Influence of Heating Temperature and pH on Acid Gelation of Micellar Calcium Phosphate-Adjusted Skim Milk.

Micellar calcium phosphate (MCP) plays an important role in maintaining the structure and stability of the casein micelle and its properties during processing. The objective of this study was to investigate how heating (10 min at 80 or 90 °C) at different pH levels (6.3, 6.6, 6.9, or 7.2) impacted the acid-induced gelation of MCP-adjusted milk, containing 67 (MCP67), 100 (MCP100), or 113 (MCP113) % of the original MCP content. The unheated sample MCP100 at pH 6.6 was considered the control. pH acidification to pH 4.5 at 30 °C was achieved with glucono delta-lactone while monitoring viscoelastic behaviour by small-amplitude oscillatory rheology. The partitioning of calcium and proteins between colloidal and soluble phases was also examined. In MCP-depleted skim milk samples, the concentrations of non-sedimentable caseins and whey proteins were higher compared to the control and MCP-enriched skim milk samples. The influence of MCP adjustment on gelation was dependent on pH. Acid gels from sample MCP67 exhibited the highest storage modulus (G'). At other pH levels, MCP100 resulted in the greatest G'. The pH of MCP-adjusted skim milk also impacted the gel properties after heating. Overall, this study highlights the substantial impact of MCP content on the acid gelation of milk, with a pronounced dependency of the MCP adjustment effect on pH variations.

Effect of pH and Shear on Heat-Induced Changes in Milk Protein Concentrate Suspensions.

The effect of shear on heat-induced changes in milk protein concentrate suspensions was examined at different pH levels, revealing novel insights into micellar dissociation and protein aggregation dynamics. Milk protein concentrate suspensions, adjusted to pH of 6.1, 6.4, 6.8, or 7.5, underwent combined heat (90 °C for 5 min or 121 °C for 2.6 min) and shear (0, 100, or 1000 s-1) treatment. The fragmentation of protein aggregates induced by shear was evident in the control MPC suspensions at pH 6.8, irrespective of the temperature. At pH 7.5, shear increased the heat-induced micellar dissociation. This effect was particularly pronounced at 121 °C and 1000 s-1, resulting in reduced particle size and an elevated concentration of κ-casein (κ-CN) in the non-sedimentable phase. At pH 6.1 or 6.4, shear effects were dependent on sample pH, thereby modifying electrostatic interactions and the extent of whey protein association with the micelles. At pH 6.1, shear promoted heat-induced aggregation, evidenced by an increase in particle size and a significant decline in both whey proteins and caseins in the non-sedimentable phase. At pH 6.4, shear-induced fragmentation of aggregates was observed, prominently due to comparatively higher electrostatic repulsions and fewer protein interactions. The influence of shear on heat-induced changes was considerably impacted by initial pH.

Effect of Protein Content on Heat Stability of Reconstituted Milk Protein Concentrate under Controlled Shearing.

Milk protein concentrates (MPCs) possess significant potential for diverse applications in the food industry. However, their heat stability may be a limitation to achieving optimal functional performance. Shearing, an inherent process in food manufacturing, can also influence the functionality of proteins. The aim of this research was to examine the heat stability of reconstituted MPCs prepared at two protein concentrations (4% and 8% w/w protein) when subjected to varying levels of shearing (100, 1000, or 1500 s-1) during heating at 90 °C for 5 min or 121 °C for 2.6 min. While the impact of shear was relatively minor at 4% protein, it was more pronounced in 8% protein MPC suspensions, leading to a considerable decline in heat stability. An increase in protein concentration to 8% amplified protein interactions, intensified by shearing. This, in turn, resulted in comparatively higher aggregation at elevated temperatures and subsequently reduced the heat stability of the reconstituted MPCs.

Structural Properties of Casein Micelles with Adjusted Micellar Calcium Phosphate Content.

Micellar calcium phosphate (MCP) content of skim milk was modified by pH adjustment followed by dialysis. Turbidity, casein micelle size and partitioning of Ca and caseins between the colloidal and soluble phases of milk were determined. Protein structure was characterised by Fourier transform infrared (FTIR) spectroscopy and proton nuclear magnetic resonance (1H NMR), whereas organic and inorganic phosphorus were studied by phosphorus-31 nuclear magnetic resonance (31P NMR). The sample with the lowest MCP content (MCP7) exhibited the smallest particle size and turbidity, measuring 83 ± 8 nm and 0.08 ± 0.01 cm-1, respectively. Concentrations of soluble caseins increased with decreasing MCP levels. At ~60% MCP removal, FTIR analysis indicated a critical stage of structural rearrangement and 31P NMR analysis showed an increase in signal intensity for Ca-free Ser-P, which further increased as MCP concentration was further reduced. In conclusion, this study highlighted the importance of MCP in maintaining micellar structure and its impact on the integrity of casein micelle.

