WO2020200984A1 - Compositions nutritionnelles comprenant des protéines de lait de vache, leurs procédés de préparation leurs utilisations - Google Patents

Compositions nutritionnelles comprenant des protéines de lait de vache, leurs procédés de préparation leurs utilisations Download PDF

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Publication number
WO2020200984A1
WO2020200984A1 PCT/EP2020/058453 EP2020058453W WO2020200984A1 WO 2020200984 A1 WO2020200984 A1 WO 2020200984A1 EP 2020058453 W EP2020058453 W EP 2020058453W WO 2020200984 A1 WO2020200984 A1 WO 2020200984A1
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Prior art keywords
casein
fat
protein
composition
whey
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Inventor
Renate Antonia Ganzevles
Urszula KUDLA
Johannes Andries Nieuwenhuijse
Christina Josephina Antonia Maria Timmer-Keetels
Andries Dirk Siemensma
Thom HUPPERTZ
Astrid Maria Hermina HORSTMAN
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FrieslandCampina Nederland BV
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FrieslandCampina Nederland BV
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/19—Dairy proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/40—Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the present invention relates to nutritional compositions comprising bovine milk proteins, methods for preparing the same and uses thereof, in particular for supporting or increasing muscle protein synthesis.
  • Milk of all mammals contains caseins, globular proteins, and some non-protein N- containing components (NPN), which in bovine milk represents about 5% of the total N-content.
  • NPN non-protein N- containing components
  • Of the true protein in bovine milk typically 75-85% is casein, and 15-25% is globular protein, also called‘globular serum protein’ or‘whey protein’.
  • Caseins are the proteins from milk that precipitate at pH 4.6, whereas whey proteins remain soluble at this pH. Caseins are non-globular proteins having little secondary structure. As a result, they cannot be denatured by heat. In milk of all mammals, caseins are associated into particles containing some 50 to 30 000 molecules - the so- called casein micelles - associated by weak, mainly electrostatic and hydrophobic, interactions. Caseins can be separated from the other milk constituents by renneting, acidification, or microfiltration. The resulting high-casein product is referred to as, respectively, cheese-curd, acid-casein and Micellar-Casein-lsolate. The remaining liquid is called cheese whey, acid-casein whey, and milk serum, respectively.
  • the whey protein fraction of cows’ milk comprises approximately ⁇ 50% b-lactoglobulin, ⁇ 15% a-lactalbumin, other globular serum proteins, and also NPN.
  • the proteins can be separated from whey or milk serum using ultrafiltration, thereby yielding Whey Protein Concentrate (WPC) or Serum Protein Concentrate (SPC).
  • WPC Whey Protein Concentrate
  • SPC Serum Protein Concentrate
  • most of the proteins in WPC and SPC are globular proteins, also called‘globular serum proteins’.
  • Cheese whey and whey protein concentrate from cheese whey further contain Casein MacroPeptide (CMP), which is not a globular protein.
  • CMP Casein MacroPeptide
  • the globular proteins from milk do not coagulate at the conditions in the stomach of humans if they are in their native (folded) state.
  • many standard processes as applied in the food (dairy) industry e.g. high-pasteurisation or high-pressure treatment, result in denaturation and aggregation of the whey proteins.
  • native whey proteins are soluble at pH 4.6
  • denatured whey proteins precipitate at this pH.
  • Caseins are known to coagulate at the conditions in the human stomach. Denatured whey proteins in heated milk tend to coagulate along with caseins, leading to less dense coagulates with faster hydrolysis. Caseins in cheese-curd are already coagulated at the moment of consumption.
  • whey protein compared to casein are mainly attributed to the amino acid composition and the faster digestion and absorption kinetics of the globular proteins. The latter results in a greater increase in postprandial plasma amino acid availability and thereby a greater increase in muscle protein synthesis.
  • nutritional compositions that mainly contain casein and/or caseinate tend to coagulate in the stomach.
  • casein is often referred to as "coagulating, or slow protein”
  • globular (whey) protein is an example of a "non coagulating, or fast protein”.
  • Whey protein and casein also markedly differ in their amino acid composition.
  • Whey protein is characterized by a considerably higher leucine content compared to casein. Because leucine has been identified as the amino acid that directly stimulates postprandial muscle protein accretion, the high leucine content may also contribute to the proposed greater anabolic properties of whey protein than of casein.
  • the present inventors especially sought for the manufacture of compositions comprising milk proteins that induces a fast, steep increase of amino acids, specifically leucine, in blood serum when orally administered to a human subject, in particular wherein the release profile also shows a sustained release of free essential amino acids as compared to that induced by conventional, heat-sterilized or UHT milk.
