Composition of volatile compounds in bovine milk heat treated by instant infusion pasteurisation and their correlation to sensory analysis

Volatile compounds in skim milk and nonstandardised milk subjected to instant infusion pasteurisation at 80°C, 100°C and 120°C were compared with raw milk, high temperature short time pasteurised milk and milk pasteurised at 85°C/30 s. The composition of volatile compounds differed between infusion pasteurisation treated samples and the reference pasteurisations. The sensory properties of skim milk subjected to instant infusion pasteurisation were described by negative attributes, such as cardboard sour and plastic flavours, which are not associated normally with fresh milk. Partial least squares modelling showed good correlation between the volatile compounds and the sensory properties, indicating the predictive and possible causal importance of the volatile compounds for the sensory characteristics.     

Kinetics of alkaline phosphatase and lactoperoxidase inactivation,and of beta-lactoglobulin denaturation in milk with different fat content

In the context of identifying intrinsic time temperature integrators (TTIs) for evaluating heat processing of milk, the extent to which milk fat content has an effect on alkaline phosphatase (ALP) and lactoperoxidase (Lpo) inactivation and on beta-lactoglobulin (beta-Ig) denaturation kinetics was studied. Inactivation and denaturation kinetics were analysed in whole, semi-skimmed and skimmed milk. In previous experiments (isothermal and non-isothermal heating conditions), heat inactivation of ALP and Lpo and heat denaturation of beta-Ig were found to follow first order kinetics. This allowed experimental design to be simplified. Data analysis was performed by non-linear regression and results were evaluated by construction of joint confidence regions. The possible effect of milk fat was illustrated by temperature time tolerance (TTT-) diagrams. Although initial ALP activity was lower in skimmed milk compared with semi-skimmed or whole milk, kinetics were comparable and fat content did not seem to substantially affect the ALP test result for pasteurized milk. Unlike ALP, Lpo inactivation and beta-Ig denaturation kinetics differed significantly in milk with different fat content. Differences between Lpo inactivation kinetics were relatively small and acceptable in the context of quantifying the process impact. Denaturation of beta-Ig, on the other hand, seemed to be enhanced at higher milk fat content (> 72 degrees C).     

Effect of Lenient Steam Injection (LSI) heat treatment of bovine milk on the activities of some enzymes,the milk fat globule and pH

This study investigated the effects of Lenient Steam Injection (LSI) treatment at temperatures 70–150 °C on the enzymatic activities of the indigenous milk enzymes alkaline phosphatase, lactoperoxidase (LPO), xanthine oxidase (XO), lipoprotein lipase (LPL) and plasmin in comparison with two reference heat treatments of 63 °C for 30 s and of 72 °C for 15 s by indirect heating. Milk fat globule (MFG) size distributions and pH were also monitored. Alkaline phosphatase, LPO, XO and LPL activities decreased with increasing LSI temperature. Plasmin activity was increased at temperatures <80 °C and decreased at temperatures above 90 °C. Milk fat globule homogenisation was observed at temperatures above 110–130 °C.     

The use of sedimentation field-flow fractionation in the size characterization of bovine milk fat globules as affected by heat treatment

Size distribution of fat globules affects the appearance, taste and stability of milk and milk-based products. Full-fat, semi-fat and chocolate bovine milk were subjected to heat treatment within a temperature range of 50–125 °C for 1 h. Sedimentation field-flow fractionation was employed to determine the changes in mean particle diameter of milk fat globules as affected by heat treatment. The mean particle diameter of fat droplets increased with increasing heating temperature for most samples. The particle size of fat globules increased on average 40 nm (4.65%) for full-fat and 72 nm (8.52%) for semi-fat milk following the heat treatment (50–125 °C). Chocolate milk exhibited considerable increase in particle size (104 nm, 12.53%) within a certain temperature range (50–110 °C), followed by a decrease in particle size when heated at 125 °C for 1 h. Heat-induced flocculation due to attractive interactions between hydrophobic sites on denatured protein molecules on different droplets was assumed to be mainly responsible for the increases in particle size observed in this study. Extensive heat-induced denaturation of milk proteins was also indicated by Native PAGE. Sedimentation field-flow fractionation proved to be a useful technique for adequately monitoring heat-induced changes in particle size distributions in milk.     

