authors |
Taha, M; Quental, MV; Silva, FAE; Capela, EV; Freire, MG; Ventura, SPM; Coutinho, JAP |
nationality |
International |
journal |
JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY |
author keywords |
Good's buffer ionic liquids; aqueous biphasic systems; Microtox; Vibrio fischeri; bovine serum albumin; purification; stability |
keywords |
BOVINE SERUM-ALBUMIN; 2-PHASE SYSTEMS; LIPASE PURIFICATION; PROTEIN SEPARATION; GLASS-ELECTRODE; COSMO-RS; EXTRACTION; STABILITY; FLUIDS; SALTS |
abstract |
BACKGROUND: Self-buffered aqueous biphasic systems (ABS) are prepared with ionic liquids (ILs) based on Good's buffers (GB-ILs) and used to purify proteins. RESULTS: A set of new GB-ILs based on the tetrabutylphosphonium cation ([P-4444](+)) was here synthesized and characterized. The self-buffering behaviour of the GB-ILs was asserted by measuring their protonation constants by potentiometry. These ionic liquids display self-buffering characteristics as well as a low toxicity towards the luminescent bacteria Vibrio fischeri. The ability of the GB-ILs to form ABS with the potassium citrate salt was investigated. These systems were then evaluated to extract and purify BSA from bovine serum samples. Extraction efficiencies of 100+/-5% of BSA to the GB-IL-rich phase were obtained in a single step. In addition, good recovery yields (59.4+/-0.8% to 80.1+/-0.7%), and purity levels of BSA (75.0+/-0.3% to 92+/-1%) were also achieved. The BSA secondary structure in the aqueous IL-rich solutions was evaluated through infrared spectroscopic studies revealing the protein-friendly nature of the synthesized GB-ILs. Dynamic light scattering (DLS), 'Conductor-like Screening Model for Real Solvents' (COSMO-RS), and molecular docking studies were carried out to better understand the main driving forces of the extraction process. CONCLUSION: The results obtained show that GB-IL based ABS can be prepared and used for protein purifications, with the GB-ILs showing a stabilizing effect on the protein. (C) 2017 Society of Chemical Industry |
publisher |
WILEY |
issn |
0268-2575 |
year published |
2017 |
volume |
92 |
issue |
9 |
beginning page |
2287 |
ending page |
2299 |
digital object identifier (doi) |
10.1002/jctb.5222 |
web of science category |
Biotechnology & Applied Microbiology; Chemistry, Multidisciplinary; Engineering, Environmental; Engineering, Chemical |
subject category |
Biotechnology & Applied Microbiology; Chemistry; Engineering |
unique article identifier |
WOS:000408909400011
|