Enzymes, lignin peroxidase (LiP), manganese-dependent
peroxidase (MnP) and laccase were purified from
the culture filtrate by acetone precipitation (66% v/v)
and Sephadex G-100 column chromatography. The LiP
activity was determined using a spectrophotometer according
to the method of Linko (1988). One unit of
enzyme activity was defined as the amount of enzyme
oxidizing one lmole of veratryl alcohol in 1 min. MnP
was assayed using a spectrophotometer by the method
of Kuwahara et al. (1984). One unit of enzyme activity
was expressed as the amount of enzyme capable of increasing
one optical density (OD) at 610 nm in a min.
The laccase activity was determined spectrophotometrically
as described by Evans (1985) with guaiacol as a
substrate. In this case, one unit of the enzyme activity
was expressed as the amount of enzyme able to increase
one OD at 440 nm.