Rennet-induced gelation of calcium and phosphate supplemented skim milk subjected to CO2 treatment.

A Doehlert design was performed to study the effect of calcium and phosphate supplementation at 0 to 25 mmol/kg and 0 to 16 mmol/kg, respectively, on the rennet gelation of reconstituted skim milk subjected to pH-reversible CO(2) acidification. Supplemented reconstituted skim milk samples were acidified to pH 5.80 by the addition of CO(2) under pressure and depressurized under vacuum to restore the initial pH value. The second-order polynomial models satisfactorily predicted the effect of salt addition on the micellar molar Ca:P ratio and the average diameter of the casein micelles, whereas only trends were used in the analysis of the rennet-clotting behavior of salt-supplemented, CO(2)-treated milk. Whether added Ca was the most determinant factor on the micellar molar Ca:P ratio, added Pi (a mixture of Na(2)HPO(4) and NaH(2)PO(4)) was the most determinant factor on the other responses studied, and its effect was most pronounced when Ca was simultaneously added. By comparison with control samples, changes observed in this study were essentially due to salt supplementation and not to the CO(2) treatment. Therefore, this CO(2) treatment could be considered as an entirely reversible treatment rather than only pH-reversible, and predictions might be applied to untreated milk. In the case of Ca-supplemented milk, the micellar molar Ca:P ratio increased, the average micellar diameter decreased, and the rennet-clotting properties were improved, whereas opposite effects were observed upon Pi supplementation. Since modification of the micellar molar ratio is the result of change in the chemical composition of micellar calcium phosphate, the effect of calcium and phosphate supplementation on the rennet clotting of milk was found to be also dependent on the nature of the interaction between caseins and colloidal calcium phosphate.

Effect of controlled kappa-casein hydrolysis on rheological properties of acid milk gels.

An experimental method based on the controlled chymosin-induced kappa-casein hydrolysis of milk was proposed to modify micellar reactivity. Milk samples with a degree of kappa-casein hydrolysis of 19, 35, and 51% were obtained. The physicochemical properties of partially converted casein micelles were determined. The net negative charge of casein micelles was reduced with increasing degree of kappa-casein hydrolysis and a small but significant decrease in hydrodynamic diameter and micellar hydration were noted. Dynamic low amplitude oscillatory rheology was used to monitor the rheological properties of acid milk gels (GDL) made with partially chymosin-hydrolyzed milks in comparison with those of strictly acid and rennet gels. An increase in the gelation pH value was observed with increasing the degree of kappa-casein hydrolysis. The moduli values (G' and G'') reached 2 h after the point of gel were, for all degrees of hydrolysis tested, significantly higher than those of strictly rennet and acid gels. Comparison of changes in delta G'/delta t with time indicated differences in gel formation that could be related to the increased values of G' obtained for acid gel made with chymosin-treated milk. At a given time after gelation (2 h), increasing the degree of kappa-casein hydrolysis in milk led also to an increase in the loss tangent and the serum holding capacity of acid milk gels suggesting a correlation between these two parameters.

Casein interactions studied by the surface plasmon resonance technique.

Surface plasmon resonance technique was investigated for the first time to study the apparent hydrophobicity and association properties of the major bovine caseins: alpha(s)-(alpha(s1)- and alpha(s2)-caseins in a 4:1 proportion), beta-, and kappa-caseins. The apparent hydrophobicities of the caseins were evaluated by a new method based on the binding level of casein on a hydrophobic sensor chip, and kinetic and equilibrium affinity constants were determined for the following casein interactions: alpha(s)/alpha(s), alpha(s)/beta, alpha(s)/kappa, beta/beta, and beta/kappa, using a sensor chip modified with covalent immobilized caseins. The study by surface plasmon resonance technology of these casein interactions under different conditions (pH, ionic strength, calcium concentration, chemical modification) demonstrated that, at neutral pH, electrostatic repulsive forces play an important role since an increase in ionic strength of the medium resulted in a stronger interaction. When charge repulsions were reduced by either acidification, increase in ionic strength, or dephosphorylation, casein interactions were reinforced, presumably due to weak attractive forces. Moreover, in this molecular model, we showed that addition of calcium greatly increased the binding response between the most phosphorylated caseins and that the added calcium (2 mM) participated directly in the formation of bridges between the phosphate groups of the casein molecules.

Effect of salt addition on the micellar composition of milk subjected to pH reversible CO2 acidification.

