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.     

High-pressure homogenisation of raw bovine milk. Effects on fat globule size distribution and microbial inactivation

Whole raw milk was processed using a 15 L h⁻¹ homogeniser with a high-pressure (HP) valve immediately followed by a cooling heat exchanger. The influence of homogenisation pressure (100–300 MPa) and milk inlet temperature T_in (4°C, 14°C or 24°C) on milk temperature T₂ at the HP valve outlet, on fat globule size distribution and on the reduction of the endogenous flora were investigated. The T_in values of 4–24°C led to milk temperatures of 14–33°C before the HP valve, mainly because of compression heating. High T_in and/or homogenisation pressure decreased the fat globule size. At 200 MPa, the d_4.3 diameter of fat globules decreased from 3.8±0.2 (control milk) to 0.80±0.08 μm, 0.65±0.10 or 0.37±0.07 μm at T_in=4, 14°C or 24°C, respectively. A second homogenisation pass at 200 MPa (T_in=4°C, 14°C or 24°C) further decreased d_4.3 diameters to about 0.2 μm and narrowed the size distribution. At all T_in tested, an homogenisation pressure of 300 MPa induced clusters of fat globules, easily dissociated with SDS, and probably formed by sharing protein constituents adsorbed at the fat globule surface. The total endogenous flora of raw milk was reduced by more than 1 log cycle, provided homogenisation pressure was ⩾200 MPa at T_in=24°C (T₂∼60°C), 250 MPa at T_in=14°C (T₂∼62°C), or 300 MPa at T_in=4°C (T₂∼65°C). At all T_in tested, a second pass through the HP valve (200 MPa) doubled the inactivation ratio of the total flora. Microbial patterns of raw milk were also affected; Gram-negative bacteria were less resistant than Gram-positive bacteria.     

Effect of high pressure homogenization on lactic acid bacteria phages and probiotic bacteria phages

To investigate the effect of high pressure homogenization on virus inactivation, phages specific for Lactobacillus delbrueckii, Lactobacillus helveticus, Streptococcus thermophilus, Lactococcus lactis, Lactobacillus paracasei and Lactobacillus plantarum were studied. The influence of pressure, number of passes, suspension medium and phage concentration were studied at 25 °C. Reductions in viability were proportional to pressure and number of passes, though the inactivation extent was phage-dependent. At 100 MPa, some bacteriophages were completely inactivated (6 log₁₀ reduction) after 3 or 5 passes, while others remained infective after 8 passes. For all phages, treatment at 60 MPa was insufficient for complete inactivation, even after 8 passes. No clear influence of suspending medium was observed. Inactivation seems to depend on phage concentration; the higher the initial load, the bigger the reduction achieved. Although these results showed that several phages studied are resistant to high-pressure homogenization, this strategy could be combined with others to control their presence in raw milk.     

Effect of soy protein substitution for sodium caseinate on the transglutaminate-induced cold and thermal gelation of myofibrillar protein

The influence of soy protein isolate (SPI) substitution for sodium caseinate (SC) on the properties of cold-set (4 °C) and heat-induced gels of pork myofibrillar protein (MP) incubated with microbial transglutaminase (TG) was investigated. The strength of cold-set MP–SC gels (formed in 0.45 M, NaCl, 50 mM phosphate buffer, pH 6.25) increased with time of TG incubation, but those gels with more than 66% SPI substituted for SC had a >26% reduced strength (P < 0.05). Upon cooking, both incubated and non-incubated protein sols were quickly transformed into highly elastic gels, showing up to 6000 Pa in storage modulus (G′) at the final temperature (72 °C). However, no differences (P < 0.05) in G′ were observed between heated samples with SPI and SC. Myosin heavy chain, casein and soy proteins gradually disappeared with TG incubation, contributing to MP gel network formation. Both cold-set and heat-induced gels had a compact protein matrix, attributable to protein cross-linking by TG.     

