ABSTRACT
OBJECTIVE: To investigate whether milk polar lipids (PL) impact human intestinal lipid absorption, metabolism, microbiota and associated markers of cardiometabolic health. DESIGN: A double-blind, randomised controlled 4-week study involving 58 postmenopausal women was used to assess the chronic effects of milk PL consumption (0, 3 or 5 g-PL/day) on lipid metabolism and gut microbiota. The acute effects of milk PL on intestinal absorption and metabolism of cholesterol were assessed in a randomised controlled crossover study using tracers in ileostomy patients. RESULTS: Over 4 weeks, milk PL significantly reduced fasting and postprandial plasma concentrations of cholesterol and surrogate lipid markers of cardiovascular disease risk, including total/high-density lipoprotein-cholesterol and apolipoprotein (Apo)B/ApoA1 ratios. The highest PL dose preferentially induced a decreased number of intestine-derived chylomicron particles. Also, milk PL increased faecal loss of coprostanol, a gut-derived metabolite of cholesterol, but major bacterial populations and faecal short-chain fatty acids were not affected by milk PL, regardless of the dose. Acute ingestion of milk PL by ileostomy patients shows that milk PL decreased cholesterol absorption and increased cholesterol-ileal efflux, which can be explained by the observed co-excretion with milk sphingomyelin in the gut. CONCLUSION: The present data demonstrate for the first time in humans that milk PL can improve the cardiometabolic health by decreasing several lipid cardiovascular markers, notably through a reduced intestinal cholesterol absorption involving specific interactions in the gut, without disturbing the major bacterial phyla of gut microbiota. TRIAL REGISTRATION NUMBER: NCT02099032 and NCT02146339; Results.
Subject(s)
Cardiovascular Diseases/blood , Lipid Metabolism/drug effects , Lipids/pharmacology , Overweight/metabolism , Sphingomyelins/metabolism , Animals , Apolipoprotein A-I/blood , Apolipoprotein B-100/blood , Cholestanol/metabolism , Cholesterol/metabolism , Cholesterol, HDL/blood , Cross-Over Studies , Dietary Supplements , Double-Blind Method , Emulsifying Agents/pharmacology , Feces/chemistry , Female , Gastrointestinal Microbiome/drug effects , Humans , Ileostomy , Intestinal Absorption/drug effects , Lipids/administration & dosage , Lipids/analysis , Middle Aged , Milk/chemistry , Postmenopause , Risk FactorsABSTRACT
Understanding how proteins stabilize amorphous calcium ortho-phosphate (ACP) phases is of great importance in biology and for pharmaceutical or food applications. Until now, most of the former investigations about ACP-protein stability and equilibrium were performed under conditions where ACP colloidal nanoclusters are surrounded by low to moderate concentrations of peptides or proteins (15-30 g L-1). As a result, the question of ACP-protein interactions in highly concentrated protein systems has clearly been overlooked, whereas it corresponds to actual industrial conditions such as drying or membrane filtration in the dairy industry for instance. In this study, the structure of an ACP phase is monitored in association with one model phosphorylated protein (casein) using solid-state nuclear magnetic resonance (ssNMR) under two conditions of high protein concentration (300 and 400 g L-1). At both concentrations and at 25 °C, it is found that the caseins maintain the mineral phase in an amorphous form with no detectable influence on its structure or size. Interestingly, and in both cases, a significant amount of the nonphosphorylated side chains interacts with ACP through hydrogen bonds. The number of these interacting side chains is found to be higher at the highest casein concentration. At 45 °C, which is a destabilizing temperature of ACP under protein-free conditions, the amorphous structure of the mineral phase is partially transformed at a casein concentration of 300 g L-1, while it remains almost intact at a casein concentration of 400 g L-1. Therefore, these results clearly indicate that increasing the concentration of proteins favors ACP-protein interactions and stabilizes the ACP clusters more efficiently.
