Formation of soluble protein complexes and yoghurt properties influenced by the addition of whey protein concentrate

The effects of whey protein concentrate (WPC) on the formation of soluble protein complexes and yoghurt texture were evaluated. Skim milk (SM) and skim milk enriched with 1% WPC (SM + 1%WPC) or 2% WPC (SM + 2%WPC) were left unheated or heated and then made into yoghurt gels. Yoghurt prepared from heated SM + 2%WPC had significantly higher storage modulus, water holding capacity and firmness values and a denser microstructure than those prepared only from skim milk. Electrophoretic analysis of the milk showed that the level of β-lactoglobulin and κ-casein in the serum phase increased with increasing WPC concentration, indicating that the content of disulfide-linked β-lactoglobulin and κ-casein was higher in SM + 2%WPC than in SM, suggesting that more soluble protein complexes had been formed. Consequently, yoghurt prepared from heated SM enriched with WPC may have more bonds and more protein complexes in the protein network than yoghurt prepared only from SM, thus resulting in firmer gels.Practical applicationsYoghurt, one of the most popular fermented milk products, is of high economic importance to the dairy industry worldwide. In particular, high-protein yoghurt, such as Greek-style or set-type yoghurt, has been driving its ongoing popularity over recent years. In current industrial production of high-protein yoghurt, protein fortification and heat treatment of milk are two of the most important processing parameters affecting yoghurt texture. Whey protein concentrate has been added to milk to reduce whey separation and to increase the firmness of the yoghurt. From a technological point of view, the interaction of the denatured whey proteins with casein micelles or with κ-casein in the serum phases is regarded as responsible for obtaining a good yoghurt structure. The present research has shown that it is possible to produce yoghurt with a range of textural properties by precisely controlling the rate of whey protein fortification during its manufacture. Therefore, this study provides a better understanding of the effect of WPC fortification and aims to extend this insight for the production of good-quality yoghurt.     

Antihypertensive activity of milk fermented by Enterococcus faecalis strains isolated from raw milk

A total of 231 microorganisms were isolated from raw cow milk samples and the angiotensin-converting enzyme-inhibitory (ACEI) activity of the resultant fermented milk produced with the isolated microorganisms was assayed. Forty-six of these microorganisms were selected on the basis of high ACEI activity. Four Enterococcus faecalis strains stood out as producers of fermented milk with potent ACEI activity (IC₅₀ (the protein concentration that inhibits 50% of ACE activity): 34–59 μg mL⁻¹). Single doses (5 mL kg⁻¹) of the whey fraction obtained from these fermented milk samples were administered to spontaneously hypertensive rats (SHR) and to normotensive Wistar-Kyoto (WKY) rats in order to investigate their possible antihypertensive activity. Highly significant decreases in the systolic blood pressure (SBP) and in the diastolic blood pressure (DBP) were observed when the fermented milk was administered to SHR. Nevertheless, the fermented milk did not modify the SBP and the DBP of the WKY rats. Raw cow milk is an excellent source of wild lactic acid bacteria able to produce fermented milk with antihypertensive activity and antihypertensive activity of milk fermented by Enterococcus faecalis strains was associated with peptides different from Ile-Pro-Pro and Val-Pro-Pro.     

Ability of conventional and nutritionally-modified whey protein concentrates to stabilize oil-in-water emulsions

The ability of a modified whey protein concentrate (MWPC), which contains relatively high proportions of phospholipid and high molecular weight protein fractions, to form and stabilize 10 wt% corn oil-in-water emulsions (pH 7.0, 5 mM phosphate buffer) was compared with that of a conventional whey protein concentrate (CWPC). The MWPC stabilized emulsions required less protein to prepare stable emulsions with monomodal particle size distributions and small mean droplet diameters (d₄₃ ≈ 0.3 μm at [WPC] ⩾ 0.5 wt%) than CWPC stabilized emulsions (d₄₃ ≈ 0.4 μm at [WPC] ⩾ 0.9 wt%) under similar homogenization conditions (5 passes at 5000 psi). In addition, the emulsions stabilized by 0.9 wt% MWPC were more stable to high salt concentration (NaCl ⩽ 200 mM), thermal processing (30–90 °C for 30 min) and pH (3, 6 and 7) than those stabilized by the same concentration of CWPC, which was attributed to polymeric steric repulsion rather than electrostatic repulsion. This study has important implications for the wide application of WPC as a natural emulsifier in food products.     

