-Whey protein fractionation based on the selective isoelectric precipitation of oe-lactalbumin has been optimised by an overall approach. Precipitation phenomena and rheological characteristics of the fluid after heat treatment have both been considered. Control of calcium concentration, a new operating parameter, has optimised the precipitation step. A reduction of this concentration leads to an increase in the oe-lactalbumin fraction precipitated at a moderate protein concentration. The use of citric acid, a calcium complexation agent, is proposed. It permits simultaneous adjustment of the pH and a reduction in the free calcium concentration. Co-precipitation of immunoglobulins and bovine serum albumin has been studied. However, there are no conditions under which oe-lactalbumin is the only prote in that precipitates. To separate precipitated and soluble fractions, centrifugation is the method proposed, as it is more efficient than microfiltration. After solubilisation, the two forms of oe-lactalbumin may be recovered: the apo-oe-lactalbumin with a calcium-free solve nt, and the native form with a calcium solvent. When a solution of calcium chloride is used, solubilisation is a fractionation step (increase of 23% in a-LA purity), as the immunoglobulins remain insoluble. whey protein / fractionation / optimisation / precipitation / solubilisation Résumé-Optimisation d'un procédé de fractionnement des protéines du lactosérum fondé sur la précipitation sélective de l'a-lactalbumine. Le procédé de fractionnement des protéines du lactosérum par précipitation isoélectrique de l'a-lactalbumine est optimisé selon une approche globale. Les phénomènes de précipitation et les caractéristiques rhéologiques du fluide après traitement thermique sont pris en compte. Le contrôle de la concentration en calcium, paramètre opératoire additionnel, permet d'optimiser l'étape de précipitation, car lorsque cette teneur est réduite, le phénomène de précipitation de l'a-lactalbumine est amplifié. L'utilisation d'acide citrique, agent de complexation du calcium, est la procédure proposée. Elle permet d'ajuster simultanément le pH et de réduire la concentration en calcium ionique. La coprécipitation des immunoglobulines et de la sérum albumine bovine dans la gamme de conditions opératoires étudiées est un phénomène inévitable. * Correspondence and reprints 412 C Bramaud et al La centrifugation, plus efficace que la microfiltration, est le moyen proposé pour séparer les phases soluble et précipitée. Selon la composition saline du solvant, l'étape de solubilisation permet de produire les deux formes de l'a-lactalbumine. L'utilisation d'une solution de chlorure de calcium apporte une nouvelle sélectivité au procédé de fractionnement (gain de 23 % de pureté en a-LA), les immunoglobulines ne se solubilisant pas.
The selective precipitation of alpha-lactalbumin (alpha-LA) at a pH around its isoelectric point (4.2) under heat treatment is the basis for a fractionation process of whey proteins. As precipitation is a phenomenon dependent on the protein hydrophobicity, and as the release of the tightly bound calcium occurring at pH around 4 modifies the alpha-LA hydrophobicity, the specific role of calcium on isoelectric precipitation is investigated. A study of the extent of alpha-LA precipitation in a whey protein concentrate under various operating conditions of pH, temperature, protein concentration, and calcium content is presented. We propose a mechanism for this phenomenon as a combination of a complexation equilibrium and of an irreversible precipitation, to account for the influence of temperature, alpha-LA concentration total ionic content, and calcium concentration, and also to estimate the complexation equilibrium constant. (c) 1995 John Wiley & Sons, Inc.
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