Effect of high hydrostatic pressure processing on the rennet coagulation kinetics and physicochemical properties of sheep milk rennet-induced gels

The effects of high hydrostatic pressure on the constituents and coagulation ability and their effect on cheese production of sheep milk have not been studied in detail. The objective of this work was to evaluate the effect of high hydrostatic pressure processing on the coagulation kinetics and physicochemical properties of sheep milk and to explore how such treatment could improve the cheesemaking process. Five batches of milk were tested: 1 untreated control batch and 4 batches each subjected to a different pressure (150, 300, 450, or 600 MPa) for 5 min at 10°C. As treatment pressure increased, values of electrical conductivity and oxidation-reduction potential were found to decrease. However, no significant reduction in pH was recorded. Treatment pressures >300 MPa produced milk with lower lightness (luminosity) and a more yellow and green hue. Pressures >150 MPa resulted in micellar fragmentation, as well as significant increases in particle size, viscosity, and water-holding capacity as a consequence of the denaturing of soluble proteins. High-pressure treatments increased the solubility of colloidal calcium phosphate, leading to a considerable increase in the concentration of minerals in the serum phase. The highest concentrations of calcium and phosphorus in the rennet whey of milk were reached at 300 MPa. Curd coagulation time was reduced by 28% at pressures >300 MPa, and an increase in the curd firming rate was observed. As treatment pressure increased to 450 MPa, the firmness, elasticity, and the percentage creep recovery of gels increased, whereas values of compliance and fracture strain were reduced. Thus, we can conclude that 300 MPa is the optimum treatment pressure for milk intended for cheesemaking by enzymatic coagulation. This pressure produced milk with optimal coagulation kinetics and water-holding properties with the least loss of fat and protein to the whey.     

High-pressure homogenization of raw and pasteurized milk modifies the yield, composition, and texture of queso fresco cheese

High-pressure homogenization (HPH) of milk was studied as an alternative processing operation in the manufacturing of queso fresco cheese. Raw and pasteurized (65°C for 30 min) milks were subjected to HPH at 0, 100, 200, and 300 MPa and then used to manufacture queso fresco. The cheeses were evaluated for yield, moisture content, titratable acidity, nitrogen content, whey protein content, yield force, yield strain, and tactile texture by instrumental or trained panel analyses. The combination of HPH and thermal processing of milk resulted in cheeses with increased yield and moisture content. The net amount of protein transferred to the cheese per kilogram of milk remained constant for all treatments except raw milk processed at 300 MPa. The highest cheese yield, moisture content, and crumbliness were obtained for thermally processed milk subjected to HPH at 300 MPa. The principal component analysis of all measured variables showed that the variables yield, moisture content, and crumbliness were strongly correlated to each other and negatively correlated to the variables yield strain, protein content (wet basis), and sensory cohesiveness. It is suggested that the combination of thermal processing and HPH promotes thermally induced denaturation of whey protein, together with homogenization-induced dissociation of casein micelles. The combined effect results in queso fresco containing a thin casein-whey matrix that is able to better retain sweet whey. These results indicate that HPH has a strong potential for the manufacture of queso fresco with excellent yield and textural properties.     

Low-fat set yogurt made from milk subjected to combinations of high hydrostatic pressure and thermal processing

The combined use of high hydrostatic pressure (300 to 676 MPa, 5 min) and thermal treatment (85 degrees C, 30 min) in milk for the manufacture of low-fat yogurt was studied. The objective was to reduce syneresis and improve the rheological properties of yogurt, reducing the need for thickeners and stabilizers. The use of high hydrostatic pressure alone, or after thermal treatment, reduced the lightness and increased the viscosity of skim milk. However, milk recovered its initial lightness and viscosity when thermal treatment was applied after high hydrostatic pressure. The MALDI-TOF spectra of skim milk presented monomers of whey proteins after a treatment of 676 MPa for 5 min. Yogurts made from skim milk subjected to 400 to 500 MPa and thermal treatment showed increased yield stress, resistance to normal penetration, and elastic modulus, while having reduced syneresis when compared to yogurts from thermally treated or raw milks. The combined use of thermal treatment and high hydrostatic pressure assures extensive whey protein denaturation and casein micelle disruption, respectively. Although reaggregation of casein submicelles occurs during fermentation, the net effect of the combined HHP and thermal treatment is the improvement of yogurt yield stress and reduction of syneresis.     

