Ultrasonic processing of dairy systems in large scale reactors

High intensity low frequency ultrasound was used to process dairy ingredients to improve functional properties. Based on a number of lab-scale experiments, several experimental parameters were optimised for processing large volumes of whey and casein-based dairy systems in pilot scale ultrasonic reactors. A continuous sonication process at 20 kHz capable of delivering up to 4 kW of power with a flow-through reactor design was used to treat dairy ingredients at flow rates ranging from 200 to 6000 mL/min. Dairy ingredients treated by ultrasound included reconstituted whey protein concentrate (WPC), whey protein and milk protein retentates and calcium caseinate. The sonication of solutions with a contact time of less than 1 min and up to 2.4 min led to a significant reduction in the viscosity of materials containing 18% to 54% (w/w) solids. The viscosity of aqueous dairy ingredients treated with ultrasound was reduced by between 6% and 50% depending greatly on the composition, processing history, acoustic power and contact time. A notable improvement in the gel strength of sonicated and heat coagulated dairy systems was also observed. When sonication was combined with a pre-heat treatment of 80 °C for 1 min or 85 °C for 30 s, the heat stability of the dairy ingredients containing whey proteins was significantly improved. The effect of sonication was attributed mainly to physical forces generated through acoustic cavitation as supported by particle size reduction in response to sonication. As a result, the gelling properties and heat stability aspects of sonicated dairy ingredients were maintained after spray drying and reconstitution. Overall, the sonication procedure for processing dairy systems may be used to improve process efficiency, improve throughput and develop value added ingredients with the potential to deliver economical benefits to the dairy industry.     

Gelation of Milk Protein Concentrates Induced by Moderate Hydrostatic Pressures

Gelation of aqueous milk protein concentrates made up of spray dried milk was monitored both at atmospheric pressure and at moderate pressures (up to 1000 bar) using rheological methods. A Paar Physica UDS 200 rheometer was used to measure the gelation time at atmospheric pressure as a function of the temperature for a protein concentration of 16% [w/w]. The temperature dependence of the gelation time is well approximated by the Ross-Murphy model, which was originally developed to describe the concentration dependence of the gelation time. Using a purpose built Haake High Pressure High Temperature Rheometer, the effects of moderate pressures upon the gelation of milk protein concentrates (13% [w/w] protein content) were investigated. It was found that the gelation time reduces significantly (by up to almost an order of magnitude), when pressures of up to 1000 bar were applied. These findings, combined with the work of others indicate that these pressures bring about two effects: (1) casein micelle dissociation and as a result facilitate reassociation of those smaller micelle fragments, and (2) denaturation of beta-lactoglobulin. However, the main contribution to enhanced gelation was attributed to the caseins. It is not clear if association of whey proteins with caseins occur at moderate pressures around 50 °C. We have modelled the dependence of the rate of gelation on pressure by an Eyring reaction rate process type equation. The major contribution to enhanced gelation upon pressurisation was attributed to a disruption of hydrophobic groups and ionisation of charged groups caused by an increase in electrostriction - however, disruption of hydrogen bonds was suggested to contribute to a lesser but still important extent.     

Functional properties of ultrasonically generated flaxseed oil-dairy emulsions

This study reports on the functional properties of 7% flaxseed oil/milk emulsion obtained by sonication (OM) using 20 kHz ultrasound (US) at 176 W for 1–8 min in two different delivery formulae, viz., ready-to-drink (RTD) and lactic acid gel. The RTD emulsions showed no change in viscosity after sonication for up to 8 min followed by storage up to a minimum of 9 days at 4 ± 2 °C. Similarly, the oxidative stability of the RTD emulsion was studied by measuring the conjugated diene hydroperoxides (CD). The CD was unaffected after 8 min of ultrasonic processing. The safety aspect of US processing was evaluated by measuring the formation of CD at different power levels. The functional properties of OM gels were evaluated by small and large scale deformation studies. The sonication process improved the gelation characteristics, viz., decreased gelation time, increased elastic nature, decreased syneresis and increased gel strength. The presence of finer sono-emulsified oil globules, stabilized by partially denatured whey proteins, contributed to the improvements in the gel structure in comparison to sonicated and unsonicated pasteurized homogenized skim milk (PHSM) gels. A sono-emulsification process of 5 min followed by gelation for about 11 min can produce gels of highest textural attibutes.     

