牛血铜锌超氧化物岐化酶规模化生产工艺研究  

Research of Large-scaled Production Technics of Copper/Zinc Superoxide Dismutase from Bovine Blood

作　　者：乌云达来[1] 孙振钧[1] 王冲[1] 侯燕军[2] 张宏[2] 谭竹钧[2] 屈冉[1] 

机构地区：[1]中国农业大学资源与环境学院,北京100094 [2]内蒙古大学生命科学学院,内蒙古呼和浩特010021

出　　处：《家畜生态学报》2006年第5期46-52,共7页Journal of Domestic Animal Ecology

基　　金：北京生态重点学科资助(XK10019440)

摘　　要：本研究利用红细胞连续分离和热变性、超滤、丙酮沉淀、离子交换等技术,建立了一套牛血铜锌超氧化物歧化酶(Cu,Zn-SOD)规模化生产的简便易行新工艺。该工艺采用碟式分离机对红细胞进行连续分离,其分离速度可达1000kg牛血/h以上;采用添加Cu2+、Zn2+的一次热变性方法除去血红蛋白,可以代替传统方法中的氯仿和乙醇,并且在超滤技术浓缩提取液后,沉淀过程中丙酮的用量减少了90%;减轻了这些有机溶剂对环境造成的压力。在规模化生产条件下,利用新工艺,投料14批,共570.5kg红细胞,得到SOD冻干粉36069.35 mg,平均比活达到6164.19U/mg.pro,平均收率为34.41万U/kg红细胞,并且主要化学试剂成本降低93.0%。此外试验结果还表明,冷冻红细胞热变性后,酶的平均总活力一般稳定在72.86万U/kg红细胞;与新鲜红细胞制备的SOD在收率、纯度、比活等方面无明显差别,即可以以冷冻红细胞为原料制备SOD,从而解决了原料长期保存问题。By adopting continuous separation of red cell from blood, heat denaturation, uhra-filtmtion, acetone deposition and ionic exchange , a new type of fast and simple technology to produce at large copper/zinc superoxide dismutase was established. By separating continuously the red cell with the disc separator, the separation speed reached 1000 kg bovine/h. The hemoglobin was removed by the heat denaturation method under the present copper zinc salt without using any chloroform and alcohol in traditional method. When ultrafiltration technology was used to concentrate extracted solution, the acetone level decreased by 90%. The pollution of organic solvent was abated. By adopting new technology and delivering 14 batches of material of 570.5 kg red cell, 36069.35 mg frozen SOD was gained, average activity was 6164.19U/mg. pro, average output was 34.41 × 104U/k9 red cell, the main cost of reagent was decreased 93.0%. It also showed that the average activity of enzyme was 72.86 × 104U/ kg red cell as these frozen red cell was denaturated. The experimental results show that there are no obvious differences between SOD between frozen red cell and fresh red cell in SOD yield, purification degree, specific activity ete. So SOD can be made from frozen red cell and the problem about reserving material for a long time was solved.

关 键 词：牛血 超氧化物岐化酶(Cu  Zn-SOD) 新工艺 

分 类 号：S811.5[农业科学—畜牧学]