Acidic beta-lactoglobulin beverage preparation

Abstract

The present invention relates to a novel packaged heat-treated beverage preparation having a pH in the range of 2.0 to 4.7. The present invention further relates to a method for producing the packaged heat-treated beverage preparation, and various uses of the packaged heat-treated beverage preparation.

Description

[Technical Field]

[0001] The present invention relates to a novel packaged heat-treated beverage preparation having a pH in the range of 2.0 to 4.7. The present invention further relates to a method for producing the packaged heat-treated beverage preparation, and various uses of the packaged heat-treated beverage preparation. [Background technology]

[0002] Dietary supplements containing whey protein are commonly used for muscle synthesis, weight management, and muscle and weight maintenance. Dietary supplements are targeted to various types of consumers, such as sportspeople, athletes, children, the elderly, and patients with or at risk of malnutrition and / or increased protein requirements.

[0003] Whey protein can be isolated from whey or milk serum. Whey typically contains a mixture of beta-lactoglobulin (BLG), alpha-lactalbumin (ALA), serum albumin, and immunoglobulins, of which BLG is the most predominant. Therefore, whey protein concentrate (WPC) contains a mixture of these proteins. Whey protein isolate (WPI) contains less fat and lactose than WPC.

[0004] Beverages containing whey protein are well known, for example, acidic heat-treated beverages containing whey protein.

[0005] Etzel 2004 (Etzel, MR, 2004, Manufacture and use of dairy protein fraction. American Society for Nutritional Science, pp. 996-1002) describes a beverage containing 2.5% by weight of WPI at a pH of 2-7. They found that the beverage, when subjected to heat treatment, was only obtainable if an anti-aggregation agent was added. Summary of the Invention

[0006] The inventors have observed that sensory attributes such as astringency and texture play an important role in consumer selection of liquid nutritional beverages.

[0007] Some of the challenges in incorporating whey proteins into acidic heat-treated beverages are the formation of an unstable precipitate that settles in the beverage, high viscosity or gel formation, and unpleasant taste due to a highly astringent and / or dry texture.

[0008] It is an object of the present invention to provide an acidic packaged heat-treated beverage preparation comprising whey protein and having improved organoleptic and / or visual properties.

[0009] Another object of the present invention is to provide a high protein beverage that has low viscosity, a pleasant taste, optionally low astringency, and may be clear or opaque.

[0010] The inventors have now discovered that such packaged thermally processed beverages can be provided within a wide acidic pH range up to pH 4.7 while still having low viscosity, and optionally low levels of astringency and dry texture. The present invention provides both clear beverages and opaque but stable beverages.

[0011] Thus, one aspect of the present invention is a packaged heat-treated beverage preparation having a pH in the range of 2.0 to 4.7, wherein the beverage a total amount of protein of 2 to 45% w / w by weight of the beverage, of which at least 85% w / w is BLG; - optionally with sweeteners and / or flavors The present invention relates to a preparation comprising:

[0012] Another aspect of the present invention is a method for producing a packaged thermally treated beverage preparation having a pH in the range of 2.0 to 4.7, comprising the steps of: a) - 2 to 45% by weight of a total amount of proteins, at least 85% of which are BLG; - optionally with sweeteners and / or flavors providing a liquid solution comprising: b) packaging the liquid solution; Including, The liquid solution of step a) and / or the packaged liquid solution of step b) are subjected to a heat treatment including at least pasteurization, Regarding the method.

[0013] Furthermore, one aspect of the present invention relates to the use of a protein solution comprising a total amount of protein of 2 to 45% w / w, based on the weight of the solution, of which at least 85 w / w% is BLG, for controlling the turbidity of a heat-treated acidic beverage preparation having a pH in the range of 2.0 to 4.7.

[0014] Yet another aspect of the present invention relates to the use of a protein solution comprising a total amount of protein of 2 to 45% w / w, of which at least 85 w / w% is BLG, based on the weight of the solution, for controlling astringency in heat-treated acidic beverage preparations having a pH in the range of 2.0 to 4.7.

