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.     

Comparison of pressure treatment and heat treatment of skim milk with added starch on subsequent acid gelation of milk

Skim milk with added starch (waxy rice starch or potato starch at levels of 0-1.5 g/100 g) was either pressure-treated (500 MPa, 20 °C, 30 min) or heat-treated (80 °C, 30 min) and subsequently acidified (using glucono-δ-lactone) to form acid milk gels. In the second part of the study, the pH of the skim milk samples was adjusted from the natural condition (pH 6.64) to pH 6.5, 6.6 or 6.9 before the pressure or heat treatment and re-adjusted back to pH 6.64 after the respective treatment. The rheological properties of the samples during acidification and of the final acid gels were studied. The storage modulus, G′, of the final acid milk gels increased as more waxy rice starch was added to milk before pressure or heat treatment. However, acid milk gels made from pressure-treated milk with added potato starch did not show significant changes in the G′ of the final acid gels whereas those made from the heat-treated counterparts showed a marked increase in the final G′ as the potato starch level increased. Waxy rice starch was gelatinised in milk by both pressure treatment and heat treatment whereas potato starch was gelatinised by heat treatment only. Increasing the pH of milk before pressure or heat treatment increased the final G′ of the acid milk gel produced on subsequent acidification of the milk and the final G′ was increased further by the addition of waxy rice starch before the pressure or heat treatment.     

Improvement of rheological properties of firm acid gels by skim milk heating is conserved after stirring

The enhancement of the strength of set acid gels by heating milk was related to rheological parameters (water retention capacity, storage modulus) of corresponding stirred gels. To obtain accurate rheological data from stirred gel it was necessary to maintain a constant granulometry of gel particles and to recognize time after stirring as a contributing factor. Two hours after stirring, the gel exhibited a higher storage modulus when milk was heated above 80 degrees C. A measurement of viscosity of just-stirred yoghurt was sufficient to predict correctly the quality of a stirred gel analysed by viscoelastic measurements. Increased resistance to syneresis of just-stirred gels was related to higher viscosity. The quantity of beta-lactoglobulin (beta-Ig) bound to casein micelles explains the improvement of these gel qualities. We have considered that the structure of the initial firm gel (mesostructure level) was conserved in fragments within the stirred gel. Consequently, the explanation given by various authors for the effect of heating milk on the properties of set gels can also be applied to stirred gels. The same mechanism, described in literature for structure formation of set gels from acidified milk is purposed to explain the role of heating milk on the recovery of gel structure after stirring. The beta-Ig association with casein micelles during heating favoured micelle connections during the acidification. It also favoured the association of gel fragments after stirring during the recovery in gel structure.     

Effect of a CO2-acidification cycle on physicochemical and acid gelation properties of skim milk reconstituted with added pectin

The influence of a CO₂-acidification cycle on the acid (glucono-δ-lactone, GDL) gelation properties of skim milk with and without added low-methoxyl pectin (LM-pectin) was assessed. Ionic calcium level, zeta potential, particle size, buffering properties and small amplitude oscillatory rheology moduli were monitored. The presence of LM-pectin in milk had an impact on the average size of the casein micelles and a large and dominant influence on its rheological behaviour during GDL acidification. The application of a CO₂–pH cycle (pH_target 4.9) as a milk pretreatment induced during GDL acidification a stabilization of the colloidal system in wide pH range (pH 6.0–5.1) with modifications of the structure of the casein micelles before the onset of gelation. These modifications induced a significant improvement on its acid gelation behaviour. The measurements of Ca²⁺ level during GDL acidification showed that an important and significant part of the Ca²⁺ released during the CO₂–pH cycling was electrostatistically trapped by pectin molecules in the serum.     

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.     

Rheological and structural properties of differently acidified and renneted milk gels

In this study we assessed the rheological and structural properties of differently acidified and renneted milk gels by controlling pH value and renneting extent. Skim milk were exactly renneted to 4 extents (20, 35, 55, and 74%) and then direct acidified to the desired pH (4.8, 5.0, 5.2, 5.5, 5.8, and 6.2), respectively. Rheological properties were assessed by dynamic rheological measurements, structural properties were studied by spontaneous whey separation and confocal laser scanning micrograph, and protein interactions were studied by dissociation test. Results showed that minimally renneted milk samples (20 and 35%) formed weak gels with low storage modulus, and the acidification range within which gels could form was narrow (pH ≤5.2). Highly renneted milk samples formed more gels with high storage modulus. The results of this study revealed that acidification determined the structural properties of highly renneted milk gels. As pH increased from 5.0 to 6.2, highly renneted milk gels had lower loss tangent, decreased spontaneous syneresis, and smaller pores. For both the low and high rennetings, divalent calcium bonds contributed less at low pH than at high pH. In conclusion, renneting increased the pH range suitable for gel formation; acidification determined the spontaneous syneresis and microstructure of highly renneted milk gels.     