Effects of high hydrostatic pressure on the physicochemical and emulsifying properties of sweet potato protein

The influence of high hydrostatic pressure (HHP) treatment on the physicochemical and emulsifying properties of sweet potato protein (SPP) at various concentrations, e.g. 2%, 4% and 6% (w/v, SPP‐2, SPP‐4 and SPP‐6), was investigated. Significant differences in hydrophobicity, enthalpy of denaturation and solubility were observed (P < 0.05). Emulsifying activity indexes (EAI) of SPP‐2 and SPP‐6 increased at 400 MPa, whereas EAI of all SPP significantly decreased at 600 MPa (P < 0.05). Emulsion stability (ESI) was significantly decreased for SPP‐2 and SPP‐6, while increase in ESI was observed for SPP‐4 above 200 MPa (P < 0.05). SPP‐2 emulsions showed sharp decrease in apparent viscosity with pressure increase, while pseudo plastic flow behaviour was not changed for all of emulsions. Sporamins A and B were well‐adsorbed in pressurised emulsion without displacement. These results suggest that HHP treatment could be used to modify the physicochemical and emulsifying properties of SPP.     

Effect of high pressure treatment on the physicochemical and functional properties of egg yolk

The effects of high-pressure (HP) treatment at 100–500 MPa on some physicochemical and functional properties of egg yolk (EY) were investigated. Protein solubility, viscosity, surface hydrophobicity (H₀), free sulfhydryl (SH) content, differential scanning calorimetry characteristics, emulsifying activities and emulsifying stability were evaluated. HP-treatment resulted in protein aggregation, as evidenced by gradual decrease in protein solubility and significantly increased in viscosity. HP-treatment at 100–500 MPa induced a gradual decrease in H₀ and SH content, possibly due to protein unfolding and subsequent aggregation/re-association of unfolded proteins. Emulsifying activity index (EAI) was slightly decreased between 100 and 300 MPa and when the pressure is above 400 MPa, EAI was significantly (P < 0.05) decreased relative to the untreated EY. HP-treatment at 100 MPa significantly (P < 0.05) increased the ESI values of EY, while a significant (P < 0.05) decrease was observed when the pressure was above 200 MPa. It was also investigated that there are significant correlations between physicochemical properties of EY, and the differences in the modification of EY protein by HP treatment at different pressure levels may be attributed to the differences in aggregation and unfolding/refolding extents of proteins.     

高压均质处理次数对肌原纤维蛋白水溶液结构及理化特性的影响

以鸡肉中的肌原纤维蛋白（myofibrillar proteins,MP）为对象,研究在103 MPa压力条件下不同高压均质（high pressure homogenization,HPH）处理次数（1～6 次）对MP水溶液结构以及理化特性的影响。结果表明：HPH可以显著提高MP在水中的溶解性（P＜0.05）。随着HPH处理次数的增加,MP在水溶液中的粒径先减小后变大;表观黏度减小,流动能力增强;表面疏水性和活性巯基含量先增加后减少;圆二色光谱结果显示不同HPH处理次数对MP水溶液的二级结构构象的影响不同;过多的HPH处理次数会导致MP水溶液中蛋白质聚集,从而影响其溶解性和稳定性;经过4 次HPH处理的MP水溶液具有较好的溶解性和稳定性。说明通过选择适当次数的HPH处理可以实现对MP在水中溶解性的调控,为其在食品加工中的应用提供新思路。     

超高压处理对豆浆感官状态和流变特性的影响

超高压处理使豆浆中蛋白质颗粒和脂肪球的存在状态发生明显地变化,蛋白质颗粒变小,而脂肪球增大。豆浆在≤２００ＭＰａ的压力处理１５ｍｉｎ后,浊度略有上升;当压力＞２００ＭＰａ时,浊度又下降,但豆浆的沉降稳定性随处理压力的增大而提高,豆浆的粘度也开始有所增大,３００～４００ＭＰａ范围内粘度的增加最为明显。豆浆的热稳定性在不同压力处理后均有所提高。豆浆的浓度在１．５％～２．５％蛋白质（ｗ／ｖ）时,压力处理后豆浆沉降稳定性的提高更为明显,粘度的增加也最多。豆浆的贮藏模量Ｇ′在≤２００ＭＰａ压力处理后增加,而压力＞２００ＭＰａ时又下降;压力处理后的损耗模量Ｇ″有所增加,且在４００ＭＰａ处理后增加的较多;相对粘弹性ｔｇδ在０～３００ＭＰａ处理后有所下降,而在４００ＭＰａ后又增加。     