[0015] A further aspect of the invention relates to a packaged thermally treated beverage preparation according to the invention for use in a method for treating a disease associated with protein malabsorption.

[0016] A further aspect of the present invention relates to the use of the packaged heat-treated beverage preparation according to the present invention as a dietary supplement. [Brief explanation of the drawings]

[0017] [Figure 1] FIG. 1 shows images of BLG and WPI beverages with pH 3.7 and protein content 6% w / w that were heat treated at 120° C. for 20 seconds and 75° C. for 15 seconds. [Figure 2] FIG. 1 shows images of WPI-B pH 3.0 to 3.7 120° C. and BLG pH 3.7 120° C. / 20 seconds. [Figure 3] FIG. 1 shows images of WPI-B pH 3.0 to 3.7 75° C. and BLG pH 3.7 75° C. / 15 seconds. [Figure 4] Figure 1 shows images of WPI-B pH 3.7 and BLG pH 3.9, 75°C / 15 seconds. [Figure 5] FIG. 1 shows the turbidity of 6% UHT treated (120° C. / 20 sec) BLG beverage preparations. [Figure 6] FIG. 1 shows the turbidity of 6% pasteurized (75° C. / 15 sec) BLG beverage compositions. [Figure 7] FIG. 1 shows the viscosity of 6% UHT treated (120° C. / 20 sec) BLG beverage formulations. [Figure 8] FIG. 1 shows the yellowness index (b*) of 6% UHT-treated (120° C. / 20 seconds) beverage compositions. [Figure 9] FIG. 1 shows the yellowness index (b*) of 6% pasteurized (75° C. / 15 seconds) beverage compositions. [Figure 10] Figure 1 shows images of 15% BLG beverage pH 3.7 (left) and 6% WPI-a pH 3.7 (right) at 75°C / 15 seconds. [Figure 11] FIG. 1 shows sensory evaluation of high protein BLG beverage compositions and images of 6 w / w% and 15 w / w% BLG samples at pH 3.7. [Figure 12] 1 shows high protein beverage formulations prepared by heating (from left to right) 30, 27.5, 25, 20% BLG for 5 minutes at 75° C. Viscosity remained low even after heating. [Figure 13] FIG. 1 shows images of different WPI and BLG samples. [Figure 14] Figure 1 shows the sensory evaluation of beverages (scale of 0-15): WPI pH 3.0 120°C / 20 sec and BLG pH 3.7 75°C / 15 sec. [Figure 15] FIG. 1 shows the effect of pH and temperature on sourness. [Figure 16] FIG. 1 shows sensory data regarding the astringency of BLG beverages at pH 3.0 (120° C. / 20 seconds) and pH 3.7 (75° C. / 15 seconds). [Figure 17] FIG. 1 shows sensory data for the dry texture of a pH 3.7 beverage at 120° C. / 20 seconds and 75° C. / 15 seconds. [Figure 18] FIG. 1 shows sensory data on whey aroma when BLG is kept in its native conformation. [Figure 19] shows images of a 6% BLG beverage with pH 3.7, heat treated at 95°C for 5 minutes, and with added minerals. [Figure 20] shows an image of a 6% BLG beverage with a pH of 3.7 that was heat treated at 75°C for 5 minutes and had minerals added. [Figure 21] shows the stability of a dairy BLG beverage at pH 4.3 with and without added sucrose, heat treated at 93°C for 4 minutes. [Figure 22] Figure 1 shows images of an opaque 6% protein BLG beverage prepared by heating at 75°C / 5 min at pH 4.2-4.5. [Figure 23] FIG. 1 shows images of BLG and SPI beverages heat-treated at 75° C. for 5 minutes at pH 3.7. [Figure 24] FIG. 1 shows images of a BLG beverage and an SPI beverage both having a pH of 3.7. DETAILED DESCRIPTION OF THE INVENTION