Inactivation of Enterobacter aerogenes in reconstituted skim milk by high- and low-frequency ultrasound

The inactivation of Enterobacter aerogenes in skim milk using low-frequency (20 kHz) and high-frequency (850 kHz) ultrasonication was investigated. It was found that low-frequency acoustic cavitation resulted in lethal damage to E. aerogenes. The bacteria were more sensitive to ultrasound in water than in reconstituted skim milk having different protein concentrations. However, high-frequency ultrasound was not able to inactivate E. aerogenes in milk even when powers as high as 50 W for 60 min were used. This study also showed that high-frequency ultrasonication had no influence on the viscosity and particle size of skim milk, whereas low-frequency ultrasonication resulted in the decrease in viscosity and particle size of milk. The decrease in particle size is believed to be due to the breakup of the fat globules, and possibly to the cleavage of the κ-casein present at the surface of the casein micelles. Whey proteins were also found to be slightly affected by low-frequency ultrasound, with the amounts of α-lactalbumin and β-lactoglobulin slightly decreasing.     

利用光谱学方法解析具备高“奶油感”发酵乳的关键质构特征

“奶油感”是最受消费者喜好的发酵乳感官属性,反映令人愉悦的特征。为开发无添加但“奶油感”增强型的发酵乳,首先必须明确具备高“奶油感”发酵乳的质构特征。目前关于发酵乳“奶油感”的定义和评价标准尚未统一,关键质构特征尚不清晰。前人研究表明,发酵乳的凝胶网络形成过程及强度、稳定性和表观黏度可能是影响发酵乳感官感知的主要因素。该研究采用多种光谱学手段,解析具有不同强度“奶油感”发酵乳的关键质构特征。以经描述型感官评价得到的五种具备不同“奶油感”强度的发酵乳为研究对象,通过多散斑扩散光谱技术测定发酵乳凝胶网络的形成过程及凝胶强度,通过多重光散射技术来研究发酵乳的稳定性,再辅以流变学分析发酵乳的表观黏度。首先采用多散斑扩散波光谱技术研究具有不同强度“奶油感”发酵乳凝胶过程粒子均方位移的变化,结果显示凝胶的弹性指数突变时间按“奶油感”由弱到强分别为108,115,106,132和143 min,表明发酵乳形成凝胶的时间基本呈逐渐增加趋势;凝胶时间的增加可能是因为酪蛋白充分重排聚集形成了更均匀的凝胶网络,从而增强了“奶油感”的感知;弹性指数的终点值代表发酵乳的凝胶强度,结果显示凝胶强度处于适中水平的发酵乳“奶油感”较强。接着利用多重光散射技术评价了发酵乳的稳定性,“奶油感”由弱到强的发酵乳稳定性动力学指数分别为2.2,2.1,1.9,2.0和1.4,表明发酵乳稳定性与发酵乳“奶油感”的感知强度呈正相关。最后利用流变学方法测定了发酵乳的表观黏度,“奶油感”由弱到强的发酵乳表观黏度分别为(0.362±0.016),(0.271±0.013),(0.251±0.021),(0.479±0.031)和(0.343±0.024) Pa·s,表明发酵乳的表观黏度和“奶油感”感知强度不存在相关性。发酵乳的凝胶时间及凝胶稳定性与“奶油感”的感知密切相关。研究为明确发酵乳“奶油感”的关键质构、开发“奶油感”增强型的发酵乳制品提供理论依据。     

