Role of Ca2+ in the pepsin-induced coagulation and in the dynamic in vitro gastric digestion behavior of casein micelles.

The effect of Ca2+ on pepsin-induced hydrolysis of κ-casein and subsequent coagulation of casein micelles was studied in a micellar casein (MC) solution at pH ≈ 6.0 at 37 °C without stirring. An NaCl-supplemented MC solution was used as a positive control to assess the effect of increased ionic strength after CaCl2 addition. Quantitative determination of the released para-κ-casein during the reaction using reverse-phase high-performance liquid chromatography showed that specific hydrolysis of κ-casein by pepsin was little affected by the addition of either CaCl2 or NaCl. However, rheological properties and microstructures of curds induced by pepsin hydrolysis depended markedly on the addition of salts. Addition of CaCl2 up to 17.5 mM facilitated coagulation, with decreases in coagulation time and critical hydrolysis degree, and increases in firming rate and maximum storage modulus (G'max); further addition of CaCl2 (22.5 mM) resulted in a lower G'max. Increased ionic strength to 52.5 mM by adding NaCl retarded the coagulation and resulted in a looser curd structure. In a human gastric simulator, MC, without the addition of CaCl2, did not coagulate until the pH decreased to ≈5.0 after ≈50 min of digestion. Addition of CaCl2 facilitated coagulation of casein micelles and resulted in more cohesive curds with dense structures during digestion, which slowed the emptying rate of caseins. At the same CaCl2 concentration, a sample with higher ionic strength coagulated more slowly. This study provides further understanding on the effect of divalent (Ca2+) ions and ionic strength on the coagulation of casein micelles and the digestion behavior of milk.

Plant oil bodies and their membrane components: new natural materials for food applications.

Plants store triacylglycerols in the form of oil bodies (OBs) as an energy source for germination and subsequent seedling growth. The interfacial biomaterials from these OBs are called OB membrane materials (OBMMs) and have several applications in foods, e.g., as emulsifiers. OBMMs are preferred, compared with their synthetic counterparts, in food applications as emulsifiers because they are natural, i.e., suitable for clean label, and may stabilize bioactive components during storage. This review focuses mainly on the extraction technologies for plant OBMMs, the functionality of these materials, and the interaction of OB membranes with other food components. Different sources of OBs are evaluated and the challenges during the extraction and use of these OBMMs for food applications are addressed.

Role of biochemical and mechanical disintegration on ß-carotene release from steamed and fried sweet potatoes during in vitro gastric digestion.

The role of biochemical and mechanical disintegration on ß-carotene release from steamed sweet potatoes (SSP) and fried sweet potatoes (FSP) during in vitro gastric digestion was investigated. Results revealed that, in the absence of mechanical forces generated by the stomach, biochemical digestion did not have a great effect on the breakdown of cell walls within the sweet potato food matrix and the release of ß-carotene was similar in both SSP and FSP. Cell wall in the plant-food may act as a physical 'barrier' towards the action of gastric juice and to the release of nutrients into the gastric digesta. However, FSP underwent quicker softening and collapse during in vitro gastric digestion compared to the compact and denser structure of SSP. This may explain the faster cell wall breakdown and subsequent ß-carotene release from FSP cellular matrix than SSP when mechanical forces are applied as in the human gastric simulator (HGS).

Structural and physicochemical changes in almond milk during in vitro gastric digestion: impact on the delivery of protein and lipids.

Almond milk (about 3% protein and 7% lipids) was prepared using wet disintegration of raw almonds and then subjected to in vitro gastric digestion using an advanced dynamic digestion model (i.e., a human gastric simulator). Microstructural changes, physicochemical behavior, and protein digestion were examined; the release of lipids and protein during digestion was quantified. Under acidic gastric conditions, almond oil bodies flocculated. Proteolysis by pepsin led to destabilization and coalescence of the oil bodies, resulting in creaming and phase separation. This phase separation significantly delayed the delivery of lipids to the small intestine. After 225 min of digestion, ∼42% of the lipids remained in the stomach. In contrast, protein release was not significantly affected by the gastric behavior of the almond oil bodies. This study provides a better understanding of how the digestive system manages plant lipids, and may be useful in the microstructural design of foods to achieve a controlled physiological response during digestion.