[0018] definition In the context of the present invention, the term "β-lactoglobulin" or "BLG" relates to β-lactoglobulin from mammalian species, e.g., in its native unfolded and / or glycosylated form, including naturally occurring genetic variants. This term further includes aggregated BLG, precipitated BLG, and crystalline BLG. When referring to the amount of BLG, reference is made to the total amount of BLG, including aggregated BLG. The total amount of BLG is determined according to Example 1.31. The term "aggregated BLG" relates to BLG that is at least partially unfolded and further aggregated with other denatured BLG molecules and / or other denatured whey proteins, typically through hydrophobic interactions and / or covalent bonds.

[0019] BLG is the most predominant protein in bovine whey and milk serum and exists in several genetic variants, the predominant ones in cow's milk being labeled A and B. BLG is a lipocalin protein and can bind many hydrophobic molecules, suggesting a role in their transport. BLG has also been shown to be able to bind iron via siderophores, which may play a role in combating pathogens. A homologue of BLG is missing in human breast milk.

[0020] Bovine BLG is a relatively small protein of approximately 162 amino acid residues with a molecular weight of approximately 18.3-18.4 kDa. Under physiological conditions, it is primarily a dimer, but dissociates into monomers below approximately pH 3 while maintaining its native state, as determined using nuclear magnetic resonance spectroscopy. Conversely, BLG also occurs in tetramers, octamers, and other multimeric aggregate forms under a variety of natural conditions.

[0021] In the context of the present invention, the term "non-aggregated β-lactoglobulin" or "non-aggregated BLG" also includes naturally occurring genetic variants of β-lactoglobulin from mammalian species, e.g., in its native unfolded and / or glycosylated form. However, the term does not include aggregated, precipitated or crystallized BLG. The amount or concentration of non-aggregated BLG is determined according to Example 1.6.

[0022] The percentage of non-aggregated BLGs to total BLGs was calculated (m 総BLG -m 非凝集BLG ) / m 総BLG * 100% determined by m 総BLG is the concentration or amount of BLG determined according to Example 1.31, and m 非凝集BLG is the concentration or amount of unaggregated BLG determined according to Example 1.6.

[0023] In the context of the present invention, the term "crystal" relates to a solid material whose constituent elements (such as atoms, molecules or ions) are arranged in a highly ordered microscopic structure, forming a crystal lattice that extends in all directions.

[0024] In the context of the present invention, the term "BLG crystal" refers to a protein crystal primarily containing non-aggregated, preferably native, BLG, arranged in a highly ordered microscopic structure, forming a crystal lattice extending in all directions. BLG crystals can be, for example, monolithic or polycrystalline, and can be, for example, intact crystals, crystal fragments, or combinations thereof. Crystal fragments are formed, for example, when intact crystals are subjected to mechanical shear during processing. Crystal fragments also have the highly ordered microscopic structure of crystals, but may lack the uniform surface and / or uniform corners or edges of intact crystals. See, for example, Figure 18 of PCT Application No. PCT / EP2017 / 084553 for examples of many intact BLG crystals and Figure 13 of PCT Application No. PCT / EP2017 / 084553 for examples of BLG crystal fragments. In either case, BLG crystals or crystal fragments can be visually identified using an optical microscope as well-defined, compact, and coherent structures. BLG crystals or crystal fragments are usually at least partially transparent. Protein crystals are also known to be birefringent, and this optical property can be used to identify unknown particles with crystalline structure. On the other hand, amorphous BLG aggregates usually appear as poorly defined, opaque, and irregularly sized open or porous masses.

[0025] In the context of the present invention, the term "crystallize" relates to the formation of protein crystals. Crystallization can, for example, occur spontaneously or can be initiated by the addition of crystallization seeds.

[0026] In the context of the present invention, the term "edible composition" relates to a composition that is safe for human consumption and use as a food ingredient and does not contain problematic amounts of toxic components such as toluene or other undesirable organic solvents.

