2.2.5. Enzyme Immobilization
The different immobilization supports were suspended in an enzyme solution (10 g agarose beads:100 mL enzyme solutions in immobilization buffers). The immobilization buffers differ for each immobilization protocol: 100 mM sodium bicarbonate buffer pH 10.2 when immobilization on a glyoxyl support was performed [18]; 5 mM sodium phosphate buffer pH 7.0 for immobilization on MANAE support; or 200 mM sodium phosphate buffer pH 7.0 for immobilization on glutaraldehyde support [17,20]. The immobilization suspensions were gently stirred at room temperature. Different enzyme concentrations were tested. For all immobilization experiments, samples of the suspensions and the supernatants were periodically withdrawn and enzyme activity of the samples was determined as described above. A reference enzyme suspension identical to each immobilization suspension was prepared, using inert agarose, as a reference.