Simultaneous detection and enumeration of viable lactic acid bacteria and bifidobacteria in fermented milk by using propidium monoazide and real-time PCR

This study describes a procedure that allows specific detection and enumeration of viable bacteria in four species of lactic acid bacteria (Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus casei subsp. casei and Lactobacillus acidophilus) and of Bifidobacterium lactis, mixed in fermented milk products. The procedure is based on the combined use of propidium monoazide (PMA), able to distinguish between viable and irreversibly damaged cells, with species-specific quantitative real-time PCR (RTi-PCR). Loss of viability of the species in a fermented milk through storage at 4 °C was similarly (P < 0.05) detected by PMA–RTi-PCR and selective plate counts. Furthermore, comparison of results obtained by both methods showed a Pearson linear correlation of 0.995. The enumeration of viable bacteria by PMA–RTi-PCR could be performed in 3 h, whereas enumeration by selective plate counts required three days. The procedure developed is a fast method for the identification, enumeration and discrimination of viability of lactic acid bacteria and bifidobacteria mixed in fermented milk products.     

Purification of angiotensin I-converting enzyme inhibitory peptides and antihypertensive effect of milk produced by protease-facilitated lactic fermentation

Fresh low-fat milk was fermented with five mixed lactic acid bacteria for up to 30 h at 42 °C. A protease, prozyme 6, was added 5 h after the beginning of fermentation. The whey was separated from the fermented milk and freeze-dried. As the fermentation time extended to 30 h, soluble protein content increased from 30.9 to 195.9 mg g⁻¹, free amino acid content increased from 2.8 to 192.8 mg g⁻¹, peptide content increased from 6.4 to 402.8 mg g⁻¹ and γ-aminobutyric acid (GABA) increased from 0 to 80.6 mg 100 g⁻¹, while inhibition of angiotensin I-converting enzyme (ACE) increased as indicated by a decrease of IC₅₀ from 1.18 to 0.24 mg mL⁻¹, respectively. The amino acid sequences of two ACE inhibitory peptides were Gly–Thr–Trp and Gly–Val–Trp, of which the IC₅₀ values were 464.4 and 240.0 μm, respectively. The systolic blood pressure and diastolic blood pressure of spontaneously hypertensive rat (SHR) were reduced 22 and 21.5 mm Hg, respectively, after 8 weeks of oral administration of diluted whey (peptide concentration 5 mg mL⁻¹) from the 30 h fermentation.     

Selection of probiotic bacteria for the fermentation of a soy beverage in combination with Streptococcus thermophilus

Lactobacillus delbrueckii subsp. lactis R0187, Lactobacillus helveticus R0052, Lactobacillus rhamnosus R0011 and Bifidobacterium longum R0175 were examined for their ability to grow in combination with Streptococcus thermophilus cultures in milk and a laboratory soy beverage (LSB; both standardized to 4.5% protein and 2.3% fat). Strains R0011 and R0187 did not rapidly acidify the soy beverage despite good growth rates on soy carbohydrates. The S. thermophilus populations in the LSB were similar to that of milk even though milk had 30% more buffering capacity. In milk but not in soy, symbiosis with respect to acidification rate was observed between S. thermophilus and L. helveticus or B. longum. The populations of L. helveticus in the fermented products were similar in pure cultures or in the presence of the streptococci. However B. longum did not compete well in the mixed culture. Fermentation conditions varied as a function of the ability of S. thermophilus strains to acidify media to a pH of 4.65 (between 8 and 24 h). The probiotic populations in the mixed culture were influenced by the S. thermophilus strain and by the time of fermentation. Variations in growth rates of the bacteria did not appear to be linked to differences in initial redox or α-amino nitrogen levels. Strain selection enabled the preparation of a mixed starter, probiotic-fermented soy beverage containing 1.1 × 10⁸ CFU/mL of L. helveticus R0052, which represented approximately 13% of the total final population.     