Stinky tofu as a rich source of bioavailable S-equol in Asian diets

The compound S-equol (4′,7-dihydroxy-isoflavandiol), a gut bacterial metabolite of isoflavone daidzein, benefits health, but only 20–60% of humans can produce equol after ingesting isoflavones, and it exists only in foods of animal origin in trace amounts. A recent study found a source of stinky tofu contained S-equol. As stinky tofu is a popular traditional fermented soy food in Taiwan, we analyzed S-equol contents of commercial samples, surveyed the intake frequency, and investigated the bioavailability of S-equol by monitoring urinary kinetics following ingestion. Our results showed 91% of the 138 stinky tofu dishes contained S-equol. The mean content per serving (average 198 g) was 2.3 ± 2.5 mg, the highest being 16.3 mg. Stinky tofu eaters on average ingested this food 3.3 times per month. S-equol from ingested stinky tofu appeared in urine within 1 h, reaching maximum excretion at 3.4 h, and 67% of the ingested S-equol was recovered in urine, indicating a rapid and high absorption. Our studies suggest stinky tofu can be a promising dietary source of S-equol for its high content and bioavailability. Further study on the S-equol producing bacteria in stinky tofu is merited for the development of other S-equol rich soy products.     

The effect of high hydrostatic pressure on the flow behaviour of skim milk–gelatin mixtures

The flow behaviour of aqueous solutions of gelatin, and skim milk–gelatin mixtures treated by high-pressure processing (HPP) were investigated. HPP was carried out at 5 °C for 15 min, at 150 MPa, 300 MPa, 450 MPa and 600 MPa, and the gelatin concentrations were varied from 0 to 1 wt.%. Viscosity measurements showed that the HPP treatment did not affect the flow behaviour of gelatin alone, nor that of the skim milk–gelatin mixtures made with < 0.4 wt.% gelatin. However, at gelatin concentration > 0.4 wt.%, the mixtures treated with 300 and 450 MPa exhibited a peculiar flow behaviour, where at intermediate shear rates the viscosity was higher than that of the non-treated mixture or the mixtures treated at 150 MPa and 600 MPa. Particle size measurements showed that for gelled mixtures (> 0.4 wt.% gelatin) 300 MPa HPP treatment resulted in an increase in the particle size, while at all other pressure treatments (> 150 MPa), a shift in particle size distribution to lower sizes was observed. Confocal microscopy showed that these skim milk–gelatin mixtures were phase-separated with a gelatin continuous phase, this was confirmed by dynamic rheological measurements which showed that qualitatively the viscoelastic properties of the mixtures were the same. A mechanism of the effect of high-pressure treatment on the casein micelle in skim milk–gelatin mixtures is proposed.Industrial relevanceThis fundamental work, dealing with the effect of high pressure on the physicochemical properties skim milk–gelatin mixtures could be relevant to the industry in several ways. Firstly, skim milk–gelatin mixtures are widely used in the dairy industry, particularly in yoghurt manufacture, where gelatine is used as a stabiliser. In addition the application of High Hydrostatic Pressure to such a system is also relevant, as this technology could be used as a substitute to the conventional heat treatment processes. Secondly, an important finding of this study is that under certain conditions of high pressure and gelatine concentration, an increase in viscosity is observed at intermediate shear-rate (between 10 and 100 s^?1). This is highly relevant to Industry if the system requires subsequent pumping. Thirdly, from a sensory view point, this range of shear rates (10 and 100 s^?1) is comparable to that experienced by a food bolus during swallowing. Thus, this effect of high pressure on the viscosity can influence sensory attribute of the skim milk–gelatin food system.     

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.     

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.     

Physical,Chemical and Sensory Qualities of Roselle Water Extract-coagulated Tofu Compared with Tofu from Two Natural Coagulants

Roselle calyces (Hibiscus sabdariffa) aqueous extracts were used in coagulating soymilk at four different concentrations (2.5%, 5%, 10% and 20%). The physical, chemical and sensory qualities of the tofu preparations were compared with fermented maize liquor and Calotropis procera extract coagulated tofus. pH of roselle extracts was acidic in nature (2.01 – 3.74) which was attributed to the ability to coagulate soy proteins. pH, titratable acidity of the roselle coagulated tofus ranged from 5.32 – 6.26 and 0.16 – 0.43% respectively and the yield and protein content ranged from 87.3 – 95.9 g and 42.6 – 46.3 g/100 g respectively. The yield of roselle coagulated tofu increased with increase in concentration of the roselle extracts. Roselle extract when utilized at 2.5% concentration will yield tofu that is acceptable and comparable to tofu coagulated with fermented maize steep water in terms of appearance, flavour and overall acceptability at p > 0.05. Roselle extract at 5% and 10% also yielded tofus that were acceptable in terms of all of the attributes tested.     

