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Laccase enzyme was estimated using 0.1 ml 10 mM Guaiacol prepared in ethanol, 0.4 ml 50 mM Phosphate buffer (pH–5.0) and 0.5 ml enzyme solution, which was collected freshly from all the flasks and centrifuged at 10,000 rpm for 20 min. O.D. was recorded at 460 nm immediately after addition of enzyme solution and after 10 min of incubation at 30 °C (Arora and Sandhu 1985).
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