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Thermodynamic insights on the liquid-liquid fractionation of gluten proteins in aqueous ethanol

Abstract : Wheat gluten includes two major proteins classes, gliadin (25–60 kg/mol) and glutenin polymers (100 to > 2,000 kg/mol) each comprising several polypeptides routinely identified by size-exclusion chromatography and electrophoresis. Gluten proteins are rich in glutamine (30%) and contain several repeated sequences, linking them to the wide class of intrinsically disordered protein (IDP). Here we showed that an ethanol/water (EtOH/W, 50/50, v/v) extract of an industrial gluten, comprising 1/3 of glutenin polymers and 2/3 of gliadin, underwent liquid-liquid phase separation (LLPS) below 14 °C, leading to two coexisting phases, respectively rich and poor in protein. As the quenching depth increased, proteins of lower and lower molecular weight joined the rich phase, akin to what would have been obtained for a polydisperse polymer sample. Within the rich phase the mass ratio of glutenin over gliadin decreased from 2.5 to 0.5 as the temperature dropped from 14 °C to −0.8 °C. Concomitantly the concentration in glutenin polymers increased up to 143 ± 6 g/L (at 9 °C) and then stopped to evolve, suggesting that the binodal line intersected the gelation line below this temperature. Applying the Flory-Huggins (FH) lattice model for each gluten protein classes, we demonstrated that their partitioning in the coexisting phases followed a same temperature dependency. However, some gliadin species joined the rich phase above their critical temperature. Here, specific interactions with the glutenin polymers through weak forces were exemplified. The study demonstrated the relevance of the Flory-Huggins (FH) lattice model in predicting phase behavior even when applied to complex protein mixtures.
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https://hal.inrae.fr/hal-03337519
Contributor : Dominique Fournier <>
Submitted on : Wednesday, September 8, 2021 - 9:51:45 AM
Last modification on : Friday, September 10, 2021 - 3:33:20 AM

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Marie-Hélène Morel, Justine Pincemaille, Laure Lecacheux, Paul Menut, Laurence Ramos, et al.. Thermodynamic insights on the liquid-liquid fractionation of gluten proteins in aqueous ethanol. Food Hydrocolloids, Elsevier, 2022, 123, pp.107142. ⟨10.1016/j.foodhyd.2021.107142⟩. ⟨hal-03337519⟩

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