A Production Technology Of Copper/zinc Superoxide Dismutase From Bovine Blood At A Large Scale

This study reported a new simple and convenient technology that can be used to realize the large-scale production of copper/zinc superoxide dwasmutase (Cu, Zn-SOD) from bovine blood. The production procedure includes many technologies such as continuous separation technology of red cell, heat denaturation, ultrafiltration, acetone deposition and ionic exchange etc. The SOD-producing cost can be reduced by 89.1% when the technology was applied at large-scale in a factory.By establishing a fast and simple technology with the disc separator, continuous separation of red cell from blood, the results showed the separation rate reached 2000kg bovine blood/h. By using freezen separated red cell as material to make SOD, it can keep the cell for a long time, which was a problem when the fresh material was used. The hemoglobin was removed by the heat denaturation method under the present of copper zinc salt, without using any chloroform and alcohol in traditional method. When ultrafiltration technology was used to concentrate extracted solution, the acetone use level decreased by 90%. The SOD can be purified further by ion-exchange process. The experimental results show that there are no obvious differences between SOD between freezen red cell and fresh red cell in SOD yield, purification degree, specific activity etc. The SOD enzyme's activity and yield was steady commonly in the technology procedures such as heat denaturation, ultrafiltration, acetone deposition etc. After freezen red cell denatures, the enzyme's average total activity was steady at the level of 72.86 ten thousand U/Kg red cell and enzyme's total activity was at an average of 59.66 ten thousand U/kg red cell after ultrafiltrating concentration. Recovery rate was 81.89%. The average of total enzyme activity was 43.95 ten thousand U/Kg red cell after acetone deposition, and recovery rate was 60.33%. The average total enzyme activity was 33.07 ten thousand U/Kg red cell after DEAE clume chromatography. The enzyme's recovery rate was 45.38%. When it comes to fresh red cell, the average total enzyme activity was 86.93 ten thousand U/Kg red cell with heat denaturation. The average total enzyme activity was 61.05 ten thousand U/Kg red cell with ultrafiltrating concentration method. Recovery rate was 73.73%. The average total enzyme activity was 51.35 ten thousand U/Kg red cell after acetone deposition , and recovery rate was 59.31%. The average total enzyme activity was 36.10 ten thousand U/Kg red cell with DEAE clume chromatography. The SOD enzyme's recovery rate was 42.66%.Under the condition of a large-scale production in Da Ban biochemical product factory, Inner Mongolia, 570.5kg of 14 sets of feeding bovine blood made of 36069.35mg of SOD freeze-drying power with this SOD production technology. The average of comparative activity of SOD was 6164.19U/mg pro and the average yield was 34.41 ten thousand U/kg red cell.