基于PEG和不同盐的双水相体系萃取牛血清白蛋白研究
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摘要
建立了PEG 4000/NaH_2PO_4和PEG4000/(NH_4)_2SO_4两种双水相体系,比较了两种双水相体系对牛血清白蛋白萃取行为的影响.考察了无机盐种类和浓度、PEG质量分数、牛血清白蛋白浓度等因素对双水相形成以及对牛血清白蛋白分配行为的影响.结果表明PEG 4000/NaH_2PO_4双水相系统的最佳萃取条件为:PEG4000=11.50%,NaH_2PO_4=20%,此时K值最小为0.45,达最大回收率.PEG4000/(NH_4)_2SO_4双水相系统最佳萃取条件为:PEG4000=12%,(NH4)_2SO_4=17.50%,此时K值最小为0.42,达最大回收率.
The PEG4000/NaH_2PO_4 and PEG4000/(NH_4)_2SO_4 aqueous two-phase system is established and the influence of different aqueous two-phase system on bovine serum albumin(BSA)extraction behavior is compared.The sorts and concentration of inorganic salt,PEG mass fraction,and the influence of the concentration of BSA on aqueous two-phase system formation as well as on the distribution behavior of BSA are also investigated.The results show that PEG PEG4000/NaH_2PO_4 optimum extraction condition of aqueous two-phase systems is:PEG4000=10.50%,NaH_2PO_4=20%.When the minimum value of Kis 0.45,the recovery rate reaches the highest.PEG4000/(NH_4)_2SO_4 aqueous two-phase system optimum separation condition is as follows:PEG4000=12%,(NH_4)_2SO_4=17.50%.When the minimum value of Kis 0.42,the recovery rate reaches the highest.
The PEG4000/NaH_2PO_4 and PEG4000/(NH_4)_2SO_4 aqueous two-phase system is established and the influence of different aqueous two-phase system on bovine serum albumin(BSA)extraction behavior is compared.The sorts and concentration of inorganic salt,PEG mass fraction,and the influence of the concentration of BSA on aqueous two-phase system formation as well as on the distribution behavior of BSA are also investigated.The results show that PEG PEG4000/NaH_2PO_4 optimum extraction condition of aqueous two-phase systems is:PEG4000=10.50%,NaH_2PO_4=20%.When the minimum value of Kis 0.45,the recovery rate reaches the highest.PEG4000/(NH_4)_2SO_4 aqueous two-phase system optimum separation condition is as follows:PEG4000=12%,(NH_4)_2SO_4=17.50%.When the minimum value of Kis 0.42,the recovery rate reaches the highest.
引文
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[2]辜鹏,谢放华,黄海艳,等.双水相萃取技术的研究现状与应用[J].化工技术与开发,2007,36(11):29-33.
[3]谢海.双水相萃取技术研究现状[J].化学工业与工程,2006,23(5):463-466.
[4]郑楠,刘杰.双水相萃取技术分离纯化蛋白质的研究[J].化学与生物工程,2006,23(10):7-9.
[5]孙晨,刘广宇,王永亮.双水相萃取茶叶中茶多酚[J].食品研究与开发,2012,33(8):69-71.
[6]刘杨,王雪青,宠广昌,等.双水相萃取法富集分离螺旋藻藻蓝蛋白的研究[J].海洋科学,2008,32(7):30-32,37.
[7]苟小军,徐文俊.生物化学实验教程[M].成都:四川科学技术出版社,2009.
[8]刘叶青.生物分离工程实验[M].北京:高等教育出版社,2007.
[9]谢云飞,常雅宁,谢志镭,等.用双水相体系分离血红蛋白(HB)[J].华东理工大学学报(自然科学版),2010,36(5):685-689.
[10]赵新颖,屈锋,董敏,等.双水相萃取结合液相色谱法分离蛋白质[J].分析化学,2012,40(1):38-42.
[11]路秀玲,赵东旭,金业涛,等.膨胀床吸附高效纯化牛血红蛋白[J].化工学报,2003,54(9):1257-1263.
[12]丁士勇,张家年.牛肉肌红蛋白和血红蛋白稳定性研究[J].食品科学,2006,27(7):93-94.