猪肺血管紧张素转化酶纯化工艺中试放大研究
Pilot-scale study on purification of angiotensin converting enzyme(ACE)from pig lung
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摘要: 采用DEAE离子交换层析-超滤法纯化猪肺血管紧张素转化酶(angiotensin converting enzyme,ACE),在小试工艺条件的基础上对DEAE离子交换层析进行中试放大。以ACE比活及酶活回收率为指标,考察工艺放大后的纯化效果,并进一步完善纯化工艺条件。结果表明,DEAE离子交换层析柱放大15倍后,最终得到的ACE比活为(1.05±0.04)U/mg,达到电泳级纯,与小试工艺的纯化结果无明显差异,酶活回收率为34.6%±0.3%,高于小试工艺。由此认为,猪肺ACE提取放大纯化工艺是可行的,且可提高纯化效率,该工艺为更大规模的纯化ACE奠定了基础。Abstract: Angiotensin converting enzyme(ACE)from pig lung was purified by DEAE ion exchange chromatography-ultrafiltration,and the DEAE ion exchange chromatography was amplified on the basis of the small-scale process. The specific activity of ACE and the recovery of enzyme activity were taken as indexes to examine the purification effect of the scale-up process and to further improve the purification conditions. The results showed that after 15 times amplification of DEAE ion exchange chromatography,the specific activity of purified ACE was(1.05±0.04)U/mg,which reached the grade of electrophoresis pure. There had no significant difference with the purification results of the small-scale process. The recovery of enzyme activity was 34.6%±0.3%,that higher than the small-scale process. This study indicated the pilot-scale of the purification was feasible and could greatly increase the purification efficiency. The process would provide a foundation for a larger scale of ACE purification.