Subject(s)
Calcium Phosphates/chemistry , Caseins/chemistry , Magnetic Resonance Spectroscopy , Protein Stability , TemperatureABSTRACT
Understanding the mechanisms that determine the diffusion and interaction of macromolecules (such as proteins and polysaccharides) that disperse through dense media is an important fundamental issue in the development of innovative technological and medical applications. In the current work, the partitioning and diffusion of macromolecules of different sizes (from 4 to 10 nm in diameter) and shapes (linear or spherical) within dispersions of casein micelles (a protein microgel) is studied. The coefficients for diffusion and partition are measured using FRAP (fluorescence recovery after photobleaching) and analyzed with respect to the structural characteristics of the microgel determined by the use of TEM (transmission electron microscopy) tomography. The results show that the casein microgel displays a nonspecific attractive interaction for all macromolecules studied. When the macromolecular probes are spherical, this affinity is clearly size-dependent, with stronger attraction for the larger probes. The current data show that electrostatic effects cannot account for such an attraction. Rather, nonspecific hydration molecular forces appear to explain these results. These findings show how weak nonspecific forces affect the diffusion and partitioning of proteins and polysaccharides in a dense protein environment. These results could be useful to better understand the mechanisms of diffusion and partitioning in other media such as cells and tissues. Furthermore, there arises the possibility of using the casein micelle as a size-selective molecular device.
Subject(s)
Caseins/chemistry , Macromolecular Substances/chemistry , Animals , Cattle , Diffusion , Gels , Micelles , Models, Molecular , Molecular Weight , Osmolar Concentration , Protein ConformationABSTRACT
We examine the internal structure of milk casein micelles using the contrast variation method in Small-Angle Neutron Scattering (SANS). Experiments were performed with casein dispersions of different origins (i.e., milk powder or fresh milk) and extended to very low q-values (â¼9 × 10(-4) Å(-1)), thus making it possible to precisely determine the apparent gyration radius Rg at each contrast. From the variation of I(q â 0) with contrast, we determine the distribution of composition of all the particles in the dispersions. As expected, most of these particles are micelles, made of casein and calcium phosphate, with a narrow distribution in compositions. These micelles always coexist with a very small fraction of fat droplets, with sizes in the range of 20-400 nm. For the dispersions prepared from fresh milk, which were purified under particularly stringent conditions, the number ratio of fat droplets to casein micelles is as low as 1 to 10(6). In that case, we are able to subtract from the total intensity the contribution of the fat droplets and in this way obtain the contribution of the micelles only. We then analyze the variation of this contribution with contrast using the approach pioneered by H. B. Stuhrmann. We model the casein micelle as a core-shell spherical object, in which the local scattering length density is determined by the ratio of calcium phosphate nanoclusters to proteins. We find that models in which the shell has a lower concentration of calcium phosphate than the core give a better agreement than models in which the shell has a higher density than the core.
Subject(s)
Caseins/chemistry , Milk/chemistry , Animals , Cattle , Micelles , Scattering, Small AngleABSTRACT
Correction for 'Structural heterogeneity of milk casein micelles: a SANS contrast variation study' by Antoine Bouchoux et al., Soft Matter, 2015, DOI: 10.1039/c4sm01705f.
ABSTRACT
Membrane filtration operations (ultra-, microfiltration) are now extensively used for concentrating or separating an ever-growing variety of colloidal dispersions. However, the phenomena that determine the efficiency of these operations are not yet fully understood. This is especially the case when dealing with colloids that are soft, deformable, and permeable. In this paper, we propose a methodology for building a model that is able to predict the performance (flux, concentration profiles) of the filtration of such objects in relation with the operating conditions. This is done by focusing on the case of milk filtration, all experiments being performed with dispersions of milk casein micelles, which are sort of â³naturalâ³ colloidal microgels. Using this example, we develop the general idea that a filtration model can always be built for a given colloidal dispersion as long as this dispersion has been characterized in terms of osmotic pressure Π and hydraulic permeability k. For soft and permeable colloids, the major issue is that the permeability k cannot be assessed in a trivial way like in the case for hard-sphere colloids. To get around this difficulty, we follow two distinct approaches to actually measure k: a direct approach, involving osmotic stress experiments, and a reverse-calculation approach, that consists of estimating k through well-controlled filtration experiments. The resulting filtration model is then validated against experimental measurements obtained from combined milk filtration/SAXS experiments. We also give precise examples of how the model can be used, as well as a brief discussion on the possible universality of the approach presented here.