Rheological Properties of Mixed Gels using Waxy and Non‐waxy Wheat Starch

Mixed starches with an amylose content of 5, 10, 18, 20, 23, and 25% were prepared by blending starches isolated from waxy and non‐waxy wheat at different ratios. The dynamic viscoelasticity of mixed 30% and 40% starch gels was measured using a rheometer with parallel plate geometry. The change in storage shear modulus (G′) over time at 5 °C was measured, and the rate constant of G′ development was estimated. As the proportion of waxy starch in the mixture increased, starch gels showed lower G′ and higher frequency dependence during 48 h storage at 5 °C. Since the amylopectin of waxy starch granules was solubilized more easily in hot water than that of non‐waxy starch granules, mixed starch containing more waxy starch was more highly solubilized and formed weaker gels. G′ of 30% and 40% starch gels increased steadily during 48 h. 30% starch gel of waxy, non‐waxy and mixed starches showed a slow increase in G′. For 40% starch gels, mixed starch containing more waxy starch showed rapidly developed G′ and had a higher rate constant of starch retrogradation. Waxy starch greatly influenced the rheological properties of mixed starch gels and its proportion in the mixture played a major role in starch gel properties.     

Effects of milk pH alteration on casein micelle size and gelation properties of milk

The effects of changes in pH above and below the natural pH of milk (ca 6.6) on the casein micelle size and the gelation properties of the pH adjusted and restored samples were investigated. The size of casein micelles increased at pH 7.0 and 7.5, then started to decrease above pH 8.5. It is postulated that at pH below 8.5 the casein micelles swell, while elevated pH causes their dissociation. Conversely, the size of casein micelles decreased on acidification to pH 5.5 and increased when the pH dropped below 5.5, due to the shrinkage of casein micelles at lower pH before their aggregation at pH below 5.5. In response to neutralising treated milk back to normal milk pH of 6.6, it was found that the casein micelle size of treated milk with a narrow range of pH change between 6.0 and 7.0 was reversible, while beyond this range the structure of casein micelles became irreversible. The restoration of casein micelle size was followed by the restoration of some parameters such as soluble calcium, ethanol stability, and milk whiteness. Acid-induced gelation did not change the elastic modulus, while rennet-induced gelation was affected by initial milk pH. In reference to the size of reversible range elastic modulus (G’) of acid or rennet gels made from restored milk, the sizes were similar to control milk after 6 h of gelation.     

Structural mechanisms leading to improved water retention in acid milk gels by use of transglutaminase

Water retention in transglutaminase (TG)-treated acid milk gels was studied and linked with the gel formation dynamics. Heat-treated skim milk with and without pre-treatment by TG was acidified at 20 °C, 30 °C and 40 °C at constant glucono-δ-lactone (GDL) level to obtain different acidification rates. Formation dynamics and structural properties of acid-induced gels were followed by rheological and near-infrared light backscattering measurements as well as microscopy. TG-treated gels showed decreased tan δ values all through the acidification, which was pronounced around the gelation point. Backscattered light intensity was lowered in TG-treated gels compared to the controls indicating that TG-treated gels were comprised of smaller aggregates. Water holding capacity (WHC) was measured by using centrifugation at selected pH points (pH 5.2, 5.0, 4.8 and 4.6) during acidification. Both acidification temperature and TG treatment had significant effects on the water retention properties of the gels. Spontaneous syneresis observed at high acidification temperatures (≥30 °C) was prevented upon TG-treatment. WHC of TG-treated gels was significantly higher compared to the control gels at all pH points. TG-treated milk gels showed a homogeneous network formed of smaller aggregate and pore sizes at the gelation point and did not show any large-scale re-organisation thereafter. Transglutaminase is likely to act as a fixative of the protein network at an early stage of gelation and thereby limiting network rearrangements that take place in acid milk gels formed at high acidification temperatures leading to contraction and subsequent wheying off.     