  • said sustained release means that the level of amino acids in blood remains higher for a longer period in time, e.g. amino acid concentrations in blood are still higher than the conventional heat-sterilized composition after 2 hours after ingestion.
  • the transfer rate of protein through the stomach appears to be the crucial, rate limiting step for uptake of amino acids in the blood, and that this transfer rate can be controlled by tuning interactions between bovine milk proteins. More specifically, steering parameters were identified which can prevent interaction between whey protein and casein, such that whey protein remains fast and casein remains slow.
  • This resulted in the development of a composition comprising casein, whey protein, and fat droplets, said whey proteins being dominantly native and the fat droplets being predominantly covered by/coated with casein.
  • the caseins in the composition are not distributed evenly across the entire composition, but they are enriched/concentrated at the surface of the fat droplets.
  • casein coagulates causes an extra delay in stomach transfer. Without wishing to be bound by theory, this may be explained by creaming of casein and fat in the stomach (depending on the protein/fat ratio), and/or by a lowered accessibility for digestive enzymes due to an increased firmness of the curd, resulting in a lower transfer rate through the stomach.
  • the invention provides a composition that has a digestive profile resembling that of raw milk yet with improved microbiological quality.
  • the present invention therefore relates to a nutritional composition
  • a nutritional composition comprising casein, whey proteins comprising a-lactalbumin (aLac) and b-lactoglobulin (bLac), and protein- coated fat droplets, wherein
  • the total protein content of the composition is more than 20% (w/w), based on total solids;
  • the fat content of the composition is in the range of 4 to 70% (w/w) based on total solids;
  • the fat droplets are coated with an average protein load of at least 2 mg/m 2 and the weight ratio of casein:(al_ac + bLac) on the fat droplets is at least 4 times higher, than the weight ratio of casein:(al_ac + bLac) in the total composition;
  • the whey protein in the composition is dominantly present in the native (undenatured) state.
  • EP1314361 discloses the manufacture of a shelf stable liquid nutritional composition with a protein content ranging from 20-90g/l, in which all or a major part of the protein is composed of whey protein in intact, unhydrolysed form. To that end, an acid phase composed of whey protein and carbohydrates is adjusted to a pH of 2.5-3.5 and subsequently UHT-sterilized. A fat phase is separately heat-sterilised and the two separately sterilized phases are combined. Due to the high temperature treatment, the whey proteins in a composition of EP1314361 are not dominantly present in a native state.
  • WO2014/01 1039 relates to a method for producing a protein-comprising composition with reduced digestive coagulation.
  • the composition comprises a mixture of at least two different proteins, of which at least one is a coagulating protein and at least one is an anti-coagulating protein, comprising the steps of: a) heat-sterilising a first liquid component comprising the coagulating protein, b) heat-sterilising a second liquid component comprising the anti-coagulating protein, and c) mixing the first component with the second component to obtain a mixture of said proteins, wherein said mixture has a weight ratio of coagulating protein to anti-coagulating protein of between 20: 1 and 1 : 1.
  • the coagulating protein (casein) is homogenised with fat and then heated.
  • the non-coagulating protein whey proteins including aLac and bLac
  • whey proteins including aLac and bLac is added raw or pasteurised such that these proteins remain native or at least not coagulate with the coagulating protein (casein).
  • WO2014/01 1039 aims at reducing casein coagulation
  • the present invention involves promoting or conserving the ability of casein to coagulate.
  • a related patent publication (WO2014/01 1030) discloses heating whey protein and caseinate separately, in order to reduce coagulation.
  • whey protein is mildly heated as one stream and casein is heated and homogenised with fat as the other stream, and coagulation (of most probably the casein part) is still observed.
  • US 8,835,383 relates to the coagulation of protein-containing nutritional compositions in the upper gastro-intestinal tract, more in particular in the stomach and provides a method for reducing such coagulation. It is proposed that a protein composition that under normal conditions coagulates in the stomach, can be made to coagulate to a far lesser extent or not at all, by including a different protein or a protein in a different form resulting in an anti-coagulating effect.
  • Preferred anti-coagulating proteins are pea and soy or a combination thereof.
  • the nutritional composition of the present invention comprises casein, whey proteins, in particular a-lactalbumin and b-lactoglobulin, and protein-coated fat droplets.
  • the total protein content of the composition is more than 20 wt%, preferably at least 22 wt%, and most preferably at least 27 wt%, based on total dry solids.
  • the protein content is chosen such that it can provide, upon reconstitution of a dry powdered composition with a liquid, a liquid nutritional composition having a protein concentration of at least 4 wt%, more preferably 4-17 wt%, and most preferably 4-15 wt% protein in order to ensure an adequate supply of proteins/amino acids to a subject.
