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Aims: To discover novel laccases potential for industrial applications. Methods and Results: Fungi were cultivated on solid media containing indicator compounds that enabled the detection of laccases as specific colour reactions. The indicators used were Remazol Brilliant Blue R (RBBR), Poly R-478, guaiacol and tannic acid. The screening work resulted in isolation of 26 positive fungal strains. Liquid cultivations of positive strains confirmed that four efficient laccase producers were found in the screening. Biochemical characteristics of the four novel laccases were typical for fungal laccases in terms of molecular weight, pH optima and pI. The laccases showed good thermal stability at 60°C. Conclusions: Plate-test screening based on polymeric dye compounds, guaiacol and tannic acid is an efficient way to discover novel laccase producers. The results indicated that screening for laccase activity can be performed with guaiacol and RBBR or Poly R-478. Significance and Impact of the Study: Laccases have many potential industrial applications including textile dye decolourization, delignification of pulp and effluent detoxification. It is essential to find novel, efficient enzymes to further develop these applications. This study showed that relatively simple plate test screening method can be used for discovery of novel laccases.

Production of mannanase by four hemicellulolytic microorganisms was studied. The highest mannanase activity was produced byBacillus subtilis. β-Mannosidase and α-galactosidase were not detected inB. subtilis culture filtrate. The hydrolysis of galactomannans was limited by the increasing degree of substitution of the substrate. No monomeric sugars were produced in the hydrolysis withB. subtilis culture filtrate.