Unraveling the Influences of Sodium, Potassium, Magnesium, and Calcium on the Crystallization Behavior of Lactose.

The inability of lactose to properly crystallize due to the presence of high amounts of salts poses significant hurdles for its downstream processing with some dairy waste streams such as acid whey. This study aimed to investigate the physicochemical and thermal behaviors of lactose in the presence of cations commonly present in acid whey. A model-based study was conducted, utilizing various cations (Mg, Ca, K, and Na) at concentrations (8, 30, 38, and 22 mM, respectively) that are typically found in acid whey. The research experiments were conducted using a factorial design. The thermal analysis of concentrated solutions revealed augmentation in the enthalpy of water evaporation in the presence of individual cations and their combinations in comparison with pure lactose (698.4 J/g). The degree of enthalpy increased following the order of Na+ (918.6 J/g), K+ (936.6 J/g), Mg2+ (987.0 J/g), Ca2+ (993.2 J/g), and their mixture (1005.4 J/g). This resulted in a substantial crystal yield decline in the exactly reversed order to that of the enthalpy. The greatest decline was observed in the presence of the salt mixture (63%) followed by Ca (67%) compared with pure lactose (79%). The yield reduction was also inversely related to the solubility of lactose. The presence of divalent cations appeared to play a role in the isomerization of lactose molecules observed using DSC and XRD diffractograms according to the disappearance of peaks related to ß lactose. The effect of salts on the crystallization of lactose was a combination of cation-lactose interactions, changes in the solubility of lactose, ion-dipole interactions between water and cations, and changes in the structure of water molecules. By deviating the composition of acid whey, the crystallization of lactose can be enhanced, leading to the improved downstream processing of acid whey.

Heat-Induced Changes in κ-Carrageenan-Containing Chocolate-Flavoured Milk Protein Concentrate Suspensions under Controlled Shearing.

Milk protein dispersions containing added cocoa powder (1.5% (w/w)) and sucrose (7% (w/w)) and varying levels of κ-carrageenan (0.01, 0.03, or 0.05% w/w) were subjected to combined heat treatment (90 °C/5 min or 121 °C/2.6 min) and shear (100 or 1000 s-1) to investigate the heat stability of milk proteins. The application of shear led to a notable reduction in non-sedimentable proteins, resulting in an increase in the average particle size and apparent viscosity of the dispersions, particularly at high concentrations of k-carrageenan and elevated temperatures. This indicates that shear forces induced prominent protein aggregation, especially at higher κ-carrageenan concentrations. This aggregation was primarily attributed to the destabilisation of micelles and presence of loosely bound caseins within the κ-carrageenan network, which exhibited increased susceptibility to aggregation as collision frequencies increased due to shear.

Influence of pH on Heat-Induced Changes in Skim Milk Containing Various Levels of Micellar Calcium Phosphate.

The present study investigated the effect of micellar calcium phosphate (MCP) content and pH of skim milk on heat-induced changes in skim milk. Four MCP-adjusted samples, ranging from 67 to 113% of the original MCP content, were heated (90 °C for 10 min) at different pH values (6.3, 6.6, 6.9, and 7.2), followed by determining changes in particle size, turbidity, protein distribution, and structure. The results demonstrate a strong effect of MCP level and pH on heat-induced changes in milk, with the MCP67 samples revealing the greatest thermal stability. Specifically, decreasing MCP content by 33% (MCP67) led to a smaller increase in non-sedimentable κ-casein and a lower decrease in αs2-casein concentrations after heating compared to other samples. Lower MCP content resulted in a moderate rise in the average particle size and turbidity, along with lower loading of ß-turn structural component after heating at low pH (pH 6.3). Notably, MCP113 exhibited instability upon heating, with increased particle size, turbidity, and a significant decrease in non-sedimentable αs2-casein concentration, along with a slight increase in non-sedimentable κ-casein concentration. The FTIR results also revealed higher loading of intermolecular ß-sheet, ß-turn, and random coil structures, as well as lower loading of α-helix and ß-sheet structures in MCP-enhanced skim milk samples. This suggests significant changes in the secondary structure of milk protein and greater formation of larger aggregates.

