| 摘要: | 研究期間：10103~10202;The present proposal is aimed at to investigate the preparation of macroporous chitosan membrane and attached metal ions incorporated membrane microspheres form for immobilization of a model enzyme (biofunctionalized enzyme). In particular, the covalent immobilization of enzymes (BSA, Trypsin, Urease, papain, tyrosin, Aminotrans ferase) on macroporous chitosan membrane with a nominal cut-off of about 0.02 to 0.4 μm which had been surface-functionalized with a thin polymer layer with reactive epoxide groups (1, 4 butanediol diglycidyl ether, glutaraldehyde and epichlorohydrin). The surface functionalization is based on a photo-initiated graft copolymerization, which has optimized for the preparation of affinity membranes for covalent immobilizations of proteins as affinity receptors. The optimization of the immobilization protocol (immobilization conditions, blocking and washing steps, storage conditions) is another topic. The resulting enzyme-membranes shall be characterized in terms of amount of immobilized enzyme and enzyme activity will be investigated. To prepare a novel immobilized biofunctional membrane evaluate of their properties and application by reversible enzyme immobilization. These support systems could provide a large specific surface area and therefore, the enzyme can be immobilized only on the surface, and would also provide less diffusion limitation problem than that of the porous supports. The resultant immobilized enzyme systems can be characterized and their activity retention, immobilization efficiency, catalytic properties, reusability and stability aspects are compared. The expected results of the present research are silica gel supported cross-linked chitosan membrane with surface macropores can be prepared by phase inversion methods. This silica gel in the core of the membrane could act as a rigid support and the macroporous chitosan layer on the bead surface could provide a sufficient amount of amino groups and suitable steric position for enzyme coupling. The product of this research, the macroporous chitosan biofunctionalized membrane could find many promise applications in biomedical, bioengineering and bioanalytical fields. Besides that, the protocol of relating enzyme immobilization method to its structure might lead to the rational design of enzyme immobilization considering the specificity of the molecular. They include synthetic and analytical purposes, bioremediation of contaminated soils and wastewater treatment. Among them, wastewater treatment, and wine and beverages stabilization appear to be very promising and, with regard to this aspect, it is worthwhile to make some considerations |