Microstructural and biochemical changes in raw and germinated cowpea seeds upon high-pressure treatment

Raw and germinated cowpea seeds (Vigna sinensis) were subjected to high-pressure treatment at 300–500 MPa for 15 min. Standard analyses, i.e., determination of protein and starch content, protein solubility, starch availability as well as microscopic observation (SEM) were used in order to study biochemical and microstructural changes in treated seeds.In opposite to germination process, the high-pressure treatment of seeds did not evoke significant changes in starch content (total and available). However, both the processes used significantly affected protein content (total and soluble). Pressurization of raw and germinated seeds evoked diversified effect on the total protein content. Whereas, the germinated seeds and/or these high pressure-treated ones demonstrated significantly higher content of soluble protein compared to raw material. The most distinct changes in microstructure of germinated seeds treated with pressure (500 MPa/15 min) were observed, and they mostly were related to the alteration in protein structure.     

Effects of combined high pressure and enzymatic treatments on the hydrolysis and immunoreactivity of dairy whey proteins

The effect of high-pressure (HP) treatment on the hydrolysis of dairy whey proteins by trypsin, chymotrypsin and pepsin was analysed. Isostatic pressure (100–300 MPa for 15 min at 37 °C) was applied to the protein substrate prior to its enzymatic hydrolysis. Digestion was also conducted at atmospheric pressure (0.1 MPa) and under high pressure. The extent of hydrolysis was measured by the o-phthaldialdehyde method, the peptide profile was analysed by reverse-phase high performance liquid chromatography (RP-HPLC) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and the residual immunochemical reactivity was assessed by an ELISA test using a pool of seven sera from children allergic to bovine milk, an individual serum also positive (positive control) and two sera from non-allergic children (negative controls). The high pressure increased the degree of hydrolysis by the three enzymes used. Chymotrypsin and trypsin showed the highest proteolysis at 100 and 200 MPa followed by pepsin at 300 MPa. The β-lactoglobulin was hydrolysed by trypsin and chymotrypsin at atmospheric and at high pressures, whereas the pepsin only hydrolysed this protein under high pressure. Pepsin and trypsin hydrolysed α-lactalbumin in all cases. In contrast, this protein was not digested by chymotrypsin, irrespective of the pressure applied. An important decrease of immunochemical reactivity was found for pepsin and trypsin hydrolysates obtained under high pressure. The pool of seven sera detected immunoreactivity in the products of chymotrypsin hydrolysis under high pressure, which was not detected when the serum of one patient was used. The results suggest that dairy whey hydrolysates obtained by pepsin and trypsin in combination with HP treatment could be used as a source of peptides in hypo-allergenic infant formulae.     

High hydrostatic pressure processing on microstructure of probiotic low-fat yogurt

The effect of milk processing on the microstructure of probiotic low-fat yogurt was studied. Skim milk fortified with skim milk powder was subjected to three treatments prior to innoculation: thermal treatment at 85 °C for 30 min, high hydrostatic pressure at 676 MPa for 5 min, and combined treatments of high hydrostatic pressure (HHP) and heat. The processed milk was then fermented by using two different starter cultures containing Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus, and Bifidobacterium longum. The microstructure of heat-treated milk yogurt had fewer interconnected chains of irregularly shaped casein micelles, forming a network that enclosed the void spaces. On the other hand, microstructure of HHP yogurt had more interconnected clusters of densely aggregated protein of reduced particle size, with an appearance more spherical in shape, exhibiting a smoother more regular surface and presenting more uniform size distribution. The combined HHP and heat milk treatments led to compact yogurt gels with increasingly larger casein micelle clusters interspaced by void spaces, and exhibited a high degree of cross-linking. The rounded micelles tended to fuse and form small irregular aggregates in association with clumps of dense amorphous material, which resulted in improved gel texture and viscosity.     

