Loss in activity and denaturation remain key challenges to the potential use of laccase in industrial
applications. One of the most important aims of enzyme technology is to enhance the stability and
reusability of enzymes through immobilization processes. Here, a purified laccase (Tplac) from the white
rot fungus Trametes pubescens was entrapped onto chitosan beads with the crosslinker glutaraldehyde, in
order to improve the stability and recovery rate of Tplac, and was applied in decolorization of various
synthetic dyes. The optimal conditions for Tplac immobilized onto chitosan beads were 0.8% (v/v)
glutaraldehyde concentration, 3 h crosslinking time, 2 mL enzyme solution (approximately 43.672 U/mL),
and 4 h immobilization time. The pH adaptability and resistance to thermal denaturation of immobilized
Tplac were considerably enhanced compared with free Tplac, and both the operational stability and
durability during multiple reuses were superior to those of free Tplac; after six cycles of continuous use,
the activity of immobilized enzyme remained above 60%. Also, immobilized Tplac was able to degrade
various synthetic dyes, especially metal-complex dye Acid Black 172. Results of this study demonstrated
that, alongside the better stability and reusability of immobilized Tplac, the immobilized enzyme could
be used in many applications.