Development of different deep eutectic solvent aqueous biphasic systems for the separation of proteins.

ABSTRACT: In this work, aqueous biphasic systems (ABSs) formed by different deep eutectic solvents (DESs) were prepared and applied to extract proteins. The five kinds of DESs comprised amino acids and polyols ([amino acids][polyols]). They were combined with another DES resulting from tetrabutylammonium chloride and polypropylene glycol 400 ([TBAC][PPG400]) to form ABSs. The phase-forming abilities of [TBAC][PPG400]/[amino acids][polyols] were compared with those of [TBAC][PPG400]/amino acids and [TBAC][PPG400]/polyols. The results exhibited that the biphasic formation ability of [amino acids][polyols] lies between those of amino acids and polyols when [TBAC][PPG400] acts as the other phase in ABSs. The systems comprising [TBAC][PPG400] and [l-proline][xylitol] ([Pro][Xyl]) were further investigated to optimize the extraction performance. It was found that 97.30% chymotrypsin tended to distribute into the [Pro][Xyl]-rich phase under optimum conditions. The practical application of the system was demonstrated by the extraction of chymotrypsin from porcine pancreas. Besides, UV-Vis spectrophotometry (UV-Vis), Fourier transform infrared spectroscopy (FT-IR), and circular dichroism (CD) spectroscopy proved that the conformation of proteins remained unchanged during the extraction process. The extraction mechanism of the formation of DES-protein aggregates was investigated via conductivity, dynamic light scattering (DLS), and transmission electron microscopy (TEM). The overall results suggest that the DES/DES-based ABSs have outstanding potential in the green extraction of proteins.

SUBMITTER: Meng J 

PROVIDER: S-EPMC9064010 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature