Production, Biochemical Characterization, and Application of Laccase from Halophilic MLK46 Recovered from Mangrove Rhizosphere.

Laccase production was evaluated in 108 fungal isolates recovered from the eastern coast of Saudi Arabia, a critical element in environmental biodegradation and biotransformation. The most active isolate was identified as MLK46 (GenBank accession no. PQ100161). It exhibited maximal productivity at pH 6.5, 30 °C, and incubation for 5 d, with 1% sodium nitrate and 1% galactose as the preferred nitrogen and carbon sources, respectively. Productivity was enhanced by NaCl, CuSO, and FeCl supplementation, with a maximum at 0.3 mM, 0.2 mM, and 61.7 mM concentrations, respectively. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for the purified enzyme through diethylaminoethyl (DEAE)-Sepharose chromatography revealed a prominent band at 71.1 kDa with maximum activity at pH 6 and stability at pH 6-9. Furthermore, it was optimally active at 50 °C and thermally stable at 50-80 °C with a half-life time () of 333.7 min to 80.6 min, respectively. Its activity was also enhanced by many metallic ions, especially Fe ions; however, it was inhibited by Hg and Ag ions. The enzyme demonstrated significant degradation of specific substrates such as 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), guaiacol, -dianisidine, and 2,6-dichlorophenol, with a kinetic efficiency constant which ranged from 40.95 mM s to 238.20 mM s. UV spectrophotometry confirmed efficient oxidation peaks by electron transition against guaiacol (at 300 nm), -dianisidine (at 480 nm), ABTS (at 420 nm), and 2,6-dichlorophenol (at 600 nm). The results collectively demonstrate the potential of laccase from MLK46 as a promising agent for the effective biodegradation of several industrial pollutants under extreme conditions.