Deciphering the segregation of proteins in high-protein dairy powders after spray-drying.

High-protein dairy powders are ingredients mainly produced by spray-drying, then subjected to aging during transport and storage. They often undergo physicochemical changes at this stage, such as the development of the Maillard reaction, primarily because of their intrinsic chemical properties, but also as a result of nonoptimal storage conditions. Components present at the particle surface are the first to be targeted by moisture and other environmental disruptions. Consequently, the identification, control, and prediction of particle surface components are useful to anticipate the effect of powder aging on product quality. Here, a new diafiltration method is proposed which fractionates proteins from a binary colloidal dispersion of 80% casein micelles and 20% whey proteins, according to their presence at the surface or core of the particle. This method shows that whey proteins are strongly enriched at the particle surface, whereas casein micelles are located at the core of the particles. This protocol also allows the identification of the rehydration kinetics for each rehydrated protein layer of the particle, revealing that 2 distinct forms of swelling occur: (1) a rapid swelling and elution of whey proteins present at the particle surface, and (2) a swelling of casein micelles located below the whey proteins, associated with a slow elution of casein micelles from the particles being rehydrated.

Semi-industrial production of a minimally processed infant formula powder using membrane filtration.

Infant formula (IF) is submitted to several heat treatments during production, which can lead to denaturation or aggregation of proteins and promote Maillard reaction. The objective of this study was to investigate innovative minimal processing routes for the production of first-age IF powder, thus ensuring microbial safety with minimal level of protein denaturation. Three nutritionally complete IF powders were produced at a semi-industrial scale based on ingredients obtained by fresh bovine milk microfiltration (0.8 and 0.1-µm pore size membranes). Low-temperature vacuum evaporation (50°C) and spray-drying (inlet and outlet temperatures of 160 and 70°C, respectively) were conducted to produce the T- formula with no additional heat treatment. The T+ formula was produced with a moderate heat treatment (75°C for 2 min) applied before spray-drying, whereas the T+++ formula received successive heat treatments (72°C for 30 s on the milk; 90°C for 2-3 s before evaporation; 85°C for 2 min before spray-drying), thus mimicking commercial powdered IF. Protein denaturation and Maillard reaction products were followed throughout the production steps and the physicochemical properties of the powders were characterized. The 3 IF powders presented satisfactory physical properties in terms of aw, free fat content, glass transition temperature, and solubility index, as well as satisfactory bacteriological quality with a total flora <103 cfu/g and an absence of pathogens when a high level of bacteriological quality of the ingredients was ensured. Protein denaturation occurred mostly during the heat treatments of T+ and T+++ and was limited during the spray-drying process. The IF powder produced without heat treatment (T-) presented a protein denaturation extent (6 ± 4%) significantly lower than that in T+++ (58 ± 0%), but not significantly different from that in T+ (10 ± 4%). Although T- tended to contain less Maillard reaction products than T+ and T+++, the Maillard reaction products did not significantly discriminate the infant formulas in the frame of this work. The present study demonstrated the feasibility of producing at a semi-industrial scale an infant formula being bacteriologically safe and containing a high content of native proteins. Application of a moderate heat treatment before spray-drying could further guarantee the microbiological quality of the IF powders while maintaining a low protein denaturation extent. This study opens up new avenues for the production of minimally processed IF powders.

Addition of calcium and magnesium chlorides as simple means of varying bound and precipitated minerals in casein micelle: Effect on enzymatic coagulation.