  • This liquid composition can be in a liquid, or semi liquid (e.g. spoonable) form.
  • the protein source may comprise intact and/or hydrolysed proteins.
  • up to 10 wt%, preferably up to 5 wt% (based on total dry solids) is hydrolysed protein.
  • the total amount of aLac and bLac is typically at least 10 wt% based on the total protein level.
  • the aLac and bLac can be derived from any suitable whey protein source.
  • a-lactalbumin and b-lactoglobulin is derived from milk, from cheese whey, from acid casein whey, from milk serum, or from concentrated, diluted, demineralized or powdered variants thereof.
  • the nutritional composition of the present invention may contain further whey proteins, such as serum albumin, lactoferrin, and/or immunoglobulins (Igs). In a specific aspect, it contains all "non-casein” proteins as found in bovine milk.
  • the aLac and bLac in the nutritional composition are preferably predominantly present in a native/undenatured state, meaning that at least 50%, preferably at least 55%, and most preferably at least 60% is in native state. This is suitably achieved by using proteins that have not been exposed to temperatures above 85°C (e.g. UHT treatment) at which >50% of the whey proteins denatures.
  • sources of aLac and bLac are used that have undergone at least one heat treatment of 15 sec. at 72°C or an equivalent "mild” heat treatment.
  • the casein can be obtained from conventional sources.
  • the casein is selected from the group consisting of micellar casein, non-micellar casein, acid casein, calcium caseinate, magnesium caseinate, sodium caseinate, potassium caseinate and ammonium caseinate, or any combination thereof.
  • the casein can for example be obtained from whole milk, skimmed milk, or milk protein concentrate.
  • the weight ratio of total whey proteins (i.e. a-lactalbumin, b-lactoglobulin, and optional further whey proteins) to caseins in the nutritional composition is preferably in the range of 10:90 to 70:30, for example 10:90 to 50:50, 20:80 to 60:40, or 20:80 to 50:50, or 25:75 to 55:45. More preferably, the weight ratio of caseins to total whey proteins is in the range of 23:77 to 50:50.
  • the fat content of the nutritional composition is in the range of 4-70 wt%, preferably 10-60 wt%, most preferably 15-50 wt%, based on total solids. It can be varied according to the type of fat, the nutritional needs, the type of formulation (dry or liquid) and/or the protein content.
  • the nutritional composition is a liquid composition (e.g. prepared from a powdered composition described herein above) having a fat content of at least 1 wt%, preferably at least 2 wt%, more preferably at least 3 wt%, based on the weight of the final composition.
  • Exemplary fat contents are in the range of 1 -20 wt%, 2-18 wt%, 2-16 wt%, 3-20 wt%, 3-18 wt%, 3-15 wt%, 3-10 wt%, 4-8 wt% or 10-15 wt%.
  • the fat content is in the range of 2 to 8 wt%, more preferably 3-6 wt%, and most preferably 4-7 wt%.
  • the fat may be a dairy milk fat, a vegetable oil, a vegetable fat, a hydrogenated vegetable oil, a marine oil, an algae oil, single cell oil, or a mixture of any of the foregoing.
  • the fat is a dairy fat, preferably selected from the group consisting of butter oil, butter fat, and milk fat.
  • milk fat can be used whole milk, cream, anhydrous milk fat, or fractions from milk fat.
  • a combination of dairy milk fat and vegetable fat is used.
  • the composition comprises a mixture of milk fat and vegetable oils.
  • vegetable oils are sunflower oil, coconut oil, and rapeseed oil.
  • the fat droplets have a core that consists of at least 90 wt%, preferably at least 95 wt% of triglycerides.
  • the fat droplets typically have an average diameter in the range of 0.2-1 .0 micron, preferably 0.3-0.8 micron.
  • the fat droplet surface may contain milk fat globule membrane (MFGM) material.
  • MFGM milk fat globule membrane
  • the weight ratio of total protein to total fat in the nutritional composition is not critical. Typically, the weight ratio of casein to fat in the composition is in the range of from 1 :3 to 3:1 , preferably 1 :2 to 2: 1.
  • the nutritional composition of the invention is among others characterized in that the protein-coated fat droplets are coated, predominantly with caseins, at an average protein load of at least 2 mg/m 2 , preferably at least 4 mg/m 2 . In one embodiment, it is above 10 mg/m 2 and/or below 25 mg/m 2 . In a specific aspect, the protein load is in the range of 10-22 mg/m 2 , like about 12, 14, 16, 18, or 20 mg/m 2 .
  • the weight ratio casein: (aLac + bLac) on the fat droplets is at least 4, preferably at least 5, more preferably at least 7, even more preferably 8-30 times higher than the weight ratio of casein: (aLac + bLac) in the total composition, for example at least 9, or at least 10, or at least 12, or at least 16, or at least 18, or at least 20 times higher.