[0027] In the context of the present invention, the term "ALA" or "alpha-lactalbumin" relates to alpha-lactoglobulin from mammalian species, e.g., in native and / or glycosylated form, including naturally occurring genetic variants. This term further includes aggregated ALA and precipitated BLG. When referring to the amount of ALA, it refers to the total amount of ALA, e.g., including aggregated ALA. The total amount of ALA is determined according to Example 1.31. The term "aggregated ALA" relates to ALA that is typically at least partially unfolded and further aggregated with other denatured ALA molecules and / or other denatured whey proteins, typically by hydrophobic interactions and / or covalent bonds.

[0028] Alpha-lactalbumin (ALA) is a protein present in the milk of almost all mammalian species. ALA forms the regulatory subunit of the lactose synthase (LS) heterodimer, and beta-1,4-galactosyltransferase (β4Gal-T1) forms the catalytic component. Together, these proteins enable LS to generate lactose by transferring a galactose moiety to glucose. One of its main structural differences from beta-lactoglobulin is that ALA lacks a free thiol group that can serve as the initiation point for the covalent aggregation reaction.

[0029] In the context of the present invention, the term "non-aggregated ALA" also includes naturally occurring genetic variants of ALA, e.g., natively unfolded and / or glycosylated, from mammalian species. However, this term does not include aggregated or precipitated ALA. The amount or concentration of non-aggregated BLG is determined according to Example 1.6.

[0030] The percentage of non-aggregated ALA to total ALA was calculated (m 総ALA -m 非凝集ALA ) / m 総ALA * 100% determined by m 総ALA is the concentration or amount of ALA determined according to Example 1.31, and m 非凝集ALA is the concentration or amount of non-aggregated ALA determined according to Example 1.6.

[0031] In the context of the present invention, the term "caseinomacropeptide" or "CMP" relates to a hydrophilic peptide, residues 106-169, derived from the hydrolysis of "κ-CN" or "kappa-casein" from mammalian species, e.g., in native and / or glycosylated form, and includes naturally occurring genetic variants by aspartic proteinases, e.g., chymosin.

[0032] In the context of the present invention, the term "BLG isolate" refers to a composition containing BLG in an amount of at least 85% w / w based on total protein. The BLG isolate preferably has a total protein content of at least 30% w / w, preferably at least 80% w / w based on total solids.

[0033] In the context of the present invention, the term "BLG isolate powder" relates to BLG isolate in powder form, preferably a free-flowing powder.

[0034] In the context of the present invention, the term "BLG isolate liquid" relates to a BLG isolate in liquid form, preferably an aqueous liquid.

[0035] The term "whey" refers to the liquid phase remaining after precipitating and removing casein from milk. Precipitation of casein can be achieved, for example, by acidifying the milk and / or by using rennet enzymes. Several types of whey exist, including "sweet whey," which is a whey product produced by rennet-based precipitation of casein, and "acid whey" or "sour whey," which is a whey product produced by acid-based precipitation of casein. Acid-based precipitation of casein can be achieved, for example, by the addition of food acids or by bacterial culture.

[0036] The term "whey" relates to the liquid remaining when casein and milk fat globules have been removed from milk, for example by microfiltration or large-pore ultrafiltration. Whey is sometimes called "ideal whey".

[0037] The term "whey protein" or "serum protein" relates to proteins present in whey.

[0038] In the context of the present invention, the term "whey protein" relates to a protein found in whey or whey. The whey protein may be a subset of the protein species found in whey or whey, or even a single whey protein species, or the whey protein may be the complete set of protein species found in whey and / or whey.