Response surface methodology was used to investigate the effect of salt supplementation on the micellar composition of reconstituted skim milk subjected to acidification by CO2 pressure to pH 5.8, followed by depressurization under vacuum. Using a Doehlert design, calcium and phosphate were added to skim milk in the range of 0 to 25 mmol/kg and 0 to 16 mmol/kg of milk, respectively, and the pH was adjusted to 6.65 +/- 0.02. After carbonation, the milk sample was depressurized, and the pH returned to its initial value without modification of the ionic strength. Micellar composition was assessed by the concentration of micellar Ca, P, Mg, and protein, and the buffering properties of milk. The second order polynomial models satisfactorily predicted the effect of salt supplementation on the micellar composition (R2adj > 0.75). Added calcium was the most determinant factor, and favored the removal of Ca, P, Mg, and proteins from the soluble phase to the micellar phase when this addition was less than 17.5 mmol/kg of milk. Above this concentration, only the concentration of micellar Ca increased. The buffering response surface showed that the amount of micellar calcium phosphate increased to a maximum upon addition of 17.5 mmol of Ca/kg. By comparison with a control sample (supplemented but untreated skim milk), changes were essentially due to salt supplementation and not to the CO2 treatment. We suggest that Ca formed micellar calcium phosphate when added at a concentration less than 17.5 mmol/kg; whereas above this concentration, Ca bound directly to micellar proteins.

Protein-polyphenol reactions. 1. Nutritional and metabolic consequences of the reaction between oxidized caffeic acid and the lysine residues of casein.

1. Studies were made on the lysine content of casein reacted with caffeic acid oxidized aerobically under alkaline conditions of enzymically with tyrosinase (EC 1. 14. 18. 1). 2. Loss of fluorodinitrobenzene (FDNB)-reactive lysine was rapid at pH 10 and increased with time and the temperature of the reaction, with concentration of caffeic acid and with the oxygenation of the mixture. In presence of the enzyme mushroom tyrosinase, maximum reduction of reactive lysine occurred at pH 7 and was dependent on the reaction time and on the concentration of caffeic acid. 3. Reaction of alpha-formyl-LO-[U-14C]lysine with caffeic acid at pH 10 showed the rapid formation of five reaction products which appeared to polymerize gradually as the reaction progressed. 4. The nutritionally available lysine content of the casein-caffeic acid mixtures, as assayed with rats, was reduced after both alkaline and enzyme reactions, as were faecal digestibility, net protein ratio and net protein utilization. Biological value however was not reduced. 5. In metabolic studies using goat milk casein labelled with L-[3H]lysine and reacted with caffeic acid in the same way, the lysine-caffeoquinone reaction products were not absorbed by the rat but were excreted directly in the faeces. 6. The importance of the reaction of proteins with caffeoquinone and chlorogenoquinone (formed by the oxidation of caffeic and chlorogenic acids respectively) is discussed in relation to the production of sunflower protein, leaf protein and other vegetable-protein concentrates.

[Use of small peptides to study the loss of available methionine, lysine and tryptophan during technological processing]. / Utilisation de petits peptides pour l'étude des pertes de disponibilité de la méthionine, de la lysine et du tryptophane au cours de traitements technologiques.

Peptides such as glycyl-L-methionyl-glycine, glycyl-L-lysine, L-lysyl-glycine and glycine-L-tryptophyl-glycine are used to study side-chain reactivity of three essential amino acids during food processing. The treatment of glycyl-L-methionyl-glycine with sodium hypochlorite resulted in the two following types of reaction: 1. Methionyl residues are oxidised to the corresponding sulfoxide at sodium hypochlorite concentrations up to 0,1 p. 100 w/v; 2. Oxidation of methionine residues to methionine sulfone and deamination reactions also occur for sodium hypochlorite concentrations over 0,2 p. 100 w/v. Sodium hypochlorite treatment of glycyl-L-lysine and L-lysine-glycine causes, probably by deamination of the epsilon-NH2 groups, a loss of lysine of 20 and 30 p. 100, for sodium hypochlorite concentrations of 0,1 and 0,2 p. 100 w/v, respectively. Treatment of glycyl-L-tryptophyl-glycine with hydrogen peroxide (0,05 M) modifies tryptophan residues in such a way that it cannot be retrieved after hydrolysis of the tripeptide with methane sulfonic acid and subsequent chromatographic analysis; six new unidentified components appear on the chromatogram. Glycyl-L-methionyl-glycine was incubated with food constituents or additives such as reducing oses, acrolein, p-benzoquinone, methyl iodide, or dichloro I, I ethylene. Thin layer chromatography and RMN show that carbonyl compounds or quinones do not react with the thioether group. Alkylating agents sometimes used in food processing only gave traces of sulfonium compounds. Thus, it appears that the only reaction liable to render methionine residues unavailable in foods would be its oxidation to methionine sulfone.

Continuous Proteolysis with a stabilized stabilized protease. I. Chemical stabilization of an alkaline protease.

Due to the loss of enzymatic activity as a function of time, an alkaline protease, selected for the continuous preparation of protein hydrolysates (J. Boudrant and C. Cheftel, Biotechnol. Bioeng., 18,1735, 1976), was chemically stabilized by a simple treatment with glutaraldehyde. Two fractions, soluble and insoluble, were obtained. The activities of these two fractions were measured with casein and N-benzoyl-L-arginine ethyl ester (BAEE) as a function of glutaraldehyde concentration used. It was noted that the insoluble fraction was practically inactive with the first substrate and that the heat stability of the soluble form was likewise enhanced. Molecular weights of these two forms were unchanged, but the uv-spectrum of the soluble form was modified. From amino acid analysis, it appears that this treatment mainly provokes a decrease in lysine content.