Construction And Effect Analysis Of Immobilization Magnetic Carrier To Recombinant Chitinase

Magnetic nanometer material is a new type of functional polymer materials have been widely used in various fields of biological technology, especially immobilization of various enzymes have became the hot issues in the study of enzyme.Chitinase can catalyzes to degrade chitin of fungal cell wall, cell wall structural damage, so that the loss of capacity infection and pathogenicity of fungi. This paper takes Fe3O4as core, to compose3different functionalized magnetic polymer microspheres, and as immobilized carrier to immobilize of recombinant chitinase. The results are as follows:1、Fe3O4magnetic particle was synthesized by the modified chemical coprecipitation method. Ferrous chloride, ferric chloride and sodium hydroxide as the main material was used to synthesize magnetic Fe3O4microspheres. Fe3O4magnetic particle size is30nm-80nm.2、Fe3O4magnetic microspheres as core, the amido magnetic carrier (G1type) was prepared by3-aminopropyl triethoxysilane (APTESA302).The surface of the magnetic microspheres introduced amino particle size is50nm～100nm.3、Fe3O4magnetic microspheres as core, carboxylic magnetic carrier (G2type) was prepared by anhydrous sodium acetate and sodium citrate dissolved in ethylene glycol. The surface of carboxylic magnetic particles is spherical, and diameter is about200nm.4、Fe3O4magnetic microspheres as core, polyethylene glycol (PEG) as the dispersion medium, glycidyl methacrylate (GMA) as the functional monomer, two ethylene glycol dimethacrylate (EGDMA) as cross linking agent, azo different nitrile (AIBN) as initiator agent, epoxy magnetic carrier (G3型)was prepared. The diameter of epoxy magnetic carrier is about100nm.5、Three kinds of functional magnetic carrier was respectively used to immobilized recombinant chitinase, and the optimum immobilization conditions were determined. The results show that, G1type carrier enzyme dosage100ml, reaction time5h is the best; G2type carrier enzyme dosage80ml, reaction time7h is the best; G3type carrier enzyme dosage120ml, reaction time9h is the best.6、The effect of G1type, G2type and G3type magnetic carrier immobilized enzyme was researched in this papers. The results showed that:G3type carrier has higher protein loading capacity; low-temperature storage5d, G3type immobilized enzyme residual activity was89.4%. Residual enzyme activity of immobilized enzyme remained at more than66%even reusing up to five times; the thermal stability of immobilized enzyme is better than free enzyme; the optimum pH of immobilized enzyme in6-8.