Calcium bioaccessibility increased during gastrointestinal digestion of α-lactalbumin and ß-lactoglobulin.

Calcium bioaccessibility depends on the amount of soluble calcium under intestinal digestion. The changes in calcium during in vitro static digestion of α-lactalbumin and ß-lactoglobulin in presence of calcium chloride (0 mM, 20 mM and 50 mM) were followed by combining electrochemical determination of free calcium with the determination of soluble calcium by inductively coupled plasma optical emission spectroscopy. α-Lactalbumin and, more evident, ß-lactoglobulin were found to increase calcium bioaccessibility with increasing intestinal digestion time by around 5% and 10%, respectively, due to the complex binding of calcium to peptides formed from protein hydrolysis by gastrointestinal enzymes. In vitro digested samples of ß-lactoglobulin in presence of CaCl2 had nearly twice as much complex bound calcium as α-lactalbumin samples. The calcium bioaccessibility decreased significantly with the increasing concentration of added calcium chloride, although the amount of calcium chloride had little effect on the extension of digestion of α-lactalbumin and ß-lactoglobulin. Simulated digestion fluids were found to have a negative effect on calcium bioaccessibility, especially the presence of hydrogen phosphate, and the amount of precipitated calcium increased significantly with increasing amount of added calcium chloride. Based on analysis and visualization by sequences of the peptides formed during digestion of α-lactalbumin and ß-lactoglobulin, it was observed that peptides containing aspartic acid and glutamic acid acting as calcium chelators, may prevent precipitation of calcium in the intestines and increase calcium bioaccessibility. These results provide knowledge for the design of new dairy based functional foods to prevent calcium deficiency.

Temperature effects on calcium binding to caseins.

The kinetics of binding of calcium ions in molar excess to individual caseins and casein ingredients was studied in pH 6.4 aqueous solutions using stopped-flow absorption spectroscopy. An initial second-order reaction, faster for ß-casein than for αs-casein due to lower energy of activation (ΔEa1,ß = 8.2 kJ∙mol-1; ΔEa1,α = 18.1 kJ∙mol-1, respectively), is followed by a slower first-order reaction with similar energies of activation (ΔEa2,ß = 25.3 kJ∙mol-1 and ΔEa2,α = 20.7 kJ∙mol-1) as determined from temperature dependence of rate between 25 °C and 50 °C. Sodium caseinate reacts faster with calcium than both αs-casein and ß-casein in the first reaction of the two consecutive reactions, while the rate of the second falls between αs-casein and ß-casein. Global spectral analysis showed the UV-visible spectra of the reaction intermediates of the caseins to be more similar to the final products than to the initial casein reactants. Dynamic and static light scattering indicated decreasing particle sizes and increasing particle surface upon calcium-binding most significantly at low temperatures. The calcium binding to casein was found endothermic by isothermal titration calorimetry. Calcium binding seems to be controlled by enthalpy/entropy compensation corresponding to an isoequilibrium temperature of 38 °C in agreement with binding of calcium to o-phosphoserine rather than to aspartate or glutamate side chains of the caseins. Binding capacity and affinity for calcium to αs-casein and sodium caseinate both increased with increasing temperature in agreement with the endothermic nature of the binding. Decreasing enthalpy of binding for each calcium indicating a decrease in heat capacity of the caseins upon calcium-binding. The small difference between binding enthalpy and energy of activation for association of calcium to αs-casein lead to the conclusion that calcium dissociation goes through an early transition state. The rate of calcium dissociation hardly depends on temperature also explaining why calcium binding to caseins is important for calcium bioaccessibility.

Increasing calcium phosphate aqueous solubility and spontaneous supersaturation combining citrate and gluconate with perspectives for functional foods.

Uptake of calcium from food depends on solubility of calcium salts in the intestines, and precipitation of calcium phosphates decreases bioaccessibility of food calcium. Citrate as a high affinity complex binder for calcium was found spontaneously to create strongly supersaturated solutions by rapid dissolution of calcium hydrogen phosphate characterized by short lag phases for precipitation. Gluconate with weaker affinity for calcium binding showed longer lag phases for precipitation from supersaturated solutions. For citrate/gluconate combinations, the highest degree of supersaturation with longest lag phases for precipitation were found by trial-and-error experiments for a citrate/gluconate ratio of 1:10 for dissolution of calcium hydrogen phosphate resulting in supersaturation factors around three and without precipitation for more than a month. The aim of the present study was to provide a physicochemical explanation of this robust supersaturation. Calcium speciation based on electrochemical calcium activity measurement identified a low [Ca2+]·[HCitr2-] product as critical for supersaturation.

Enthalpy-entropy compensation in calcium binding to acid-base forms of glycine tyrosine dipeptides from hydrolysis of α-lactalbumin.

