Binding of retinyl acetate to whey proteins or phosphocasein micelles: Impact of pressure-processing on protein structural changes and ligand embedding

A whey protein isolate (WPI) and native phosphocaseins (PC) at pH 6.5–6.6 were processed with retinyl acetate (RAC) using pressure-assisted technological tools to improve RAC embedding through processing-induced protein structural changes. To this end, protein-RAC dispersions were submitted to ultra-high pressure homogenisation (UHPH) at 300 MPa and an initial fluid temperature (T_in) of 14 °C or 24 °C, or isostatic high-pressure at 300 MPa and 14 °C or 34 °C for 15 min. A short-time thermal treatment (STTT, 73 °C for 4 s) able to generate WPI aggregates was assessed for comparison. Processing effects were investigated in terms of protein particle sizes and molecular weights (M_w). M_w calculated using protein size determination obtained from light scattering measurements were in agreement with the known values. The amounts of RAC retained in WPI particles (unfolded and/or aggregated proteins) or in PC assemblies were quantitated after protein precipitation by ammonium sulphate. A 2.3–3.7 nmol RAC was carried per mg of pressure-denatured whey proteins, significantly less than after STTT (6.3 nmol RAC per mg of heat-denatured whey proteins) indicating that RAC embedding varied according to the technological tool, pressure or temperature. A 3.8–5.4 nmol RAC was carried per mg of PC assemblies through pressure-induced dissociation/reassociation of PC micelles. Combined pressure and mild temperature increased RAC embedding in PC assemblies.     

High-pressure-induced changes in the rennet coagulation properties of bovine milk

The mechanism of high-pressure (HP)-induced changes in rennet coagulation properties of milk, particularly the role of whey protein-casein micelle associations, was studied. Treatment at 100 or 250 MPa reduced the rennet coagulation time (RCT) of raw skimmed bovine milk, compared with untreated milk. Treatment at 400 MPa had little effect, but at 600 MPa, RCT increased considerably. HP-induced increases in RCT did not occur in serum protein-free milk or milk treated with the sulphydryl-oxidising agent KIO₃, which prevents association of denatured β-lactoglobulin with casein micelles. Treatment at 5 or 10 °C at 250–600 MPa resulted in shorter RCT than treatment at 20 °C. In milk without KIO₃, coagulum strength was highest after treatment at 250 or 400 MPa, whereas in milk with KIO₃ it was highest after treatment at 400 MPa. These results indicate the significance of HP-induced association of whey proteins with casein micelles for rennet coagulation properties of milk.     

Microbial shelf-life of high-pressure-homogenised milk

The anti-bacterial effect of high pressure homogenisation (HPH) on milk is widely reported but the shelf-life of HPH-treated milk, as reported in this communication has not been studied thus far. Raw whole milk was homogenised at 200 or 250 MPa at 55 or 70 °C and counts of total bacteria (TBC), psychrotrophs, pseudomonads, coliforms, lactobacilli, Bacillus cereus and Staphylococcus aureus were determined throughout subsequent storage for 14 days at 4 °C. Immediately after HPH treatment, counts of all bacteria were below the level of detection but after storage for 14 days at 4 °C, TBC, psychrotroph and pseudomonad counts had reached ∼10⁸ cfu mL⁻¹ in all samples treated with HPH. The limited shelf-life obtained indicates that HPH of milk at these processing parameters it is not a suitable alternative to pasteurisation for extending the shelf-life of milk.     

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.     

