Effect of pH at pressure treatment on the acid gelation of skim milk

Skim milk at pH between 6.4 and 7.3 was pressure treated at 200–600 MPa for 30 min and then slowly acidified with glucono-δ-lactone to form acid gels. Milks at low pH produced acid gels with low elastic moduli (final G′) and yield stresses and those at higher pH produced acid gels with higher final G′ and yield stresses. Pressure treatment disrupted the casein micelles at all pH and transferred high levels of casein to the serum phase. Denaturation of α-lactalbumin occurred at a pressure of 600 MPa only, and the level of denaturation increased with increasing pH. Denaturation of β-lactoglobulin (β-LG) occurred at all pressures, with the level of denaturation increasing with the magnitude of the pressure treatment and with pH. The denaturation of the whey proteins and the disruption of the casein micelles could not entirely account for the changes in the rheological properties of the acid gels, as denaturation of up to 50% of the whey proteins produced acid gels with very low final G′ and yield stresses. It is proposed that the pH and the magnitude of the pressure treatment affect the interactions of the denatured β-LG with the casein proteins in the pressure-treated milks, and that this affects the ability of the denatured β-LG to participate in the acid gel structures.Industrial relevanceThe control and manipulation of the firmness of acid skim milk gels is important in many dairy food applications such as yogurts and some types of cheeses. This study has demonstrated that acid gel firmness can be substantially manipulated when the milk is pH adjusted and pressure treated before acidification, and that these effects are different to those obtained through heating. The commercial uptake of high pressure processing in the dairy industry is dependent on this technology producing unique functional properties in milk when compared with traditional processing. The results of this study indicates that high pressure processing of milk may offer unique functional properties in acid gel applications which could be used for the development of new or improved dairy products.     

Effect of milk solids concentration on the gels formed by the acidification of heated pH-adjusted skim milk

Reconstituted skim milk of 10–25% total solids was adjusted to pH values between about 6.2 and 7.1 and heated at 80 °C for 30 min. Gels were formed from the heated milks by slow acidification to pH 4.2 and the gelation process and final gels were analyzed for their rheological properties. At each milk concentration, the final acid gel firmness (final G′) and breaking stress could be changed markedly by manipulation of the pH during heating. The final gel firmness and breaking stress could also be modified by changing the concentration of the milk solids prior to heating and acidification. The results indicated that similar gel firmness and breaking stress could be achieved over a range of milk concentrations by control of the pH of the milk during heating. When expressed as a percentage change in final G′ or breaking stress relative to that obtained at the natural pH, plots of the change in final G′ or breaking stress versus pH fell close to a single curve, indicating that the same mechanism may influence the gelation properties at all milk concentrations. The final G′ and breaking stress were related to the denaturation and interaction of the whey proteins with the casein micelles, and the formation of non-sedimentable casein when the milk was heated.     

Effect of pH adjustment at heating on the rheological properties of acid skim milk gels with added potato starch

The rheological properties of acid skim milk gels, prepared from milk with added potato starch and pH adjusted (pH 6.5–7.1) prior to heat treatment and acidification, were investigated. The storage modulus, G′, of the final acid gels was increased by heating the milk at higher pH and further increased by adding starch. The effect of pH at heating and addition of starch appeared to be additive and independent of each other up to a starch addition level of 1%. Above this starch level, the pH at heating had a lesser effect. This may have been due to the increased viscosity of the aqueous phase as a result of starch gelatinization or to direct contributions of the starch to the gel network structure. Confocal microscopy showed that milk proteins developed fewer but broader protein clusters at higher pH than at lower pH. Starch addition resulted in an increased density of the protein network.     