Droplet-Stabilized Oil-in-Water Emulsions Protect Unsaturated Lipids from Oxidation.

Droplet-stabilized emulsions use fine protein-coated lipid droplets (the shell) to emulsify larger droplets of a second lipid (the core). This study investigated the oxidation resistance of polyunsaturated fatty acid (PUFA) oil within droplet-stabilized emulsions, using shell lipids with a range of melting points: olive oil (low melting), trimyristin (high-melting), and palmolein oil (intermediate melting point). Oxidation of PUFA oil was accelerated with a fluorescent lamp in the presence of ferrous iron (100 µM) for 9 days, and PUFA oxidation was monitored via conjugated dienes, lipid hydroperoxides, and hexanal levels. Oxidation was slower in droplet-stabilized emulsions than in conventional emulsions or control emulsions of the same composition as droplet-stabilized emulsions but different structure, and trimyristin gave the greatest oxidation resistance. Results suggest the structured interface of droplet-stabilized emulsions limits contact between pro-oxidants and oxidation-sensitive bioactives encapsulated within, and this antioxidative effect is greatly enhanced with solid surface lipids.

Free fatty acid profiles of emulsified lipids during in vitro digestion with pancreatic lipase.

Individual free fatty acids released from milk protein-stabilized emulsions prepared with milk fat, soya bean oil or tuna fish oil during in vitro digestion with pancreatic lipase were monitored using gas chromatography. The results showed that saturated fatty acids (C16:0 and C18:0) were released faster than unsaturated fatty acids (C18:1n9, C18:2n6 and C18:3n3) from soya bean oil emulsions; short chain fatty acids were released faster than long chain fatty acids from milk fat emulsions; long chain polyunsaturated fatty acids, such as eicosapentaenoic acid and docosahexaenoic acid, were released more slowly than other fatty acids from fish oil emulsions. The results confirm that the release behaviour of fatty acids from emulsions during digestion is related not only to the position of the fatty acids in the triglycerides in the fat/oil, but also to the length of the carbon chain of the fatty acid. The rates and the extents of the digestion of lipids consisting of short chain fatty acids are higher than those of lipids consisting of long chain fatty acids.

Formation of interfacial milk protein complexation to stabilize oil-in-water emulsions against calcium.

Oil-in-water emulsions (30% soya oil) stabilized by sodium caseinate or whey protein were destabilized in the presence of calcium, whereas lactoferrin-stabilized emulsion was stable under same conditions. At pH7, addition of lactoferrin to the caseinate or whey protein-stabilized emulsions led to association of lactoferrin with adsorbed proteins by electrostatic interactions, hence resulted in the formation of lactoferrin-caseins/whey protein interfacial complexes at the interface of the emulsion droplets. The stability of the interfacial complexes-coated emulsion droplets in the presence of Ca(2+) was examined by determination of particle size, zeta-potential and microstructure as a function of Ca(2+) concentration and added lactoferrin concentration. For emulsions made with 1% w/w caseinate or WPI, the stability of emulsions in the presence of 20 mM CaCl(2) has been improved markedly after addition of 0.2% w/w lactoferrin, suggesting the small amount of lactoferrin on the interface can largely enhance the calcium stability of milk proteins-coated emulsion droplets. Steric repulsion between the interfaces of droplets produced by the large lactoferrin molecules on the interface was considered to contribute the preventing the flocculation caused by Ca(2+) binding and reduction in the electrostatic repulsion between the emulsion droplets.

Adsorption behaviour of lactoferrin in oil-in-water emulsions as influenced by interactions with beta-lactoglobulin.