Inactivation of Saccharomyces cerevisiae and Lactobacillus plantarum in orange juice using ultra high-pressure homogenisation

Yeasts and lactic acid bacteria are the usual contaminants in orange juice and responsible for decreasing the shelf life of the product. Ultra high-pressure homogenisation has been shown to be an alternative to the traditional thermal pasteurisation of pumpable foods. The product was pumped through a homogeniser valve at 100 MPa, 150 MPa, 200 MPa, 250 MPa and 300 MPa using two synchronized overlapped intensifiers at a flow rate of approximately 270 mL/min. The inlet temperature was kept at 10 °C, pH at 4.1 and soluble solids at 10.0 °Bx. After processing, the product was immediately cooled to 4 °C and the microbiological count was determined. The study showed that Lactobacillus plantarum and Saccharomyces cerevisiae are sensible to ultra high-pressure homogenisation treatment. The results indicated that pressures higher than 250 MPa were able to completely destroy initial loads of 1.2 × 107 UFC/mL of L. plantarum and 2.9 × 105 UFC/mL of S. cerevisiae in orange juice, making this technology a promising way to nonthermally process orange juices.Industrial relevanceThis paper deals with inactivation of microorganism contaminants of orange juice using dynamic ultra high process technology. It is of industrial interest and relevance to evaluate the use of this non-thermal emerging technology to process fluid foods that may result into better taste, optimum product functionality, safety and quality characteristics.     

The effect of sonication and high pressure homogenisation on the properties of pure cream

The homogenisation of milk and cream has been widely studied but the effect of sonication on the structural and functional properties of cream is not well known. In this study, raw milk, ultrafiltration retentate and cream samples were sonicated at 20 kHz and the rennet and acid gelation properties of these sonicated samples investigated. High pressure homogenisation at 80 bar was also performed for comparison. Sonication of raw milk and retentate samples led to a decrease in the fat globule size. Conversely, the fat globules in cream samples sonicated at < 10 °C flocculated to form grapelike structures whereas the cream samples sonicated at 50 °C did not form such aggregates. High pressure homogenisation at 50 °C led to similar flocculated structures, but these were not observed at low temperatures. This suggests a potential benefit of sonication technology in allowing low temperatures to be utilised for cream homogenisation, reducing energy demand. However, a gel made using cheese-milk with sonicated cream resulted in separation of a fat layer rather than the incorporation of the fat globules into the gel matrix. Rennet gelation properties of both the sonicated or homogenised samples were significantly superior to a native control sample where the resultant gels had shorter coagulation times and decreased syneresis.Industrial RelevanceHomogenisation of dairy cream is normally carried out at temperatures of around 50 °C, to ensure that the fat is in the liquid state. In this work, we show that we can achieve comparable changes to the fat globules within the cream using ultrasound at much lower temperatures (< 10 °C). The ability to form flocculated fat particles at lower temperatures could lead to reduced costs through reduced energy demand.     

Effect of high pressure,temperature, and storage on the color of porcine longissimus dorsi

The color of pork longissimus dorsi high pressure (HP) treated at 200 to 800 MPa at 5 and 20 °C for 10 min was determined to a high degree by pressure level and to a lesser degree by temperature. Severe color changes appeared up to a threshold pressure at 400 MPa. HP treatment at 20 °C compared to 5 °C resulted in meat, which was less red and slightly lighter. Storage at 2 °C for 6 days had no effect on lightness due to no further protein denaturation, but meat HP treated above 300 MPa became significantly less red and more yellow within the first day of storage. Reflectance spectra showed that a short-lived ferrohemochrome myoglobin species was formed during HP treatment at 300 to 800, but transformed into a brown, ferric form of the pigment within the first day of storage. This explains the observed changes in the redness and yellowness after one day of storage.     

Enrichment of the conjugated linoleic acid content of bovine milk fat by dry fractionation

This study investigated the effect of fat fractionation on the conjugated linoleic acid (cis-9, trans-11-C_18 : 2) content of bovine milk fat. Anhydrous milk fat was fractionated into hard and soft fractions using controlled cooling and agitation. Fractionation of milk fat pre-melted at 60°C using a temperature programme of 33–10°C and a cooling rate of 0.58°C h⁻¹ yielded a soft fraction containing 63.2% more conjugated linoleic acid (2.22 g 100 g⁻¹ FAME), which was also enriched in polyunsaturated fatty acids and vaccenic acid (trans-11-C_18 : 1) compared with the parent fat. Agitation following fractionation was found to have a negative effect on the conjugated linoleic acid content of the soft fraction. Refractionation of the soft fraction did not increase the yield of conjugated linoleic acid. The conjugated linoleic acid and trans fatty acid content of 26 selected food products ranging in milk fat content from 0 to 100% is reported. Conjugated linoleic acid concentrations ranged from 0 to 16.2 mg g⁻¹ fat and were generally lower than the trans fatty acid content which ranged from 0 to 155.7 mg g⁻¹ fat. Spreads containing vegetable oils contained higher trans fatty acid and lower conjugated linoleic acid contents than milk fat-containing products. This study highlights that a milk fat fraction enriched in conjugated linoleic acid may be achieved by dry fractionation.     

