SELF-IMMOBILIZED, CROSS-LINKED ENZYME AGGREGATES (CLEAs) FOR BIOCATALYSIS

Abstract

In this study, we show that enzymes can be immobilized as cross-linked enzyme aggregates (CLEAs) without using any solid carriers to improve their stability. Firstly, millifluidic reactors with two laminar flows of an enzyme solution and an organic solvent were employed to prepare uniform CLEAs of cellulase and laccase. CLEA had a size between 200 and 400 nm and were highly uniform. They can be collected on silica gels, entrapped in a membrane and alginate beads as practical biocatalysts to hydrolyze cellulose with an activity up to 86% of the free cellulase. Similarly, h-CLEA laccase were prepared and entrapped in a membrane for degrading azo dye. Finally, glucose oxidase (GOx) and horseradish peroxidase (HRP) were co-immobilized as combi-CLEA. The combi-CLEA was used to catalyze cascade chemical reactions and minimize the decomposition of H2O2 in the presence of catalase. The combi-CLEA was further exploited to develop colorimetric glucose assays.