Indexed on: 30 Aug '14Published on: 30 Aug '14Published in: Journal of Physical Chemistry B
We report interaction between bovine serum albumin ([BSA] = 1% (w/v)) and gelatin B ([GB] = 0.25-3.5% (w/v)) occurring close to their common isoelectric pH (pI). This interaction generated distinguishable multiple soft matter phases like opaque coacervates (phase I) and transparent gels (phase II), where the former are composed of partially charge neutralized intermolecular complexes (zeta potential, ζ ≤ 0) and the latter of overcharged complexes (ζ ≥ 0) that organized into a network pervading the entire sample volume. These phase states were completely governed by the protein mixing ratio r = [GB]:[BSA]. Coacervates, when heated above 32 °C, produced thermoirreversible turbid gels (phase III), stable in the region 32 ≥ T ≤ 50 °C. When the transparent gels were heated to T ≥ 34 °C, these turned into turbid solutions that did form a turbid fragile gel (phase IV) upon cooling. Mechanical and thermal behaviors of aforesaid coacervates (phase I) and gels (phase II) were examined; coacervates had lower storage modulus and melting temperature compared to gels. Cole-Cole plots attributed considerable heterogeneity to coacervate phase, but gels were relatively homogeneous. Raman spectroscopy data suggested differential microenvironment for these phases. Coacervates were mostly hydrated by partially structured water with degree of hydration dependent on gelatin concentration whereas for gels hydration was invariant of [GB]. Small-angle neutron scattering (SANS) data gave static structure factor profiles, I(q), versus wavevector q, that were remarkably different. For transparent gels, data could be split into two distinct regions: (i) 0.01 < q < 0.1 Å(-1), I(q) = IOZ(0)/(1 + q(2)ζgel(2))(2) (Debye-Bueche function) with ζgel = 9-13 nm, and (ii) 0.1 < q < 0.35 Å(-1), I(q) = IOZ(0)/(1 + q(2)ξgel(2)) (Ornstein-Zernike function) with ξgel = 3.1 ± 0.6 nm. Similarly, for coacervate, the aforesaid two q-regions were described by (i) I(q) = IPL(0)q(-α) with α = 1.7 ± 0.1 and (ii) I(q) = IOZ(0)/(1 + q(2)ξcoac(2)) with ξcoac = 1.6 ± 0.2 nm, a value close to the persistence length of gelatin chain (lp ≈ 2 nm). Phase transition from one equilibrium state to another, i.e., phase I to II, was hierarchical in the charge state of the protein-protein complex. Within the same charge state, transition from phase I to III and from phase II to IV was thermally activated. The aforesaid mechanisms are captured in a unique ζ-T phase diagram.