The effect of the glucono-δ-lactone/caseinate ratio on sodium caseinate gelation

The influence of the acidification rate and the final pH on the properties of sodium caseinate gels (2–6%, w/v) prepared by acidification with glucono-δ-lactone (GDL) at 10 °C was investigated. The rheological properties of the systems were analysed under shear at incipient gelation and under uniaxial compression throughout the entire gelation process. The water holding capacity (WHC) of these gels and their amount of soluble protein in different media were also evaluated. Up to ∼50% of the total κ-casein content was soluble in gels with a higher amount of GDL, and this contributed to the high WHC observed under this condition. The acidification rate did not affect the rheological properties measured under shear during gelation, but the GDL/caseinate ratio used had a significant effect on the steady-state properties. Fast acidification resulted in lower Young's modulus and stress at fracture values, but higher residual stress values and relaxation times. In contrast, slow acidification produced a more interconnected network probably because of the extensive reorganization or rearrangement within the segments near the isoelectric point.     

Combined milk gel generated with a novel coagulating enzyme by Virgibacillus sp. SK37, a moderately halophilic bacterium

The hydrolysis of milk proteins by the recombinant AprX‐SK37 protease and the changes in the rheological properties of the milk gel generated with AprX‐SK37 and glucono‐δ‐lactone (GDL) were investigated. The AprX‐SK37 and rennet selectively hydrolysed κ‐casein to yield a 16‐kDa band, while subtilisin hydrolysed all of the casein components. Milk treated only with AprX‐SK37 formed softer gel. Storage modulus (G′) values of the combined gels increased with GDL concentrations up to 7 g/L. High tan δ was observed in the combined gel at 8.75 g/L GDL alongside syneresis. AprX‐SK37 is a promising milk‐clotting enzyme when combined with an optimal GDL concentration.     

RHEOLOGY OF ACID-INDUCED SODIUM CASEINATE STABILIZED EMULSION GELS

The storage modulus G', loss modulus G", and phase angle δ of acidinduced sodium caseinate emulsion gels were measured at 25C for a certain period of time after the addition of glucono-δ-lactone (GDL). Comparison between pure protein gels and emulsion gels revealed that the presence of emulsion droplets greatly enhanced the gel strength. Acidification by mixing an emulsion with a GDL solution caused immediate gelation but the emulsion gel had similar mechanical properties to the gel formed by direct addition of GDL granules. The viscoelasticity of the gel was strongly related to the pH value of the system. There was no evidence for a three-dimensional network when the pH value was higher than 5.8 or lower than 3.2. The largest storage and loss moduli were observed for gels formed at pH values near the isoelectric point of sodium caseinate (pH 4.6). Rheological differences for gels made at different pH values became distinguishable at low frequencies, where a much smaller phase angle was determined for a gel made at a pH value below the isoelectric point. Partial recovery of the three-dimensional gel network was observed for disrupted gels formed at a pH near the isoelectric point.     

OSCILLATORY PROBE RHEOMETRY AS A TOOL FOR DETERMINING THE RHEOLOGICAL PROPERTIES OF STARCH-WATER SYSTEMS 1

An oscillatory probe rheometer was effective at measuring the viscosity of starch pastes and the viscoelastic properties of starch gels. Because low shear strains were applied, the integrity of the gel was not disrupted during the testing. As the starch concentration of the systems increased, the complex modulus (G*) and storage modulus (G′) increased. At 20°C, the strength of the resulting gels, as measured by log G′, was a linear function of starch concentration. Loss tangent values (G″/G′) could be used to determine the approximate temperature at which the sol became a gel. As the starch concentration of the systems increased, the temperature of the sol to gel transformation increased from approximately 35°C (5% starch) to 60°C (10% starch).     

Comparison of heat and pressure treatments of skim milk, fortified with whey protein concentrate, for set yogurt preparation: effects on milk proteins and gel structure

Heat (85 degrees C for 20 min) and pressure (600 MPa for 15 min) treatments were applied to skim milk fortified by addition of whey protein concentrate. Both treatments caused > 90 % denaturation of beta-lactoglobulin. During heat treatment this denaturation took place in the presence of intact casein micelles; during pressure treatment it occurred while the micelles were in a highly dissociated state. As a result micelle structure and the distribution of beta-lactoglobulin were different in the two milks. Electron microscopy and immunolabelling techniques were used to examine the milks after processing and during their transition to yogurt gels. The disruption of micelles by high pressure caused a significant change in the appearance of the milk which was quantified by measurement of the colour values L*, a* and b*. Heat treatment also affected these characteristics. Casein micelles are dynamic structures, influenced by changes to their environment. This was clearly demonstrated by the transition from the clusters of small irregularly shaped micelle fragments present in cold pressure-treated milk to round, separate and compact micelles formed on warming the milk to 43 degrees C. The effect of this transition was observed as significant changes in the colour indicators. During yogurt gel formation, further changes in micelle structure, occurring in both pressure and heat-treated samples, resulted in a convergence of colour values. However, the microstructure of the gels and their rheological properties were very different. Pressure-treated milk yogurt had a much higher storage modulus but yielded more readily to large deformation than the heated milk yogurt. These changes in micelle structure during processing and yogurt preparation are discussed in terms of a recently published micelle model.     

