Aqueous two-phase system formed by thermoreactive vinyl imidazole/vinyl caprolactam copolymer and dextran for partitioning of a protein with a polyhistidine tail |
| |
| Authors: | Teixeira Franco,Telma,Galaev,Igor Yu,Hatti-Kaul,Rajni,Holmberg,Niklas,Bu¨ low,Leif,Mattiasson,Bo |
| |
| Affiliation: | (1) Chemical Engineering Faculty, State University of Campinas (UNICAMP), PO Box 6066, 13081–970 Campinas, Brazil;(2) Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, PO Box 124, S-22100 Lund, Sweden;(3) Department of Pure and Applied Biochemistry, Center for Chemistry and Chemical Engineering, Lund University, PO Box 124, S-22100 Lund, Sweden |
| |
| Abstract: | A new type of aqueous two-phase system (ATPS) has been developed for application combining two attractive concepts in downstream processing: the immobilised metal affinity partitioning and the use of thermoprecipitating polymers. ATPS consisting of the thermoprecipitating copolymer of N-vinyl caprolactam/1-vinyl imidazole loaded with Cu ions (Cu-poly-VI-VCL) in the top phase and dextran T70 in the bottom phase was used for purification of recombinant lactate dehydrogenase carrying an affinity tag of 6 histidine residues (His-LDH ) from a crude E. coli extract. The enzyme partitioned preferentially into the top Cu-poly-VI-VCL-rich phase. After phase separation, the latter was mixed with EDTA. Temperature increase to 45°C resulted in thermoprecipitation of VCL/VI-polymer, which could subsequently be recycled. His-LDH remained solubilized in the aqueous phase resulting in 8-fold purification and 80 % recovery in a single step. |
| |
| Keywords: | |
| 本文献已被 SpringerLink 等数据库收录！ |
|