In casein micelle (CM), Ca is either precipitated in the colloidal calcium phosphate (CCP) stabilized by clusters of phosphoserine (SEP) residues, or is directly bound to SEP (or glutamic and aspartic acids) of caseins without inorganic phosphate. However, it is currently not possible to titrate separately the different micellar Ca forms, making it difficult to assess their respective importance for CM properties and behavior. Both Ca2+ and Mg2+ have the same binding constants with SEP. Moreover, MgHPO4 is more soluble than CaHPO4, and its natural concentration in milk is lower. Thus, upon addition of MgCl2, Mg is mainly exchanged with CM in the bound form, whereas upon addition of CaCl2, Ca is mainly exchanged in the precipitated form. Our objective was to assess the role of the 2 forms of micellar cations (bound and precipitated) during the enzymatic coagulation of cow milk. Magnesium chloride, CaCl2, or KCl (10 mM) were added to milk and pH was adjusted to 6.6 after overnight equilibration. The KCl-supplemented milk was a positive control to assess the effect of the increased ionic strength after MgCl2 and CaCl2 addition. Mineral partition between soluble and colloidal phases after salt addition was assessed both experimentally and by using computer simulation. Enzymatic coagulation was proceeded at 30°C. Hydrolysis of κ-casein was followed by the quantitative determination of caseinomacropeptide released by RP-HPLC, aggregation of para-κ-casein micelles was measured through the evolution of backscattering properties of milk and the gel time and gel firming kinetics were determined using a Chymograph (Chr. Hansen, Horsholm, Denmark). The KCl addition did not affect mineral partition. As expected, CaCl2 addition mainly increased the CCP content, whereas the addition of MgCl2 mainly increased the bound divalent cations content. The kinetics of κ-casein hydrolysis was slowed down after CaCl2 and MgCl2 addition, and was negatively correlated with the concentration of divalent cations in the soluble phase of milk. Aggregation and gel firming curves plotted versus the progress of κ-casein hydrolysis were similar for both CaCl2- and MgCl2-supplemented milk. In view of the dual-binding model for CM assembly, this means that both Ca forms reduce electronegative repulsions between para-micelles by specific charge shielding. Inclusion of 2 Ca forms in structural models for CM allows a more detailed comprehension of how mineral equilibria affect CM properties.

Mg2+ improves the thermotolerance of probiotic Lactobacillus rhamnosus GG, Lactobacillus casei Zhang and Lactobacillus plantarum P-8.

Food-related carbohydrates and proteins are often used as thermoprotectants for probiotic lactobacilli during industrial production and processing. However, the effect of inorganic salts is rarely reported. Magnesium is the second-most abundant cation in bacteria, and commonly found in various foods. Mg2+ homeostasis is important in Salmonella and has been reported to play a critical role in their thermotolerance. However, the role of Mg2+ in thermotolerance of other bacteria, in particular probiotic bacteria, still remains a hypothesis. In this study, the effect of Mg2+ on thermotolerance of probiotic lactobacilli was investigated in three well-documented probiotic strains, Lactobacillus rhamnosus GG, Lactobacillus casei Zhang and Lactobacillus plantarum P-8, in comparison with Zn2+ and Na+ . Concentrations of Mg2+ between 10 and 50 mmol l-1 were found to increase the bacterial survival upon heat challenge. Remarkably, Mg2+ addition at 20 mmol l-1 led to a 100-fold higher survival of L. rhamnosus GG upon heat challenge. This preliminary study also showed that Mg2+ shortened the heat-induced extended lag time of bacteria, which indicated the improvement in bacterial recovery from thermal injury. SIGNIFICANCE AND IMPACT OF THE STUDY: In order to improve the productivity and stability of live probiotics, extensive investigations have been carried out to improve thermotolerance of probiotics. However, most of these studies focused on the effects of carbohydrates, proteins or amino acids. The roles of inorganic salts in various food materials, which have rarely been reported, should be considered when incorporating probiotics into these foods. In this study, Mg2+ was found to play a significant role in the thermotolerance of probiotic lactobacilli. A novel strategy may be available in the near future by employing magnesium salts as protective agents of probiotics during manufacturing process.

Structural markers of the evolution of whey protein isolate powder during aging and effects on foaming properties.

The market for dairy powders, including high added-value products (e.g., infant formulas, protein isolates) has increased continuously over the past decade. However, the processing and storage of whey protein isolate (WPI) powders can result in changes in their structural and functional properties. It is therefore of great importance to understand the mechanisms and to identify the structural markers involved in the aging of WPI powders to control their end use properties. This study was performed to determine the effects of different storage conditions on protein lactosylations, protein denaturation in WPI, and in parallel on their foaming and interfacial properties. Six storage conditions involving different temperatures (θ) and water activities (aw) were studied for periods of up to 12mo. The results showed that for θ≤20°C, foaming properties of powders did not significantly differ from nonaged whey protein isolates (reference), regardless of the aw. On the other hand, powders presented significant levels of denaturation/aggregation and protein modification involving first protein lactosylation and then degradation of Maillard reaction products, resulting in a higher browning index compared with the reference, starting from the early stage of storage at 60°C. These changes resulted in a higher foam density and a slightly better foam stability (whisking) at 6mo. At 40°C, powders showed transitional evolution. The findings of this study will make it possible to define maximum storage durations and to recommend optimal storage conditions in accordance with WPI powder end-use properties.

