WANG Bao-quan1,ZHANG Yong-zhou1,PING Juan1,LI Zhao-hua2,LI Mei-ji4,DONG Xian-zhi2,PANG Xiao-bin3
1.The Huaihe Hospital of Henan University, Kaifeng 475001, China; 2.Institue of Biophsics Chinese Academy of Sciences, Beijing 100101, China; 3.The Pharmaceutical College of Henan University, Kaifeng 475001, China; 4.The Bioengineering Institute of Lanzhou University Gansu, Lanzhou 273000, China
Abstract��
OBJECTIVE To purify superoxide dismutase from fresh pig blood by using definite molecular weight polyacylic acid sodium (PAAS) as dispersant, explore the effect of different molecular weight of polyacylic acid sodium on SOD activity, and compare the new process with the traditional process to determine the optimum molecular weight of polyacylic acid sodium for purification of SOD from blood. METHODS Low, medium, and high molecular weight polyacylic acid sodium were used as highly efficient electrolyte with copper chloride as enzyme activator agent to improve the traditional blood purification technology via the key steps such as hemolysis, thermal alteration, cold acetone precipitation, ultrafiltration concentration, filtration chromatography and ion exchange chromatography. RESULTS High purity enzyme was obtained, the enzyme activity was up to 5 585 and 6 148 U��mg-1, and the product yields were 15.2% and 11.5%, respectively. The activity recoveries were 55.2% and 45.8% resectively after purification. SDS-PAGE showed a single stripe,and the subunit��s molecular was around 16��103. Different molecular weight of PAAS played inconsistent role in the process of purification of SOD. PAAS with a solid content of 3.45% and a hemolysis volume of 20% was suitable for the purification of blood SOD. CONCLUSION Low molecular weight PAAS is suitable as a polymer electrolyte to purify blood SOD, while the medium and high molecular weight PAAS are not appropriate for industrialized purification.
WANG Bao-Quan-,
ZHANG Yong-Zhou-,
PING Juan- etc
.Separation and Purification of Superoxide Dismutase from Blood by Polyacrylic acid Sodium and the Influential Factors[J] Chinese Pharmaceutical Journal, 2013,V48(23): 1985-1989
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2013, Vol. 48 