The density of protein precipitates and its effect on centrifugal sedimentation

Isoelectric soya-protein precipitate densities were measured for mean particle sizes ranging from 3.4-65 mum by gradient centrifugation, centrifugation in water-immiscible solvents, tracerdilution, gravity sedimentation of isolated particles. Coulter counter volume determination, and a comparison of Coulter counter and centrifugal sedimentation size distributions. The immiscible system and tracer dilution methods were both found to be unreliable due to experimental uncertainties. The Coulter counter volume measurement indicated the existence of a density-size relationship with the aggregate density decreasing as the size increased. Comparison with sedimentation measurements showed that the Coulter counter measures 80% of the total aggregate volume for 6-mum particles. The relation between aggregate density (rho(a), kg m (-3)) and size (d, mum) was measured for isoelectric soya protein and casein precipitated by ammonium sulfate, using a comparison of the Coulter counter size distribution and centrifugal sedimentation. The functions were described for soya by \documentclass{article}\pagestyle{empty}\begin{document}$$ \rho _a - 1004 = 246d;{ - 0.408} $$\end{document} and for casein by \documentclass{article}\pagestyle{empty}\begin{document}$$ \rho _a - 1136 = 31d;{ - 0.441} $$\end{document} The gradient centrifugation method measured the buoyant density of hydrated protein precipitate which was independent of size, and is consistent with an aggregate structure consisting of primary particles. However, the aggregate structure was not described for all sizes by the theoretical cubic packing of hard-sphere primary particles, nor by the successive random addition of primary particles. The density-size functions indicated up to a fivefold difference in Stokes settling velocities compared to those calculated assuming a constant density difference.