Oil-in-water emulsions (pH 7.0 or pH 3.0) containing 30 wt% soya oil and various concentrations of lactoferrin were made in a two-stage valve homogenizer. The average droplet size (d32), the surface protein coverage (mg/m2) and composition, and the zeta-potential of the emulsions were determined. The value of d32 decreased with increasing lactoferrin concentration up to 1%, and then was almost independent of lactoferrin concentration beyond 1% at both pH 7.0 and pH 3.0. The surface protein coverage of the emulsions made at pH 7.0 increased almost linearly with increasing lactoferrin concentration from 0.3 to 3%, but increased only slightly in emulsions made at pH 3.0 at lactoferrin concentrations >1%. The surface protein coverage of the emulsions made at pH 3.0 was lower than that of the emulsions made at pH 7.0 at a given protein concentration. The emulsion droplets had a strong positive charge at both pH 7.0 and pH 3.0, indicating that stable cationic emulsion droplets could be formed by lactoferrin alone. When emulsions were formed with a mixture of lactoferrin and beta-lactoglobulin (beta-lg) (1:1 by weight), the charge of the emulsion droplets was neutralized at pH 7.0 suggesting the formation of electrostatic complexes between the two proteins. The composition of the droplet surface layer showed that both proteins were adsorbed, presumably as complexes, from the aqueous phase at pH 7.0 in equal proportions, whereas competitive adsorption occurred between lactoferrin and beta-lg at pH 3.0. At this pH, beta-lg was adsorbed in preference to lactoferrin at low protein concentrations (1%), whereas lactoferrin appeared to be adsorbed in preference to beta-lg at high protein concentrations.

Flocculation and coalescence of droplets in oil-in-water emulsions formed with highly hydrolysed whey proteins as influenced by starch.

The effects of added unmodified amylopectin starch, modified amylopectin starch and amylose starch on the formation and properties of emulsions (4 wt.% corn oil) made with an extensively hydrolysed commercial whey protein (WPH) product under a range of conditions were examined. The rate of coalescence was calculated based on the changes in the droplet size of the emulsions during storage at 20 degrees C. The rates of creaming and coalescence in emulsions containing amylopectin starches were enhanced with increasing concentration of the starches during storage for up to 7 days. At a given starch concentration, the rate of coalescence was higher in the emulsions containing modified amylopectin starch than in those containing unmodified amylopectin starch, whereas it was lowest in the emulsions containing amylose starch. All emulsions containing unmodified and modified amylopectin starches showed flocculation of oil droplets by a depletion mechanism. However, flocculation was not observed in the emulsions containing amylose starch. The extent of flocculation was considered to correlate with the rate of coalescence of oil droplets. The different rates of coalescence could be explained on the basis of the strength of the depletion potential, which was dependent on the molecular weight and the radius of gyration of the starches. At high levels of starch addition (>1.5%), the rate of coalescence decreased gradually, apparently because of the high viscosity of the aqueous phase caused by the starch.

Influence of polysaccharides on the rate of coalescence in oil-in-water emulsions formed with highly hydrolyzed whey proteins.

The objective of this study was to examine the effects of added xanthan gum, guar gum, or kappa-carrageenan on the formation and properties of emulsions (4 wt % corn oil) formed with an extensively hydrolyzed commercial whey protein (WPH) product under a range of conditions. The rate of coalescence was calculated on the basis of the changes in the droplet size of emulsions during storage of the emulsions at 20 degrees C. Compared with the emulsion made without the addition of polysaccharides, the rate of creaming and coalescence in emulsions containing xanthan gum, guar gum, or kappa-carrageenan was markedly enhanced with increasing concentration of polysaccharides during storage for up to 7 days. At a given concentration, the rate of coalescence was highest in the emulsions containing guar gum, whereas it was lowest in the emulsions containing kappa-carrageenan. All emulsions containing xanthan gum, guar gum, or kappa-carrageenan showed flocculation of oil droplets by a depletion mechanism. This flocculation was considered to enhance the coalescence of oil droplets. The different rates of coalescence could be explained on the basis of the strength of the depletion potential, which was dependent on the molecular weight and the radius of gyration of the polysaccharides.