Modifying the physicochemical properties,solubility and foaming capacity of milk proteins by ultrasound-assisted alkaline pH-shifting treatment

This study investigated the effects of different treatment of alkaline pH-shifting on milk protein concentrate (MPC), micellar casein concentrate (MCC) and whey protein isolate (WPI) assisted by the same ultrasound conditions, including changes in the physicochemical properties, solubility and foaming capacity. The solubility of milk proteins had a significant increase with gradual enhancement of ultrasound-assisted alkaline pH-shifting (p < 0.05), especially for MCC up to 99.50 %. Also, treatment made a significant decline in the particle size of MPC and MCC, as well as the turbidity of the proteins (p < 0.05). The foaming capacity of MPC, MCC, and WPI was all improved, especially at pH 11, and at this pH, the milk protein also showed the highest surface hydrophobicity. The best foaming capacity at pH 11 was the result of the combined effect of particle size, potential, protein conformation, solubility, and surface hydrophobicity. In conclusion, ultrasound-assisted pH-shifting treatment was found to be effective in improving the physicochemical properties and solubility and foaming capacity of milk proteins, especially MCC, with promising application prospect in food industry.     

Effects of microfiltration combined with ultrasonication on shelf life and bioactive protein of skim milk

To extend the shelf life and retain bioactive proteins in milk, this study utilized microfiltration (MF) combined with ultrasonication to treat skim milk and investigated its efficiency in removing bacteria and retaining bioactive proteins compared with HTST pasteurization and microfiltration alone. Results showed that microfiltration combined with ultrasonication at 1296 J/mL could completely remove the bacteria in skim milk. Ultrasonication further extended the shelf life (4 °C) of microfiltered skim milk, which could reach at least 40 days when MF was combined with ˃1296 J/mL ultrasonication. In addition, ELISA showed that HTST pasteurization significantly decreased the levels of IgG by ~30%, IgA by ~ 50%, IgM by ~60%, and lactoferrin by ~40%, whereas the activity of the enzymes lactoperoxidase and xanthine oxidase were also decreased by ~ 20%. Compared with HTST, MF alone or combined with ultrasonication retained these bioactive proteins to a larger degree. On the other hand, proteomics indicated both damage to casein micelle and fat globule structures in milk when ultrasonication at >1296 J/mL was applied, as shown by increases in caseins and milk fat globular proteins. Simultaneously, this ultrasound intensity also decreased levels of bioactive proteins, such as complement factors. Taken together, this study provided new insights that may help to implement this novel combination of non-thermal technologies for the dairy industry aimed at improving milk quality and functionality.     

Dissolution and reconstitution of casein micelle containing dairy powders by high shear using ultrasonic and physical methods

The effect of shear on the solubilization of a range of dairy powders was investigated. The rate of solubilization of low solubility milk protein concentrate and micellar casein powders was examined during ultrasonication, high pressure homogenization and high-shear rotor–stator mixing and compared to low-shear overhead stirring. The high shear techniques were able to greatly accelerate the solubilization of these powders by physically breaking apart the powder agglomerates and accelerating the release of individual casein micelles into solution. This was achieved without affecting the structure of the solubilized proteins. The effect of high shear on the re-establishment of the mineral balance between the casein micelles and the serum was examined by monitoring the pH of the reconstituted skim milk powder after prior exposure to ultrasonication. Only minor differences in the re-equilibration of the pH were observed after sonication for up to 3 min, suggesting that the localized high shear forces exerted by sonication did not significantly affect the mass transfer of minerals from within the casein micelles.     

