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目的以新鲜猪血为原料,首次利用不同相对分子质量聚丙烯酸钠作为分散剂来提纯血液超氧化物歧化酶(SOD),探索不同相对分子质量聚丙烯酸钠对血液超氧化物歧化酶提纯过程中酶活性的影响,进而确立最佳相对分子质量,并和传统血液提纯工艺进行比较,以期改进后的工艺能简化传统提纯流程。方法利用低,中,高相对分子质量的聚丙烯酸钠作为高效电解质和氯化铜为酶激活剂来改进传统血液提纯工艺,经溶血,热变,冷丙酮沉淀,超滤浓缩,分别经过凝胶过滤和离子交换色谱等步骤,得到高活性酶,比活力达5 585和6 148 U·mg~(-1),产品得率分别为15.2%和11.5%,纯化后活性回收分别为55.2%和45.8%,其聚丙烯酰胺凝胶电泳凝胶电泳显示酶呈现明显的单一条带,单亚基相对分子质量在16×10~3附近。结果通过实验现象和酶活性对照,不同相对分子质量的聚丙烯酸钠在血液提纯过程中所起的作用并不一致,固含3.45%低相对分子质量聚丙烯酸钠用量为溶血体积20%时适合作为超高效分散剂来提纯血液超氧化物歧化酶,中高相对分子质量的聚丙烯酸钠不适合工业化提纯应用。结论低相对分子质量聚丙烯酸钠在血液超氧化物歧化酶提纯工艺中更为合适,该工艺大大简化了传统的血液提纯工艺,适合工业化大规模生产.
OBJECTIVE To purify blood superoxide dismutase (SOD) with fresh pig blood as the raw material for the first time using different molecular weight sodium polyacrylate as dispersant to explore the effect of different molecular weight sodium polyacrylate on blood purification of superoxide dismutase Enzyme activity, and then establish the best relative molecular mass, and compared with the traditional blood purification process, in order to improve the process to simplify the traditional purification process. Methods By using low, medium and high relative molecular weight sodium polyacrylate as efficient electrolytes and copper chloride as enzyme activator, the traditional blood purification process was improved. After hemolysis, thermal shock, cold acetone precipitation and ultrafiltration, Filtration, and ion-exchange chromatography. The specific activity was 5858 and 6 148 U · mg ~ (-1), the yields were 15.2% and 11.5%, respectively, and the activity recoveries were 55.2% and 45.8%, the polyacrylamide gel electrophoresis gel electrophoresis showed a clear single band showed a single subunit relative molecular mass of 16 × 10 ~ 3 in the vicinity. Results Through the comparison of experimental phenomena and enzymatic activity, sodium polyacrylate with different relative molecular weights played an inconsistent role in the process of blood purification. Solid dosage of 3.45% sodium polyacrylate with low relative molecular mass of 20% Highly effective dispersant to purify blood superoxide dismutase, high molecular weight sodium polyacrylate is not suitable for industrial purification applications. Conclusion Low relative molecular weight sodium polyacrylate is more suitable for the purification of blood superoxide dismutase, which greatly simplifies the traditional blood purification process and is suitable for industrial mass production.