Instant infusion pasteurisation of bovine milk. I. Effects on bacterial inactivation and physical–chemical properties

Two types of milk, skim milk and non-standardised raw milk, were heat treated using direct heating by instant infusion pasteurisation with treatment temperatures in the range from 72°C to 120°C and with holding times of less than 1 second. Indirect heating by HTST pasteurisation (72°C for 15 seconds) was used for comparison. The inactivation of microorganisms reached at least the same level when using instant infusion pasteurisation compared to HTST pasteurisation. Changes in the physical-chemical properties were observed in the skim milk fractions of instant infusion pasteurised non-standardised milk, whereas for instant infusion pasteurised skim milk less influence from the treatments was observed.     

Cheese made from instant infusion pasteurized milk: Rennet coagulation,cheese composition,texture and ripening

Milk subjected to instant infusion pasteurization (IIP) at 72 °C, 100 °C and 120 °C (holding time 0.2 s) exhibited increased rennet coagulation time and decreased curd firming rate for increasing heat treatment temperature, when compared with raw or high temperature short time pasteurized (HTST) milk. However, addition of 4.5 mm or 9.0 mm of calcium restored the impaired rennet coagulation ability. Open texture cheeses produced from IIP milk (100 °C and 120 °C) contained significantly more moisture, had lower pH and shorter texture than similar cheese from IIP at 72 °C and HTST pasteurized milk. Cheese ripening was also affected by heat treatment, and different patterns of casein breakdown and peptide formation resulted from cheeses made from milk treated to IIP at 100 °C and 120 °C compared with cheeses made using IIP at 72 °C or HTST.     

In vitro immunogenicity of various native and thermally processed bovine milk proteins and their mixtures

In vitro immunogenicity of various native and thermally processed (72°C/15 s and 100°C/30 s) bovine milk protein fractions, their mixtures, whey, and skim milk, was studied by analyzing the immune response of T helper (Th) cells in human peripheral blood mononuclear cells. The secretion of Th type cytokines induced by the protein stimulants was quantified while determining the heat-induced protein denaturation. Purified whey proteins, caseins and whey fraction, and skim milk provoked substantial immune responses at various degrees, indicating their potent immunogenicity. The protein mixtures prepared using the fractionated whey proteins with or without caseins appeared less immunogenic in both native and heat-treated forms, implying their potential of producing less immunogenic dairy products. The 100°C/30 s treatment significantly altered the immunogenicity of most of the potent protein stimulants, which mostly coincided with their levels of protein denaturation. The 72°C/15 s treatment caused the least protein denaturation but altered the immunogenicity of several protein stimulants notably, including heat-stable caseins and α-lactalbumin.     

Effect of pH and heat treatment conditions on physicochemical and acid gelation properties of liquid milk protein concentrate

Milk protein concentrates (MPC) are typically dried high-protein powders with functional and nutritional properties that can be tailored through modification of processing conditions, including temperature, pH, filtration, and drying. However, the effects of processing conditions on the structure-function properties of liquid MPC (fluid ultrafiltered milk), specifically, are understudied. In this report, the pH of liquid MPC [13% protein (70% protein DM basis), pH 6.7] was adjusted to 6.5 or 6.9, and samples at pH 6.5, 6.7, and 6.9 were subjected to heat treatment at either 85°C for 5 min or 125°C for 15 s. Sodium dodecyl sulfate PAGE was used to determine the distribution of caseins and denatured whey proteins in the soluble and micellar phases, and HPLC was used to quantify native whey proteins as a measure of denaturation, based on the processing conditions. Both heat treatments resulted in substantial whey protein denaturation at each pH, with β-lactoglobulin denatured more extensively than α-lactalbumin. Changes in liquid MPC physicochemical properties were monitored at d 1, 5, and 8 during storage at 4°C. Viscosity increased after heat treatment and also over time, regardless of pH and heating conditions, suggesting the role of whey protein denaturation and aggregation, and their interactions with casein micelles. The MPC samples processed at pH 6.9 had a significantly higher viscosity than those heated at pH 6.5 or 6.7, for both temperature and time conditions; and samples processed at 85°C for 5 min had higher viscosity than those heated at 125°C for 15 s. Particle size analysis indicated the presence of larger particles after 5 and 8 d of MPC storage after heating at pH 6.9. Acid-induced gelation of the liquid MPC led to significantly higher gel firmness after processing at 85°C for 5 min, compared with 125°C for 15 s. Also, gels made from MPC adjusted to pH 6.5 had higher storage moduli, with both time and temperature combinations, demonstrating the role of pH-dependent association of denatured whey proteins with casein micelles in gel network formation. These findings enable a better understanding of the processing factors contributing to structural and functional properties of liquid MPC and can be helpful in tailoring milk protein ingredient functionality for a variety of food products.     

