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脱磷脂牛血红蛋白的制备
引用本文:赵东旭,韦新桂,谷振宇,张贵锋,苏志国.脱磷脂牛血红蛋白的制备[J].生物工程学报,2002,18(5):609-613.
作者姓名:赵东旭  韦新桂  谷振宇  张贵锋  苏志国
作者单位:中国科学院过程工程研究所,生化工程国家重点实验室,北京,100080
基金项目:国家自然科学基金的资助 (No .2 0 136 0 2 0 )~~
摘    要:研究了一种新的脱磷脂血红蛋白制备方法。在2%、5%PEG4000或2%、5%PEG10000作共溶剂的情况下,利用疏水相互作用色谱基本上完全除去了新鲜牛血红细胞裂解液中的磷脂类成分,其中以5%PEG4000为共溶剂时,血红蛋白的回收率最高,达85.0%,在血红蛋白存在下所用的苯基琼脂糖-6B型疏水介质的吸附容量为86.6 mg磷脂/mL介质。经过疏水作用色谱后,血红蛋白的P50是33864Pa,Hill系数是2.54,较好地保存了血红蛋白的生物活性。对疏水作用色谱的脱磷脂机理以及PEG的保护机制进行了讨论。  血红蛋白,疏水作用色谱,PEG,磷脂,共溶剂

关 键 词:血红蛋白,疏水作用色谱,PEG,磷脂,共溶剂
文章编号:1000-3061(2002)05-0609-05
修稿时间:2002年4月8日

Preparation of Bovine Lipid-free Hemoglobin
ZHAO Dong-Xu,WEI Xin-Gui GU Zhen-Yu ZHANG Gui-Feng SU Zhi-Guo.Preparation of Bovine Lipid-free Hemoglobin[J].Chinese Journal of Biotechnology,2002,18(5):609-613.
Authors:ZHAO Dong-Xu  WEI Xin-Gui GU Zhen-Yu ZHANG Gui-Feng SU Zhi-Guo
Affiliation:National Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100080, China. zhaodx1@yahoo.com
Abstract:A new method for preparation of Hb solution free of stromal lipid was described. Almost all the lipid in fresh hemolysate of bovine red blood was removed with hydrophobic interaction chromatography (HIC) in the presence of 2%PEG4000, 5%PEG4000, 2%PEG10000 or 5%PEG10000. With the adding of 5%PEG4000, the 80% of recovery of Hb in HIC was obtained and the maximum lipid absorbed by hydrophobic medium, butyl agarose -6B was 86.6mg/mL. The activity (P50) of hemoglobin preparation was 3386.4Pa torrs, and the Hill number was 2.54, which were near to that of the native red blood cells. The mechanism of removing lipid by HIC and the function of PEG in the process were discussed.
Keywords:hemoglobin    hydrophobic interaction chromatography    PEG    lipid  co-solvent
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