Production and Purification of Laccase enzyme by Trichoderma viride

Abstract

The ability of Trichoderma viride to synthesize extracellular enzymes (Lacase enzyme) using a substrate that contains media was examined. This isolate generated laccase at its greatest level (0.166U/ml) in liquid medium. This study was demonstrated the production, purification, and characterisation of the laccase enzyme from T.viride. The results showed that 96 hours was the optimum period of time for Laccase to incubate from this fungus. Laccase displayed the maximum level of activity at pH 4.5 and temperature 30 °C. The results revealed that the best ratio for laccase precipitation was 90% by using ammonium sulphate. In addition, for the purification of laccase enzyme, one peak of Laccase was appeared in gel filtration purified from T.viride, while 2 peaks of Laccase were seen in ion exchange chromatography. According to the findings, the molecular weight of T. viride Laccase using SDS-PAGE was roughly 66KD under denaturation conditions.

Kinetic studies on optimized extracellular laccase from Trichoderma harzianum PP389612 and its capabilities for azo dye removal

“…The highest laccase production was 2.89 U/ml (predicted value of 2.71 U/ml) obtained in the run (6) with shaking fermentation at 25 °C and pH 7 after 8 days using a 2% inoculum size of 50 ml fermentation medium involving 1.5% glucose, 0.6% peptone, 0.1% KH 2 PO 4 , 0.01% K 2 HPO 4, and 50 μg/l guaiacol. After optimization, the laccase activity recorded in our study was higher than those recorded by Mohsen et al [62], who obtained 1.03 U/ml laccase from Trichoderma viride after 96 h of incubation at 30 °C, Umar [43] found that the highest laccase activity from T. atroviride was 2.62 U/ml. After the laccase optimization process, Jenefar et al [73] obtained 2.3 U/ml laccase from Perenniporia subtephropora using medium initial pH 6 and incubation at 30 °C, while Sadhasivam et al [74] utilized Trichoderma harzianum by augmented 1 mM CuSO 4 and found that the highest yield of laccase activity reached 4.36 U/ml after 4 days.…”

“…Umar [43] evaluated ten Trichoderma species for laccase production and found that T. atroviride was the highest producer with 2.62 U/ml. Mohsen et al [62] obtained 0.266 U/ml laccase from Trichoderma viride using 0.02% guaiacol after 96-h incubation at 28 °C. Abd El Monssef et al [63] reported that Trichoderma harzianum produced 1.286 U/ml after 6 days of incubation at 30 °C using a medium supplemented by 0.04% guaiacol and an initial pH of 5.5.…”