Influence of Actinidin-Induced Hydrolysis on the Functional Properties of Milk Protein and Whey Protein Concentrates.

The main aim of the study was to establish the impact of limited proteolysis by actinidin on the functionality of selected milk protein systems. The plant protease actinidin was used to produce hydrolysates (MPHs) from milk protein concentrate (MPC) and whey protein concentrate (WPC) to 0, 5, 10 or 15% of the degree of hydrolysis (DH) at an enzyme-to-substrate ratio of 1:100 (5.21 units of actinidin activity g-1 of protein). The functionalities assessed included solubility, heat stability, emulsification and foaming properties. In general, significant changes in the functionalities of MPH were associated with the extent of hydrolysis. Solubility of hydrolysates increased with increasing %DH, with WPC showing about 97% solubility at 15% DH. Emulsifying properties were negatively affected by hydrolysis, whereas heat stability was improved in the case of WPC (~25% of heat stability increased with an increase in DH to 15%). Hydrolysates from both WPC and MPC had improved foaming properties in comparison to unhydrolysed controls. These results were also supported by changes in the FTIR spectra. Further adjustment of hydrolysis parameters, processing conditions and pH control could be a promising approach to manipulate selected functionalities of MPHs obtained using actinidin.

Milk Protein Hydrolysis by Actinidin-Kinetic and Thermodynamic Characterisation and Comparison to Bromelain and Papain.

Plant proteases, including actinidin, papain and bromelain, have been widely used in the food industry but with limited application in dairy systems. This research aimed to establish and compare operational parameters (kinetics, temperature, enzyme type, time and thermodynamics) relevant to the applications of these enzymes in the hydrolysis of whey protein isolates (WPI), whey protein concentrates (WPC) or milk protein concentrates (MPC). The degree of hydrolysis (DH) increased with the rise in temperature, and the maximum DH was achieved at 60 °C for all three dairy systems. The addition of papain resulted in a greater %DH of whey proteins in comparison to bromelain. The cleavage of proteins was clearly time-dependent (p < 0.05), while the pH did not change significantly (p > 0.05) during this time. PAGE analysis revealed that all three enzymes mainly acted on α-lactalbumin and αs-casein in WPI and MPC, respectively. Kinetic parameters from the Lineweaver-Burk plot at 60 °C using WPC and MPC as a substrate varied widely, establishing that WPC hydrolysis was characterised by a lower KM, higher kcat, kcat/KM and Vmax compared to MPC in the case of all three enzymes. The difference in kcat/KM values amongst all enzymes (actinidin > papain > bromelain) indicated the difference in the strength of substrate binding sites. The thermodynamic parameters of these enzymes with MPC and WPC were also determined at a temperature range of 15-60 °C, and the results indicate the potential application of papain and actinidin in the dairy industry.

What is the impact of amino acid mutations in the primary structure of caseins on the composition and functionality of milk and dairy products?

The impact of amino acid mutations within the peptide structure of bovine milk protein is important to understand as it can effect processability and subsequently effect its physiological properties. Genetic polymorphisms of bovine caseins can influence the chemical, structural, and technological properties, including casein micelle morphology, calcium distribution, network creation upon gelation, and surface activity. The A1 and A2 genetic variants of ß-casein have recently acquired growing attention from both academia and industry, prompting new developments in the area. The difference between these two genetic variants is the inclusion of either proline in ß-casein A2 or histidine in ß-casein A1 at position 67 in the peptide chain. The aim of this review was to examine the extent to which milk and ingredient functionality is influenced by ß-casein phenotype. One of the main findings of this review was although ß-casein A1 was found to be the dominant variant in milks with superior acid gelation and rennet coagulation properties, milks comprised of ß-casein A2 possessed greater emulsion and foam formation capabilities. The difference in the casein micelle assembly, hydrophobicity, and chaperone activity of caseins may explain the contrast in the functionality of milks containing ß-casein from either A1 or A2 families. This review provides new insights into the subtle variations in the physicochemical properties of bovine milks, which could potentially support dairy producers in the development of new dairy products with different functional properties.

