Cloud-point temperatures for lysozyme in electrolyte solutions: effect of salt type, salt concentration and pH.
Liquid-liquid phase-separation data were obtained for aqueous saline solutions of hen egg-white lysozyme at a fixed protein concentration (87 g/l). The cloud-point temperature (CPT) was measured as a function of salt type and salt concentration to 3 M, at pH 4.0 and 7.0. Salts used included those from mono and divalent cations and anions. For the monovalent cations studied, as salt concentration increases, the CPT increases. For divalent cations, as salt concentration rises, a maximum in the CPT is observed and attributed to ion binding to the protein surface and subsequent water structuring. Trends for sulfate salts were dramatically different from those for other salts because sulfate ion is strongly hydrated and excluded from the lysozyme surface. For anions at fixed salt concentration, the CPT decreases with rising anion kosmotropic character. Comparison of CPTs for pH 4.0 and 7.0 revealed two trends. At low ionic strength for a given salt, differences in CPT can be explained in terms of repulsive electrostatic interactions between protein molecules, while at higher ionic strength, differences can be attributed to hydration forces. A model is proposed for the correlation and prediction of the CPT as a function of salt type and salt concentration. NaCl was chosen as a reference salt, and CPT deviations from that of NaCl were attributed to hydration forces. The Random Phase Approximation, in conjunction with a square-well potential, was used to calculate the strength of protein-protein interactions as a function of solution conditions for all salts studied.
Grigsby
J J
JJ
Chemical Engineering Department, University of California, Berkeley, 94720, USA.
Blanch
H W
HW
Prausnitz
J M
JM
D000465:Algorithms; D002412:Cations; D004573:Electrolytes; D006863:Hydrogen-Ion Concentration; D009113:Muramidase / Q000737:chemistry*; D012996:Solutions; D013696:Temperature
D002412:Cations; D004573:Electrolytes; D012996:Solutions; D009113:Muramidase