Sensory profile,acceptability, and their relationship for diabetic/reduced calorie chocolates

The purpose of this study was to determine the sensory properties and acceptability of lab developed prototypes of conventional, diabetic (with no sugar), and diabetic/reduced calorie milk chocolates (no sugar and 25% calorie reduction) with high-intensity sweeteners, sucralose and stevioside, and partial fat replacement with whey protein concentrate (WPC). PLS was performed in order to relate sensory properties and consumer acceptability and to determine drivers of liking and disliking. There was no difference between conventional, diabetic and diabetic/reduced calorie milk chocolates for brightness, cocoa aroma, cocoa butter aroma, and cocoa flavor (p > 0.05). Acceptability was higher for sucrose substitution by sucralose than by stevioside and partial fat replacement reduced acceptability of flavor even more (p ? 0.05). Crucial attributes which determine consumer acceptability in samples are sweet aroma, melting rate, and sweetness, whereas bitterness, bitter aftertaste, adherence, and sandiness were drivers of disliking.     

Proliferation and intracellular glutathione in Jurkat T cells with concentrated whey protein products

The effect of two whey protein concentrates (WPCs) and three whey protein isolates (WPIs) on the growth and intracellular glutathione concentration of Jurkat T cells was determined. Standard RPMI 1640 media containing foetal calf serum with no WPC or WPI supplementation was used as the control, while supplementation with N-acetyl cysteine—a known glutathione promoter—was included as a positive control. Both WPCs lowered the cell count-adjusted glutathione concentration following a 24 h incubation period and one significantly (p<0.05) increased cell proliferation. Only one of the three WPIs significantly (p<0.05) inhibited cell proliferation although its composition with respect to β-lactoglobulin, glycomacropeptide, α-lactalbumin, IgG, proteose peptone and BSA content was almost identical to another WPI, as determined by HPLC. Based on co-migration with standards under two different modes of chromatography, lactoferrin was detected in the WPI showing the inhibitory effect at a level of 0.4 mg mL⁻¹, but not in any of the other concentrated whey protein products. None of the whey protein products tested increased cell-adjusted intracellular glutathione concentration.     

Effect of milk fat concentration and gel firmness on syneresis during curd stirring in cheese-making

An experiment was undertaken to investigate the effect of milk fat level (0%, 2.5% and 5.0% w/w) and gel firmness level at cutting (5, 35 and 65 Pa) on indices of syneresis, while curd was undergoing stirring. The curd moisture content, yield of whey, fat in whey and casein fines in whey were measured at fixed intervals between 5 and 75 min after cutting the gel. The casein level in milk and clotting conditions was kept constant in all trials. The trials were carried out using recombined whole milk in an 11 L cheese vat. The fat level in milk had a large negative effect on the yield of whey. A clear effect of gel firmness on casein fines was observed. The best overall prediction, in terms of coefficient of determination, was for curd moisture content using milk fat concentration, time after gel cutting and set-to-cut time (R² = 0.95).     

High hydrostatic pressure processing on microstructure of probiotic low-fat yogurt

The effect of milk processing on the microstructure of probiotic low-fat yogurt was studied. Skim milk fortified with skim milk powder was subjected to three treatments prior to innoculation: thermal treatment at 85 °C for 30 min, high hydrostatic pressure at 676 MPa for 5 min, and combined treatments of high hydrostatic pressure (HHP) and heat. The processed milk was then fermented by using two different starter cultures containing Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus, and Bifidobacterium longum. The microstructure of heat-treated milk yogurt had fewer interconnected chains of irregularly shaped casein micelles, forming a network that enclosed the void spaces. On the other hand, microstructure of HHP yogurt had more interconnected clusters of densely aggregated protein of reduced particle size, with an appearance more spherical in shape, exhibiting a smoother more regular surface and presenting more uniform size distribution. The combined HHP and heat milk treatments led to compact yogurt gels with increasingly larger casein micelle clusters interspaced by void spaces, and exhibited a high degree of cross-linking. The rounded micelles tended to fuse and form small irregular aggregates in association with clumps of dense amorphous material, which resulted in improved gel texture and viscosity.     