Influence of enzymatic cross-linking on milk fat globules and emulsifying properties of milk proteins

The enzyme transglutaminase (TGase) can modify dairy protein functionality through cross-linking of proteins. This study examined the effects of TGase treatment on milk fat globules and the emulsifying properties of milk proteins. The extent of TGase-induced cross-linking of caseins increased with incubation time, with no differences between whole and skim milk. Extensive clustering of fat globules in extensively cross-linked raw whole milk occurred on homogenisation at 400 or 800 bar. Considerably less clustering of fat globules was observed when recombined milk (90 g fat L^–1) was prepared from TGase-treated skim milk and homogenised at 400 or 800 bar. TGase treatment did not affect fat globule size in cream, but prevented coalescence of fat globules therein, possibly through cross-linking of milk fat globule membrane components. TGase-induced cross-linking of milk proteins affected their emulsifying properties and may increase the stability of natural milk fat globules against coalescence.     

Properties of low-fat stirred yoghurts made from high-pressure-processed skim milk

Physical properties of stirred yoghurt made from reconstituted skim milk that was high-pressure (HP)-treated at 100, 250 or 400 MPa, at 25, 70 or 90 °C, for 10 min, prior to inoculation with yoghurt cultures, were studied; portions of milk HP-treated at 25 °C were also heat-treated at 90 °C for 10 min before or after pressure treatment. Control yoghurts were made from skim milk given a heat treatment at 90 °C for 10 min. Fermentation time was not affected by treatment applied to the milk. HP treatment of skim milk at 25 °C, before or after heat treatment, gave stirred yoghurts of similar viscosities to that made from conventionally heat-treated milk. Lower viscosities were obtained when stirred yoghurts were made with milk HP-treated at elevated temperatures. A model is proposed to correlate properties of yoghurt with HP/heat-induced changes in interactions and structures of protein in the milk samples.Industrial relevanceTo meet end user expectations, the dairy industry needs to diversify its product range by tailoring specific functionalities. To meet these expectations, new processing methods such as high-pressure processing are of interest for their potential to achieve specific and/or novel functionalities and/or improve efficiencies, including reduced chemical and water use. In this paper, an investigation of the use simultaneous pressurization and heating of milk before the manufacture of stirred yoghurt is presented.     

High-pressure calorimetric evaluation of ice crystal ratio formed by rapid depressurization during pressure-shift freezing of water and pork muscle

The amount of ice nuclei formed during the pressure release is important for the final formation and development of ice crystals in pressure shift freezing (PSF) frozen products. In this study, a high-pressure (HP) calorimeter was used to evaluate the ratio of ice crystals instantaneously formed by rapid depressurization during PSF of pure water and pork muscle tissue. Experiments were carried out initial pressure levels of 62, 115, 157 and 199 MPa, with corresponding phase change temperatures of −5, −10, −15 and −20 °C, respectively (slightly higher than phase change point of water–ice I). The ice crystal ratio was determined based on calorimetric peak measured and heat balance. The evaluated regression relationship between observed ice crystal ratio (R_ice in %) and pressure (P, MPa) was R_ice–water = 0.115P + 0.00013P² (R² = 0.96, n = 9) for pure water, and R_ice–pork = 0.080P + 0.00012P² (R² = 0.95, n = 11) for pork muscle. Compared to other methods, the calorimetric evaluation does not require any of the pressure-related properties of the test sample. HP calorimetry can thus be used to evaluate ice crystal ratio for PSF of foods even though their pressure related properties may be unknown.     