Disruption and sedimentation of casein micelles and casein micelle isolates under high-pressure homogenization

Native casein micelles were isolated from raw skim milk by ultrafiltration (< 30 kDa) or microfiltration (< 0.2 μm) and subjected to high-pressure homogenization (HPH) at 100, 200, 250, 300, and 350 MPa. Of particular interest was the effect of HPH on casein micelle size in solutions varying in ionic strength (0, 5, 10, and 15 mM CaCl₂) and micelle size populations. Particle size distribution reflected an initial decrease in micelle diameter in all samples at 100 MPa. In samples containing 10 and 15 mM CaCl₂, there was an abrupt increase in particle size and subsequent casein precipitation followed by sedimentation upon centrifugation at elevated pressures (300 and 350 MPa). The amount of sedimentable casein protein increased as CaCl₂ concentration (10 and 15 mM) and pressure (300 and 350 MPa) increased as determined by UV absorbance of sample supernatant. SDS-PAGE indicated extensive micellar disruption at elevated pressures (300 and 350 MPa) and confirmed that the sedimented portion of the samples contained casein proteins and minimal amounts of whey proteins. Results indicated that through HPH treatment casein micelle size can be modified based on CaCl₂ concentration and pressure applied. Based on these findings, HPH in combination with an appropriate suspending medium has the ability to modify micelles to a desired size for a number of potential applications.Industrial relevanceThe modification of structure-function properties of the casein micelle from bovine milk by using high-pressure homogenization is relevant in (1) the development of new ingredients to change rheological/textural properties of dairy based foods, and (2) the discovery of new and/or improved functionalities for protein quaternary structures.     

超高压对罗非鱼肌动球蛋白物化特性的影响

本文研究了超高压(100、300、500 MPa处理15、30、45 min)作用下罗非鱼肌动球蛋白物化特性的变化。结果发现,随着压力的增加和保压时间的延长,罗非鱼肌动球蛋白的溶解度降低,其中500 MPa处理45 min降至最低,为89.17%,表明超高压有利于蛋白质的聚集变性。超高压作用下罗非鱼肌动球蛋白的Ca2+-ATPase活性消失,表明肌球蛋白发生了变性。随压力与保压时间的增加,肌动球蛋白的表面疏水性增加,且300 MPa处理45 min增至最大,表明超高压作用使更多的疏水性基团暴露。随压力的增加,肌动球蛋白的总巯基含量降低,且500 MPa处理30 min降为最低值,二硫键含量升高,在300 MPa处理45 min增至最高,表明肌动球蛋白中巯基发生氧化形成了二硫键。以上物化特性的改变表明经过超高压处理的罗非鱼肌动球蛋白构象发生了改变。     

Using physical processes to improve physicochemical and structural characteristics of fresh and frozen/thawed sheep milk

We assessed the impact of stirring (ST), high shear dispersing (HSD) and low (LPH, 3.5 MPa) and high pressure homogenization (HPH, 50 MPa) on physicochemical and structural characteristics of whole and skimmed sheep milk fresh or previously frozen and thawed (FT). Freezing affected the size of the fat globules, their interaction with caseins, reduced calcium solubility (10%) and buffering capacity (5–11%). Amongst the studied processes, HSD was the only one unable to improve the milk stability. The other ones reduced the size of the fat globules and increased fat and casein interactions, favoring milk stability and reducing the creaming occurrence (>22%). LPH and HPH also reduced the sedimentation in skimmed milk (>37%). Moreover, all processes recovered the buffering capacity of FT samples. The effectiveness of the processes can be ordered as ST < LPH < HPH, but the final choice will depend on the stability improvement required for milk vs. acquisition and operational equipment costs.Practical applicationSheep milk is normally not homogenized because it has a lower fat globule size than cow milk, which reduces the creaming occurrence. However, creaming happens in some instances and it can be intensified if the milk is preserved frozen (to accumulate enough volume) prior to the dairy production, causing defects in the final products (mainly yogurts). The studied physical processes can be strategically used to solve this problem, increasing the milk emulsion stability, reducing the sedimentation occurrence and changing the buffering capacity to reach the same value of fresh milk.     