  • the protein load on the fat droplets and the relative amounts of casein: (aLac + bLac) in the composition can be determined by methods known in the art using fractionation and taking into account the fat droplet particle size, the protein content, and the fat content. If the composition is in dry form, it is first rehydrated with water to a fat content in the range of 2-10 wt%.
  • Separation of the protein-coated fat droplets from the remainder of the composition advantageously comprises increasing the density of the liquid phase surrounding the fat droplets. This is suitably done by adding sucrose to the reconstituted composition.
  • the size distribution of the fat droplets can be determined using a Malvern Mastersizer.
  • the refractive index of the dispersed phase is set at 1 .46 and absorption at 0.01 .
  • the invention also provides a method for providing the nutritional composition.
  • This method is herein also referred to as "split stream process”; reflecting that casein and fat are homogenized, and optionally heated together, prior to be mixed with native whey protein.
  • the method comprises the steps of: i) mixing an aqueous composition comprising casein with a fat source, and homogenizing the mixture to obtain casein-coated fat droplets and ii) blending said casein-coated fat droplets with a source of native whey protein comprising aLac and bLac.
  • Homogenization serves to obtain a stable and homogeneous composition wherein the fat is predominantly present in the form of protein-coated fat droplets.
  • Homogenization conditions can be chosen and optimized by a person skilled in the art. Good results are obtained with a two-step homogenization method, wherein the first step is used to reduce the size of the sample to coarse particles, while the second step further reduces or obliterates those particles.
  • the mixture of casein and fat can be homogenized at 100-200 bar in the first stage and then at 15-25 bar in the second stage.
  • Homogenization temperatures are preferably in the range 45-70°C.
  • step i) a heating or preservation step can be included in step i), such as a pasteurization, sterilization, or UHT treatment, optionally followed by drying.
  • step i) further comprises spray drying of the casein-coated fat droplets.
  • the source of native whey protein can be cheese whey, acid whey or milk serum obtained via membrane filtration;
  • the source of native whey protein may have been subjected to a mild heating, microfiltration treatment, ceramic membrane filtration, or any other mild preservation technique.
  • At least 60% of the a-lactalbumin and b-lactoglobulin in the whey protein stream is native. This can be accomplished by avoiding exposure of whey protein to denaturing conditions, for example the whey protein is preferably not subjected to a treatment at a temperature of above 80°C. However, some degree of mild heating, e.g. in the range of from 70 to 72°C, for 10 to 20 seconds, is permitted.
  • the source of native whey protein can be in the form of an aqueous solution or it can be in a dry state. Accordingly, the two streams in the split-stream process can be blended in either a liquid/liquid form, in a dry/dry form (dry blending), or a dry/liquid form, e.g. by dispersing dried casein-coated fat droplets in an aqueous solution comprising native a-lactalbumin and b-lactoglobulin. The resulting product can be dried if so desired.
  • the method comprises (a) providing a mixture of (e.g. 2-10 w%) casein and (e.g. 2-10w%) fat and homogenizing said mixture in an aqueous medium (in the absence of added whey protein to maximize casein coating) to obtain an emulsion of casein particles in the form of casein deposited on fat droplets; (b) providing an aqueous solution of (e.g.
  • a native whey protein source comprising a-lactalbumin and b-lactoglobulin (and possibly other whey proteins), optionally mildly heating or microfiltrating said solution, optionally followed by drying; and (c) mixing the casein and fat stream of step (a) with the whey protein stream of step (b) in either wet or dry form, if desired under aseptic conditions.
  • the emulsion of fat and casein is UHT treated. Microfiltration or other mild preservation techniques may be applied to the whey protein fraction before it is aseptically mixed with the fat and casein stream.
  • the emulsion of fat and casein is UHT treated and subsequently spray dried or freeze dried. After this, it is dry blended with a whey protein powder.
  • the nutritional composition as provided herein finds many uses in oral nutrition as well as enteral nutrition.
  • the invention therefore also relates to the use of the nutritional composition as or in an oral or an enteral nutritional composition.
  • This composition is administered to a subject.
  • the subject can be a mammalian subject, preferably a human subject.
  • the human subject can be of any age. For example, it is an adult, or an elderly person, e.g. a subject that is frail and aging.
  • the subject may be a healthy subject or a subject that is in a disease state, a subject that is recovering from a disease state/sickness/illness, or malnourished, obese, recovering from an injury, etc.
