Bio-imprinting Of Protein/DNA Macromolecules And Electro-driven Catalysis Of Enzyme

Mechanism of proteins as enzymes was studied in the thesis. The research can be divided into two aspects: (1) the proteins and nucleic acid are tailor-off by bio-imprinting to create new active sites, so as to explore the contribution of enzyme activity by static protein/DNA-ligand shape fit; (2) intelligent polymer-immobilizsed enzymes are applied by alternating electric field, and the influence of electric frequency on catalytic power is investigated.Using the flexibility of proteins, egg white albumin was pore-formed and heat-denatured in the presence of substrate template. BSA protein and herring sperm DNA were trasted by precipitation method in organic solvent. The results of the experiments demonstrated that the protein which was bioimprinted with reactant has high catalyzed activity, and bioimprinted egg white has activity in aqueous as well as in organic environment. Bioimprinted bio-macromolecules have catalytic activity, and the catalytic activity of bioimprinted BSA with BAEE would increased as we increased concentration of BAEE, but the relation between catalyzed activity of bio-imprinted BSA with Ac-Phe-OH and concentration of Ac-Phe-OH was indefinite. Bio-imprinted herring sperm DNA has poor catalyzed activity, it is possible that bio-imprinted host molecule was only protein what herring sperm DNA contained, because the ratio of A260 to A280 for herring sperm DNA was 1.9, which expressed that purity quotient of herring sperm DNA what the experiment applied was low.In order to study the change of trypsin's activity in alternating electric field, trypsins were immobilized in polymers, electric frequency of reaction system was changed in order to stimulate polymer to shrink, and changed trypsin's activity by controlled their movement. Immobilized enzyme could can be obtained in the condition of AMPS or MAA was applied as monomer, BIS as crosslinking agent, APS as initiator, TEMED as accelerator, and optimally ratio for dosage of monomer to crosslinking agent. The results of the experiments expressed that PAMPS was superior to PMAA when they were used to embed enzyme. There was not obvious rule for amplitude of fluctuation of absorption in every frequency when trypsin catalyzed BAPA in alternating electric field. Immobilized enzyme has prominent improvement when the frequency was 100 Hz, but it would be not as good as in 10 Hz and 300 Hz than there was not electrostimulation. The results of the experiments demonstrated that the activity of trypsin could be altered as the change of frequency of alternating electric field of reaction system.