Rheological properties of milk gels formed by a combination of rennet and glucono-delta-lactone

The effects of heat treatment of milk, and a range of rennet and glucono-delta-lactone (GDL) concentrations on the rheological properties, at small and large deformation, of milk gels were investigated. Gels were made from reconstituted skim milk at 30 degrees C, with two levels each of rennet and GDL. Together with controls this gave a total of sixteen gelation conditions, eight for unheated and eight for heated milk. Acid gels made from unheated milks had low storage moduli (G') of < 20 Pa. Heating milks at 80 degrees C for 30 min resulted in a large increase in the G' value of acid gels. Rennet-induced gels made from unheated milk had G' values in the range approximately 80-190 Pa. However, heat treatment severely impaired rennet coagulation: no gel was formed at low rennet levels and only a very weak gel was formed at high levels. In gels made with a combination of rennet and GDL unusual rheological behaviour was observed. After gelation, G' initially increased rapidly but then remained steady or even decreased, and at long ageing times G' values increased moderately or remained low. The loss tangent (tan delta) of acid gels made from heated milk increased after gelation to attain a maximum at pH approximately 5.1 but no maximum was observed in gels made from unheated milk. Gels made by a combination of rennet and GDL also exhibited a maximum in tan delta, indicating increased relaxation behaviour of the protein-protein bonds. We suggest that this maximum in tan delta was caused by a loosening of the intermolecular forces in casein particles caused by solubilization of colloidal calcium phosphate. We also suggest that in combination gels made from unheated milk a low value for the fracture stress and a high tan delta during gelation indicated an increased susceptibility of the network to excessive large scale rearrangements. In contrast. combination gels made from heated milk formed firmer gels crosslinked by denatured whey proteins and underwent fewer large scale rearrangements.     

Effect of insoluble calcium concentration on rennet coagulation properties of milk

Rennet-induced gels were made from milk acidified to various pH values or milk at pH 6.0 that had added EDTA. The objective was to examine the effect of removing insoluble Ca (INS Ca) from casein micelles (CM) on rennet gelation properties. For the pH trial, diluted lactic acid was added to reconstituted skim milk to decrease the pH to 6.4, 6.0, 5.8, 5.6, and 5.4. For the EDTA trial, EDTA was slowly added (0, 2, 4, and 6 mM) to reconstituted skim milk, and the final pH values were subsequently adjusted to pH 6.0. Dynamic low amplitude oscillatory rheology was used to monitor gel development. The Ca content of CM and rennet wheys made from these milks was measured using inductively coupled plasma spectroscopy. The INS Ca content of milk was altered by the acidification pH values or level of EDTA added. In all samples, the storage modulus (G′) exhibited a maximum (GM), with a decrease in G′ during longer aging times. Gels made at pH 6.4 had higher GM compared with gels made at pH 6.7 probably due to the reduction in electrostatic repulsion, whereas the INS Ca content only slightly decreased. The highest GM value of gels was observed at pH 6.4 and the GM value decreased with decreasing pH from 6.4 to 5.4. This was due to an excessive loss of INS Ca from CM. There was a decrease in GM with the increase in the concentration of added EDTA, which was probably due to the loss of colloidal calcium phosphate, which weakens the integrity of CM. Loss tangent (LT) values at GM increased with a reduction in milk pH and the addition of EDTA to milk. Rennet gels at the point of the GM were subjected to constant low shearing to fracture the gels. With a reduction in INS Ca content, the yield stress decreased, whereas LT values increased indicating a weaker, more flexible casein network. Microstructure of rennet-induced gels near the GM point and 2 to 10 h after this point was studied using fluorescence microscopy. At GM, gels made from milk acidified to pH 6.4 exhibited more branched, interconnected networks, whereas strands and clusters became larger with a reduction in milk pH to 5.4. Gels made from milk with EDTA added had more finely dispersed protein clusters compared with gels made from milk with no EDTA added. These microscopic observations supported the effect of loss of INS Ca on GM and LT. There was a decrease in apparent interconnectivity between strands in gel microstructure during aging, which agreed with the decrease in G′ after GM. It can be concluded that low levels of solubilization of INS Ca and the decrease in milk pH resulted in an increase in GM. With greater losses of INS Ca there was excessive reduction in cross-linking within CM, which resulted in weaker, more flexible rennet gels. This complex behavior cannot be explained by adhesive hard sphere models for CM or rennet gels made from these CM.     