Effect of pH at heat treatment on the hydrolysis of κ-casein and the gelation of skim milk by chymosin

Skim milk was adjusted to pH values between 6.5 and 7.1 and heated at 90 °C for times from 0 to 30 min. After heat treatment, the samples were re-adjusted to the natural pH (pH 6.67) and allowed to re-equilibrate. High levels of denatured whey proteins associated with the casein micelles during heating at pH 6.5 (about 70-80% of the total after 30 min of heating). This level decreased as the pH at heating was increased, so that about 30%, 20% and 10% of the denatured whey protein was associated with the casein micelles after 30 min of heating at pH 6.7, 6.9 and 7.1, respectively. Increasing levels of κ-casein were transferred to the serum as the pH at heating was increased. The loss of κ-casein and the formation of para-κ-casein with time as a consequence of the chymosin treatment of the milk samples were monitored by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The loss of κ-casein and the formation of para-κ-casein were similar for the unheated and heated samples, regardless of the pH at heating or the heat treatment applied. Monitoring the gelation properties with time for the chymosin-treated milk samples indicated that the heat treatment of the milk markedly increased the gelation time and decreased the firmness (G^′) of the gels formed, regardless of whether the denatured whey proteins were associated with the casein micelles or in the milk serum. There was no effect of pH at heat treatment. These results suggest that the heat treatment of milk has only a small effect on the primary stage of the chymosin reaction (enzymatic phase). However, heat treatment has a marked effect on the secondary stage of this reaction (aggregation phase), and the effect is similar regardless of whether the denatured whey proteins are associated with the casein micelles or in the milk serum as nonsedimentable aggregates.     

Heat and/or high-pressure treatment of skim milk: changes to the casein micelle size, whey proteins and the acid gelation properties of the milk

Skim milk was subjected to heat, pressure or combined processes. In general, higher levels of whey protein denaturation were observed for milk subjected to combined processes than those heat- or pressure-treated only. Heat treatment caused small changes to the casein micelle size. Pressure treatment decreased the casein micelle size; however, the effect was less marked when heat and pressure treatments were combined. Acidification of the skim milks produced gels with a range of firmness, yield stresses and yield strains depending on the treatments applied. These changes in acid gel properties were not related only to whey protein denaturation levels in the milks.     

Gelatinization of waxy starches under high pressure as influenced by pH and osmolarity: Gelatinization kinetics, final structure and pasting properties

We investigated the influence of pH and osmolarity on the high-pressure-induced gelatinization of waxy corn and waxy rice starches in salt solutions, and the properties of the resulting gels. Gelatinization kinetics, the gel swelling power of starches, their structure and their rheological properties were studied for starch suspensions treated at 500 MPa. Gelatinization took place mostly in the first 15 min of the pressure treatment and both the gelatinization speed and the maximal level of gelatinized starch decreased with increasing osmolarity. pH had a minor influence on gelatinization kinetics differing from one starch to another. The resulting gels appeared as a mix of a gel and starch granules with a higher proportion of native granules with increasing osmolarity. Gel strength and swelling were positively correlated to their proportion of gelatinized starch. Thus, gels with different structures and gelatinization levels can be obtained under pressure depending on pH and osmolarity.     

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.     

Effect of potato starch addition on the acid gelation of milk

Potato starch was added to skim milk at levels of 0–1.5%. The milks were heated and then acidified to form acid milk gels. The properties of the milks during acidification and the final properties of the acid gels were studied. The addition of starch resulted in a higher storage modulus, G′, in the final acid gels, and increasing the level of starch caused a linear increase in the final G′. Compared with acid gels prepared with no starch, the gelation time was reduced and the gelation pH was increased. However, the temperature and frequency dependences of the acid gels were not affected by the addition of starch. Furthermore, the breaking strain of the acid gels was not markedly affected by the addition of starch, whereas the breaking stress was dependent on the level of starch added. Confocal microscopy showed that the acid gels contained swollen starch granules embedded in a protein network, and that the protein network increased in density as the level of starch added increased.     

Effects of combined high-pressure and heat treatment on the textural properties of soya gels

SPI, 7S, and 11S globulin at 12% (w/v) protein concentration, at neutral pH, did not form gels when heat-treated (90 °C, 15 min) or when high pressure-treated (300–700 MPa), except for the 11S, which formed a gel when heat-treated. The combination of heat and pressure (that is heating the solutions in a water bath and then pressure-treating at room temperature or the reverse sequence), led to differences: when heat-treatment was before high-pressure treatment, only the 11S fraction formed a self-standing gel; however, when the solutions were pressurised before heat treatment, all the proteins formed self-standing gels. The textural and water-holding properties were measured on the gels formed with the three different soy proteins.     

