Characterization of a Recombinant Laccase B from Trametes hirsuta MX2 and Its Application for Decolorization of Dyes

Trametes hirsuta is able to secrete laccase isoenzymes including constitutive and inducible forms, and has potential application for bioremediation of environmental pollutants. Here, an inducible group B laccase from T. hirsuta MX2 was heterologously expressed in Pichia pastoris, and its yield reached 2.59 U/mL after 5 days of methanol inducing culture. The optimal pH and temperature of recombinant laccase (rLac1) to 2,2 '-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) were 2.5 and 60 degrees C, respectively. Metal ions showed different effect on rLac1 which Mg2+, Cu2+, and K+ increased enzyme activity as their concentration increased, whereas Zn2+, Na+, and Fe2+ inhibited enzyme activity as their concentration increased. rLac1 showed good tolerance to organic solvents, and more than 42% of its initial activity remained in 10% organic solvents. Additionally, rLac1 exhibited a more efficient decolorization ability for remazol brilliant blue R (RBBR) than for acid red 1 (AR1), crystal violet (CV), and neutral red (NR). Molecular docking results showed RBBR has a stronger binding affinity with laccase than other dyes by interacting with substrate binding cavity of enzyme. The results indicated rLac1 may be a potential candidate for dye removal from textile wastewater.

Automated summary

Trametes hirsuta secretes laccase isoenzymes, and an inducible laccase from this species was successfully expressed in Pichia pastoris. The recombinant laccase showed optimal pH and temperature for the oxidation of ABTS, and its activity was influenced by metal ions and organic solvents. Furthermore, the laccase exhibited efficient decolorization ability for certain dyes, suggesting its potential application in dye removal from wastewater.