Physical properties of acid milk gels: Acidification rate significantly interacts with cross-linking and heat treatment of milk

Rheological properties and microstructure of acid milk gels largely depend on the pre-treatment of the milk, including heating and enzymatic modification of the gel-forming proteins. It was the aim of our study to investigate the impact of the acidification rate on gels which were produced from milk reconstituted from powder containing casein cross-linked by microbial transglutaminase (mTGase), and on gels which were produced from cross-linked raw milk. 3–7% Glucono-δ-lactone (GDL) was used as acidulant. Gel stiffness was determined by dynamic small-deformation rheometry and penetration experiments, and forced syneresis and gel permeability served as indicators for gel microstructure.     

Evaluation of a fluorescence and infrared backscatter sensor to monitor acid induced coagulation of skim milk

A prototype sensor that employs both ultraviolet excited fluorescence and infrared light backscatter was evaluated as an in-line process analytical technology (PAT) tool to monitor acid induced coagulation kinetics of skim milk. Coagulation experiments were carried out at 32 °C using three concentrations of glucono-delta-lactone (GDL). Measurement of storage modulus (G′) of acidified skim milk gel was used as a reference rheological method to monitor the coagulation kinetics. Prediction models were developed to predict the times required for acidified skim milk coagulum to reach selected G′ values (0.5 Pa, 1 Pa, 5 Pa, 10 Pa and 15 Pa) using time parameters extracted from the ultraviolet excited fluorescence and infrared light backscatter profiles. A strong correlation was observed between the predicted times developed using time parameters extracted from the prototype sensor profiles and the measured G′ times extracted from the rheometer (R² = 0.97, standard error of prediction = 2.8 min). This study concluded that the prototype fluorescence and infrared backscatter sensor investigated combined with the developed rheological prediction model can be used as a potential PAT tool for in-line monitoring of coagulation kinetics in the manufacture of acid induced milk gels.Industrial relevanceThe prototype fluorescence and infrared backscatter sensor investigated in this study combined with the developed rheological prediction model can be employed to monitor and control coagulation kinetics in a wide range of dairy processing applications including fresh cheese varieties and yoghurt manufacture.     

Casein gelation under simultaneous action of transglutaminase and glucono-delta-lactone

Casein solutions (5% w/v) were treated with microbial transglutaminase (MTG) and glucono-delta-lactone (GDL) under varying conditions in order to obtain gels. Storage modulus (G') and gelation time of the gels were measured by oscillation rheometry, while protein cross-linking was determined by gel permeation chromatography. The addition of only GDL to milk resulted in very weak gels, while MTG on its own was not able to create gel networks. Simultaneous action of both ingredients led to gels, the firmness of which was linearly related to the added amount of MTG, but passed through a maximum with rising GDL concentrations. Using chromatographical analysis, increasing G' values were interrelated with the formation of MTG-induced oligomers. The gelation time was directly proportional to the GDL concentration but not influenced by the addition of MTG within the studied range of concentration.     

Casein gelation under simultaneous action of transglutaminase and glucono‐δ‐lactone

Casein solutions (5% w/v) were treated with microbial transglutaminase (MTG) and glucono‐δ‐lactone (GDL) under varying conditions in order to obtain gels. Storage modulus (G ′) and gelation time of the gels were measured by oscillation rheometry, while protein cross‐linking was determined by gel permeation chromatography. The addition of only GDL to milk resulted in very weak gels, while MTG on its own was not able to create gel networks. Simultaneous action of both ingredients led to gels, the firmness of which was linearly related to the added amount of MTG, but passed through a maximum with rising GDL concentrations. Using chromatographical analysis, increasing G ′ values were interrelated with the formation of MTG‐induced oligomers. The gelation time was directly proportional to the GDL concentration but not influenced by the addition of MTG within the studied range of concentration.