  • the nutritional composition Because of the presence of a fraction of protein/amino acids with a high transfer rate through the stomach and a fraction of proteins/amino acids with a low transfer rate through the stomach, the nutritional composition has interesting digestibility properties. Since a composition provided herein preserves properties that are considered important to gastric digestion of protein in combination with microbiological safety, it can be considered as an alternative for raw milk with respect to its gastric behaviour.
  • the composition induces a bimodal release profile of free essential amino acids (in particular leucine) in blood serum when orally administered to a human subject. In particular, the release profile shows a double peak, or a steep increase with a‘shoulder’ on the right (see Figure 7).
  • the release profile shows a delayed and/or sustained release of essential amino acids after an initial steep increase in blood amino acids as compared to that induced by UHT milk or sterilized milk.
  • the composition of the present invention is advantageously used to quickly meet a leucine threshold concentration in blood in order to trigger muscle protein synthesis, which is then followed by a long and sustained delivery of amino acids to prolong increased muscle protein synthesis rates. Therefore, the invention provides a nutritional composition that is especially suitable for overnight recovery of muscles or in periods of bed rest or limb immobilization.
  • the invention provides a nutritional composition or a composition obtainable by the method according to the invention for use in one or more of: (i) a method of preventing a condition linked to loss of muscle mass and/or muscle strength; (ii) a method of treating and/or preventing a condition linked to loss of muscle mass and/or muscle strength; (iii) a method of increasing muscle mass and/or muscle strength.
  • the subject is in need of supporting or enhancing muscle protein accretion and/or increasing muscle mass, quality, strength and function, and/or preventing or attenuating loss thereof.
  • the subject may be a physically active subject, e.g. an athlete in need of enhanced muscle growth and/or enhanced muscle recovery.
  • a composition provided herein is advantageously used in a subject involved in weight training or endurance exercise in order to promote muscle growth.
  • the combination of exercise and the consumption of a protein-rich composition of the invention allows to build new (contractile or mitochondrial) muscle tissue.
  • the composition is consumed at nighttime before going to bed.
  • the invention provides a nutritional composition for use in a method of treating/counteracting and/or preventing a condition linked to loss of muscle mass and/or strength in a subject.
  • the subject is recovering from a sports performance or injury, or the subject suffers from a decline of lean body mass (disuse) atrophy e.g. by hospitalization, space flight, sickness/illness, etc.), muscle and bone mass, quality, strength and function decline, sarcopenia, cachexia, osteoporosis and/or osteosarcopenia.
  • Figure 1 Analysis of different compositions in an in-vitro stomach model.
  • Figure 2 Illustration of coagulation in the in-vitro stomach at 15 minutes after start digestion.
  • Figure 3 Analysis of different compositions in an in-vitro stomach model.
  • Figure 5 Serum amino acid concentration after oral ingestion of various dairy compositions. Panel A: leucine. Panel B: total essential amino acids. For details see Example 5.
  • Protein digestion data were obtained using an established in vitro digestion stomach model. Given that the transit time of liquids through the stomach is much faster than that of (semi)solids, the amount of protein in the - fast emptying - liquid fraction (or liquid phase) is key for a fast digestion.
  • a Simulated Gastric Fluid (380 mL) is prepared from:
  • the Applikon ADI1010 (Controller) and ADI1025 (fermenter) were started and 350 mL of the sample was pipetted in the fermenter. The sample was heated to and equilibrated at 37°C. The stirring bar was set at 100 rpm.
  • the first sample was taken and the pH profile started. This gave the start signal for the ADI1010 to add 1 M HCI following the set pH curve as depicted in Table 1. The slope of the decrease is the same for all the tested samples. The amount of HCI added was recorded.
  • samples typically 1 * 40 ml per sampling point
  • samples were taken for analysis during the test series. Since the pH can differ a little, samples were taken based on time points, not based on pH. Pictures of the fermenter were taken at the moments the 40 ml samples are taken. After the sampling, samples were immediately filtered on a gauze (mesh size 1 -2 mm). From each sample the protein content (nitrogen content multiplied by 6.38) was analysed using the Kjeldahl nitrogen analysis method.
  • MCI80L is a liquid micellar casein isolate, obtained from FrieslandCampina; protein content ⁇ 15%, casein content ⁇ 13%
  • Nutriwhey 800F is a whey protein concentrate from acid whey with about 80 wt% protein on dry matter obtained from FrieslandCampina DMV in powder form - Cream (43% fat) obtained from FrieslandCampina production plant in
  • Premixes 1 a and 1 b (casein/fat) were prepared by blending water with MCI liquid in the correct proportions and subsequently adding the cream under continuous stirring
  • Premix 2 whey protein
  • whey protein whey protein
  • the whey protein powder was slowly added, avoiding formation of lumps, to water at 40°C in the bucket under continuous stirring.
  • the dispersion was left to hydrate for 1 hour, while stirring.