Enhanced creaming of milk fat globules in milk emulsions by the application of ultrasound and detection by means of optical methods

The effects of application of ultrasonic waves to recombined milk emulsions (3.5% fat, 7% total solids) and raw milk on fat destabilization and creaming were examined. Coarse and fine recombined emulsions (D[4,3]=9.3 μm and 2.7 μm, respectively) and raw milk (D[4,3]=4.9 μm) were subjected to ultrasound for 5 min at 35°C and 400 kHz or 1.6 MHz (using a single transducer) or 400 kHz (where the emulsion was sandwiched between two transducers). Creaming, as calculated from Turbiscan measurements, was more evident in the coarse recombined emulsion and raw milk compared to that of the recombined fine emulsion. Micrographs confirmed that there was flocculation and coalescence in creamed layer of emulsion. Coalescence was confirmed by particle size measurement. These results imply that ultrasound has potential to pre-dispose fat particles in milk emulsions to creaming in standing wave systems and in systems with inhomogeneous sound distributions.     

Gel forming behaviour of milk protein concentrates at moderate pressures

Abstract

Gel formation is a time dependent process, which is a function of many external variables. Here we report the effect of pressure on the gelation of certain milk proteins. It is found that hydrostatic pressure (1000 bar) decreases the gelation time of concentrated reconstituted milk (13%w/w protein) at around 50°C by more than sevenfold.     

Intensified recovery of valuable products from whey by use of ultrasound in processing steps – A review

The current review focuses on the analysis of different aspects related to intensified recovery of possible valuable products from cheese whey using ultrasound. Ultrasound can be used for process intensification in processing steps such as pre-treatment, ultrafiltration, spray drying and crystallization. The combination of low-frequency, high intensity ultrasound with the pre-heat treatment minimizes the thickening or gelling of protein containing whey solutions. These characteristics of whey after the ultrasound assisted pretreatment helps in improving the efficacy of ultrafiltration used for separation and also helps in preventing the blockage of orifice of spray dryer atomizing device. Further, the heat stability of whey proteins is increased. In the subsequent processing step, use of ultrasound assisted atomization helps to reduce the treatment times as well as yield better quality whey protein concentrate (WPC) powder. After the removal of proteins from the whey, lactose is a major constituent remaining in the solution which can be efficiently recovered by sonocrystallization based on the use of anti-solvent as ethanol. The scale-up parameters to be considered during designing the process for large scale applications are also discussed along with analysis of various reactor designs. Overall, it appears that use of ultrasound can give significant process intensification benefits that can be harnessed even at commercial scale applications.     

Effects of ultrasound synergized with microwave on structure and functional properties of transglutaminase-crosslinked whey protein isolate

In the present study, ultrasound (400 W, U), microwave heating (75 ℃ for 15 min, M) and ultrasound synergized with microwave heating (UM) pretreatments of whey protein isolate (WPI) were applied to investigate and compare their influence on structure, physicochemical and functional characteristic of transglutaminase (TGase)-induced WPI. From the results of size exclusion chromatography, it could be seen that all three physical pretreatments could promote the formation of polymers in TGase cross-linked WPI, whose polymer amounts were increased by the order of U, UM and M pretreatment. Among three physical methods, M pretreatment had the strongest effect on structure and functional characteristics of TGase-induced WPI. Furthermore, compared with TGase-induced WPI, α-helix and β-turn of M−treated TGase-induced WPI (M−WPI−TGase) were reduced by 7.86% and 2.93%, whereas its β-sheet and irregular curl were increased by 15.37% and 7.23%. Zeta potential, emulsion stability and foaming stability of M−WPI−TGase were increased by 7.8%, 59.27% and 28.95%, respectively. This experiment exhibited that M was a more effective pretreatment method than U, UM for WPI, which could promote its reaction with TGase and improve its functional properties.     