Influence of heat treatment of goat milk on casein micelle size,rheological and textural properties of acid gels and set type yoghurts

Acid gels and yoghurts were made from goat milk that was heated at 72°C/30 s, 85°C/5 min, and 95°C/5 min, followed by acidification with starter culture at 43C until pH 4.6. The rheological and textural properties of acid gels and yoghurts were analyzed using dynamic low amplitude oscillatory rheology and back extrusion texture analysis, respectively. The effect of goat milk heat treatment on the mean casein micelle diameter and protein profile was also determined by dynamic light scattering and SDS PAGE electrophoresis, respectively. The shortest gelation and fermentation time was recorded for yoghurt prepared from milk heated at 85°C/5 min. Also, the pH of gelation, the storage moduli (G′) and yield stress were higher for this yoghurt, compared with the other two. Textural properties of goat milk yoghurts such as firmness and consistency were strongly affected by milk heat treatment, and the highest values were recorded for yoghurt produced from milk preheated at 85°C/5 min, as well. The largest casein micelles were measured after 85°C/5 min treatment and their size decreased at higher temperature, despite higher denaturation of whey proteins at the most intense heat regime, indicating the structure changes that influence on the acid gelation.     

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 a_w, 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.     

Effects of milk heat treatment and solvent composition on physicochemical and selected functional characteristics of milk protein concentrate

Milk protein concentrate (MPC) powders (～81% protein) were made from skim milk that was heat treated at 72°C for 15 s (LHMPC) or 85°C for 30 s (MHMPC). The MPC powder was manufactured by ultrafiltration and diafiltration of skim milk at 50°C followed by spray drying. The MPC dispersions (4.02% true protein) were prepared by reconstituting the LHMPC and MHMPC powders in distilled water (LHMPC_w and MHMPC_w, respectively) or milk permeate (LHMPC_p and MHMPC_p, respectively). Increasing milk heat treatment increased the level of whey protein denaturation (from ～5 to 47% of total whey protein) and reduced the concentrations of serum protein, serum calcium, and ionic calcium. These changes were paralleled by impaired rennet-induced coagulability of the MHMPC_w and MHMPC_p dispersions and a reduction in the pH of maximum heat stability of MHMPC_p from pH 6.9 to 6.8. For both the LHMPC and MHMPC dispersions, the use of permeate instead of water enhanced ethanol stability at pH 6.6 to 7.0, impaired rennet gelation, and changed the heat coagulation time and pH profile from type A to type B. Increasing the severity of milk heat treatment during MPC manufacture and the use of permeate instead of water led to significant reductions in the viscosity of stirred yogurt prepared by starter-induced acidification of the MPC dispersions. The current study clearly highlights how the functionality of protein dispersions prepared by reconstitution of high-protein MPC powders may be modulated by the heat treatment of the skim milk during manufacture of the MPC and the composition of the solvent used for reconstitution.     