[0039] In the context of the present invention, the major non-BLG proteins of a standard whey protein concentrate from sweet whey are ALA, CMP, bovine serum albumin, immunoglobulins, osteopontin, lactoferrin and lactoperoxidase. In the context of the present invention, the weight percentages of the major non-BLG whey proteins of a standard whey protein concentrate from sweet whey are as follows: ALA in an amount of 18% w / w relative to total protein, CMP in an amount of 18% w / w relative to total protein; BSA in an amount of 4% w / w relative to total protein, Casein species in an amount of 5% w / w of total protein, immunoglobulins in an amount of 6% w / w relative to the total protein, osteopontin in an amount of 0.5% w / w of total protein; Lactoferrin in an amount of 0.1% w / w of total protein, and Lactoperoxidase in an amount of 0.1% w / w of total protein.

[0040] In the context of the present invention, the term "mother liquor" refers to the whey protein solution remaining after crystallizing BLG and at least partially removing the BLG crystals. The mother liquor may still contain some BLG crystals, but typically only small BLG crystals that have escaped separation.

[0041] In the context of the present invention, the term casein relates to the casein proteins found in milk and includes both the naturally occurring micellar casein found in raw milk, the individual casein species, and the caseinates.

[0042] In the context of the present invention, a liquid that is "supersaturated" or "supersaturated with respect to BLG" contains dissolved non-aggregated BLG at a concentration that exceeds the saturation point of non-aggregated BLG in that liquid under given physical and chemical conditions. The term "supersaturation" is well known in the field of crystallization (see, for example, Gerard Coquerela, "Crystallization of molecular systems from solution: phase diagrams, supersaturation, and other basic concepts," Chemical Society Reviews, pp. 2286-2300, No. 7, 2014), and supersaturation can be determined by several different measurement techniques (e.g., by spectroscopy or particle size analysis). In the context of the present invention, supersaturation with respect to BLG is determined by the following procedure.

[0043] Procedure for testing whether a liquid under a particular set of conditions is supersaturated with respect to BLG: a) Transfer a 50 ml sample of the liquid to be tested into a centrifuge tube (VWR Cat. No. 525-0402) with a height of 115 mm, an internal diameter of 25 mm, and a capacity of 50 ml. During steps a) to h), care should be taken to maintain the sample and subsequent fractions in the original physical and chemical conditions of the liquid. b) The sample is immediately centrifuged at 3000 g for 3.0 minutes with a maximum acceleration of 30 seconds and a maximum deceleration of 30 seconds. c) Immediately after centrifugation, transfer as much of the supernatant as possible (without disturbing the pellet, if one has formed) to a second centrifuge tube (same type as in step a). d) Take a 0.05 mL aliquot of the supernatant (aliquot A). e) 10 mg of BLG crystals (at least 98% pure, non-agglomerated BLG based on total solids) with particle size of at most 200 microns is added to the second centrifuge tube and the mixture is vortexed. f) Leave the second centrifuge tube at the original temperature for 60 minutes. g) Immediately after step f), centrifuge the second centrifuge tube at 500 g for 10 minutes, then take another 0.05 mL aliquot of the supernatant (aliquot B). h) If present, recover the centrifugation pellet of step g), resuspend it in milliQ water and immediately inspect the suspension for the presence of visible crystals under a microscope. i) Determine the concentration of unaggregated BLG in portion samples A and B using the method outlined in Example 1.6 - express the results as % BLG w / w relative to the total weight of the portion sample. The concentration of unaggregated BLG in portion sample A is C BLG,A The concentration of non-aggregated BLG in the small sample B is called C BLG,B It is called. j) The liquid sampled in step a) is BLG,B is c BLG,A and crystals were observed in step i), there was supersaturation (for the particular conditions).

[0044] In the context of the present invention, the terms "liquid" and "solution" encompass both compositions that do not contain particulate matter and compositions that contain a combination of liquid and solid and / or semi-solid particles, such as, for example, protein crystals or other protein particles. Thus, a "liquid" or "solution" can be a suspension or even a slurry. However, a "liquid" or "solution" is preferably pumpable.

[0045] In the context of the present invention, the terms "whey protein concentrate" (WPC) and "serum protein concentrate" (SPC) relate to dry or aqueous compositions containing a total amount of protein of 20 to 89% w / w relative to the total solids.