The effect of malting on phenolic compounds and radical scavenging activity in grains and breakfast cereals.

Breakfast cereals are popular grain foods and sources of polyphenols. Malting alters polyphenol content and activity; however, effects are varied. The total polyphenol content (TPC), radical scavenging activity (RSA), and polyphenol profile were analyzed in unmalted and malted grains (wheat, barley, and sorghum) and breakfast cereals (wheat, barley) by Folin Ciocalteu Reagent (FCR), % inhibition of the free radical 2,2-diphenyl-1-picryl-hydrazyl, and high performance liquid chromatography. Higher TPC was observed in all malted grains and breakfast cereals compared with unmalted samples (p < 0.05). Higher RSA was also observed in all malted samples compared to unmalted samples (p < 0.05) except for wheat grain to malted wheat grain. In this study, malting induced additional polyphenols and antioxidant activity in grains and cereal products. Malted grain breakfast cereals may be practical sources of polyphenol antioxidants. PRACTICAL APPLICATION: This study utilized malting in a unique way to investigate potential health benefits of polyphenols and antioxidant activity in grains (wheat, barley, and sorghum) and ready-to-eat breakfast cereals (wheat and barley). This study found that grains and breakfast cereals are important sources of antioxidant polyphenols, and these were significantly increased in malted varieties. Understanding this is important as grains and breakfast cereals are widely consumed staple foods. Consuming healthier grain products may be a practical strategy in reducing the risk of noncommunicable diseases such as colorectal cancer and type-2 diabetes, where wholegrain consumption may be important in prevention.

Properties of sodium caseinate as affected by the ß-casein phenotypes.

The aim of the study was to investigate the properties of sodium caseinate dispersions and oil-in-water emulsions obtained from cows' milk of either A1/A1, A1/A2, or A2/A2 ß-casein phenotype. Protein structural characterisation was examined using Fourier Transform Infrared and Nuclear Magnetic Resonance spectroscopies, with physicochemical and interfacial properties assessed by analysing adsorbed protein content, hydrophobicity, solubility, and emulsion stability of the samples. Results showed variations in the secondary structure of all samples dependent of the presence of A1 or A2 ß-caseins. The main differences included greater amounts of α-helix and ß-sheet in A1/A1 and A1/A2 sodium caseinate dispersions that influenced their lower solubility, while random coils/polyproline II helixes were found only in A2/A2 sodium caseinate dispersion. In contrast, upon adsorption on the interface of A2/A2 sodium caseinate emulsion, the protein adopted ordered conformational motifs. This conformational shift supposedly arose from structural differences between the two ß-casein proteoforms, which most likely enhanced the emulsion properties of A2/A2 sodium caseinate compared to either A1/A1 or A1/A2 sodium caseinates. The A2 ß-casein in both, A1/A2 and A2/A2 sodium caseinates, appears to be able to more rapidly reach the oil droplet surface and was more efficient as emulsifying agent. The current results demonstrated that the conformational rearrangement of proteins upon adsorption to emulsion interfaces was dependent not only on hydrophobicity and on solubility, but also on the conformational flexibility of A1/A1, A1/A2, and A2/A2 ß-casein phenotypes. These findings can assist in predicting the behaviour of sodium caseinates during relevant industrial processing.

Anti-salmonella properties of kefir yeast isolates: An in vitro screening for potential infection control.

The rise of antibiotic resistance has increased the need for alternative ways of preventing and treating enteropathogenic bacterial infection. Various probiotic bacteria have been used in animal and human. However, Saccharomyces boulardii is the only yeast currently used in humans as probiotic. There is scarce research conducted on yeast species commonly found in kefir despite its claimed potential preventative and curative effects. This work focused on adhesion properties, and antibacterial metabolites produced by Kluyveromyces lactis and Saccharomyces unisporus isolated from traditional kefir grains compared to Saccharomyces boulardii strains. Adhesion and sedimentation assay, slide agglutination, microscopy and turbidimetry assay were used to analyze adhesion of Salmonella Arizonae and Salmonella Typhimurium onto yeast cells. Salmonella growth inhibition due to the antimicrobial metabolites produced by yeasts in killer toxin medium was analyzed by slab on the lawn, turbidimetry, tube dilution and solid agar plating assays. Alcohol and antimicrobial proteins production by yeasts in killer toxin medium were analyzed using gas chromatography and shotgun proteomics, respectively. Salmonella adhered onto viable and non-viable yeast isolates cell wall. Adhesion was visualized using scanning electron microscope. Yeasts-fermented killer toxin medium showed Salmonella growth inhibition. The highest alcohol concentration detected was 1.55%, and proteins with known antimicrobial properties including cathelicidin, xanthine dehydrogenase, mucin-1, lactadherin, lactoperoxidase, serum amyloid A protein and lactotransferrin were detected in yeasts fermented killer medium. These proteins are suggested to be responsible for the observed growth inhibition effect of yeasts-fermented killer toxin medium. Kluyveromyces lactis and Saccharomyces unisporus have anti-salmonella effect comparable to Saccharomyces boulardii strains, and therefore have potential to control Salmonella infection.