High-intensity ultrasound pretreatment influence on whey protein isolate and its use on complex coacervation with kappa carrageenan: Evaluation of selected functional properties

The aim of this work was to evaluate the influence of high-intensity ultrasound (HIUS) treatment on whey protein isolate (WPI) molecular structure as a previous step for complex coacervation (CC) with kappa-carrageenan (KC) and its influence on CC functional properties. Protein suspension of WPI (1% w/w) was treated with an ultrasound probe (24 kHz, 2 and 4 min, at 50 and 100% amplitude), non HIUS pretreated WPI was used as a control. Coacervation was achieved by mixing WPI and KC dispersions (10 min). Time and amplitude of the sonication treatment had a direct effect on the molecular structure of the protein, FTIR-ATR analysis detected changes on pretreated WPI secondary structure (1600–1700 cm⁻¹) after sonication. CC electrostatic interactions were detected between WPI positive regions, KC sulfate group (1200–1260 cm⁻¹), and the anhydrous oxygen of the 3,6 anhydro-D-galactose (940–1066 cm⁻¹) with a partial negative charge. After ultrasound treatment, a progressive decrease in WPI particle size (nm) was detected. Rheology results showed pseudoplastic behavior for both, KC and CC, with a significant change on the viscosity level. Further, volume increment, stability, and expansion percentages of CC foams were improved using WPI sonicated. Besides, HIUS treatment had a positive effect on the emulsifying properties of the CC, increasing the time emulsion stability percentage. HIUS proved to be an efficient tool to improve functional properties in WPI-KC CC.     

Recent advances in the application of ultrasound in dairy products: Effect on functional,physical, chemical,microbiological and sensory properties

Alternative methods for improving traditional food processing have increased in the last decades. Additionally, the development of novel dairy products is gaining importance due to an increased consumer demand for palatable, healthy, and minimally processed products. Ultrasonic processing or sonication is a promising alternative technology in the food industry as it has potential to improve the technological and functional properties of milk and dairy products. This review presents a detailed summary of the latest research on the impact of high-intensity ultrasound techniques in dairy processing. It explores the ways in which ultrasound has been employed to enhance milk properties and processes of interest to the dairy industry, such as homogenization, emulsification, yogurt and fermented beverages production, and food safety. Special emphasis has been given to ultrasonic effects on milk components; fermentation and spoilage by microorganisms; and the technological, functional, and sensory properties of dairy foods. Several current and potential applications of ultrasound as a processing technique in milk applications are also discussed in this review.     

Ultrasound impact on whey protein concentrate-pectin complexes and in the O/W emulsions with low oil soybean content stabilization

Consumers’ preference for products with reduced levels of fat increased in the last years. Proteins and polysaccharides have an important role due to their functional and interaction properties because, when combined in ratios and pH of higher potential for electrostatic interactions they may act as emulsifiers or stabilizers. This study evaluated the ultrasound impact on the electrostatic interaction between pectin (PEC) and whey protein concentrate (WPC) at different WPC:PEC ratios (1:1 to 5:1), and its effect on the emulsification and stability of emulsions formulated with WPC:PEC blends (1:1, 4:1) at low soybean oil contents (5 to 15%). Zeta potential analysis showed greater interactions between biopolymers at pH 3.5, which was proven in FTIR spectra. Rheology and turbidimetry showed that the ultrasound reduced the suspension viscosity and the size of the biopolymer complexes. Suspensions were Newtonian, whereas the emulsions showed shear-thinning behavior with slight increase in apparent viscosity as a function of oil content, and remained stable for seven days, with small droplets (<8 μm) stabilized and entrapped in a pectin network evidenced by confocal laser microscopy. Sonication was successfully applied to emulsion stabilization, improving the functional properties of WPC:PEC blends and enabling their application as low-fat systems, providing healthier products to consumers.     

