J.L. Maubois

Peptides du lait à activité biologique

HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés. Peptides du lait à activité biologique J.L. Maubois, J. Léonil

Two-step chromatographic procedure for the purification of hen egg white ovomucin, lysozyme, ovotransferrin and ovalbumin and characterization of purified proteins.

An improved procedure is described involving gel permeation and anion-exchange chromatography for the purification of four major hen egg white proteins. The procedure involves a first-step purification of ovomucin and lysozyme by gel permeation on a Superose 6 Prep Grade column. In the second step, anion-exchange chromatography on Q Sepharose Fast Flow led to the isolation of ovotransferrin and ovalbumin from a gel permeation chromatographic peak. The purities were estimated as ca. 80, 100, 80 and 100% for ovomucin, lysozyme, ovotransferrin and ovalbumin, respectively. The purification yield was over 60% for each protein. Further characterization of purified lysozyme revealed that it was fully active and homogeneous in relation to the electrospray ionization mass spectrum. The electrospray ionization mass spectrum showed different ovotransferrin species. The amino acid composition of purified ovomucin was compared to those published previously.

Selective separation of amino acids with a charged inorganic nanofiltration membrane: Effect of physicochemical parameters on selectivity

A charged organic-inorganic nanofiltration (NF) membrane prototype was used to separate a mixture of nine amino acids (AA) on the basis of differential electrostatic interactions with the membrane because, for a given pH, some of them were positively charged, some were negative, and some were zwitterions. Effect of pH, amino acid concentration (C(r)), and added ionic strength ([NaCI]) on the process selectivity was studied. A global statistical study revealed that pH was the dominant parameter regarding fractionation. C(r) and [NaCI] had a weaker effect, but the ratio C(r)/[NaCI] demonstrated a pronounced effect on system selectivity. Two split-ups of the mixture were obtained at pH 2 and at pH 12, for a 1-g/L total AA concentration and a C(r)/[NaCI] ratio of 0.16. Under these conditions, the differences in transmissions between basic and acid AA were higher than 70%. Interpretation of the results according to the Donnan theory allows us to foresee the potentialities of charged nanofiltration membranes for the fractionation of a complex mixture, such as peptidic hydrolysate to streams containing peptides and amino acids having different isoelectric points. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 291-302, 1997.

Ionic interactions in nanofiltration of β casein peptides

Nanofiltration (NF) membrane technology shows interesting potentials for separating organic components on the basis of solute charge and size in the range of 300–1000 g mol−1. Separation properties of two inorganic NF membranes were studied with a set of 10 small peptides (molecular mass range: 300–900 g mol−1; 3 < pI < 10) contained in a well-characterized tryptic β casein hydrolysate. Peptides transmission strongly depended on ionic interactions in the system. Physicochemical conditions such as ionic strength and especially pH were crucial to the separation, because the membrane and peptides showed amphoteric properties. Thus, the three categories of peptides (acid, basic, neutral) were separated according to their pI because of presumed concentration gradients of charged peptides at the membrane: positive for basic peptides and negative for acid peptides. At optimum pH 8 this led to high transmissions of basic peptides (even over 100%), intermediate transmissions for neutral peptides, and low transmissions for acid peptides. The addition of multicharged cationic and anionic species in the hydrolysate induced a markedly enhanced selectivity when the polyelectrolyte was a membrane coion and a complete reversion of selectivity when it was a membrane counterion.

DEHYDRATION BY DESORPTION AND BY SPRAY DRYING OF DAIRY PROTEINS: INFLUENCE OF THE MINERAL ENVIRONMENT

ABSTRACT A drying method by desorption in a water activity meter was used to simulate the conditions of spray drying and to determine the water transfer inside dairy concentrates towards the surface and from the surface to the drying air. The concentrates were also spray dried and solubility index of powders were determined during reconstitution. Whey protein concentrates (WPC) and native phosphocaseinate suspensions (NPCS) were used to study the effect of NaCl (420 mM), CaCl2 (222 raM), sodium phosphate (173 mM) and sodium citrate (238 mM) on the water transfers. The decrease in water transfer during drying was explained by the high hygroscopicity of added mineral salts to WPC. NaCl addition to NPCS decreased the water transfer during drying, but increased the solubility index. Citrate and phosphate addition to NPCS increased the water transfer during drying and reconstitution. CaCl2 increased the water transfer during drying but the solubility index was always low. Results are discussed as a function of...

Selective determination of phosphopeptide β-CN(1–25) in a β-casein digest by adding iron: characterization by liquid chromatography with on-line electrospray-ionization mass spectrometric detection

A beta-casein tryptic digest has been analysed by reversed-phase high-performance liquid chromatography (RP-HPLC) with on-line electrospray-ionization mass spectrometry (ESI-MS). Analyses of peptides were carried out before and after addition of iron(II) to the peptides in solution. In both cases, the majority of peptides were identified by the determination of molecular masses by ESI-MS and by prior knowledge of the amino acid sequence of beta-casein, and thus of its corresponding tryptic peptides. In the presence of iron(II), only phosphopeptide beta-CN(1-25) was able to bind iron to form different complexes that have increased retention times on the RP-HPLC column and that also absorbed at 280 nm. The method presented here appears to be selective for peptides containing phosphoseryl cluster(s).