摘要
脱离了红细胞的血红蛋白(Hb)在溶液中易被氧化成高铁血红蛋白(MetHb),失去载氧活性.实验发现,当对红细胞裂解液进行微孔膜分离时,高铁血红蛋白增加不多;但在用层析法去除超氧化物歧化酶等其他红细胞蛋白后再进行超滤膜浓缩血红蛋白时,则出现较多的血红蛋白被氧化成高铁血红蛋白的现象,其氧化反应随超滤过程膜表面流体切向速度的增加而加快,随溶液温度的增加而加快.抗氧化剂的存在能有效地降低高铁血红蛋白的生成,谷胱甘肽(GSH)、半胱氨酸、N-乙酰半胱氨酸、亚硫酸钠、抗坏血酸(Vc)在溶液中和超滤过程中都能起到对血红蛋白载氧活性的保护作用.其中Vc的效果最好,最适加入量(摩尔比)Vc/Hb=8,pH8.将抗氧化的优化条件整合到从红细胞裂解液开始到超滤浓缩血红蛋白的整个流程,在离子交换层析后,添加Vc作为抗氧化剂进行超滤浓缩,Hb活性得到了很好的保护,MetHb的含量控制在2.3%,成功地制备了低MetHb含量的纯化血红蛋白.
The effect of different antioxidation agents on the reduction of methemmoglobin （MetHb） was investigated. Five antioxidation agents, i.e. vitamin C （Vc）, reduced glutathione （GSH）, cysteine, n-acetul-cyateine and sodium sulfite were used and compared according to different changes of MetHb content. The experimental results showed that all the antioxidation agents could protect the bioactiviW of Hb from oxidation, while Vc had strong ability to reduce MetHb. The optimal conditions [Vc/Hb=8 （mole ratio）, pH=8] obtained were applied to the membrane filtration process following the ion-exchange chromatography step. Bioactivity of the hemoglobin was well maintained when Vc was added as antioxidation agent, and the content of MetHb was successfully controlled to 2.3% in the final purified hemoglobin product.
出处
《过程工程学报》
EI
CAS
CSCD
北大核心
2005年第6期659-664,共6页
The Chinese Journal of Process Engineering
基金
国家自然科学基金资助项目(编号:20136020)
关键词
血红蛋白
膜分离
载氧活性
高铁血红蛋白
抗氧化剂
hemoglobin
membrane separation
bioactivity
methemoglobin
antioxidation agent