  • premix 1 a was mixed with premix 2 at a weight ratio of 0.87:0.13 and subsequently simultaneously homogenised (Bos MG2-13B two- stage homogenizer, 150 bar and 20 bar for the first and second homogenisation step, flow around 100 l/h and temperature 50°C) and pasteurised in a water bath for 5 minutes at 85°C
  • premix 1 a was first homogenised (Bos MG2-13B two-stage homogenizer, 150 bar and 20 bar for the first and second homogenisation step, flow around 100 l/h and temperature 50°C) and subsequently mixed with premix 2 at a weight ratio of 0.87:0.13
  • premix 1 b was first homogenised (Bos MG2-13B two-stage homogenizer, 150 bar and 20 bar for the first and second homogenisation step, flow around 100 l/h and temperature 50°C) and subsequently mixed with premix 2 at a weight ratio of 0.7:0.3.
  • This example describes the digestibility analysis of the nutritional compositions of Example 2 using the in-vitro stomach model of Example 1 .
  • Test compositions (450 ml) standardized at 3.3 wt% protein were prepared by combining 247.5 gram of each of the nutritional compositions of Example 2 with 202.5 gram water. A volume of 350 ml of each of the 3 test compositions was subjected to the in-vitro stomach model.
  • Figure 1 depicts the amount of protein in the liquid phase in the in-vitro stomach as a function of time.
  • the reference sample R25:75
  • 100% of the protein was found in the liquid phase at all times; the protein concentration in the liquid phase was the same as the start sample. Since liquids, compared to solids, have a shorter transit time through the stomach, it is likely that this composition leads to a fast increase in protein in the small intestine and hence a fast appearance in blood.
  • the protein concentration in the liquid phase was significantly decreased: after 15 mins of digestion time, the concentration is only 45% of the start sample. The remaining fraction is present in the stomach in the form of coagulates, presumably formed by the casein fraction, that do not pass through the gauze. It took 3 hours until all the protein was in the liquid phase again. This will most likely lead to an overall slower digestion of the protein.
  • the SS50:50 sample showed a decrease of protein concentration in the liquid phase. The concentration decrease was presumably less, because there is less casein in the sample and hence less coagulation.
  • Figure 2 shows photographic images of the samples in the in-vitro stomach taken after 15 minutes digestion time.
  • split stream samples SS25:75 and SS50:50; pictures A and B
  • coagulation was observed, with a relatively clear serum phase, while this was not the case in the reference sample (picture A).
  • Example 2 the same compositions as in Example 2 were prepared, with two additional samples. The last two samples were obtained by heating the SS25:75 and SS50:50 samples in a water bath for 5 minutes at 85°C.
  • a 350 ml volume of each of the 5 samples was analysed for digestibility using the in- vitro stomach model of Example 1 .
  • MCI88 is a powdered micellar casein isolate, obtained from FrieslandCampina; protein content ⁇ 86%, casein content >77%
  • Nutriwhey 800F is a whey protein concentrate from acid whey with about 80 wt% protein on dry matter obtained from FrieslandCampina DMV in powder form Anhydrous Milk Fat (AMF) obtained from FrieslandCampina production plant in Noordwijk. AMF was filled in cans and sterilised further before use.
  • AMF Anhydrous Milk Fat
  • Premixes 1 a and 1 b (casein/fat) were prepared by adding MCI powder slowly to water while stirring and leaving to hydrate for 2 to 3 hours. Subsequently, the MCI dispersion was heated to 40°C and preheated AMF (in a water bath to 50°C) was blended with MCI in water in the correct proportions under continuous stirring
  • Premixes 1 a and 1 b were then homogenised using a Sterideal two-stage homogenizer, at 150 bar and 20 bar for the first and second homogenisation step, a flow around 100 l/h, and a temperature of 50°C.
  • Premix 2 whey protein
  • whey protein whey protein
  • the whey protein powder was slowly added, avoiding formation of lumps, to water at 40°C in the bucket under continuous stirring.
  • the dispersion was left to hydrate for at least 1 hour, while stirring.
  • premix 1 a and premix 2 were mixed at a weight ratio of 0.87:0.13 and for SS50:50, premix 1 b was mixed with premix 2 at a weight ratio of 0.68:0.32. All compositions were cooled and stored at 4°C until further use.
  • EXAMPLE 6 Characterization of protein-coated fat droplets.
  • This example describes the analysis of the average fat droplet size and the specific surface protein load of the fat droplets.
  • Exemplary liquid compositions were prepared as described in Example 5. Full fat commercial UFIT milk was used as reference sample.
  • a Malvern Mastersizer 3000 was used to measure the fat globule size distribution at 24°C.