Starch–cassia gum interactions: A microstructure – Rheology study

We present for the first time the interactions of starch and cassia gum – a novel galactomannan recently approved for use in food processing. Viscoelastic, pasting and microstructural characterization of various starches (waxy; high amylose; normal; cross-linked waxy corn starch; potato starch) containing different levels of the cassia gum was carried out. Significant changes were observed in the morphology of granule remnants formed during gelatinization in the starch pastes prepared with and without the addition of cassia gum. The freeze-dried starch–cassia gum pastes presented a shrunken and tight arrangement of the starch granule remnants, when studied by scanning electron microscopy. A significant reduction in the granule remnant size was also calculated using laser diffraction particle size analysis. The extent of interaction with cassia gum differed significantly among the various starch types. All the unmodified corn starches recorded an increase in peak viscosity at all levels of the cassia gum addition. An increase in the final viscosity of these starches was also observed by the addition of cassia gum, with high amylose and normal corn starch showing the maximum. Similarly, the extent of breakdown and setback viscosity also differed among the different starch types. Ranges of dynamic rheological measurements (temperature, time and frequency sweeps) were performed within the viscoelastic zones. Rheological parameters, such as storage modulus (G′), loss modulus (G″) and the gelatinization temperature (T_gel), of the corn starches during the heating cycle were observed to increase, when cassia gum was present at lower levels. The starch–gum systems also exhibited higher tan δ values during both the heating and the cooling cycles, indicating the dominance of the viscous modulus. The G′ and G″ of all the corn starch gels containing cassia gum showed higher values throughout the frequency sweep range. However, the increase in G′ and G″ of different starches was not always consistent with the increase in cassia gum levels. The changes in rheological behaviour during storage of the starch gels, aged on the plate of the rheometer and then studied through time sweeps at 5 °C and frequency sweeps at 25 °C, suggested that the starch gels containing cassia gum had less pronounced changes in the rheological parameters than had their control counterparts.     

Sol gel transitions during acid gelation of milk containing modified waxy maize starch. Differences between chemical and bacterial acidification measured using rheological and spectroscopic techniques

Modified waxy maize starch (1%, w/v) was added to skim milk and the mixtures were heated and homogenized. Acidification was conducted at 40 °C, using either glucono-δ-lactone (GDL) or a commercial starter culture. The physico-chemical changes occurring during acidification were monitored using small oscillatory rheology, diffusing wave spectroscopy and ultrasound spectroscopy. A similar gelling behaviour was noted for GDL and bacterial-induced gels; however, a difference was noted in the values of storage modulus (G′). The presence of starch did not seem to affect the development of the gel structure, nor the mobility and positional correlations of the casein micelles during acidification. On the other hand, starch increased the final storage modulus, G′ of the acid milk gels. These results indicate the absence of direct interactions between micelles and the modified starch granules.     

Paste viscosity of rice starches of different amylose content and carboxymethylcellulose formed by dry heating and the physical properties of their films

Starch modified by combination with sodium carboxymethylcellulose (CMC) has been reported to have improved film properties. In this study, rice starches with different amylose content were heat-treated in a dry state after being impregnated with low or medium-viscosity CMC. Noticeable change was found in pasting properties of the starches after dry heat treatment with CMC. It indicated that crosslinkage occurred between the starch and CMC. The waxy starch showed significant change in viscosity throughout pasting after dry heating with CMC, suggesting that the ester bonds were mostly formed between the hydroxyl groups in amylopectin branches of rice starch and carboxylate acid groups of CMC. Particle size also increased after heat treatment with CMC. The modified starch-based films showed improvement in the tensile strength. Both water vapor and oxygen permeability reduced for the modified starch-based films. Dynamic mechanical analysis (DMA) study showed that the values of G′ of modified starch-based film were higher than those of native starch-based film over the temperature range −40 to 60 °C. The heating process with CMC could be used as a modification method for starch and provide desirable properties of starch-based films.     