Effects of homogenisation pressures on physicochemical changes in different layers of ultra‐high temperature whole milk during storage

The objective of this study was to investigate the effects of homogenisation pressures (10, 20, and 30 MPa) on the physicochemical changes in top, middle and bottom layers of ultra‐high temperature whole milk stored at 25 °C for 6 months. Fat separation, fat globule sizes, fat and protein contents, viscosity, free amino acids content and pH were analysed at an interval of 15 days. Results showed that higher homogenisation pressure retarded the differentiations of milk fat globule sizes, fat and protein contents and viscosity in different milk layers, but did not affect the final fat separation, proteolysis and pH.     

Effect of high-pressure treatment on denaturation of bovine β-lactoglobulin and α-lactalbumin

The effect of high-pressure treatment on denaturation of β-lactoglobulin and α-lactalbumin in skimmed milk, whey, and phosphate buffer was studied over a pressure range of 450–700 MPa at 20 °C. The degree of protein denaturation was measured by the loss of reactivity with their specific antibodies using radial immunodiffusion. The denaturation of β-lactoglobulin increased with the increase of pressure and holding time. Denaturation rate constants of β-lactoglobulin were higher when the protein was treated in skimmed milk than in whey, and in both media higher than in buffer, indicating that the stability of the protein depends on the treatment media. α-Lactalbumin is much more baroresistant than β-lactoglobulin as a low level of denaturation was obtained at all treatments assayed. Denaturation of β-lactoglobulin in the three media was found to follow a reaction order of n = 1.5. A linear relationship was obtained between the logarithm of the rate constants and pressure over the pressure range studied. Activation volumes obtained for the protein treated in milk, whey, and buffer were −17.7 ± 0.5, −24.8 ± 0.4, and −18.9 ± 0.8 mL/mol, respectively, which indicate that under pressure, reactions of volume decrease of β-lactoglobulin are favoured. Kinetic parameters obtained in this work allow calculating the pressure-induced denaturation of β-lactoglobulin on the basis of pressure and holding times applied.     

Effects of ultra-high pressure homogenization on microbial and physicochemical shelf life of milk

The effect of ultra-high pressure homogenization (UHPH) on microbial and physicochemical shelf life of milk during storage at 4°C was studied and compared with a conventional heat preservation technology used in industry. Milk was standardized at 3.5% fat and was processed using a Stansted high-pressure homogenizer. High-pressure treatments applied were 100, 200, and 300 MPa (single stage) with a milk inlet temperature of 40°C, and 200 and 300 MPa (single stage) with a milk inlet temperature of 30°C. The UHPH-treated milks were compared with high-pasteurized milk (PA; 90°C for 15 s). The microbiological quality was studied by enumerating total counts, psychrotropic bacteria, lactococci, lactobacilli, enterococci, coliforms, spores, and Pseudomonas. Physicochemical parameters assessed in milks were viscosity, color, pH, acidity, rate of creaming, particle size, and residual peroxidase and phosphatase activities. Immediately after treatment, UHPH was as efficient (99.99%) in reducing psychrotrophic, lactococci, and total bacteria as was the PA treatment, reaching reductions of 3.5 log cfu/mL. Coliforms, lactobacilli, and enterococci were eliminated. Microbial results of treated milks during storage at 4°C showed that UHPH treatment produced milk with a microbial shelf life between 14 and 18 d, similar to that achieved for PA milk. The UHPH treatments reduced the L* value of treated milks and induced a reduction in viscosity values of milks treated at 200 MPa compared with PA milks; however, these differences would not be appreciated by consumers. In spite of the fat aggregates detected in milks treated at 300 MPa, no creaming was observed in any UHPH-treated milk. Hence, alternative methods such as UHPH may give new opportunities to develop fluid milk with an equivalent shelf life to that of PA milk in terms of microbial and physicochemical characteristics.     