Predicting the distribution of whey protein fouling in a plate heat exchanger using the kinetic parameters of the thermal denaturation reaction of ß-lactoglobulin and the bulk temperature profiles.

Fouling of plate heat exchangers (PHE) is a severe problem in the dairy industry, notably because the relationship between the build-up of protein fouling deposits and the chemical reactions taking place in the fouling solution has not yet been fully elucidated. Experiments were conducted at pilot scale in a corrugated PHE, and fouling deposits were generated using a model ß-lactoglobulin (ß-LG) fouling solution for which the ß-LG thermal denaturation reaction constants had been previously determined experimentally. Then 18 different bulk temperature profiles within the PHE were imposed. Analysis of the fouling runs shows that the dry deposit mass per channel versus the ratio R=kunf/kagg (with kunf and kagg representing, respectively, the unfolding and aggregation rate constants computed from both the identification of the ß-LG thermal denaturation process and knowledge of the imposed bulk temperature profile into the PHE channel) is able to gather reasonably well the experimental fouling mass data into a unique master curve. This type of representation of the results clearly shows that the heat-induced reactions (unfolding and aggregation) of the various ß-LG molecular species in the bulk fluid are essential to capture the trend of the fouling mass distribution inside a PHE. This investigation also illustrates unambiguously that the release of the unfolded ß-LG (also called ß-LG molten globule) within the bulk fluid (and the absence of its consumption in the form of aggregates) is a key phenomenon that controls the extent of protein fouling as well as its location inside the PHE.

Physicochemical characterization of mozzarella cheese wheys and stretchwaters in comparison with several other sweet wheys.

To better understand the origins of the problems occurring during mozzarella cheese whey concentration, lactose crystallization, and spray-drying steps, a physicochemical characterization was achieved. For this purpose, mozzarella cheese wheys were sampled and their content in different compounds such as total nitrogen, noncasein nitrogen, nonprotein nitrogen, lactate, citrate, chloride, sulfate, phosphate anions, calcium, magnesium, potassium, sodium cations, and the sugars glucose and galactose were measured. In a second step, the results were compared with the corresponding content in cheddar cheese wheys, raclette cheese wheys, soft cheese wheys, and Swiss-type cheese wheys. At the end of this survey, it was shown that mozzarella cheese wheys were more concentrated in lactate and in minerals--especially phosphate, calcium, and magnesium--than the other cheese wheys and that they contained galactose. These constituents are known to be hygroscopic. Complementary surveys are now necessary to compare the hygroscopicity of galactose and lactate and discover whether the amounts of these compounds found in mozzarella cheese wheys are a factor in the problems encountered during the concentration, lactose crystallization, and spray-drying steps.

Fat properties during homogenization, spray-drying, and storage affect the physical properties of dairy powders.

Changes in fat properties were studied before, during, and after the drying process (including during storage) to determine the consequences on powder physical properties. Several methods were combined to characterize changes in fat structure and thermal properties as well as the physical properties of powders. Emulsion droplet size and droplet aggregation depended on the homogenizing pressures and were also affected by spray atomization. Aggregation was usually greater after spray atomization, resulting in greater viscosities. These processes did not have the same consequences on the stability of fat in the powders. The quantification of free fat is a pertinent indicator of fat instability in the powders. Confocal laser scanning microscopy permitted the characterization of the structure of fat in situ in the powders. Powders from unhomogenized emulsions showed greater free fat content. Surface fat was always overrepresented, regardless of the composition and process parameters. Differential scanning calorimetry melting experiments showed that fat was partially crystallized in situ in the powders stored at 20 degrees C, and that it was unstable on a molecular scale. Thermal profiles were also related to the supramolecular structure of fat in the powder particle matrix. Powder physical properties depended on both composition and process conditions. The free fat content seemed to have a greater influence than surface fat on powder physical properties, except for wettability. This study clearly showed that an understanding of fat behavior is essential for controlling and improving the physical properties of fat-filled dairy powders and their overall quality.