Structural Changes of ß-Casein Induced by Temperature and pH Analysed by Nuclear Magnetic Resonance, Fourier-Transform Infrared Spectroscopy, and Chemometrics.

This study investigated structural changes in ß-casein as a function of temperature (4 and 20 °C) and pH (5.9 and 7.0). For this purpose, nuclear magnetic resonance (NMR) and Fourier-transform infrared (FTIR) spectroscopy were used, in conjunction with chemometric analysis. Both temperature and pH had strongly affected the secondary structure of ß-casein, with most affected regions involving random coils and α-helical structures. The α-helical structures showed great pH sensitivity by decreasing at 20 °C and diminishing completely at 4 °C when pH was increased from 5.9 to 7.0. The decrease in α-helix was likely related to the greater presence of random coils at pH 7.0, which was not observed at pH 5.9 at either temperature. The changes in secondary structure components were linked to decreased hydrophobic interactions at lower temperature and increasing pH. The most prominent change of the α-helix took place when the pH was adjusted to 7.0 and the temperature set at 4 °C, which confirms the disruption of the hydrogen bonds and weakening of hydrophobic interactions in the system. The findings can assist in establishing the structural behaviour of the ß-casein under conditions that apply as important for solubility and production of ß-casein.

Effects of pressurized thermal processing on native proteins of raw skim milk and its concentrate.

Heating, pressurization, and shearing can modify native milk proteins. The effects of pressurized heating (0.5 vs. 10 MPa at 75 or 95°C) with shearing (1,000 s-1) on proteins of raw bovine skim milk (SM, ∼9% total solids) and concentrated raw skim milk (CSM, ∼22% total solids) was investigated. The effects of evaporative concentration at 55°C and pressurized shearing (10 MPa, 1,000 s-1) at 20°C were also examined. Evaporative concentration of SM resulted in destabilization of casein micelles and dissociation of αS1- and ß-casein, rendering CSM prone to further reactions. Treatment at 10 MPa and 1,000 s-1 at 20°C caused substantial dissociation of αS1- and ß-casein in SM and CSM, with some dissociated caseins forming shear-induced soluble aggregates in CSM. The pressure applied at 10 MPa induced compression of the micelles and their dissociation in SM and CSM at 75 or 95°C, resulting in reduction of the micelle size. However, 10 MPa did not alter the mineral balance or whey proteins denaturation largely, except by reduction of some ß-sheets and α-helices, due to heat-induced conformational changes at 75 and 95°C.

Denaturation of selected bioactive whey proteins during pasteurization and their ability to modulate milk immunogenicity.

This research communication relates to the hypothesis that the consumption of raw or unprocessed cow's milk contributes to lowered prevalence of allergies. Thermal pasteurization of bovine milk can result in denaturation of minor whey proteins and loss of their bioactivity. Denaturation of bovine serum albumin (BSA), immunoglobulin G (IgG) and lactoferrin (LF) in skim milk was studied under different temperature (72, 75 or 78°C) and time (0-300 s) combinations. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) results revealed that denaturation of all 3 proteins occurred at 72°C and progressed with increase in temperature and holding time. About 59% of LF and 12% of IgG denatured under high-temperature short-time (72°C/ 15 s) pasteurization, while BSA was least impacted. To assess modulation of milk immunogenicity, secretion of selected T helper (Th)-type cytokines by human peripheral blood mononuclear cells (PBMCs) was studied in vitro in response to different concentrations of BSA (0.4-1.0 mg/ml) and IgG (0.8-1.6 mg/ml) in unheated skim milk. Addition of IgG at 1.6 mg/ml induced a prominent Th1-skewed cytokine profile that may not trigger a Th2-skewed allergic reaction. BSA did not appear to modulate milk immunogenicity to any significant extent.

