Could anynoe tell some experience of Fe3O4@SiO2 immobilization of cellulase? The immobilization efficiency of my experiment was below 50%. Thanks a lot.
We have immobilized lipase on iron particles. You may look at the following methodology and may use cellulase instead of lipase;
Surface modification of magnetic NPs
In brief, the NPs (Fe3O4 or γ Fe2O3) were set in the form of a suspension by dropping 1.5 g of NPs in 150 ml of ethanol. The suspension was ultrasonicated (35% amplitude for 20 min at room temperature). Deoxygenated water (1 ml) and 750 μl of TEOS were added to above NPs suspension. The suspension was stirred for 12 h at 37°C and NPs were collected by using a magnet, The NPs were ethanol washed to remove traces of unbound silane. These NPs dried in a rotary-evaporator were stored under N2 to get finer particles with reduced particle size.
Binding of lipolase on silane-coated NPs
A commercial Lipolase 100L (Novozymes, Denmark) was covalently immobilized on to silane-coated modified magnetic NPs. For lipase immobilization [14], 30 mg of modified NPs were suspended in sodium phosphate buffered saline (PBS, pH 7.4) and the suspension was sonicated for 5 min at room temperature. Lipase-PBS solution (5 ml of certain concentration) was added to it in the presence of N2 and suspension was kept under shaking for 24 h at 37°C for efficient immobilization of lipase onto magnetic NPs. Lipase-immobilized NPs were separated by a magnet and supernatant was collected. These NPs were given several PBS washing and were finally dispersed into PBS for further use.
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