Calcium citrate insolubilization drives the fouling of falling film evaporators during the concentration of hydrochloric acid whey.

When dairy powders are produced, the mineral fraction undergoes strong modifications during the vacuum concentration step, leading to the fouling of falling film evaporators. The objective of this study was to determine the nature of the deposits formed during the vacuum concentration of two fouling and highly mineralized products: hydrochloric acid whey and lactic acid whey. These products mainly differ in terms of their mineral composition: lactic acid whey contains a high level of lactic acid and traces of citrate, whereas hydrochloric acid whey contains citrate and no lactic acid. Concentrates at different concentration factors were produced using a pilot-scale falling film evaporator. The compositions of the fouling deposits as well as the precipitates present in the concentrates were deduced from the analytical determination of the composition of the concentrates and their respective diffusible phases. The behavior of the mineral fraction of both acid wheys during concentration was shown to be very different. In the case of hydrochloric acid whey, experimental results suggested a deposition of calcium and citrate ions in the evaporator as well as their precipitation in the highly concentrated products. On the contrary, neither mineral deposition nor precipitation occurred during the concentration of lactic acid whey. This study underlined the key role of citrate ions in the fouling of evaporators during the concentration of hydrochloric acid wheys.

High intensity pulsed electric fields applied to egg white: effect on Salmonella Enteritidis inactivation and protein denaturation.

High-intensity electric fields have been successfully applied to the destruction of Salmonella Enteritidis in diaultrafiltered egg white. The effects of electric field strength (from 20 to 35 kV x cm(-1)), pulse frequency (from 100 to 900 Hz), pulse number (from 2 to 8), temperature (from 4 to 30 degrees C), pH (from 7 to 9), and inoculum size (from 10(3) to 10(7) CFU x ml(-1)) were tested through a multifactorial experimental design. Experimental results indicate that, for Salmonella inactivation, the electric field intensity is the dominant factor with a strongly positive effect, strengthened by its positive interaction with pulse number. Pulse number, temperature, and pH have also significant positive effects but to a lesser extent. In the most efficient conditions, the pulsed electric field (PEF) treatment is capable of 3.5 log10 reduction in viable salmonellae. Simultaneously, the measure of surface hydrophobicity does not indicate any increase after PEF treatment. These results suggest that no protein denaturation occurs, unlike what is observed after comparable heat treatment in terms of Salmonella inactivation (55 degrees C for 15 min).

Advances in food powder agglomeration engineering.

Food powders are used in everyday life in many ways and offer technological solutions to the problem of food production. The natural origin of food powders, diversity in their chemical composition, variability of the raw materials, heterogeneity of the native structures, and physicochemical reactivity under hydrothermal stresses contribute to the complexity in their behavior. Food powder agglomeration has recently been considered according to a multiscale approach, which is followed in the chapter layout: (i) at the particle scale, by a presentation of particle properties and surface reactivity in connection with the agglomeration mechanisms, (ii) at the mechanisms scale, by describing the structuration dynamics of agglomerates, (iii) at the process scale, by a presentation of agglomeration technologies and sensors and by studying the stress transmission mode in the powder bed, and finally (iv) by an integration of the acquired knowledge, thanks to a dimensional analysis carried out at each scale.

How surface composition of high milk proteins powders is influenced by spray-drying temperature.

High milk proteins powders are common ingredients in many food products. The surface composition of these powders is expected to play an essential role during their storage, handling and/or final application. Therefore, an eventual control of the surface composition by modifying the spray-drying temperature could be very useful in the improvement of powder quality and the development of new applications. For this purpose, the influence of five spray-drying temperatures upon the surface composition of the powders was investigated by X-ray photoelectron spectroscopy. The major milk proteins were studied: native micellar casein and native whey, both more or less enriched in lactose. The results show a surface enrichment in lipids for all the powders and in proteins for many powders. Whatever the drying temperature, lipids and proteins are preferentially located near the surface whereas lactose is found in the core. This surface enrichment is also highly affected by the spray-drying temperature. More lipids, more proteins and less lactose are systematically observed at the surface of powders spray-dried at lower outlet air temperatures. The nature of proteins is also found essential; surface enrichment in lipids being much stronger for whey proteins containing powders than for casein containing powders. Additionally, we found a direct correlation between the lipids surface concentration and the wetting ability for the 25 powders studied.