  • the refractive index of the dispersed phase was set as 1 .46, whereas the refractive index of the continuous phase was set as 1 .33.
  • the absorption coefficient was set as 0.01 .
  • Prior to fat globule size measurements a 2 ml sample was mixed with 40 ml water and 5 ml of a solution consisting of 0.125% Tween-20 and 1.5% ETDA to dissociate the casein micelles, and the mix was incubated for 10 minutes.
  • sucrose was added to increase the density difference between the fat globules and the serum phase. After sucrose addition, the solution contained 10.0 w/w% carbohydrates. This solution was then centrifuged for 60 min at 100,000 x G at 21 °C in a Beckman/Coulter Avanti J-310 using a swing out rotor JS-24,38.
  • the fat globule volume and surface area were calculated from the d3,2 [pm] obtained from the particle size measurement, and a fat density of 0.91515 g/ml was used to convert the fat volume in fat mass.
  • the protein load (PL) [mg/m 2 ] was determined according to the following equation: 1000
  • Table 6 shows the results of protein load analyses of the three samples. Table 6: Particle size characteristics and specific protein surface load
  • EXAMPLE 7 Determining the casein to whey protein ratio.
  • the ratios between caseins and whey proteins (a-lactalbumin + b-lactoglobulin) in the exemplary compositions as described in Example 6 and the corresponding fat discs as obtained in Example 6 were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) making use of a Laemmli based buffer system and the stain free imaging methodology using a stain free enabled Bio-Rad ChemiDoc XRS+ Documentation System Bio-Rad unit provided with ImageLab software. Isolated fat discs were dissolved in prewarmed 2% SDS / 10 mM citrate solution in demiwater.
  • An unstained molecular weight marker (Bio-rad, catno. 1610363) was included to identify the major milk proteins. Electrophoresis was performed in 1x Tris/Glycine/SDS electrophoresis buffer (Bio-Rad, cat. no. 1610732) at room temperature at constant Voltage (V): 100V for 10 minutes followed by another 45 minutes at 150V. After electrophoresis the gel was removed from the frame, rinsed for 5 seconds in demi water and immediately imaged starting with 5 minutes of UV activation and followed by several exposure times (intense, faint, 10s). Results are shown in Figure 4.
  • the final images were analyzed with the Analysis Tool Box from the ImageLab software. Lanes were selected to discriminate between the samples on a gel and bands were selected within the lanes for the specific casein and whey proteins. Per specific protein the volume intensities were determined and automatically adjusted for background (adjusted volume intensity). Infant milk samples and its fat fraction were always examined on the same gel. For each product, the total intensities of all casein bands (cas) were calculated, as well as the total intensities of the a-lactalbumin (aLac) + b-lactoglobulin (bLac) bands. The ratio of the first value was then divided by the second one to obtain a casein : (aLac + bLac) ratio.
  • EXAMPLE 8 Amino acid profiles in blood after ingestion of exemplary split stream compositions.
  • Table 8 Composition and sample size of the clinical trial samples
  • a group of 10 subjects received a set of different dairy products , with about 1 week washout period between treatments.
  • Blood samples were collected in the fasting state and 15, 30, 45, 60, 75, 90, 105, 120, 150, 180, 210, 240 and 300 min after ingestion of the product.
  • Blood serum samples were later analysed for 20 amino acid concentrations using the Phenomenex EZTaast amino acid kit (https://www.phenomenex.com/Products/AminoAcidDetail/EZfaast). Data were corrected for baseline values (values in blood before ingestion of the test products) are shown in Figure 5A.
  • composition SS25:75 according to the invention was found to give a much higher maximum leucine concentration in blood serum ( ⁇ 23 mg/L) compared to full fat UHT milk ( ⁇ 12 mg/L), while both have a roughly similar amino acid composition. Furthermore, during 2 to 3 hours after ingestion, the leucine content remained higher. The SS50:50 composition gave an even higher maximum leucine concentration ( ⁇ 32 mg/L), while also the leucine concentration remained higher for longer period of time than observed for UHT milk.
  • DB25:75 was prepared by dry blending powdered premix 1a with Nutriwhey 800F at a weight ratio of 0.86:0.14.
  • DB50:50 was prepared by dry blending powdered premix 1 b with Nutriwhey 800F at a weight ratio of 0.69:0.31.