Microstructural and physicochemical properties of heat-moisture treated waxy and normal starches

Starches from normal rice (21.72% amylose), waxy rice (1.64% amylose), normal corn (25.19% amylose), waxy corn (2.06% amylose), normal potato (28.97% amylose) and waxy potato (3.92% amylose) were heat-treated at 100 °C for 16 h at a moisture content of 25%. The effect of heat-moisture treatment (HMT) on morphology, structure, and physicochemical properties of those starches was investigated. The HMT did not change the size, shape, and surface characteristics of corn and potato starch granules, while surface change/partial gelatinization was found on the granules of rice starches. The X-ray diffraction pattern of normal and waxy potato starches was shifted from B- to C-type by HMT. The crystallinity of the starch samples, except waxy potato starch decreased on HMT. The viscosity profiles changed significantly with HMT. The treated starches, except the waxy potato starch, had higher pasting temperature and lower viscosity. The differences in viscosity values before and after HMT were more pronounced in normal starches than in waxy starches, whereas changes in the pasting temperature showed the reverse (waxy > normal). Shifts of the gelatinization temperature to higher values and gelatinization enthalpy to lower values as well as biphasic endotherms were found in treated starches. HMT increased enzyme digestibility of treated starches (except waxy corn starch); i.e., rapidly and slowly digestible starches increased, but resistant starch decreased. Although there was no absolute consistency on the data obtained from the three pairs of waxy and normal starches, in most cases the effects of HMT on normal starches were more pronounced than the corresponding waxy starches.     

Effect of heat treatment of skim milk and final fermentation pH on graininess and roughness of stirred yogurt

The effects of heat treatment of skim milk (95°C/80 s, 95°C/256 s, 110°C/40 s, 110°C/180 s, 130°C/20 s and 130°C/80 s) and final fermentation pH of yogurt (4.8 and 4.4) on physical characteristics of stirred yogurt were investigated. Physical properties, including graininess and roughness, of stirred yogurt were determined during storage at 4°C for 15 days. Number of grains, perimeter of grains, visual roughness, storage modulus and yield stress decreased, when heating temperature or final fermentation pH increased. For practical applications, processing parameters such as heat treatment and fermentation pH can be optimized to improve quality or modified to create fermented milk products with different physical properties.     

Effect of non-starch polysaccharides on the in vitro digestibility and rheological properties of rice starch gel

The starch digestibility and rheological properties of gels were evaluated in the presence of three non-starch polysaccharides (agar, xanthan gum and konjac glucomannan) with rice starch. Each polysaccharide was added to 30% (w/w) rice starch suspension at defined concentrations and starch gels were prepared. The extent of starch gel digestibility was determined by an in vitro method and rheological properties by a dynamic oscillatory test and a compression test. The added polysaccharides suppressed starch hydrolysis in the gels compared with the control, and a concentration dependency of this suppressive effect was observed. Adding agar and xanthan gum increased the storage shear modulus (G′) of starch gels, while adding konjac glucomannan decreased G′ values. The results indicate that the suppressive effect of non-starch polysaccharides on starch digestibility appears to be not only due to the rigidity of the gel, but also the interaction between starch and non-starch polysaccharides.     

Amylopectin structure of heat–moisture treated starches

The effects of heat–moisture treatment (HMT; moisture content of 25%, at 100°C for 24 h) on starch chain distribution and unit chain distribution of amylopectin in normal rice, waxy rice, normal corn, waxy corn, normal potato, and waxy potato starches were investigated. After HMT, starch chain distribution (amylose and amylopectin responses) of waxy corn and potato starches were identical to those of untreated starches, whereas the chromatographic response of waxy rice starch showed a slight decrease, but with a slight increase in peak tailing. This result indicated that HMT had no (or very limited) effect on the degradation of amylopectins. Analysis of unit chain distribution of amylopectins revealed that waxy characteristics affected the molecular structure of amylopectin in untreated starches, i.e., the CL of normal‐type starches was greater than that of waxy‐type starches. After HMT, the CL and unit chain distribution of all starches were no different than those of untreated starches. The results implied that changes in the physico‐chemical properties of HMT starches would be due to other phenomena rather than the degradation of amylopectin molecular structure. However, the thermal degradation of amylopectin molecules of waxy starches could occur by HMT at higher treatment temperatures (120 and 140°C).     