Microstructure and texture of white fresh cheese made with canola oil and whey protein concentrate in partial or total replacement of milk fat

A control white fresh cheese was prepared from milk containing 24 g milk fat (MF) L⁻¹, and nine white fresh cheese-like products were made by partial or complete substitution of milk fat by whey protein concentrate (WPC) and/or canola oil (CO) emulsified with an emulsifiers blend (EB) made of polyoxyethylene sorbitan monostearate (P), sorbitan monostearate (S) and glycerol monostearate (G) in 0.5:0.2:0.3 ratio. The textural characteristics and microstructure of the cheeses were assessed by Instrumental Texture Profile Analysis and Scanning Electron Microscopy. Polynomial models were obtained that estimated the composition and texture characteristics of the cheeses as function of the MF, EB (indirectly CO) and WPC concentrations in the cheese milk. CO incorporation promoted an open microstructure in the cheese, while WPC favoured a close and compact network made of short linking strands of milk proteins.MF, EB and WPC contributed positively to all the textural characteristics of the cheeses.     

Small and large deformation rheology and microstructure of κ-carrageenan gels containing commercial milk protein products

The rheological behaviour of commercial milk protein/κ-carrageenan mixtures in aqueous solutions was studied at neutral pH. Four milk protein ingredients; skim milk powder, milk protein concentrate, sodium caseinate, and whey protein isolate were considered. As seen by confocal laser microscopy, mixtures of κ-carrageenan with skim milk powder, milk protein concentrate, and sodium caseinate showed phase separation, but no phase separation was observed in mixtures containing whey protein isolate. For κ-carrageenan concentrations up to 0.5 wt%, the viscosity of the mixtures at low shear rates increased markedly in the case of skim milk powder and milk protein concentrate addition, but did not change by the addition of sodium caseinate or whey protein isolate. For κ-carrageenan concentrations from 1 to 2.5 wt%, small and large deformation rheological measurements, performed on the milk protein/κ-carrageenan gels, showed that skim milk powder, milk protein concentrate or sodium caseinate markedly improved the strength of the resulting gels, but whey protein isolate had no effect on the gel stength.     

The effect of container type on the growth of yeast and lactic acid bacteria during production of Sethemi,South African spontaneously fermented milk

Sethemi, a South African traditional fermented milk, was prepared in the laboratory by spontaneously fermenting raw milk in four different types of containers; namely, a clay pot, gourd, nickel jar and plastic bowl. The development of bacteria and yeasts in the different containers was followed by taking samples at 24 h intervals for 7 days. The predominating microorganisms in all cases were the aerobic mesophilic bacteria, lactobacilli–leuconostocs (LAB on MRS) and lactococci (LAB on M17). No significant (p > 0.05) differences were found in the number of microorganisms between the different containers. However, there were significant (p < 0.05) differences with respect to the types of dominant yeast species, and the diversity of yeast species. The least time to attain maximum LAB counts was recorded for the nickel container. In milk fermented in the clay pot and gourd the most dominant yeast species were Debaryomyces hansenii and Saccharomyces cerevisiae, respectively, while Cryptococcus curvatus predominated in both the nickel and plastic containers. The highest yeast diversity was obtained from milk samples fermented in the clay pot represented by 15 species. Of the 23 species isolated from all the containers and raw milk, only D. hansenii, Cryptococcus humicola and Kluyveromyces marxianus were isolated from both raw and naturally fermented milks of all containers. The use of different types of containers therefore influenced the diversity of yeasts in the fermented milk and may influence the characteristics of the fermented milk product. The clay pot offered the greatest yeast diversity suggesting that it could be the best suited for preparing traditional fermented milk.     

