Production of Fungal Laccase under Solid State Bioprocessing of Agroindustrial Waste and Its Application in Decolourization of Synthetic Dyes

Abstract

Fungal laccases are preferred due to high redox potentials and low substrate specificity to xenobiotics including synthetic dyes. For large-scale applications, low enzyme yield and high cost of production has remained the challenge. Agroindustrial waste such as saw-dust of Terminalia superba abounds locally and was utilized as low-cost alternative substrate for laccase production in Solid State Bioprocessing (SSB) using Trametes sp. isolate G31. The study optimized laccase production using various parameters. Optimal production of laccase was at pH 5.0 - 7.0 with 2356 U/mL - 2369 U/mL and 25°C (2336 U/mL). Among the sources of nitrogen and carbon tested, laccase production in ammonium sulphate and sucrose supplemented media were higher. The effect of activators on laccases production showed that Cu2+ and Ca2+induced high titres of laccase at 4 -5 mM and 2 - 4 mM respectively, while production of laccase by Mn2+ was significantly high at 40 mM. The effect of 2, 2-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS), guaiacol and varatryl alcohol on laccase production was significantly different especially in glycerol. Optimum production for laccase was on day 14 with 2356 U/mL followed by steady declined up to day 34. The purified laccase had a specific activity of 5008 µmol/min/mg, purification factor of 3.85, and a molecular mass of ~40 kDa using N-PAGE. The potential of crude laccase to decolourize diverse dyes was tested. Phenol red achieved 40% decolourization for 1hr, while RBBR (93%), Crystal violet (60%), Methylene blue (53%) and Congo red (51%) for 24 hr, 72 hr, 48 hr and 120 hr respectively. Methyl red and Malachite green attained 42% (72 hr) and 32% (48 hr) decolourization. The enzyme ability to oxidize Phenol red and other synthetic dyes without mediators made it eco-friendly in treating dye wastewaters.