| Sol-gel approach of immobilized biological macromolecules is the progress of physical encapsulated progress essentially. Organic precursor composed of organic silane compound(TEOS or TMOS) is mixed with biological macromolecule uniformly in liquid phase. The mixed solution will form stable sol system through hydrolysis and condensation. Sol system gradually solidified to three-dimension network structure. Sol-gel approach follows several steps below in commom ways: hydrolysis, condensation, gelation, aging and drying. then the gel around and embedded the biological macromolecules. Given Lipase immobilised by TEOS have the problem of relatively poor biocompatibility, so this paper use glycerol to modify TEOS and obtained silane-gel precursor with better biocompatibility. Then studied the immobilization process; obtained high esterification activity of biological blot lipase.First, this paper obtain a hydrophilic silane-gel precursor which was modified by TEOS, solubility of product as selection standard, IR and 13C-NMR and 1H-NMR as chemical characterization, and the best preparation process of silane-gel precursor as followed: For solvent-free reaction system, TEOS: glycerol is 1:0.25(mol : mol), 3g of type 732 strong acidic cation-exchange resin, reaction at 180 rpm for 72 h at 50°C. The result showed that silane-gel precursor has good hydrophilicity and solubility can be up to 0.58 g/mL.Then, the process of the immobilized Candida rugosa lipase was researched, hydrophilic silane precursor gel used as a carrier, the activity of immobilized enzyme catalytic esterification as selection standard. The optimal immobilization condition as followed: modified silane-gel : lipase is 3:1(w/w), 80 mg of PEG600, 300 ul of 3- amino propyl triethoxy silane(APTES), at 0℃, phosphate buffer(p H 7.8). The result showed that the catalytic esterification activity of silane gel immobilized lipase increased to 1.42 fold, compared with free lipase.Finally, the biological imprinted lipase was immobilized by the modi fied silane gel precursor, which is by the way of immobilization to mainta in the bond of enzyme and ligand blotting at best catalytic conformation t o study the effect of biological imprinting process on the activity of lipase catalyzed esterification. Discussed lipase immobilized biological imprinting imprinting process optimum: Choose olive oil and methyl hexadecanoate imprinting template was used as a double-dip, lipase and imprinting template mass ratio of 5: 2, ethanol as a co-solvent(imprinting template and co-solvent mass ratio of 1: 2), Tween80 as a surfactant in an amount of 15 mg / mL, 90 W power ultrasound assisted blots 6min minutes, choose n-hexane as eluent and the template is removed by ultrasonic template. Modified silane gel blots of immobilized lipase catalyzed esterification biological activity under these conditions was 3.84 times the prepared free enzyme, is 1.65 times the biological imprinting lipase. |
