earticle

영어

Recently, we reported the successful entrapment of lipase into cellulose-based composite hydrogel beads by using biocompatible ionic liquid.1 However, the entrapped lipase was easily leached during long-term storage. Therefore, in this work, cellulose-binding domain (CBD) was used to enhance the protein loading into cellulose and stability of entrapped protein by increasing the interaction between protein and cellulose. As a model system, CBD was fused to the N-terminal of GFP. Protein-entrapped cellulose hydrogel beads were simply prepared by dissolution of microcrystalline cellulose in [Emim] [Ac] and dispersion of GFP or CBD-GFP in cellulose solution followed by formation of cellulose hydrogel bead using pH 6.0 citrate buffer. The GFP entrapped into cellulose showed good activity under blue light irradiation. Protein loading of CBD-GFP into cellulose was 50% higher than that of original GFP. Leaching rate of protein from cellulose bead was drastically decreased with CBD-GFP. About 15% of entrapped GFP was released after 2h incubation under desorption condition, while only 1% of CBD-GFP was released. Retained protein content of CBD-GFP and original GFP was about 83% and 67% after 10 days incubation, respectively. These results show that CBD efficiently increase the content of loaded protein into cellulose and stability of entrapped protein by increasing the interaction between cellulose and protein fused to CBD.