Production of lignocellulases enzymes from Trichoderma viride M5-2 in wheat bran (Triticum aestivum) and purification of their laccases

Abstract

The present study describes the production of lignocellulases enzymes from Trichoderma viride M5-2 in wheat bran (Triticum aestivum) and the purification of their laccases. Fermentation process was determined in wheat bran during 7 days and samples were taken every 24h. The enzymatic assay (endo, exo β1, 4-glucanase, xylanases and laccases) was performed. Laccase activity was determined with a spectrophotometer, by syringaldazine oxidation under aerobic conditions and lignin peroxidase was determined by oxidative dimerization dependent on H2O2 of 2.4-dichlorophenol. The enzyme crude was concentrated by membrane filtration with nitrogen stream and purified by ion exchange chromatography with DEAE matrix. Enzyme yield and purification parameters were measured. With the fermentation conditions in wheat bran, a sustained increase in the production of endo β1,4 glucanase and xylanases was obtained after 72 h and exo β1, 4 glucanase at 48 h and laccase enzymatic activity was checked at 24 h and lignin peroxidase
after 48h of fermentation. The fungus T. viride M5-2 reached its maximum production of laccases after two days of fermentation and through the proposed purification system, an enzymatic product with a purification factor of 12 was obtained, without affecting the enzyme yield. It is concluded that the T viride strain has the capacity to produce the lignocellulolytic enzyme complex in wheat bran. The separation method used to purify laccase enzymes is effective. It is recommended to add successive steps of purification depending on the degree of purity.
Key words: ligninase, fungi, lignin, cellulases, xylanases