Stickiness curves of high fat dairy powders using the particle gun

High fat (>42%) dairy powders are inherently sticky due to their high levels of liquid surface fat. Incorrect operating conditions when spray drying these powders can rapidly lead to blockages. The particle gun was used to characterise the stickiness curves of high fat cream and cheese powders. Stickiness was shown to increase with increasing temperature to a maximum at 50 °C after which it decreased until no stickiness was observed above 68 °C. A dramatic increase in stickiness for the powders was found when the relative humidity of the air was increased past a certain critical point for each temperature. This was attributed to the lactose component of the powder exceeding its glass transition temperature by a critical amount. Best estimates of the (T−T_g)_crit. values for White Cheese Powder, Low Fat Cream Powder and High Fat Cream Powder were 28, 37 and 38 °C, respectively.     

The influence of lactoperoxidase,heat and low pH on survival of acid-adapted and non-adapted Escherichia coli O157:H7 in goat milk

In hot climates where quality of milk is difficult to control, a lactoperoxidase (LP) system can be applied in combination with conventional preservation treatments at sub-lethal levels to inhibit pathogenic microbes. This study investigated the effect of combined heat treatments (55 °C, 60 °C and 72 °C) and milk acidification (pH 5.0) on survival of acid-adapted and non-adapted Escherichia coli O157:H7 strains UP10 and 1062 in activated LP goat milk. Heat treatment at 72 °C eliminated E. coli O157:H7. Acid-adapted strains UP10 and 1062 cells showed resistance to combined LP and heat at 60 °C in fresh milk. The inhibition of acid-adapted and non-adapted E. coli O157:H7 in milk following combined LP-activation, heat (60 °C) and milk acidification (pH 5.0) suggests that these treatments can be applied to reduce E. coli O157:H7 cells in milk when they occur at low numbers (<5 log₁₀ cfu mL^?1) but does not eliminate E. coli O157:H7 to produce a safe product.     

Effect of high pressure (HP) on the quality and shelf life of red mullet (Mullus surmelutus)

The effect of different temperature/time/pressure high hydrostatic pressure (HP) treatment on the quality and shelf life of red mullet were studied. Different high pressure treatments (at 3, 7, 15 and 25 °C, 5 to 10 min and 220, 250 and 330 MPa) were tested to establish the best processing conditions for the quality of red mullet. The effect of the process on the quality of the sample was examined by colour, Trimethylamine nitrogen (TMA-N) and Thiobarbituric acid number (TBA) analysis. Based on the results of the parameters, the best combinations of HP treatments were determined as 220 MPa/5 min/25 °C and 330 MPa/5 min/3 °C for red mullet. The effects of this combination treatment on sensory, chemical and microbiological properties of red mullet stored at 4 °C were studied. The results obtained from this study showed that the shelf life of untreated and HP treated stored at 4 °C, as determined by overall acceptability of sensory and microbiological data, are 12 days for untreated red mullet and 14 days for treated red mullet at 220 MPa for 5 min at 25 °C and 15 days for treated red mullet at 330 MPa for 5 min at 3 °C.Industrial relevanceFresh fish have short shelf life. HP treatment has shown to be an effective method to control pathogen and spoilage microorganisms in fish and fish products. However, high pressure treatment can promote colour and oxidation changes that could modify their sensory characteristics. The main objective of the first part of this study was to detect the best combination among the applied pressure (220, 250 and 330 MPa), temperature (3, 7, 15 and 25 °C) and time (5 and 10 min) combinations. The treatment ranges were chosen according to the unchanging colour, lower TBA value and TMA stability by HP and considering the economical aspects of HP processing. In the second part of the study, HP was applied on the selected samples and a shelf-life study was performed by measuring the changes in the quality parameters, of the samples throughout their storage. The storage conditions were set so as to achieve refrigeration handling (4 °C). Shelf-life estimation was performed according to the data obtained. HP (at 220 MPa for 5 min at 25 °C and at 330 MPa for 5 min at 3 °C) treatment is the most effective treatment for shelf-life extension as compared to non-treated red mullet.     

