The production of porous cross-linked enzyme aggregates (p-CLEAs) is a simple and effective methodology
for laccase immobilization. A three-phase partitioning technique was applied to co-precipitate laccase
and starch, followed by cross-linking with glutaraldehyde and removal of starch by a-amylase to
create pores in the CLEAs. Scanning electron microscopy revealed a very smooth spherical structure with
numerous large pores. The half-life of free laccase at 55 C was calculated to be 1.3 h, while p-CLEAs did
not lose any activity even after 14 h. p-CLEAs also exhibited improved storage stability, catalytic effi-
ciency and could be recycled 15 times with 60% loss of activity. The catalysts decolorized triphenylmethane
and reactive dyes by 60–70% at initial dye concentrations of 2 and 0.5 g L1
, respectively, without any
mediators. These results suggest the potential of CLEA technology in waste-water treatment.