Properties of oregano (Origanum vulgare L.), citronella (Cymbopogon nardus G.) and marjoram (Majorana hortensis L.) flavors encapsulated into milk protein-based matrices

Encapsulating properties of skimmed milk powder (SMP) and whey protein concentrate (WPC) for the coating of the essential oil (EO) of oregano (Origanum vulgare L.) and aroma extracts (AE) of citronella (Cymbopogon nardus G.) and sweet marjoram (Majorana hortensis L.) by spray-drying were evaluated. The efficiency of microencapsulation expressed as a percentage of flavoring entrapped into the microcapsules varied from 54.3% (marjoram in WPC) to 80.2% (oregano in SMP). The content of flavoring remaining on the surface of encapsulated oregano EO was remarkably lower (1.1% and 1.4%) as compared with citronella (11.2% and 15.2%) and marjoram (16.7% and 22.1%) AEs encapsulated in SMP and WPC matrixes, respectively. Consequently, the changes in the composition of individual flavor compounds during encapsulation were considerably smaller for oregano EO as compared with citronella and marjoram AEs. The release of aroma compounds from the encapsulated products was assessed by solid phase microextraction (SPME) of headspace volatiles and their analysis by gas chromatography; some differences were observed between the analysed products. However, the effect of SPME fiber polarity was another important factor affecting the amount of extracted aroma compounds from encapsulated flavors. The percentages of nonpolar aliphatic terpenes were higher in the extracts obtained by nonpolar polydimethylsiloxane or bipolar polydimethylsiloxane–divinylbenzene fibers, while the content of oxygenated constituents in most cases was higher on the polar polyacrylate fiber. The latter extracted lower amounts of volatiles during 10 min exposure. The scanning electron microscopy and particle size analysis revealed that microcapsules were well-formed spherically shaped particles; however, SMP coated products had smoother surface as compared to WPC, containing more dents and wrinkles on the capsule surface. Particle size varied from 6 to 280 μm for SMP and from 2 to 556 μm for WPC microencapsulated products.     

Economic aspects of cheese making as influenced by whey processing options

Economic consequences of the cheese making process are illustrated through several sample calculations concerning processing of whey in relation to cheese making throughput and several whey processing alternatives. Small cheese plants with daily milk throughput of approximately 100 000 kg cannot economically justify the capital for water removal equipment. For small plants that have to convert whey to a dry product, alternatives include pre-concentrating with a reverse osmosis unit or a small plate evaporator and drying on a double roller dryer. The economics are evaluated at several price levels. At the upper scale of cheese plant size (2–3 million kg d⁻¹ of milk), the investment for whey processing is about half the total investment. Cash flows are calculated for electricity, natural gas and whey powder prices. Increased investment for further processing into whey protein concentrate and dried whey solubles or lactose is evaluated at several price levels.     

Available lysine,protein digestibility and lactulose in commercial infant formulas

Available lysine, in vitro protein digestibility and lactulose values were determined in 23 commercial infant formulas. The mean available lysine content of the formulas based on dairy proteins was 66.7±9.5 mg g⁻¹ protein, similar to that of human milk, while that of soy based formulas was considerably lower (45.0±8.3 mg g⁻¹ protein). In vitro protein digestibility values ranged 85.5–88.9% for soy-based formulas and 90.5–98.3% for formulas based on dairy proteins. Formulas based on milk enriched with whey had higher lactulose content than those based on cow's milk. However, all values were below the limit of 600 mg L⁻¹ recommended for UHT milk.     

