characterising laccase obtained by two production methods using pleurotus ostreatus

This project was designed for characterising and evaluating different methods of chromatographic separation and purification regarding a laccase enzyme-rich broth produced by Pleurotus ostreatus, a variety of basidiomycetes. SSF (Solid State Fermentation) and Submerged Fermentation production processes were employed. The SSF process was based on producing a raw broth rich in laccase enzyme from mycelium grown on bran with 1:1 vinasse w/v for 20 days. A raw broth was obtained having an average 20 U/ml activity. The medium reported by Hublick and Schinner (2000) was used, with a few modifications, for the Submerged Fermentation process; the growth of pellet-shaped fungus was obtained within 15 days, having an average 10 U/ml activity in the raw broth. The isoenzymes isolated in the chromatographic processes were characterised according to their molecular and kinetic properties; their molecular weight was determined by SDS-PAGE vertical electrophoresis and their kinetic parameters (such as stability) expressed in a range of temperatures and pH.