of the pollution issues in several decades (Antošová et. al., 2018;Kant, 2012).…”

Living materials with programmable functionalities grown from engineered microbial co-cultures

“…To demonstrate the generalizability of our approach, we grew functionalised BC with two other enzymes: alpha-galactosidase enzyme Mel1 (a native S. cerevisiae secreted protein) and laccase enzymes from either Myceliophthora thermophila (MtLcc1) and Coriolopsis troggi (CtLcc1) 36 . All were cloned with a C-terminal CBD and either their native secretion signal peptide or the S. cerevisiae MFα signal peptide at the N-terminus (Supplementary Fig.…”

Decolorization and detoxification of textile wastewaters by recombinant Myceliophthora thermophila and Trametes trogii laccases

“…The yeast strain Saccharomyces cerevisiae BW31a transformed with expression plasmid pVT-100U-MtL [T2 mutant of MtL; MtL gene was mutated by directed evolution to enhance its expression in S. cerevisiae, Bulter et al (2003)] or pVT-100U-TtL (LCC1 isoform of TtL; TtL gene was synthetized with optimized codons for S. cerevisiae) was used for recombinant production of secreted laccases MtL and TtL from Myceliophthora thermophila and Trametes trogii, respectively (Antosova et al 2018). The yeast cells were cultivated in 500 mL (in a 2 L shake flask) of minimal YNB medium (6.7 g/L) without amino acids (ForMedium™) supplemented with 2% (w/v) glucose, 0.8% (w/v) alanine, 0.6 mM CuSO 4 and BSM-Ura (Brent supplement mixture drop-out uracil, ForMedium™) at the recommended concentration according to the provider's manual, at 30 °C for 3 days.…”

“…Our results suggest that decolorization efficiency can be affected not only by the type of laccase, but also by the type of mediator, pH value, reaction temperature, and presence of other inhibitors in the reaction. The decolorization of industrial WWs by different laccase systems has been studied by Antosova et al (2018);Bello-Gil et al (2018) and Sondhi et al (2018); these studies have reported decolorization with up to 90% efficiency.…”