The impact of high pressure on glucosinolate profile and myrosinase activity in seedlings from Brussels sprouts

The potential of high pressure (HP) to control bioactive components using seedlings of Brussels sprouts as a simple non-chopped vegetable system was examined. Enzyme activity in situ compared to purified enzyme and residual enzyme substrate in situ are used as three complementary measures for the HP effect. Purified myrosinase and seedlings of Brussels sprouts were submitted to HP 200–800 MPa at 5 °C for 3 min. The myrosinase activity decreased for both myrosinase systems upon increasing pressure to 800 MPa. Applying first-order kinetic to determine activation volumes revealed a linear relationship from 400 to 600 (ΔV^# = − 19.04 mL/mol) and 450–600 MPa (ΔV^# = − 37.79 mL/mol) for seedlings and purified myrosinase, respectively, indicating a protective effect of the plant matrix against enzyme inactivation. Purified myrosinase was activated at 200 MPa but at 800 MPa the glucosinolate degradation due to pressure induced disruption of the plant matrix seems to be partly counter-acted by myrosinase inactivation.Industrial relevanceHigh Pressure (HP) processing is an effective non-thermal preservation treatment for liquid and solid food. Moreover, over the last years, the potential of this technology to improve health and safety attributes of foods has been demonstrated. In particular, the ability of HP to preserve bioactive compounds has been established. There are only few studies evaluating the impact of HP on the complex bioactive glucosinolates-myrosinase. Therefore, this study opens the doors through the application of HP to preserve the bioactive glucosinolates in cruciferous vegetables by creating new processing solutions through controlled enzyme inactivation. Thus, HP could be an effective tool to achieve more effective solutions to obtain the new generation of convenient food and meet the need for new bioactive food products.     

Direct measurement of phase transitions in milk fat during cooling of cream—a low-field NMR approach

Low-field ¹H nuclear magnetic resonance (NMR) transverse relaxation (T₂) was measured during cooling (31–4 °C) of cream with low or high content of long-chain fatty acids (FA). Distributed analysis of the T₂ relaxation data revealed marked differences in the T₂ relaxation characteristics of the liquid fat population (10–100 ms), which was ascribed to differences in the size of the fat globules in the two types of cream. Principal component analysis (PCA) of the obtained T₂ relaxation decay data showed clear shifts in NMR relaxation behavior at 17 and 22 °C for cream with low or high content of long-chain FA, respectively. Differential scanning calorimetric measurements on the creams revealed that these shifts took place at the onset of crystallization of fat. Consequently, the present study demonstrated that low-field NMR can be used to measure phase transitions in cream.     

Kinetics of destruction ofEscherichia coliandPseudomonas fluorescensinoculated in ewe's milk by high hydrostatic pressure

Ewe's1999199919991999 Academic PressAcademic PressAcademic PressAcademic Pressmilk standardized to 6% fat was inoculated with Escherichia coliand Pseudomonas fluorescensat a concentration of 10⁷and 10⁸ cfu ml⁻¹respectively, and treated by high hydrostatic pressure. Treatments consisted of combinations of pressure (50-300 MPa), temperature (2, 10, 25 and 50°C), and time (5, 10 and 15 min). Violet red bile agar and crystal-violet-tetrazolium count were used to determineE. coliandP. fluorescensrespectively. Pressurization at low and moderately high temperatures produced higher P. fluorescensinactivation than treatments at room temperature, while pressurization at only moderately high temperature produced high E. coliinactivation; low and room temperatures produced similar reductions. On E. coli,reductions of 6.055 log units were produced with 300 MPa for 15 min at 50°C, while on P. fluorescens,reductions of 5.059 log units were produced with 250 MPa for 15 min at 50°C. Both micro-organisms showed a first-order kinetics of destruction in the range 0-30 min, with D-values (at different temperatures and pressures from 150 to 300 MPa) between 2.055 and 18.058 min for E. coli,and 2.058-23.053 min for P. fluorescens.Abaroprotective effect of ewe's milk (6% fat) on both micro-organisms was observed, in comparison with other studies using different means and similar pressurization conditions.     

