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Covalent binding of α-chymotrypsin on the magnetic nanogels covered by amino groups
Authors:Jun Hong  Dongmei Xu  Peijun Gong  Hanwen Sun  Li Dong  Side Yao  
Institution:aShanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800, PR China;bGraduate school of Chinese Academy of Sciences, Beijing 100039, PR China
Abstract:A new aminated carrier—magnetic nanogels covered by amino groups, was obtained by Hoffman degradation of polyacrylamide-coated Fe3O4 nanoparticles prepared by photochemical polymerization. α-Chymotrypsin (CT) was covalently bound to the magnetic nanogels by use of 1-ethyl-3-(3-dimethylaminepropyl) carbodiimide and N-hydroxysuccinimide at room temperature. Immobilization time, pH value of the reaction mixture and proportion of CT to the magnetic nanogels were investigated to obtain the optimum condition for CT immobilization. The maximal specific activity of the bound CT was determined to be 0.93 U/(mg min), 59.3% of free counterpart. The maximal binding capacity was measured to be 102 mg enzyme/g nanogel. Furthermore, the bound CT exhibited good thermal stability, storage stability and reusability.
Keywords:Magnetic nanogels  Immobilized enzyme  Photochemical polymerization  α  -Chymotrypsin
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