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  • Life Sciences & Earth Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Mycology (AREA)
  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne des compositions nutritionnelles comprenant des protéines de lait de vache, leurs procédés de préparation et leurs utilisations, en particulier pour supporter ou augmenter la synthèse de protéines musculaires. L'invention concerne une composition nutritionnelle comprenant de la caséine, des protéines de lactosérum comprenant de l'alpha-lactalbumine (aLac) et de la bêta-lactoglobuline (bLac) et des gouttelettes de graisse enrobées de protéines, où (i) la teneur totale en protéines de la composition est supérieure à 20 % (p/p), sur la base des solides totaux ; (ii) la teneur en graisse de la composition est dans la plage de 4 à 70 % (p/p) sur la base de solides totaux ; (iii) les gouttelettes de graisse sont revêtues d'une charge de protéine d'au moins 2 mg/m2 et le rapport en poids de la caséine : (aLac + bLac) sur les gouttelettes de graisse est au moins 4 fois supérieur au rapport pondéral de la caséine : (aLac + bLac) dans la composition totale ; et (iv) la protéine de lactosérum dans la composition est présente de manière dominante dans l'état natif.
PCT/EP2020/058453 2019-03-29 2020-03-26 Compositions nutritionnelles comprenant des protéines de lait de vache, leurs procédés de préparation leurs utilisations Ceased WO2020200984A1 (fr)

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Publication number Priority date Publication date Assignee Title
WO2023093672A1 (fr) * 2021-11-26 2023-06-01 安琪酵母股份有限公司 Protéine de levure, sa composition, son procédé de préparation, et utilisation de protéine de levure et composition
WO2024231449A1 (fr) * 2023-05-11 2024-11-14 Frieslandcampina Nederland B.V. Compositions nutritionnelles présentant une séparation de phase dans des conditions gastriques et comprenant des phospholipides, leurs procédés de préparation et leurs utilisations

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EP1314361A1 (fr) 2001-11-26 2003-05-28 Nestec S.A. Produit alimentaire nutritionnel de longue conservation contenant du petit-lait, procede de preparation et utilisation
WO2010027259A1 (fr) 2008-09-02 2010-03-11 N.V. Nutricia Compositions nutritionnelles à globules lipidiques enrobés
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WO2014011039A1 (fr) 2012-07-09 2014-01-16 N.V. Nutricia Procédé de production d'une composition comprenant des protéines avec une coagulation digestive réduite
US8835383B2 (en) 2009-04-27 2014-09-16 N.V. Nutricia Method for controlling the digestive coagulation of proteins
WO2015048646A1 (fr) * 2013-09-30 2015-04-02 Abbott Laboratories Protéines en poudre
US20160278413A1 (en) * 2013-09-13 2016-09-29 N. V. Nutricia Improved process for preparing infant formula using a static mixer
WO2016163882A1 (fr) 2015-04-10 2016-10-13 N.V. Nutricia Nutrition avec de grands globules lipidiques comprenant des graisses végétales revêtues de phospholipides de lait pour améliorer la vidange gastrique

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Publication number Priority date Publication date Assignee Title
EP1314361A1 (fr) 2001-11-26 2003-05-28 Nestec S.A. Produit alimentaire nutritionnel de longue conservation contenant du petit-lait, procede de preparation et utilisation
WO2010027259A1 (fr) 2008-09-02 2010-03-11 N.V. Nutricia Compositions nutritionnelles à globules lipidiques enrobés
US20110257089A1 (en) * 2008-10-24 2011-10-20 N.V. Nutricia Liquid high-fat protein composition
US8835383B2 (en) 2009-04-27 2014-09-16 N.V. Nutricia Method for controlling the digestive coagulation of proteins
US20130115258A1 (en) * 2010-04-26 2013-05-09 Massey University Emulsion
WO2014011039A1 (fr) 2012-07-09 2014-01-16 N.V. Nutricia Procédé de production d'une composition comprenant des protéines avec une coagulation digestive réduite
WO2014011030A1 (fr) 2012-07-09 2014-01-16 N.V. Nutricia Procédé de production d'une composition comprenant des protéines avec une coagulation digestive réduite
US20160278413A1 (en) * 2013-09-13 2016-09-29 N. V. Nutricia Improved process for preparing infant formula using a static mixer
WO2015048646A1 (fr) * 2013-09-30 2015-04-02 Abbott Laboratories Protéines en poudre
WO2016163882A1 (fr) 2015-04-10 2016-10-13 N.V. Nutricia Nutrition avec de grands globules lipidiques comprenant des graisses végétales revêtues de phospholipides de lait pour améliorer la vidange gastrique

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2023093672A1 (fr) * 2021-11-26 2023-06-01 安琪酵母股份有限公司 Protéine de levure, sa composition, son procédé de préparation, et utilisation de protéine de levure et composition
WO2024231449A1 (fr) * 2023-05-11 2024-11-14 Frieslandcampina Nederland B.V. Compositions nutritionnelles présentant une séparation de phase dans des conditions gastriques et comprenant des phospholipides, leurs procédés de préparation et leurs utilisations

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