Development of innovative biodegradable films based on biomass of Saccharomyces cerevisiae

Biodegradable alternatives to petroleum-based polymers are being developed for food packaging. Saccharomyces cerevisiae has been widely used for the production of fermented beverages and leavened foods. In this work, the application of high pressure homogenisation and a thermal treatment to the yeast biomass was studied to develop biodegradable films. The highest dispersibility indexes of protein (84.5 ± 3.3%), carbohydrates (24.3 ± 1.1%), RNA (40.6 ± 0.9%) and soluble solids released were found at high homogenisation pressure (125 MPa). Combinations of one or two homogenisations and a thermal treatment at 90 °C during 20 minutes were applied and dispersions were fully characterised, focusing on their film-forming capacity. The best combination was homogenisation, heat treatment and a further homogenisation, since it produced films that presented good attributes, great continuity and homogeneity without small cracks. However, hydration of films was increased from 0.31 to 0.48 gH₂O/g.d.m with the number of homogenisation processes applied. Results revealed that yeast biomass is a viable source to be used in biodegradable films.Industrial relevanceSaccharomyces cerevisiae yeasts have many applications in food industry. The development of biodegradable films based on yeast biomass carries many advantages such as, the possibility of using commercial pressed baker's yeast or an industrial residue from brewing industry, the use of a low-cost sources and the application of environmentally friendly procedures. The methodologies applied for the development of the film forming dispersion, high pressure homogenisation and thermal treatment, are able to be scaled-up to an industrial level.     

Modeling the inactivation kinetics of fruit bromelain in pineapple during high-pressure and thermal treatments

The stability of fruit bromelain (FBM) in pineapple pulp was studied within a high-pressure domain of 0.1–600 MPa/30–70 °C/1 s–30 min. The pulse effect was quantified as a function of pressure, temperature, pressure build-up and decompression times. A maximum of 60% reduction in FBM activity was obtained after a single pulse of 600 MPa/70 °C. Upon applying nth order model, the obtained reaction order (n) for thermal (0.1 MPa/30–70 °C) and high-pressure (100–600 MPa/30–70 °C) inactivation was 1.1 and 1.2, respectively. The inactivation rate constant (k) ranged from 1.2 to 45.0 × 10^− 3 U^n − 1 min^− 1. The activation energy was nonlinearly dependent on pressure (P); whereas, the activation volume was linearly related to temperature (T). The nonlinear dependence of k on P and T was modeled by an empirical equation. The D-values obtained from the empirical model appeared to be more realistic than those from the log-linear kinetics.Industrial relevancePineapple fruit bromelain (FBM) has numerous health benefits and therapeutic effects. It is a protease enzyme that helps in digestion. Processing of pineapple pulp needs attention towards retaining the maximum FBM activity in it. A detailed kinetic study of FBM within a broad range of pressure–temperature–time domain will help in designing a high-pressure process for the pineapple pulp with respect to its bromelain stability.     

Stickiness of skim milk powder using the particle gun technique

The particle gun method for investigating the initiation of stickiness of dairy powders was examined using skim milk powder (SMP). The point at which significant stickiness occurred was at a constant temperature above the glass transition temperature (T_g) of amorphous lactose regardless of the temperature and relative humidity (RH) conditions of the air in the particle gun. This point has been called (T–T_g)_critical. The initial water activity of SMP did not significantly affect (T–T_g)_critical, confirming that stickiness is a surface phenomenon. Changing the particle feed rate did not effect (T–T_g)_critical or the rate at which the powder accumulated on the target plate. The errors in measuring the value of (T–T_g)_critical and the rate of stickiness development [slope of the deposition line versus (T–T_g)] for SMP were found to be 33.6 ± 0.8 °C and 3.1 ± 0.7% deposition °C^?1, respectively, after standardisation of the ambient air conditions around the powder feeder and the initial water activity of the SMP.