High intensity ultrasound treatment of protein isolate extracted from dephenolized sunflower meal: Effect on physicochemical and functional properties

The influence of high intensity ultrasound (HIUS) on physicochemical and functional properties of sunflower protein isolates was investigated. Protein solutions (10% w/v) were treated with ultrasound probe (20 kHz) and ultrasound bath (40 kHz) for 5, 10, 20 and 30 min. Thermal stability of protein isolates was reduced as indicated by differential scanning calorimetry. Minimum thermal stability was observed at 20 min of sonication and increased further with increase in treatment time indicating aggregation at prolonged sonication. SDS-PAGE profile of proteins showed a significant reduction in molecular weight. Further, surface hydrophobicity and sulfhydryl content increased after HIUS treatment indicating partial unfolding of proteins and reduction in the intermolecular interactions. The particle size analysis showed that HIUS treatment reduced the particle size. Less turbid solution were observed largely due to reduction in particle size. HIUS decreased the available lysine content in protein isolates. Solubility, emulsifying capacity, emulsion stability, foaming capacity, foam stability and oil binding capacity were improved significantly, while as, water binding capacity was decreased. The effect of HIUS on physicochemical and functional properties of sunflower protein isolates was more pronounced in probe sonication rather than bath sonication. Protein isolates with improved functional properties can be obtained using high intensity ultrasound technology.     

Effects of ultrasound on the thermal and structural characteristics of proteins in reconstituted whey protein concentrate

The sonication-induced changes in the structural and thermal properties of proteins in reconstituted whey protein concentrate (WPC) solutions were examined. Differential scanning calorimetry, UV-vis, fluorescence and circular dichroism spectroscopic techniques were used to determine the thermal properties of proteins, measure thiol groups and monitor changes to protein hydrophobicity and secondary structure, respectively. The enthalpy of denaturation decreased when WPC solutions were sonicated for up to 5 min. Prolonged sonication increased the enthalpy of denaturation due to protein aggregation. Sonication did not alter the thiol content but resulted in minor changes to the secondary structure and hydrophobicity of the protein. Overall, the sonication process had little effect on the structure of proteins in WPC solutions which is critical to preserving functional properties during the ultrasonic processing of whey protein based dairy products.     

Detection of total protein in milk using phosphomolybdic acid‐mediated surface‐enhanced Raman spectroscopy

Although many surface‐enhanced Raman scattering (SERS)‐based methods for detecting specific proteins have been studied, simple and direct detection of total protein in liquid using a SERS‐based method remains difficult. In this study, a distinguishable effect on the SERS spectra from pre‐mixture of phosphomolybdic acid (PMA) with protein was found, indicating that PMA could be used as a SERS reporter for total protein detection in a liquid sample. Further experiments confirmed a good linear relationship between a premixed concentration of protein (casein, whey protein or bovine serum albumin) and the SERS intensity of PMA in our SERS system. Using casein as a reference, a PMA‐mediated SERS method was proposed that can quantitatively analyze protein at 2.5–25 µg/ml with a limit of detection of 1.5 µg/ml. Our PMA‐mediated SERS method is a simple and rapid method for quantitative analysis of total protein in milk. Copyright © 2015 John Wiley & Sons, Ltd.     

Enhanced gelling properties of myofibrillar protein by ultrasound-assisted thermal-induced gelation process: Give an insight into the mechanism

Effects of the incorporation of ultrasound with varied intensities (0–800 W) into the thermal-induced gelation process on the gelling properties of myofibrillar protein (MP) were explored. In comparison with single heating, ultrasound-assisted heating (<600 W) led to significant increases in gel strength (up to 17.9%) and water holding capacity (up to 32.7%). Moreover, moderate ultrasound treatment was conducive to the fabrication of compact and homogenous gel networks with small pores, which could effectively impair the fluidity of water and allow redundant water to be entrapped within the gel network. Electrophoresis revealed that the incorporation of ultrasound into the gelation process facilitated more proteins to get involved in the development of gel network. With the intensified ultrasound power, α-helix in the gels lowered pronouncedly with a simultaneous increment of β-sheet, β-turn, and random coil. Furthermore, hydrophobic interactions and disulfide bonds were reinforced by the ultrasound treatment, which was in support of the construction of preeminent MP gels.