Manufacture of acid gels from skim milk using high-pressure homogenization

The effect of high-pressure homogenization (HPH) alone or in combination with a thermal treatment (TT) was investigated for the manufacture of acid gels from skim milk. Raw skim milk was subjected to HPH (0 to 350 MPa) or a TT (90°C, 5 min), or both, in the following processing combinations: 1) HPH, 2) HPH followed by TT, 3) TT followed by HPH, 4) TT, and 5) raw milk (control). After treatments, L* (lightness) values were measured, and then skim milk was acidified with 3% glucono-δ-lactone and rheological properties (G′ and gelation time), and whey holding capacity was evaluated. Treatments in which HPH and TT were combined showed greater L* values than those in which just HPH was applied. In all treatments, the L* values decreased as the pressure was increased up to 300 MPa with little change afterward. Gelation times were lower when HPH was combined with TT compared with the acid skim milk gels that were just pressure treated. The final G′ in gels obtained from skim milk subjected to the combined process (HPH and TT) was greater and pressure-dependent compared with all other gels. A maximum G′ (∼320 Pa) was observed with skim milk subjected to a combination of thermal processing before or after HPH at 350 MPa. Acid gels obtained from HPH milk at 350 MPa showed a linear decrease in whey holding capacity over time, retaining 20% more whey after centrifugation for 25 min compared with samples treated at lower pressures and all other treatments. Our results suggest that HPH in combination with TT can be used to improve the rheological properties and stability of yogurt, thus decreasing the need for additives.     

Effects of high hydrostatic pressure on physicochemical and functional properties of walnut (Juglans regia L.) protein isolate

BACKGROUND: Walnut (Juglans regia L.) is a good source of protein that has potential application in new product formation and fortification. The main objectives of this study were to investigate the effects of high hydrostatic pressure (HHP) treatment (300–600 MPa 20 min) on physicochemical and functional properties of walnut protein isolate (WPI) using various analytical techniques at room temperature. RESULTS: The results showed significant modification of solubility, free sulfhydryl content and surface hydrophobicity with increased levels of HHP treatment, indicating partial denaturation and aggregation of proteins. Differential scanning calorimetry and fluorescence spectrum analyses demonstrated that HHP treatment resulted in gradual unfolding of protein structure. Emulsifying activity index was significantly (P < 0.05) increased after HHP treatment at 400 MPa, but significantly decreased (P < 0.05) relative to the untreated WPI with further increase in pressure. HHP treatment at 300–600 MPa significantly decreased emulsion stability index. Additionally, HHP‐treated walnut proteins showed better foaming properties and in vitro digestibility. CONCLUSION: These results suggest that HHP treatment could be applied to modify the properties of walnut proteins by appropriate of pressure levels, which will help in using walnut protein as a potential food ingredient. © 2012 Society of Chemical Industry     

Effect of high pressure treatment on ovotransferrin

High pressure processing of ovotransferrin was carried out to study the structural and physiochemical changes of ovotransferrin under various pressure levels. At pH 8 and pressures higher than 200 MPa, a decrease in total sulfhydryl groups and an increase in surface hydrophobicity were observed along with a partial aggregation. A gradual shift of denaturation peak towards higher temperature was noticed up to 500 MPa, leading to a total loss of the enthalpy of denaturation at pressures of 600 and 700 MPa, where a significant decrease in intrinsic fluorescence was also observed. At pH 3, the ovotransferrin adopted a molten globule state, associated with a significant increase in surface hydrophobicity and reactive sulfhydryl content; structurally, no clear denaturation peaks in differential scanning calorimetry (DSC) were detected at any level of pressure treatment whereas a noticeable decrease in intrinsic fluorescence was evidenced up to 600 MPa and then increased at 700 MPa pressure treatment. Fourier transform infrared spectroscopy (FT-IR) revealed that the conformational structure were changed from helices, sheets, turns, and aggregated strand to mostly intermolecular β-sheets or aggregated strands at pH 8 at 200 MPa but switched back to original structure at higher pressures.     

Inactivation of Aspergillus niger in Mango Nectar by High-Pressure Homogenization Combined with Heat Shock