Bovine milk procathepsin D: Presence and activity in heated milk and in extracts of rennet-free UF-Feta cheese

Milk samples were heat-treated at 72, 85 and 99°C for 15 or 60 s, and the effect on the stability of the milk acid proteinase zymogen procathepsin D was studied by combining immunoblotting using antibodies directed against bovine cathepsin D and its propeptide and by measuring residual procathepsin D-derived activity. Approximately half of the procathepsin D-derived activity detected in milk serum remained after heat treatment at 72°C/15 s or 72°C/60 s, while heat treatment at increased temperature further reduced the detectable activity. In accordance, immunoreactive procathepsin D was detected in serum from milk heated at 72°C/15 s and 72°C/60 s, while very low amounts of immunoreactivity were observed after treatment at higher temperatures. Contrary to the decrease in milk serum, the amount of procathepsin D antigen associated with casein micelles slightly increased with the temperature of the heat treatment, but still the measurable proteolytic activity derived from procathepsin D in the casein micelle samples decreased with temperature treatment. Moreover, the presence of procathepsin D and derived proteolytic activity was demonstrated in rennet free UF-Feta cheese. These results correlated with the finding of α_s1-I and para-κ-caseins in rennet free cheese. This is the first demonstration of procathepsin D in cheese, and of activity derived from indigenous procathepsin D in milk contributing to the proteolysis process in UF-products.     

Fourier transform infrared spectroscopy analysis of physicochemical changes in UHT milk during accelerated storage

The feasibility of using Fourier transform infrared spectroscopy (FTIR) to detect heat induced conformational rearrangements of proteins, protein–protein and protein–lipid interactions was studied with accelerated shelf-life protocols. Ultra-high temperature treated whole (WM) and skim milk (SM) were stored at 20, 30, 40 and 50 °C for 28 days. The changes leading to increased sedimentation in SM and WM at higher temperatures (≥40 °C) were observed during first 14 days of the storage period. Milk samples stored at 40 and 50 °C showed marked changes in the bands corresponding to conformations of milk lipids and formation of intermolecular β sheet of proteins, indicating protein–lipid interactions and aggregation. Dried sediment contained fat confirming protein–lipid participation in the sedimentation. FTIR was also able to detect changes that led to increased sedimentation in SM at temperatures lower than 40 °C, but only after 28 days.     

Heat-induced coagulation of concentrated skim milk heated by direct steam injection

Direct steam injection heat treatment on pilot scale was applied to investigate heat stability of concentrated skim milk across a broad range of temperatures from 117 °C to 153 °C and from 0.5 to 13 s holding time, assessing options for heat treat treatment without a significant amount of protein sediment formation. The relationship between total solids content of concentrated skim milk and temperature–time combinations of heat treatment could be established using minimal heat-induced coagulation as a criterion. Coagulation of destabilised casein micelles was shown to proceed non-linear over heating temperature. Transition of critical temperature–time combinations resulted in a marked increase in sediment formation indicating that preceding reactions, noticeable as the formation of dissociated material, need to take place to some extent to induce coagulum formation. UHT pre-heat treatment of skim milk prior to concentration was shown to increase heat stability in terms of possible temperature–time combinations without coagulation.     

Biochemical Characterization of an Extracellular Heat‐Stable Protease from Serratia liquefaciens Isolated from Raw Milk

The protease Ser2 secreted by the psychrotrophic strain Serratia liquefaciens L53, a highly proteolytic strain isolated from Brazilian raw milk was purified and characterized. Using azocasein as substrate, Ser2 exhibited activity in a wide range of pH (5 to 10) and temperature (4 to 60 °C). The optimal activity was detected at pH 8.0 and at a temperature of 37 °C. This protease, still active at 4, 7, and 10 °C, was strongly inhibited by chelating agents and by dithiothreitol, a reducing agent. These results confirmed that Ser2 belongs to the peptidase family M10 and requires Ca²⁺, Zn²⁺, and disulfide bridges for stability. This protease is able to hydrolyze three kinds of casein in the preferential order of κ→ β→ α‐casein. Highly heat‐stable in skimmed, semi‐skimmed, and whole milk at 140°C with D‐values of 2.8, 3.9, and 4.5 min, respectively, Ser2 showed a residual activity between 87 and 100 percent after heat‐treatment of 65 °C for 30 min, 72 °C for 20 s, and 140 °C for 4 s that are commonly used in dairy industries. As the protease AprX that is mainly secreted by Pseudomonas genus, Ser2 could be one of the main causes of UHT milk destabilization during storage.     

