abstract
Precipitation experiments of electro-positive albumin by the action of a wide number of salts, and at different concentrations, were performed at a constant temperature (25 degrees C). The pH range studied covered extreme acidic conditions up to hydronium concentrations where the dissociation of the protein carboxyl groups becomes noticeable. The time required for the clouding phenomenon to occur and the quantity of salted-out protein were also ascertained. The results here reported show that the salt anion is the main salting-out species for the positively charged protein, where their efficacy in salting-out albumin from aqueous solution increases in the order: F- < Cl- < Br- < NO3- < I- < SCN- similar to CIO4- < SO42-. Although at extreme pH conditions the salt cation has no significant influence on the protein salting-out, experiments performed at higher pH values, where the carboxyl groups starts to dissociate, revealed a non-monotonic effect of the salt upon protein precipitation. We interpret this observation as a result of the presence of different protein forms, with which the salt cation participates in chemical equilibrium. Overall, the proteins salting-out phenomenon induced by salt can be rationalized by a general mechanism driven by electrostatic interactions and chemical equilibrium concepts. (C) 2019 Elsevier B.V. All rights reserved.
keywords
BOVINE SERUM-ALBUMIN; ION; PH; AGGREGATION; PROTEINS; WATER
subject category
Chemistry; Physics
authors
Madeira, PP; Freire, MG; Coutinho, JAP
our authors
Mara Guadalupe Freire Martins
Coordinating Researcher
Pedro António Palma Madeira
Junior ResearcherGroups
G4 - Renewable Materials and Circular Economy
G5 - Biomimetic, Biological and Living Materials
acknowledgements
This work was developed within the scope of the project CICECO-Aveiro Institute of Materials, FCT Ref. UID/CTM/50011/2019, financed by national funds through the FCT/MCTES. P.P.M. acknowledges University of Aveiro for funding in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree -Law 57/2016, of August 29, changed by Law 57/2017, of July 19.