Method of increasing the activity of immobilized horseradish peroxidase

摘要

Method of increasing the activity of chrysalis carried out in three stages, the first stage of the process of synthesis of the carrier with the iron magnesium core, the second stage of activation of the carrier and the third stage of immobilisation of the chrysalis on the prepared medium. In the first stage, a mixture of MgFe2O3, sodium dodecylbenzenesulphonate and toluene is prepared and subjected to ultrasonic treatment, toluene and two polymers are re-added: bis [2-methacryloiloxy) ethyl] phosphate as a binder, trimethyllopropane triacrylate as a sieving agent and methacrylic acid,and after careful mixing of ingredients, benzoyl peroxide is added as an initiator, magnetic particles are separated and the carrier is rinsed in acetone, ethanol and distilled water. The vehicle is then activated in the second stage by incubation into the 1-ethyl-3-(3dimethylaminopropyl) carbidimide solution in the MES buffer for 30 minutes at 4-oC temperature, then three times the vehicle is washed with the MES buffer, after which, in the third stage, the vehicle prepared for 1:2 (v/v) is incubated with a solution of chrysodide peroxide (0,01 mg/ml, in a phosphate buffer with pH 6,2; 50 mM),at room temperature for one hour,then rinse with a phosphate buffer of pH 6,2 with Nacl. The essence of the invention is that the system thus obtained is exposed to a rotating magnetic field, with the frequency of rotation 50-Hz, the magnetic induction 18-mT, at the temperature 30-C in the time 3-15 minutes, the first stage of the vehicle's synthesis is carried out in the oxygen atmosphere, at the temperature 70-C for two hours mixing continuously, after which the temperatur e rises to 75oC and the reaction takes place for the next six hours, and the mixture of MgFe2O3, sodium dodecylbenzenesulphonate and toluene is subjected to ultrasound for fifteen minutes,and after re-adding the toulin is mixed with the mechanical mixture for 20 minutes.