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 treatment on the tofu fatty acids and acylglycerols content

Three batches of tofu were purchased from the local market. The samples were packaged under vacuum, and half of them subjected to a pressure in a range of 100 to 350 MPa at ambient temperature (22 °C) for 15 min, the rest were considered as control. The soy fat was extracted with hexane and the characterisation of fatty acids composition as methyl esters (FAME) and their diacyl–triacylglycerols (DAG, TAG) were analysed by Gas chromatography (GC). The tofu fat is low in saturated fatty acids (SFA, ∼ 16%) and high in polyunsaturated fatty acids (PUFA, ∼ 64%). The DAG content was lower than 10% and the highest value obtained for TAG was for the CN54 (∼ 60%), followed for the CN52 (∼ 28%) and CN50 (∼ 4%). No significant changes on the profile fat pattern have been found after the high pressure treatment in the assayed conditions. The ratios of PUFA/SFA and n-6/n-3 are 4 and 7 respectively. Tofu can be subjected to HP of 350 MPa without significant changes in its fat pattern.Industrial relevanceTofu is becoming an increasingly important international product outside its traditional markets. The use of high pressure processing for preservation purposes is a potential to retain or increase the nutritional value of foods of high relevance. This paper concentrates on the impact of high pressure on polyunsaturated acid and di-triacylglycerols. Interestingly no significant changes due to high pressure treatment could be observed marking this an attractive processing option.     

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.     

Effects of transglutaminase treatment on the thermal properties of soy protein isolates

The effects of covalent cross-linking of microbial transglutaminase (MTGase) on the thermal properties of soy protein isolates (SPI), including the thermal denaturation and glass transition were investigated by conventional and modulated differential scanning calorimetry (DSC). The MTGase treatment significantly increased the thermal denaturation temperatures (including the on-set temperature of denaturation, T_m and the peak temperature of denaturation, T_d) of glycinin and β-conglycinin components of SPI (P ⩽ 0.05), and the thermal pretreatment of SPI further increased the extent of this improvement. The MTGase treatment also improved the ability of SPI to resist the urea-induced denaturation. Modulated DSC analysis showed that there were two glass transition temperatures (T_g) in the reversible heat flow signals of native SPI (about 5% moisture content), approximately corresponding to 45 and 180 °C, respectively. These T_g values of SPI were significantly decreased by the MTGase treatment (at 37 °C for more than 2 h) (P ⩽ 0.05). The improvement in the hydration ability of protein and the formation of high molecular biopolymers may account for the changes of thermal properties of soy proteins caused by the MTGase cross-linking.     

Selection of probiotic bacteria for the fermentation of a soy beverage in combination with Streptococcus thermophilus

Lactobacillus delbrueckii subsp. lactis R0187, Lactobacillus helveticus R0052, Lactobacillus rhamnosus R0011 and Bifidobacterium longum R0175 were examined for their ability to grow in combination with Streptococcus thermophilus cultures in milk and a laboratory soy beverage (LSB; both standardized to 4.5% protein and 2.3% fat). Strains R0011 and R0187 did not rapidly acidify the soy beverage despite good growth rates on soy carbohydrates. The S. thermophilus populations in the LSB were similar to that of milk even though milk had 30% more buffering capacity. In milk but not in soy, symbiosis with respect to acidification rate was observed between S. thermophilus and L. helveticus or B. longum. The populations of L. helveticus in the fermented products were similar in pure cultures or in the presence of the streptococci. However B. longum did not compete well in the mixed culture. Fermentation conditions varied as a function of the ability of S. thermophilus strains to acidify media to a pH of 4.65 (between 8 and 24 h). The probiotic populations in the mixed culture were influenced by the S. thermophilus strain and by the time of fermentation. Variations in growth rates of the bacteria did not appear to be linked to differences in initial redox or α-amino nitrogen levels. Strain selection enabled the preparation of a mixed starter, probiotic-fermented soy beverage containing 1.1 × 10⁸ CFU/mL of L. helveticus R0052, which represented approximately 13% of the total final population.     

