Comparison of Two Laccases from Trametes versicolor for Application in the Decolorization of Dyes

Abstract

It has been previously demonstrated that laccases exhibit great potential for use in several industrial and environmental applications. In this paper, two laccase isoenzyme genes, lccB and lccC, were cloned and expressed in Pichia pastoris GS115. The sequence analysis indicated that the lccB and lccC genes consisted of 1,563 and 1,584 bp, and their open reading frames encoded 520 and 527 amino acids, respectively. They had 72.7% degree of identity in nucleotides and 86.7% in amino acids. The expression levels of LccB and LccC were up to 32,479 and 34,231 U/l, respectively. The recombinant laccases were purified by ultrafiltration and (NH4)2SO4 precipitation, showing a single band on SDS-PAGE, which had a molecular mass of 58 kDa. The optimal pH and temperature for LccB were 2.0 and 55°C with 2,2'-azino-bis-[3-ethylbenzthiazolinesulfonic acid (ABTS) as a substrate, whereas LccC exhibited optimal pH and temperature at 3.0 and 60°C. The apparent kinetic parameters of LccB were 0.43 mM for ABTS with a Vmax value of 51.28 U/mg, and the Km and Vmax values for LccC were 0.29 mM and 62.89 U/mg. The recombinant laccases were able to decolorize five types of dyes. Acid Violet 43 (100 g/ml) was completely decolorized by LccB or LccC (2 U/ml), and the decolorization of Reactive Blue KN-R (100 g/ml) was 91.6% by LccC (2 U/ml). Thus, the study characterizes useful laccase isoenzymes from T. versicolor that have the capability of being incorporated into the treatment of similar azo and anthraquinone dyes from dyeing industries.

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

“…Only TtL, which has already been shown to have decolorization abilities in several works, was confirmed to have potential for dye decolorization in our experiments. This was surprising as decolorizing capacity of TvL for several dyes has been previously described . MtL did not decolorize synthetic dyes in our study, which was in accordance with the work by Claus et al In their study, the dye decolorization ability of a native laccase from the ascomycete M. thermophila has been compared to that of the basidiomycetes T. versicolor and Polyporus pinisitus .…”

Lignin‐degrading enzymes

“…reaching an expression level of 34 kUÁL À1 [83]. The two isoenzymes show similar kinetic properties: the specific activity was 51 UÁmg À1 and 63 UÁmg À1 and the K m for ABTS was 0.43 mM and 0.29 mM for LccB and LccC, respectively.…”

“…T. versicolor , a typical white‐rot fungus, secretes several laccase isoenzymes. The LccB and LccC isoenzymes were recently expressed in P. pastoris GS115, reaching an expression level of 34 kU·L −1 . The two isoenzymes show similar kinetic properties: the specific activity was 51 U·mg −1 and 63 U·mg −1 and the K m for ABTS was 0.43 m m and 0.29 m m for LccB and LccC, respectively.…”

Extraction and biodegradation of ginkgolic acidsfrom Ginkgo biloba sarcotestae

“…A series of laccase isozymes (GS115-LacA, GS115-LacB, GS115-LacC, GS115-Lcc1 and GS115-Lcc2 from Trametes versicolor) was prepared and preserved by the Microbial Technology Research Laboratory of Nanjing Forestry University [16][17][18] . YPD and BMGY media were prepared as described in Pichia expression manual (Invitrogen, San Diego, CA, USA).…”