The Study On The Preparation Of Porous Poly (GMA-co-DVB) And Its Immobilization Of β-Galactosidase From Aspergillus Oryzae

Macroporous polymer particles containing surface epoxy groups used immobilising β-galactosidase were firstly synthesized simultaneously with dodecyl alcohol and cyclohexanol as liquid pore-forming agents and nano-calcium carbonate as solid porogenic agent by suspension copolymerization. The synthesis were completed with lycidyl methacrylate(GMA) as monomer, divinylbenzene(DVB) as crosslinking agent, azobisisobutyronitrile(AIBN) as initiator, polyvinyl alcohol (PVA) and gelatine as dispersant. We expore the acount of divinylbenzene(DVB), the optimum time of the immobilized enzyme, pH and the optimal enzyme concentration. At the optimum conditons, the property of two immobilized enzyme were respective determined, such as the optimum temperature of immobilized enzyme of the enzymatic reaction, the optimum pH, pH stability of the immobilized enzyme, thermal stability, operational stability, kinetic parameters and so on, and were compared with the free enzyme. The results showed that carrier obtained from the immobilized edzyme with solid-liquid dual pore-forming agents, the enzyme activity yield was80.35%, and the enzyme activity reached a maximum of540.01U/g dry carrer. That was much higher than the enzyme activity of the immobilized edzyme obtained using the pure liquid pore-forming agent.In the role of FeCl3and FeCl2, the magentic poly(GMA-DVB) beads were firstly syntheszed with dodecyl alcohol and cyclohexanol as pore-forming agents by suspension polymerization and co-precipitation. The sanning electon microscopy(SEM) micrographs wre done to characterize the suiface structure of the magentic poly(GMA-DVB) beads, and the X-ray spectroscopy and the magnetic scales verified the magentic character of the beads. The β-galactosidase was immobilized on the magentic poly(GMA-DVB) beads and compared with the non-magnetic immobilized enzyme in the pH stability, thermal stability, operational stability, etc. The results show that the magnetic polymer carrier immobilized enzyme and the non-magnetic polymer carrier immobilized enzyme had the similar kinetic properties. Due to the magnetic polymer carrier immobilized enzyme has magnetic response, the magnetic polymer carrier could be convenient and rapid separation, which had a wider application prospects in the biochemistry field.