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Calorimetric (from both isothermal micro-calorimetry and DSC), chiro-optical, viscometric and rheological data on aqueous solutions of pectic acid and low-methoxyl pectin (LMP), published over decades from different laboratories, have been comparatively revisited. The aim was to arrive at a consistent and detailed description of the behavior of galacturonan as a function of pH, i.e., of the degree of charging (as degree of dissociation, α) of the polyanion. The previously hypothesized pH-induced transition from a 31 to a 21 helix was definitely confirmed, but it has been shown, for the first time, that the transition is always coupled with loosening/tightening effects brought about by an increase in charge. The latter property has a twofold effect: the former effect is a purely physical one (polyelectrolytic), which is always a loosening one. However, in the very low range of pH and before the beginning of the transition, an increase in charge tightens the 31 helix by strengthening an intramolecular—but inter-residue—hydrogen bond. The value of the enthalpy change of 31 → 21 transition—+0.59 kcal·mol−1—is bracketed by those provided by theoretical modeling, namely +0.3 and +0.8 kcal·mol−1; the corresponding entropy value is also positive: +1.84 cal·mole r.u.−1·K−1. The enthalpic and the entropic changes in chain loosening amount only to about 23% of the corresponding 31 → 21 changes, respectively. Much like poly(galacturonic acid), the 31 conformation of LMP also stiffens on passing from pH = 2.5 to 3.0, to then start loosening and transforming into the 21 one on passing to pH = 4.0. Lowering the pH of a salt-free aqueous solution of LMP down to 1.6 brings about a substantial chain–chain association, which is at the root of the interchain junctions stabilizing the acid pH gels, in full agreement with the rheological results. A comparison of the enthalpic data reveals that, at 85 °C, LMP in acidic pH conditions has lost its initial order by about 2.3 times more than pectic acid brought from low charging to full neutralization (at α = 1.0) at 25 °C. A proper combination of experiments (enthalpic measurements) and theory (counterion condensation polyelectrolyte theory) succeeded in demonstrating, for the first time ever, a lyotropic/Hofmeister effect of the anion perchlorate in stabilizing the more disordered form of the 21 helix of galacturonan. The viscometric results in water showed that the 31 helix is capable of forming longer rheologically cooperative units compared with the 21 helix. Extrapolation to infinite ionic strength confirmed that, once all electrostatic interactions are cancelled, the elongation of the two helical forms is practically the same. At the same time, however, they indicated that the flexibility of the two-fold helix is more than fifteen times larger than that of the three-fold one. The result is nicely corroborated by a critical revisiting of 23Na relaxation experiments.

Pectate and alginate are among the most important biopolymers able to give rise to ionotropic gelation upon the addition of di- or multivalent counterions. The two ionic polysaccharides exhibit several common aspects of the gelation mechanism with calcium ions, the physiologically and commercially most relevant counterion type. The first one pertains to the role that specific Ca2+/polyion interactions play in the establishment of the ion-mediated chain/chain cross-links. Such interactions include both a specific affinity of the territorially condensed Ca2+ counterions for the polyuronate(s) and the formation of long-lasting chemical bonding (inner ion–sphere complex) of specific interchain sites accompanied by high conformational ordering. As to the first mechanism, it is dominated by the strong desolvation of the interacting ionic species, with concomitant positive variations in both enthalpy and entropy, the contribution of the latter prevailing over the former due to the favorable liberation of a very large number of water molecules of hydration. Both dilatometric and microcalorimetric data point to the higher affinity of Ca2+ for pectate than for alginate. The selective accumulation of calcium ions close to the polyanion(s) favors the onset of the second—chemical bonding—mode, which is associated with charge neutralization at the bonding site. This mode coincides with the largely accepted “egg-box” model for the calcium-mediated interchain junction of pectate and alginate. A new approach was devised for the calculation of the fraction of chemically bound divalent ions; it was based on the available circular dichroism data (further supported by scattering and viscosity results) and successfully tested by comparison with an independently determined fraction in the case of pectate. In detail, the strong bonding mode manifests in two sequential bonding modes. The first one (at low concentrations of added Ca2+ ions) entails a cross-link in which only one calcium ions is bracketed in a “twisted” egg-box between two chains; upon further counterion addition, a series of nearest-neighboring “perfect” egg-box structures develops. Both dilatometric and microcalorimetric changes associated with the latter chemical bonding modes are quantitatively larger for pectate than for alginate; clearly the latter polyuronate suffers from the relevant presence of the weakly calcium-binding mannuronic acid repeating units. Light-scattering experiments provided a clear-cut demonstration of the intermolecular bonding of calcium ions from the very beginning of the linker addition.