Selection of a pH- and temperature-stable laccase from Ganoderma australe and its application for bioremediation of textile dyes

• DOI: 10.1016/j.jenvman.2021.113619
• Publication Date: 2021-08-28T22:00:05Z
• Authors: Jing Si; Yi Wu; Hong-Fei Ma; Yong-Jia Cao; Yi-Fei Sun; Bao-Kai Cui
• Keywords: Kinetics; 0303 health sciences; 03 medical and health sciences; Biodegradation, Environmental; Textiles; Enzyme Stability; Laccase; Temperature; Ganoderma; Hydrogen-Ion Concentration; Coloring Agents

By virtue of screening, purification, and properties characterization, this study captures a new pH- and temperature-stable laccase, designated Galacc-F, from Ganoderma australe for dye bioremediating applications. The enzyme was purified to homogeneity by salt precipitation, ionic exchange, and size exclusion chromatography with a final specific activity of 22.214 U mg-1, yielding a purification fold of 23.989 and recovery of 38.44%. Its molecular weight was estimated to be 48.0 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, zymography, Sephadex G-100 column, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, which confirmed its monomeric nature. Galacc-F exhibited high levels of activity and stability over wide ranges of pH (5.0-8.0) and temperature (10-60 °C), which are highly valuable properties in industrial processes. Broad substrate specificity was observed, wherein a better affinity was found for 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) with a low value of Km (164.137 μM) and higher kcat/Km ratio (1.663 s-1 μM-1). Activity was stimulated by Cu2+ and β-mercaptoethanol but inhibited by ethylenediaminetetraacetic acid, diethylpyrocarbonate, iodoacetic acid, phenylmethylsulfonyl fluoride, and Hg2+, indicating that Galacc-F is a metalloprotease containing a typical histidine-cysteine-serine catalytic triad. It had high tolerance to surfactants, oxidants, and salts. Additionally, a fabricated protocol for native Galacc-F immobilization onto Fe3O4@Chitosan composite nanoparticles using glutaraldehyde as a crosslinker was developed. Most importantly, the enzyme was determined to be ideal for use in efficient treatment of dye effluents as compared with the laccases requiring redox mediators.