Department of Chemical Engineering, Iowa State University, Ames, Iowa 50011, USA.
Biotechnol Bioeng. 1993 Jul;42(3):333-8. doi: 10.1002/bit.260420310.
We explored the use of charged fusions for selective recovery of beta-galactosidase from cell extract using a low-cost, easily scaled, fast, charge-based separation technique-ion exchange on hollow fiber ion-exchange membranes (HFIEMs). The additional charges carried by a series of anionic fusion tails allowed selective binding and release of beta-galactosidase from Escherichia coli cell extract using the HFIEM cartridge. The purification factors increased with fusion length. The beta-galactosidase was recovered in active form. For the longest fusion studied, more than sixfold enrichment in specific activity was attained. The specific activity of the recovered fraction is comparable with that of commercial wild-type beta-galactosidase and affinity-purified fusion protein.
我们探索了使用带电融合的方法，通过一种低成本、易于放大、快速、基于电荷的分离技术——中空纤维离子交换膜（HFIEM），从细胞提取物中选择性回收β-半乳糖苷酶。一系列阴离子融合尾巴所携带的额外电荷允许β-半乳糖苷酶通过 HFIEM 试剂盒从大肠杆菌细胞提取物中选择性结合和释放。纯化因子随融合长度的增加而增加。β-半乳糖苷酶以活性形式回收。对于研究中最长的融合体，比活度的富集超过六倍。回收部分的比活度与商业野生型β-半乳糖苷酶和亲和纯化的融合蛋白相当。