Subject(s)
Filtration , Milk/chemistry , Animals , Caseins/chemistry , Colloids/chemistry , Micelles , Models, Chemical , Reproducibility of Results , Surface PropertiesABSTRACT
Food plays a significant role in the environmental impacts of human activities. However, many agro-industrial processes are multi-product systems and their impacts need to be distributed between the different co-products in order to properly address two major issues: (1) prevention of food spoilage and food losses and (2) the eco-design of food systems, from processing up to recommendations for changes in Western diets. As a culturally and nutritionally central component of most human diets, milk is critical because processing is a preservation issue and most dairy products follow from separations, thereby generating co-products. Life Cycle Assessment (LCA) is a reference and standard method that allows quantification of the potential environmental impacts of a manufactured product throughout its life cycle. Application of the method requires foreground information on the system considered, as well as input and output flows that feed and exit the system. This data paper provides data related to the fractionation of milk into cream, casein, lactose and two whey protein ingredients at industrial scale, using up-to-date technologies used in French dairy factories in years 2000-2010s. Cleaning is included. Transcription of these input and output flows into a selection of processes in the Agribalyse 3.0.1 and Ecoinvent 3.8 databases is also provided. Application of the LCA method in its attributional approach leaves methodological choices up to the practitioner, such as subdivision of the system, allocation of the environmental burden where subdivision is not applied or not possible, and aggregation of the impacts. Therefore, this data paper also provides the allocation factors that are necessary to apply mass, dry matter, protein or economic allocation at every separation operation throughout the processing itinerary. Using the characterization method EF 3.0, this data paper provides the potential environmental impacts of the 5 co-products obtained with an initial input of 600 tons of raw milk, i.e., 63 tons of cream, 183 tons of wet casein, 90 tons of lactose, 1.7 ton of dried ß-lactoglobulin and 0.3 ton of dried α-lactalbumin. The respective shares of the 5 co-products are calculated for each allocation rule. Finally, this data paper provides the potential environmental impacts for the manufacture of 1 kg of α-lactalbumin enriched ingredient, as the co-product with the longest process itinerary, with details of all intermediate input contributions as well as two possible aggregation rules: by step or by input type. The dataset participates in providing often confidential industrial-scale LCI data to the public. It will be helpful for the eco-design of future itineraries. In particular, it contributes to taking the fate of the co-products into account when using LCA for such eco-design.
ABSTRACT
Milk microfiltration process plays a key role in the dairy industry. Crossflow microfiltration of skimmed milk using a membrane with 0.1 µm mean pore size is widely used to fractionate the two main groups of dairy proteins: casein micelles (~150 nm) and serum proteins (~2-15 nm). Retentate, containing mainly casein micelles, is generally used to enrich vat milk for cheese making. Permeate, containing serum proteins, lactose and minerals, is usually ultrafiltered in order to produce protein-rich concentrate with a high nutritional value dedicated to specific populations such as infants and seniors. The great interest in these protein fractions explains the increasing number of microfiltration equipments in the dairy industry. This data article contains data associated with milk microfiltration process experiments and properties of the resulting dairy fractions annotated from a collection of scientific documents. These data are stored in INRAE public repository (see Data accessibility in the Specification Table for direct links to data). They have been structured using MILK MICROFILTRATION ontology and are replicated in @Web data warehouse providing additional querying tools (https://www6.inrae.fr/cati-icat-atweb/).
ABSTRACT
By combining the osmotic stress technique with small-angle x-ray scattering measurements, we followed the structural response of the casein micelle to an overall increase in concentration. When the aqueous phase that separates the micelles is extracted, they behave as polydisperse repelling spheres and their internal structure is not affected. When they are compressed, the micelles lose water and shrink to a smaller volume. Our results indicate that this compression is nonaffine, i.e., some parts of the micelle collapse, whereas other parts resist deformation. We suggest that this behavior is consistent with a spongelike casein micelle having a triple hierarchical structure. The lowest level of the structure consists of the CaP nanoclusters that serve as anchors for the casein molecules. The intermediate level consists of 10- to 40-nm hard regions that resist compression and contain the nanoclusters. Those regions are connected and/or partially merged with each other, thus forming a continuous and porous material. The third level of structure is the casein micelle itself, with an average size of 100 nm. In our view, such a structure is consistent with the observation of 10- to 20-nm casein particles in the Golgi vesicles of lactating cells: upon aggregation, those particles would rearrange, fuse, and/or swell to form the spongelike micelle.