ABSTRACT:  This research evaluated the inactivation of a heat-resistant  Aspergillus niger  conidia in mango nectar by high-pressure homogenization (HPH) combined with heat shock.  A.niger  were inoculated in mango nectar (10⁶ conidia mL⁻¹) and subjected to HPH (300 to 100 MPa) and heat shock (80 °C for 5 to 20 min) before or after HPH. Processes were evaluated according to number of decimal reductions reached by each isolated or combined process. Scanning electron microscopy was performed to observe conidia wall after pressure treatment. Pressures below 150 MPa did not inactivate  A. niger  while pressures of 200 and 300 MPa resulted in 2 and more than 6 log reductions, respectively. D_{80 °C} of  A. niger  was determined as 5.03 min. A heat shock of 80 °C/15 min, reaching 3 decimal conidia reductions, was applied before or after a 200 MPa pressure treatment to improve the decimal reduction to 5 log cycles. Results indicated that HPH inactivated  A. niger  in mango nectar at 300 MPa (>6.24 log cycles) and that, with pressure (200 MPa) combined with post heat shock, it was possible to obtain the same decimal reduction, showing a synergistic effect. On the other hand, pre heat shock associated with HPH resulted in an additive effect. The observation of  A. niger  conidia treated by HPH at 100 and 200 MPa by scanning electron microscopy indicated that HPH promoted intense cell wall damage, which can sensitize the conidia to post heat shock and possibly explain the synergistic effect observed.
Practical Application : The results obtained in this paper are relevant to elucidate the mechanism of conidia inactivation in order to develop the application of HPH as an alternative pasteurization process for the fruit nectar industry.     

Acid coagulation properties and suitability for yogurt production of cows’ milk treated by high-pressure homogenisation

The effects of high-pressure homogenisation (HPH) of cows’ milk were investigated for suitability for yogurt manufacture, compared with the processes currently applied in industry. Milk at different inlet temperatures (30 °C or 40 °C) was subjected to HPH treatment at 100, 200 or 300 MPa (one stage) and 130, 230 or 330 MPa (two-stage). HPH-treated milk was compared with milk heat-treated (90 °C for 90 s) and homogenised at 15 MPa, and with milk treated under the same thermal conditions and also fortified with 3% skim milk powder. Milk treated at 300 or 200 MPa showed higher gel strengths on coagulation, higher gel firmness in texture analysis, less syneresis and lower titratable acidity compared with conventionally treated milk fortified with 3% skim milk powder.     

Application of TOPSIS methodology to determine optimum hazelnut cake concentration and high pressure homogenization condition for hazelnut milk production based on physicochemical,structural and sensory properties

Vegetable milks containing antioxidants, fatty acids and vitamins can be recommended as an alternative to animal milks cause some health problems like lactose intolerance and milk protein allergy. Hazelnut oil cake which is a waste from hazelnut oil production cannot be used for human nutrition. Cold press hazelnut cake must be evaluation as food materials. The aim of this study was to evaluate the effect of cold pressed hazelnut cake concentration (5, 10, 15% w/v) and high pressure homogenization (HPH) (up to 100 MPa) on the physicochemical, structural and sensory properties of hazelnut milk and also determined the best conditions for hazelnut milk production by TOPSIS approach. Hazelnut milks produced from 15% hazelnut cake had the best physicochemical properties and physical stability, but the sensory properties of the milks were not acceptable due to viscoelastic behaviors. Physicochemical properties, physical stability and sensory characteristics of hazelnut milks were significantly affected by homogenization pressure (P?<?0.05). Colloidal stability and sensory properties of hazelnut milks were improved by increasing homogenization pressure. The viscosity values of hazelnut milks with 10 and 15% hazelnut cakes were significantly decreased by increasing the homogenization pressure. TOPSIS approach showed that 10% hazelnut cake concentration and 100 MPa homogenization pressure was the best condition for an acceptable hazelnut milk production.     

High pressure treatment of bovine milk: effects on casein micelles and whey proteins

Effects of high pressure (HP) on average casein micelle size and denaturation of alpha-lactalbumin (alpha-la) and beta-lactoglobulin (beta-lg) in raw skim bovine milk were studied over a range of conditions. Micelle size was not influenced by treatment at pressures <200 MPa, but treatment at 250 MPa increased micelle size by approximately 25%, while treatment at > or = 300 MPa irreversibly reduced it to approximately 50% of that in untreated milk. The increase in micelle size after treatment at 250 MPa was greater with increasing treatment time and temperature and milk pH. Treatment times > or = 2 min at 400 MPa resulted in similar levels of micelle disruption, but increasing milk pH to 7.0 partially stabilised micelles against HP-induced disruption. Denaturation of alpha-la did not occur < or = 400 MPa, whereas beta-lg was denatured at pressures >100 MPa. Denaturation of alpha-la and beta-lg increased with increasing pressure, treatment time and temperature and milk pH. The majority of denatured beta-lg was apparently associated with casein micelles. These effects of HP on casein micelles and whey proteins in milk may have significant implications for properties of products made from HP-treated milk.