Heat‐induced coagulation of whole milk by high levels of calcium chloride

This study investigated the interaction of calcium ions and milk proteins during heat‐induced coagulation of milk. Addition of 20–200 mM calcium chloride to milk caused coagulation on heating to 70 °C. Preheating milk at 90 °C for 10 min or ultra‐high temperature treatment at 140 °C for 6 s increased the sensitivity of milk proteins to coagulation. The former treatment was more effective than the latter in coagulating proteins. A maximum of 98% of the protein in milk preheated at 90 °C for 10 min was coagulated by 50 mM added calcium chloride at 70 °C with holding for 5 min.     

Chemical characteristics,oxidation and proteolysis in cheese produced from fresh or stored milk subjected to heat treatments

Cheese produced from fresh, stored or heat pretreated (65°C for 15 s) milk subjected to pasteurisation (72°C or 77°C for 15 s) was studied for chemical characteristics and proteolysis after 4 and 21 days of storage. Antioxidant activity was higher in cheese from fresh milk than in cheese from stored and heat pretreated milk. Malondialdehyde content as an index of lipid oxidation increased in cheese made with pretreated and preserved milk than in cheese made with fresh milk. Antioxidant activity and lipid oxidation were not significantly affected by pasteurisation temperature. Proteolysis increased during cheese maturation regardless of milk heat treatments.     

Concentration of proteins as a result of the phase separation of water-protein-polysaccharide systems Part 2. Concentration of milk proteins

The process of concentration of skimmed milk proteins as a result of the phase separation of the waterskimmed milk proteins-polysaccharide systems has been studied. The two-phase systems were obtained by mixing and laying-up skimmed milk and polysaccharide solutions. Used as polysaccharides were pectin, gum arabic and arabinogalactan. The establishment of the phase equilibrium in all the systems under study involves the concentration of the skimmed milk proteins and dilution of the polysaccharide solution. For the water-skimmed milk proteins-pectin system one can observe a transition of the lactose and calcium ions from the milk to the pectin solution. Depending on the concentration conditions, the concentration of the skimmed milk proteins in the protein phase increases by a factor of 5-12 as compared with that in milk. The dispersed phases of the water-skimmed milk proteins-gum arabic and water-skimmed milk proteinsarabinogalactan systems are represented by gel-like particles with skimmed milk concentrations of 30 and 45 wt %, respectively. A value of 30 wt % for the skimmed milk concentration seems to correspond to the critical concentration of the lyotropic gelation of the skimmed milk proteins. The largest involvement of the skimmed milk proteins in the protein phase (91%) of the water-skimmed milk proteins-polysaccharide system was observed to take place at 10°C. α-Lactalbumin is the major protein component of the polysaccharide phase. This concentration process can be defined as a membraneless osmosis. The rate of the process was studied and the time of the establishment of equilibrium in the two-phase systems obtained by mixing and laying-up the solutions was estimated. The greater effectiveness of the membraneless osmosis was shown to be mainly ensured by a fairly large size of the interphase surface of the systems under study.     

Kinetic and Thermodynamic Parameters for Heat Denaturation of Bovine Milk IgG, IgA and IgM

For heat denaturation of bovine milk immunoglobulins (IgG, IgA and IgM) in the temperature range 62 to 81°C, the D values were IgG > IgA > IgM at any temperature. The Z values were 6.29, 4.00 and 5.17°C for IgG, IgA and IgM, respectively. Heat denaturation of bovine milk immunoglobulins followed a reaction kinetics in the order of n= 1.5. The highest value for apparent energy of activation was observed for IgA, and the lowest for IgG. Bovine milk immunoglobulins could resist the HTST pasteurization treatment at 72°C for 15 sec without affecting their structure.