Calcium binding to peptides formed by hydrolysis of whey proteins during digestion is important for calcium uptake in the intestines and affects the antioxidant function of the peptides. For the two dipeptides, Gly-Tyr and Tyr-Gly, potential hydrolysis products of α-lactalbumin, calcium binding to the three forms of each dipeptide in acid-base equilibrium at intestinal pH was determined electrochemically and compared to binding to tyrosine for aqueous 0.16 M NaCl for 5 < pH < 9 at 15 °C, 25 °C, and 37 °C. At milk pH at 25 °C, binding of calcium to the zwitterion of GlyTyr dominates, with an association constant Kass2 = 22 M-1 with ΔH0 = -46 kJ·mol-1, while binding to the mononegative TyrGly dominates for TyrGly with Kass3 = 32 M-1 and ΔH0 = -38 kJ·mol-1. At intestinal conditions, pH = 7 and 37 °C, binding of calcium has similar affinity for GlyTyr and TyrGly, while at higher pH and lower temperature, GlyTyr binds stronger. Density Functional Theory calculations confirmed a stronger binding to the zwitterion of GlyTyr than of TyrGly and an increasing affinity with increasing pH for both. Calcium binding to the acid/base forms of the dipeptides is at neutral pH strongly exothermic with ΔH0 becoming less negative at higher pH, and a linear enthalpy-entropy compensation (r2 = 0.99) results in comparable binding important for calcium bioavailability along the changing distribution among acid-base forms. Calcium binding decreases radical scavenging rate and antioxidative activity of both dipeptides.

Lime Juice Enhances Calcium Bioaccessibility from Yogurt Snacks Formulated with Whey Minerals and Proteins.

Yogurt-based snacks originally with a calcium content between 0.10 and 0.17 mmol/g dry matter were enriched with a whey mineral concentrate and whey protein isolate or hydrolysate. Whey mineral concentrate was added to increase the total amount of calcium by 0.030 mmol/g dry matter. Calcium bioaccessibility was determined following an in vitro protocol including oral, gastric, and intestinal digestion, with special focus on the effect of lime juice quantifying calcium concentration and activity. Calcium bioaccessibility, defined as soluble calcium divided by total calcium after intestinal digestion amounted to between 17 and 25% for snacks without lime juice. For snacks with lime juice, the bioaccessibility increased to between 24 and 40%, an effect attributed to the presence of citric acid. Citric acid increased the calcium solubility both from whey mineral concentrate and yogurt, and the citrate anion kept supersaturated calcium soluble in the chyme. The binding of calcium in the chyme from snacks with or without lime juice was compared electrochemically, showing that citrate increased the amount of bound calcium but with lower affinity. The results indicated that whey minerals, a waste from cheese production, may be utilized in snacks enhancing calcium bioaccessibility when combined with lime juice.

Processing of tomato: impact on in vitro bioaccessibility of lycopene and textural properties.

BACKGROUND: Human studies have demonstrated that processing of tomato can greatly increase lycopene bioavailability. However, the difference between processing methods is not widely investigated. In the current study different thermal treatments of tomato were evaluated with regard to their impact on in vitro bioaccessibility and retention of lycopene and beta-carotene as well as textural properties. Thermal treatments used were low (60 degrees C) and high (90 degrees C) temperature blanching followed by boiling. RESULTS: Lycopene was relatively stable during thermal treatment, whereas beta-carotene was significantly (P < 0.05) reduced by all heat treatments except for low temperature blanching. In vitro bioaccessibility of lycopene was significantly increased from 5.1 +/- 0.2 to 9.2 +/- 1.8 and 9.7 +/- 0.6 mg kg(-1) for low and high temperature blanching, respectively. An additional boiling step after blanching did not further improve lycopene bioaccessibility for any treatment, but significantly reduced the consistency of low temperature treated samples. CONCLUSION: Choice and order of processing treatments can have a large impact on both lycopene bioavailability and texture of tomato products. Further investigations are needed, but this study provides one of the first steps towards tomato products tailored to optimise nutritional benefits.

Mechanical and thermal pretreatments of crushed tomatoes: effects on consistency and in vitro accessibility of lycopene.

The effects of mechanical and thermal treatments on the consistency and in vitro lycopene accessibility of crushed tomatoes were evaluated. Different crushing intensities and a subsequent heat treatment carried out as a heat shock (95 degrees C for 8 min) or a boiling step (100 degrees C for 20 min) were examined. Additional homogenization was compared with milder crushing regarding the effect on lycopene content and in vitro accessibility. Textural properties, polygalacturonase and pectinmethylesterase activity, pectin degree of methoxylation, lycopene content, and in vitro lycopene accessibility were evaluated. Microstructure was studied using both light and transmission electron microscopy. Crushing and subsequent heating affected the pectin degree of methoxylation and the consistency of the crushed tomatoes. The mechanical and thermal treatments did not affect the lycopene content to any great extent; however, in vitro accessibility seemed to improve with extensive crushing followed by heating. Crushing or homogenization in itself was not enough to increase in vitro lycopene accessibility.