High-pressure treatment of milk: effects on casein micelle structure and on enzymic coagulation

High isostatic pressures up to 600 MPa were applied to samples of skim milk before addition of rennet and preparation of cheese curds. Electron microscopy revealed the structure of rennet gels produced from pressure-treated milks. These contained dense networks of fine strands, which were continuous over much bigger distances than in gels produced from untreated milk, where the strands were coarser with large interstitial spaces. Alterations in gel network structure gave rise to differences in rheology with much higher values for the storage moduli in the pressure-treated milk gels. The rate of gel formation and the water retention within the gel matrix were also affected by the processing of the milk. Casein micelles were disrupted by pressure and disruption appeared to be complete at treatments of 400 MPa and above. Whey proteins, particularly beta-lactoglobulin, were progressively denatured as increasing pressure was applied, and the denatured beta-lactoglobulin was incorporated into the rennet gels. Pressure-treated micelles were coagulated rapidly by rennet, but the presence of denatured beta-lactoglobulin interfered with the secondary aggregation phase and reduced the overall rate of coagulation. Syneresis from the curds was significantly reduced following treatment of the milk at 600 MPa, probably owing to the effects of a finer gel network and increased inclusion of whey protein. Levels of syneresis were more similar to control samples when the milk was treated at 400 MPa or less.     

Changes in kernel properties,in situ gelatinization,and physicochemical properties of waxy rice with inhibition of starch branching enzyme during cooking

The kernel properties, gelatinisation and physicochemical properties of rice were investigated in a waxy rice Guang-ling-xiang-nuo (GLXN) and its transgenic line GLXN-RNAi with inhibition of starch branching enzyme I/IIb. The volume swelling, water content and leached material of cooked kernels increased with increasing cooking time, but they were lower in GLXN-RNAi than in GLXN. The kernels of GLXN-RNAi were more difficultly gelatinised and disrupted than those of GLXN during cooking. The starch in the exterior of GLXN-RNAi endosperm was not completely gelatinised. The C_A-type starch of GLXN-RNAi was more resistant to cooking than A-type starch of GLXN. The cooked kernels of GLXN-RNAi had lower rapidly digestible starch and greater slowly digestible starch than those of GLXN. Brown rice flour had higher peak, hot, final and setback viscosities and lower breakdown viscosity in GLXN than GLXN-RNAi. These results indicated that GLXN-RNAi kernels exhibited great potential in applications as health foods.     

Effect of ozone treatment on physicochemical properties of waxy rice flour and waxy rice starch

The effect of ozone treatment on physicochemical properties of waxy rice flour and waxy rice starch was investigated. Results showed that ozone treatment increased the pasting viscosity of waxy rice flour. Compared with untreated waxy rice flour, the peak viscosities of waxy rice flour for 0.5, 1 and 2 h of ozone treatments were increased by 27.4%, 32.8% and 45.5%, respectively. The alpha‐amylase in waxy rice flour was inactivated during the treatment. The gelatinisation temperature and enthalpy of waxy rice flour were kept unchanged after the treatment. For waxy rice starch, pasting viscosity, swelling power and molecular weight were increased after 0.5 h of treatment, but decreased as treatment time extended. The ozone treatment decreased gelatinisation temperature and enthalpy of waxy rice starch.     

The fate of Listeria monocytogenes during the manufacture of Camembert-type cheese: A comparison between raw milk and milk treated with high hydrostatic pressure

Camembert-type cheese was produced from: raw bovine milk; raw milk inoculated with 2 or 4 log CFU/ml Listeria monocytogenes; raw milk inoculated with L. monocytogenes and subsequently pressure-treated at 500 MPa for 10 min at 20 °C; or uninoculated raw milk pressure-treated under these conditions. Cheeses produced from both pressure-treated milk and untreated milk had the typical composition, appearance and aroma of Camembert. Curd and cheese made from inoculated, untreated milk contained large numbers of L. monocytogenes throughout production. An initial inoculum of 1.95 log CFU/ml in milk increased to 4.52 log CFU/g in the curd and remained at a high level during ripening, with 3.85 log CFU/g in the final cheese. Pressure treatment inactivated L. monocytogenes in the raw milk at both inoculum levels and the pathogen was not detected in any of the final cheeses produced from pressure-treated milk. Therefore high pressure may be useful to inactivate L. monocytogenes in raw milk that is to be used for the production of soft, mould-ripened cheese.

Industrial relevance

This paper demonstrates the potential of high pressure (HP) for treatment of raw milk to be used in the manufacture of soft cheeses. HP treatment significantly reduced the level of Listeria monocytogenes in the raw milk and so allowed the production of safer non-thermally processed camembert-like soft cheese.