Optimization of sour milk fermentation for the production of ACE-inhibitory peptides and purification of a novel peptide from whey protein hydrolysate

The effect of fermentation conditions on the production of angiotensin-I converting enzyme (ACE) inhibitory peptide in sour milk fermented by Lactobacillus helveticus LB10 was investigated using response-surface methodology. Optimal conditions to produce the maximum production of ACE-inhibitory peptides were found to be 4% (v/w) inoculum, 7.5 initial pH of medium and 39.0 °C. The fermented milk resulted in 75.46% inhibition in ACE activity. The cell-envelope proteinase, assisted by X-prolyldipeptidyl aminopeptidase of Lb. helveticus LB10 produced the ACE-inhibitory peptides. A novel ACE-inhibitory peptide from whey protein hydrolysate produced by crude proteinases of Lb. helveticus LB10 was purified. The separations were performed with Sephadex^® G-75 and Sephadex G-15 gel filtration chromatography and reversed-phase, high-performance liquid chromatography. The peptide with the RLSFNP sequence was isolated from β-lactoglobulin hydrolysate and its IC₅₀ while inhibiting ACE activity was 177.39 μm.     

Effect of cook temperature on starter and non-starter lactic acid bacteria viability,cheese composition and ripening indices of a semi-hard cheese manufactured using thermophilic cultures

Semi-hard cheeses were manufactured using Streptococcus thermophilus and Lactobacillus helveticus cultures and their ripening was characterised. During cheese manufacture, curds were cooked to a maximum temperature of 47, 50 or 53 °C, pre-pressed under whey at pH 6.15, moulded, pressed and brined. Increased cook temperature resulted in increased manufacture time, a significantly reduced growth rate of S. thermophilus during manufacture in the order 47≈50 °C>53 °C and in significantly lower mean viable cell counts of S. thermophilus up to 56 d of ripening. Increasing cook temperature had no significant effect on mean viable cell numbers of L. helveticus or non-starter lactic acid bacteria (NSLAB). Cheeses produced from curds cooked to 47 °C had significantly higher levels of moisture in non-fat substances (MNFSs), salt-in-moisture and a significantly lower pH and levels of butyrate compared with cheeses produced from curds cooked to 50 or 53 °C.     

Potential use of whey concentrate and prebiotics as carrier agents to protect Bifidobacterium-BB-12 microencapsulated by spray drying

Bifidobacterium BB-12 was microencapsulated by spray drying using liquid whey or whey retentate obtained from nanofiltration and inulin or polydextrose. The microcapsules were then characterized and the viability of the bifidobacteria was determined for 90 days at 4 °C and at − 20 °C. All the microcapsules showed high count of bifidobacteria, low moisture content, and low water activity. The microcapsules showed similar morphologies and particle sizes. The density of the microcapsules decreased with the increase of the total solids of the feed solutions. The highest hygroscopicity was noted in the microcapsules produced with polydextrose. The time of dissolution in water was longer than it was in oil for all the samples. The presence of prebiotics in the microcapsules increased the L* values and decreased the b* values; meanwhile, the samples produced with whey retentate showed lower a* values. The results of the thermal analysis suggested that prebiotics improved the stability of the microcapsules.     

Optimization of the rheological properties of probiotic yoghurts supplemented with milk proteins

This study aimed to optimize the rheological properties of probiotic yoghurts supplemented with skimmed milk powder (SMP), whey protein concentrate (WPC) and sodium caseinate (Na-Cn) by using an experimental design type simplex-centroid for mixture modeling. It included seven batches/trials: three were supplemented with each type of the dairy protein used, three corresponding to the binary mixtures and one to the ternary one in order to increase protein concentration in 1 g 100 g⁻¹ of final product. A control experiment was prepared without supplementing the milk base. Processed milk bases were fermented at 42 °C until pH 4.5 by using a starter culture blend that consisted of Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, and Bifidobacterium animalis subsp. lactis. The kinetics of acidification was followed during the fermentation period as well the physico-chemical analyses, enumeration of viable bacteria and rheological characteristics of the yoghurts. Models were adjusted to the results (kinetic responses, counts of viable bacteria and rheological parameters) through three regression models (linear, quadratic and cubic special) applied to mixtures. The results showed that the addition of milk proteins affected slightly acidification profile and counts of S. thermophilus and B. animalis subsp. lactis, but it was significant for L. delbrueckii subsp. bulgaricus. Partially-replacing SMP (45 g/100 g) with WPC or Na-Cn simultaneously enhanced the rheological properties of probiotic yoghurts taking into account the kinetics of acidification and enumeration of viable bacteria.