Purification and characterization of laccase from Marasmius species BBKAV79 and effective decolorization of selected textile dyes.
A novel laccase-producing white-rot fungus, Marasmius sp. BBKAV79 (Genbank Accession Number-KP455496, KP455497), was isolated and subjected to purification, characterization and dye decolorization study. The purified enzyme was obtained with a specific activity of 0.226 U mg-1protein and a final yield of 13.5 %. The enzyme was found to be a monomeric protein with a molecular mass of ~75 kDa as estimated by non-denaturing polyacrylamide gel electrophoresis (PAGE) and further confirmed with zymogram analysis. The optimal pH and temperature of the laccase was recorded to be 5.5 and 40 °C, respectively. The metal ions Hg2+and Ag+were found to drastically inhibit the activity of laccase at the rate of 96.6 and 96.5 %, respectively. Nevertheless, Fe3+was found to inhibit laccase activity at 40 %. Phenylmethanesulfonyl fluoride (PMSF) strongly inhibited the laccase activity, and additives viz, sodium dodecyl sulfate (SDS), hydrogen peroxide (H2O2) and sodium chloride (NaCl) were known to follow the earlier pattern of enzyme inhibition. The values of kinetic parameters Kmand Vmaxfor purified laccase were noted at 3.03 mM and 5 μmol min-1, respectively, for guaiacol as substrate. The textile dyes were decolorized at a range of 72-76 % and 88-93 % when treated with Marasmius sp. BBKAV79 and purified laccase, respectively. Based on the outcome of the present investigation, it could be, therefore, inferred that laccase isolated from Marasmius sp. BBKAV79 effectively decolorizes the textile dyes; however, the metal ions Hg2+, Ag+and Fe3+and agents like PMSF, SDS, H2O2and NaCl pose an effective inhibitory potential under specified physicochemical conditions.
Department of Studies and Research in Biotechnology and Microbiology, Karnatak University, Dharwad, 580003, India.
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Vantamuri
Department of Studies and Research in Biotechnology and Microbiology, Davangere University, Davangere, 577 002, India. adivesh79biotech@gmail.com.
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Kaliwal
Decolorization; Laccase; Marasmius sp. BBKAV79; Purification