Subject(s)
Caseins/chemistry , Micelles , Scattering, Small Angle , Stress, Mechanical , X-Ray Diffraction , Models, Molecular , Osmotic Pressure , Protein Structure, QuaternaryABSTRACT
Milk pre-processing steps-storage at 4 °C (with durations of 48, 72 or 96 h) and methods for microbiological stabilization of milk (1.4 µm microfiltration, thermization, thermization + bactofugation, pasteurization) are performed industrially before 0.1 µm-microfiltration (MF) of skimmed milk to ensure the microbiological quality of final fractions. The objective of this study was to better understand the influence of these pre-processing steps and their cumulative effects on MF performances (i.e., transmembrane pressure, and transmission and recovery of serum proteins (SP) in the permeate). Results showed that heat treatment of skimmed milk decreased ceramic MF performances, especially after a long 4 °C storage duration (96 h) of raw milk: when milk was heat treated by pasteurization after 96 h of storage at 4 °C, the transmembrane pressure increased by 25% over a MF run of 330 min with a permeation flux of 75 L.h-1.m-2 and a volume reduction ratio of 3.0. After 48 h of storage at 4 °C, all other operating conditions being similar, the transmembrane pressure increased by only 6%. When milk was 1.4 µm microfiltered, the transmembrane pressure also increased by only 6%, regardless of the duration of 4 °C storage. The choice of microbiological stabilization method also influenced SP transmission and recovery: the higher the initial heat treatment of milk, the lower the transmission of SP and the lower their recovery in permeate. Moreover, the decline of SP transmission was all the higher that 4 °C storage of raw milk was long. These results were explained by MF membrane fouling, which depends on the load of microorganisms in the skimmed milks to be microfiltered as well as the rate of SP denaturation and/or aggregation resulting from pre-processing steps.
ABSTRACT
Casein micelles dispersions have been concentrated and equilibrated at different osmotic pressures using equilibrium dialysis. This technique measured an equation of state of the dispersions over a wide range of pressures and concentrations and at different ionic strengths. Three regimes were found. i), A dilute regime in which the osmotic pressure is proportional to the casein concentration. In this regime, the casein micelles are well separated and rarely interact, whereas the osmotic pressure is dominated by the contribution from small residual peptides that are dissolved in the aqueous phase. ii), A transition range that starts when the casein micelles begin to interact through their kappa-casein brushes and ends when the micelles are forced to get into contact with each other. At the end of this regime, the dispersions behave as coherent solids that do not fully redisperse when osmotic stress is released. iii), A concentrated regime in which compression removes water from within the micelles, and increases the fraction of micelles that are irreversibly linked to each other. In this regime the osmotic pressure profile is a power law of the residual free volume. It is well described by a simple model that considers the micelle to be made of dense regions separated by a continuous phase. The amount of water in the dense regions matches the usual hydration of proteins.
Subject(s)
Caseins/chemistry , Micelles , Osmotic Pressure , Animals , Caseins/analysis , Cattle , Dialysis , Ions/chemistry , Milk/chemistry , Phase Transition , WaterABSTRACT
The objective of this work is to bring new information about the influence of temperatures (7 °C and 20 °C) on the equation of state and sol-gel transition behavior of casein micelle dispersions. Casein micelle dispersions have been concentrated and equilibrated at different osmotic pressures using equilibrium dialysis at 7 °C and 20 °C. The osmotic stress technique measured the osmotic pressures of the dispersions over a wide range of concentrations. Rheological properties of concentrated dispersions were then characterized, respectively at 7 °C and at 20 °C. The essential result is that casein micelle dispersions are less compressible at 7 °C than at 20 °C and that concentration of sol-gel transition is lower at 7 °C than at 20 °C, with compressibility defined as the inverse to the resistance to the compression, and that is proportional to the cost to remove water from structure. From our interpretations, these two features were fully consistent with a release of soluble ß-casein and nanoclusters CaP and an increased casein micelle hydration and apparent voluminosity at 7 °C as compared with 20 °C.