Unravelling Conformational Aspects of Milk Protein Structure-Contributions from Nuclear Magnetic Resonance Studies.

Changes in the molecular structure and association of milk proteins lead to many desirable (under controlled conditions) or undesirable characteristics of dairy products. Several methods have been used to study the structure of milk proteins and changes therein in different environments. Whey proteins are an excellent model for secondary structure studies using circular dichroism (CD), Fourier-transform infrared spectroscopy (FTIR) and tertiary structure studies using X-ray crystallography and nuclear magnetic resonance (NMR). However, caseins, the most abundant protein class in milk, are far more difficult to characterize. The tertiary structure of caseins cannot be observed by X-ray crystallography due to the inability to crystallize caseins. However, NMR is an appropriate approach for structural elucidation. Thus far, NMR was applied on specific peptides of individual caseins of the molecules including phosphoserine centers and colloidal calcium phosphate. The literature focuses on these parts of the molecule due to its importance in building the sub-unit particles involving individual caseins and calcium phosphate nanoclusters. This review focuses on present structural studies of milk proteins using NMR and their importance in dairy processing.

Impact of controlled shearing on solubility and heat stability of pea protein isolate dispersed in solutions with adjusted ionic strength.

Pea protein isolates (PPI) have sustained an increasing demand in the food industry as a substitute for animal-origin proteins. Shearing is an integral part of food processing that can change properties of proteins and their functionality. PPI dispersions prepared at 4 or 8% concentration (w/w protein), pH 6.8 or 7.5 and under ionic strength (IS) 100, 200 mM or non-adjusted, were subjected to controlled shearing at two levels (100 or 1500 s-1) during heating at 90 °C for 5 min. All main factors had substantial effects on the tested dependent variables. Shearing at 1500 s-1 significantly improved the solubility and heat stability of 4% PPI at pH 6.8 or 7.5 and IS-100 or 200 mM by 27-43% in comparison to 100 s-1. Following 1500 s-1 treatment, all PPI dispersions showed >85% solubility and heat stability except 4% PPI at pH 6.8 and IS 100 mM (60%). Shearing appeared to alter structural and physicochemical properties of pea proteins as well nature of protein aggregation. Heating accompanied with 100 s-1 shearing mostly resulted in insoluble covalent aggregates while shearing at 1500 s-1 mainly contributed to formation of soluble hydrophobic aggregates.

Influence of lactic, citric and phosphoric acids on the properties of concentrated lactose solutions.

Physicochemical and thermal characteristics of concentrated lactose solutions containing 0.05, 1, or 4% (w/w) of three acids commonly used in the food industry, i.e., lactic, citric, and phosphoric acid, were studied. Properties of both lactose and water were affected by all acids studied. Thermographic analysis showed that interactions between water and acids hindered evaporation of water from most of lactose solutions. This effect was mostly related to the formation of a strong hydration layer around lactose molecules by hydrogen bonds. Acid-induced hydrolysis of lactose into glucose and galactose varied depending on the concentration, hydrolytic power of acids and molecular interactions in the system. The study concluded that the varying physical, chemical, structural and thermal characteristics of lactose as affected by the presence of different acids was mainly due to the manipulation of water-lactose interactions, whereas the hydrolysis of lactose by the acids plays a smaller role.

Altering allergenicity of cow's milk by food processing for applications in infant formula.

Cow's milk-based infant formulas have a long tradition in infant nutrition, although some infants are unable to use them due to presence of several known allergens. Various processing methods have been identified capable of reducing cow's milk protein allergenicity including thermal and non-thermal methods and their combinations. Heat treatment and enzymatic hydrolysis have been in production of hypoallergenic infant formulas. However, modulation of allergenic epitopes depends on the extent of heat treatment applied, which consequently may also reduce a nutritional value of these proteins. In addition, enzymatic hydrolysis may not target allergenic epitopes thus allergenicity may persist; however released peptides may have detrimental impact on taste and functional properties of final products. Modulation of allergenicity of milk proteins appears to require a concerted effort to minimize detrimental effects as clinical studies conducted on commercial hypoallergenic formulas demonstrated persistence of allergic symptoms. This article covers traditional and novel processing methods and their impact on reduction of cow's milk allergenicity in milk-based infant formulas.