CATALYTIC ACTIVITY AND PURIFICATION OF THERMOSTABLE LIGNASES FROM GANODERMA SP. ADE468 AND ITS POTENTIAL APPLICATION IN THE DECOLOURIZATION OF DYES
Abstract
Catalytic activity and purification of thermostable lignases from Ganoderma sp. ADEB468 and the potential of the enzyme in dye decolourization were investigated. The highest catalytic activity of laccase (Lac) (175.56 U/L), lignin peroxidase (LiP) (1313.98 U/L), and manganese peroxidase (MnP) (92.27 U/L) was supported by 0.1mM Na+, 5.0 mM Fe3+, and 1.0 mM benzoic acid. The optimum temperatures of 30o, 25o, and 30oC supported the highest catalytic activity. The enzymes were stable up to 70oC for the Lac and 90oC for the LiP and MnP. At 80oC and 90oC, 118.83 U/L and 74.55 U/L residual activities of the LiP and MnP were retained after 60 and 30 min of incubation. This implies that the Ganoderma sp. ADEB468 LiP and MnP were thermostable enzymes. The highest catalytic activity and stability of the Lac and LiP was supported by pH 6 and pH 3, respectively. The calculated Km (mM) and Vmax (U/L) for the Lac, LiP, and MnP were 0.391 and 2.5 × 10o, 0.401 and 5.3 × 10-3, and 0.118 and 2.0 × 10-4, respectively. The purification step using ammonium sulphate precipitation, dialysis, and column chromatography (Sephadex – G100) was 6.25 fold and specific activity of 2.229 U/mg for the Lac, 4.97 and 1.946 U/mg for the LiP, and 7.47 and 1.343 U/mg for the MnP. The highest decolourization using crude lignases and a cell culture of Ganoderma sp. ADEB468 was 74.29% and 84.80% for Dylon green (26) 500 mg/L at 72 h and Dylon red (32)1000 mg/L at day 9. Ganoderma sp. ADEB468 under the studied conditions produced thermostable lignases that decolourized different dyes and, therefore, can be useful in the textile industry.
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