Manufacture of Cheddar cheese from high-pressure-treated whole milk

The cheese-making characteristics of high-pressure (HP)-treated milk were examined. The rennet coagulation time of pasteurised milk decreased after HP treatment at 400 MPa but increased after treatment at 600 MPa. The L-value (whiteness) of milk decreased directly after HP treatment but, over the course of coagulation, whiteness of HP-treated milk increased to the same level as in the control. Cheddar cheese was then manufactured from raw whole milk or whole milk treated by high-pressure (HP) at 400 MPa (HP400) or 600 MPa (HP600) for 10 min at 20 °C. HP treatment of raw milk at 600 MPa resulted in a 3.66 log reduction in the initial counts of non-starter lactic acid bacteria (NSLAB), decreased protein and fat content, as well as a lower pH compared to the control. Furthermore, higher treatment pressures resulted in increased incorporation of β-lactoglobulin into the cheese curd, with parallel increases in yield by 1.23% and 7.78% for HP400 and HP600 cheeses, respectively. Overall, this study showed that the effects of HP treatment on milk proteins increased rennet coagulation times and changes in cheese composition at day 1.Industrial relevanceHigh-pressure treatment is a novel technology which has been applied to a number of commercial food products. In this study, HP-induced changes in milk proteins resulted in increased cheese yields and increased cheese whiteness. In addition, HP treatment significantly reduced the microflora of raw milk cheese. Those attributes could be of interest for both industry and consumer.     

Inactivation of bacteriophages by thermal and high-pressure treatment

Dairy companies commonly experience fermentation failures due to bacteriophages that are spread mainly by milk, whey or air. Heat or high-pressure treatment may potentially reduce the phage titre, but further knowledge about the inactivation kinetics is desirable. Inactivation experiments were carried out with the commonly occurring lactococcal phages P001 and P008. Phage suspensions in calcium-enriched M17-broth were heated at 55–80 °C, or high-pressure treated at up to 600 MPa. Kinetic analysis showed that the order of inactivation reaction was above 1; thus, inactivation kinetics were approximated by a non-linear regression model. The Arrhenius parameters, rate constant, k_p,T, and activation energy, E_A (for heat treatments), and the volume of activation, ΔV^# (for pressure treatments) were calculated. Both measured and calculated results indicate that phage P008 was the more heat- and pressure-resistant of the two. By combining the results from heat and pressure inactivations, a pressure–temperature diagram for phage P008 was established.     

Retinol binding to β-lactoglobulin or phosphocasein micelles under high pressure: Effects of isostatic high-pressure on structural and functional integrity

Embedding of bioactive molecules in food matrices is of interest for food industry to develop new foods or ingredients. The present study aims at deepening understanding of ligand binding to dairy proteins regarded as nano-vehicles, using pressure-assisted technological tools. We compared effects of isostatic high-pressure up to 350–400 MPa on the stability and the retinol-binding capacity of β-lactoglobulin (β-LG) or phosphocasein (PC) at pH 6.5–6.6. Retinol binding to β-LG changed the intrinsic protein and retinol fluorescence, characterised by fluorescence resonance-energy-transfer (FRET). After correction for inner filter effects, the FRET level estimated for the β-LG–retinol complex at pH 6.5 and P_atm was compatible with a retinol position at the β-LG surface cleft. Pressurisation of the β-LG–retinol complex induced an irreversible loss of the retinol-ligand above 150 MPa. Pressurisation of PC dispersion up to 200 MPa induced a marked decrease in the PC intrinsic fluorescence intensity with a concomitant red-shift of the spectral mass centre (CSM) explained by Trp residues exposure to aqueous solvent. The pressure-induced CSM half-transition (P_1/2) was 116 MPa for PC instead of 183 MPa for β-LG. Retinol binding to PC also resulted in important FRET, but PC assemblies partly retained retinol after pressure-release from 400 MPa. Interestingly, the pressure-induced dissociation rate of PC assemblies in the 15–35 °C temperature range was significantly accelerated at lower temperature, reflecting non-Arrhenius behaviour. PC particle sizes evaluated by dynamic light-scattering, confirmed that the pressure-induced dissociation was practically achieved at 200 MPa. Thus PC dissociation paralleled the structural changes determined by fluorescence spectroscopy.