Characterization of recombinant laccase from Trametes versicolor synthesized by Arxula adeninivorans and its application in the degradation of pharmaceuticals

Abstract

Recent years have seen an increasing interest in laccase enzymes. Due to their ability of oxidizing various substrates, they are nowadays applied in multiple industrial fields including pulp delignification, textile dye bleaching, and bioremediation. In contrast to laccase production from native sources, with its generally low yield and high cost, heterologous laccase expression is far better suited to meet the growing industrial demands. TVLCC5 gene encoding Trametes versicolor laccase 5 was overexpressed in Arxula adeninivorans using the strong constitutive TEF1 promoter. Recombinant Tvlcc5 protein was purified by immobilized-metal ion affinity chromatography and biochemically characterized using 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) as substrate for standard activity assays. The enzyme showed the highest activity at 50 °C between pH 4.5–5.5. The half-life of Tvlcc5 at 60 °C was around 20 min. The negative effect of chloride anions on enzyme activity was demonstrated. A fed-batch cultivation of Tvlcc5 producing strain A. adeninivorans G1212/YRC102-TEF1-TVLCC5-6H was performed and resulted in a laccase activity of 4986.3 U L −1 . To improve the expression level of recombinant laccase in A. adeninivorans , cultivation conditions were optimized by single factor experiments. Recombinant Tvlcc5 proved to be a promising agent for degradation of pharmaceuticals that are an important source of environmental pollution. Concentration of diclofenac and sulfamethoxazole decreased to 46.8% and 51.1% respectively after 24 h incubation with Tvlcc5. When 1 mM redox mediator ABTS was added complete degradation was obtained within 1 h. Electronic supplementary material The online version of this article (10.1186/s13568-019-0832-3) contains supplementary material, which is available to authorized users.

Yeasts of the Blastobotrys genus are promising platform for lipid-based fuels and oleochemicals production

“…Cultivation mode such as shake flask (Thomas et al 2019, Sanya et al 2020) and fed-batch cultivation (Bernacki et al 2017) alongside culture media encompassing selective media (Litwińska et al 2019;Theron et al 2014;Friedlander et al 2016;Sanya et al 2020) and nonselective media (Samsonova et al 1996;Bernacki et al 2017) are usually applied to monitor the growth of these yeasts, with the main focus on nitrogen (NH 4 Cl) and carbon sources (glucose) (Sanya et al 2020).…”

Optimization, purification and characterization of laccase from Ganoderma leucocontextum along with its phylogenetic relationship

“…Vantamuri and Kaliwal 101 supplemented 20 mM EDTA in the purified enzyme of Marasmius species BBKAV79, while Das and associates 36 determined the laccase activity from Pleurotus florida . This was interesting to know that 1 mM EDTA represented 116% inhibition rate of recombinant laccase from T. versicolor 20 .…”

“…This enzyme plays a vital role in the delignification of plant products, fungal sporulation, conidial pigmentation and fruiting body formation of mushrooms 15 , 16 . This mushroom secreted metalloenzymes (laccase) possess “Janus-faced” range of activities e.g., humification, azodye oxidation, xenobiotic compounds degradation 17 , 18 , phenolics, non-phenolics, pollutants detoxification 19 , wine and water discoloration, paper processing, steroid transformation, polymerization or depolymerization processes, biochemical bleaching of pulp, pharmaceutical products synthesis and degradation 20 , producer of other enzyme, biosensors constructor, and also play an important role in nanotechnology 21 .…”

Study of Production and Characterization of Laccase Enzyme from Klebsiellapneumoniae K7 Isolate

“…The optimum pH on laccase activity was at 4.5, which purified fromTrametes versicolor . [17] Compared to phenolic substrates, the redox potential of ABTS does not depend on the pH, oxidation does not involve the protons . So, the effect of pH on the laccase activity towards the different substrates could be related to the balance between the two opposing effects, first the redox potential difference between the substrates and type 1 copper site and second binding of hydroxide anion (OH -) to the type 2/type 3 copper site [18] .…”