ABSTRACT
This data article contains annotation data characterizing Multi Criteria Assessment (MCA) Methods proposed in the agri-food sector by researchers from INRA, Europe's largest agricultural research institute (INRA, http://institut.inra.fr/en). MCA can be used to assess and compare agricultural and food systems, and support multi-actor decision making and design of innovative systems for crop production, animal production and processing of agricultural products. These data are stored in a public repository managed by INRA (https://data.inra.fr/; https://doi.org/10.15454/WB51LL).
ABSTRACT
Bioactive lipids of the milk fat globule membrane become concentrated in two co-products of the butter industry, buttermilk and butterserum. Their lipid composition is detailed here with special emphasis on sphingolipid composition of nutritional interest, determined using GC, HPLC and tandem mass spectrometry. Butterserum was 2.5 times more concentrated in total fat than buttermilk, with 7.7±1.5vs 19.5±2.9wt% and even more concentrated in polar lipids, with 1.4±0.2vs 8.5±1.1wt%. Both ingredients constitute concentrated sources of sphingomyelin (3.4-21mg/g dry matter) and contained low amounts of bioactive ceramides in a ratio to sphingomyelin of 1:5mol% in buttermilk and 1:10mol% in butterserum. Compared to other natural lecithins, these two co-products are rich in long and saturated fatty acids (C22:0-C24:0), contain cholesterol and could have interesting applications in neonatal nutrition, but also as brain-protective, hepatoprotective and cholesterol lowering ingredients.
Subject(s)
Buttermilk/analysis , Ceramides/analysis , Milk/chemistry , Sphingolipids/analysis , Animals , Fatty Acids , Heterotaxy Syndrome , HumansABSTRACT
Understanding how proteins behave in highly concentrated systems is a major issue in many fields of research, including biology, biophysics, and chemical engineering. In this paper, we provide a comprehensive (1)H NMR study of molecular mobility in dilute to highly concentrated dispersions of the exact same protein (casein) but organized in two distinct supramolecular forms: spongelike casein micelles or soft casein aggregates. Both relaxometry and diffusometry experiments were performed, so that three different parameters are reported: spin-spin relaxation rates of non-water protons (1/T(2,ne)), spin-spin relaxation rates of water protons (1/T(2,e+w)), and water self-diffusion coefficients (D(w)). The results are discussed in an effort to understand the respective effects of protein crowding and protein supramolecular organization on each mobility indicator. We also examine if connections exist between the observed changes in molecular mobility and the already documented changes in rheological and osmotic properties of casein dispersions as concentration is increased.
Subject(s)
Caseins/chemistry , Diffusion , Micelles , Nuclear Magnetic Resonance, Biomolecular , Protons , Water/chemistryABSTRACT
In the dairy industry re-use and multi-use cleaning-in-place (CIP) systems are operated by circulating chemicals and water without taking the equipment apart. The solutions, which become polluted due to the removal of fouling compounds, are drained periodically when they are considered to be too polluted. This work shows the large variations in composition (pollution, surface tension, etc) of the industrial caustic solutions coming from milk standardization and pasteurization plant CIP throughout their life time (7 days) and from 1 week to another. The work is also intended to show how nanofiltration (1 kg mol(-1) molecular weight cut-off) was robust and performed well, with good recovery of caustic solutions, even when faced with large variations of solutions composition: high caustic yield, permeation flux (J) in the range 42-110 l h(-1) m(-2), average chemical oxygen demand (COD) reduction equal to 0.58 and low surface tension change. Equations have been established for the prediction of J as a function of initial membrane hydraulic resistance (Rm) caustic concentration, volume reduction ratio (VRR) and initial soluble COD. When VRR increased, both J and pollution retention decreased despite the increase in irreversible fouling induced by the increase of soluble pollution concentration in retentate. The higher the initial soluble COD, the sharper the decrease in J vs. VRR. Since irreversible fouling was usually small (0.1-3.4 x 10(13) m(-1), that is to say of the same order of magnitude as Rm), the membrane cleaning could be efficiently performed by using single phase sodium hypochlorite alternately with a more expensive acid-base cleaning sequence. The obtained permeate was a clear regenerated cleaning solution with low soluble COD (0.2-3.5 g/l) and surface tension (56-30 mJ m(-2)) which could be successfully exploited owing to its cleaning potential.