[0046] The WPC or SPC preferably contains: 20-89% w / w protein based on total solids; BLG, 15-70% w / w of total protein; 8-50% w / w ALA based on total protein, and CMP at 0-40% w / w relative to protein.

[0047] Alternatively, but also preferably, the WPC or SPC may contain: 20-89% w / w protein based on total solids; BLG, 15-90% w / w of total protein; 4-50% w / w ALA based on total protein, and CMP at 0-40% w / w relative to protein.

[0048] Preferably, the WPC or SPC contains: 20-89% w / w protein based on total solids; BLG, 15-80% w / w of total protein; 4-50% w / w ALA based on total protein, and CMP at 0-40% w / w relative to protein.

[0049] More preferably, the WPC or SPC contains: 70-89% w / w protein based on total solids; BLG, 30-90% w / w of total protein; 4-35% w / w ALA based on total protein, and 0-25% w / w CMP relative to protein.

[0050] SPC typically contains no CMP or only trace amounts of CMP.

[0051] The terms "whey protein isolate" (WPI) and "serum protein isolate" (SPI) relate to dry or aqueous compositions containing a total amount of protein of 90-100% w / w relative to the total solids.

[0052] The WPI or SPI preferably contains: 90-100% w / w protein based on total solids, BLG, 15-70% w / w of total protein; 8-50% w / w ALA based on total protein, and CMP of 0–40% w / w of total protein.

[0053] Alternatively, but also preferably, the WPI or SPI may contain: 90-100% w / w protein based on total solids, BLG, 30-95% w / w of total protein; 4-35% w / w ALA based on total protein, and CMP of 0–25% w / w of total protein.

[0054] More preferably, the WPI or SPI may contain: 90-100% w / w protein based on total solids, BLG, 30-90% w / w of total protein; 4-35% w / w ALA based on total protein, and CMP of 0–25% w / w of total protein.

[0055] SPI typically contains no CMP or only trace amounts of CMP.

[0056] In the context of the present invention, the term "additional protein" refers to a protein that is not BLG. Additional proteins present in a whey protein solution typically include one or more of the non-BLG proteins found in whey or milk serum. Non-limiting examples of such proteins are alpha-lactalbumin, bovine serum albumin, immunoglobulins, caseinomacropeptide (CMP), osteopontin, lactoferrin, and milk fat globule membrane proteins.

[0057] The terms "consisting essentially of" and "consisting essentially of" mean that the claim or feature in question includes the specified materials or steps as well as materials or steps that do not materially affect the basic and novel characteristics of the claimed invention.

[0058] In the context of the present invention, the phrase "Y and / or X" means "Y" or "X", or "Y and X". Along the same lines of logic, "n1, n2, ..., n i-1 and / or n i " The phrase "n1" or "n2" or ... or "n i-1 " or "n1" or components: n1, n2, ... n i-1 and n i means any combination of

[0059] In the context of the present invention, the term "dry" or "dried" means that the composition or product in question contains at most 10% w / w water, preferably at most 6% w / w water, more preferably even less.

[0060] In the context of the present invention, the term "physical microbial reduction" refers to a physical interaction with a composition that results in a reduction in the total amount of viable microorganisms in the composition. This term does not include the addition of chemicals that result in the killing of microorganisms. This term also does not include the heat exposure to which the sprayed droplets are exposed during spray drying, but does include possible preheating before spray drying.

[0061] In the context of the present invention, the pH of a powder refers to the pH of 10 g of powder mixed with 90 g of demineralized water, measured according to Example 1.16.

[0062] In the context of the present invention, the weight percent (% w / w) of an ingredient of a certain composition, product or material means the weight percent of that ingredient relative to the weight of the particular composition, product or material, unless another measure (e.g., total solids or total protein) is specifically stated.

[0063] In the context of the present invention, the process step "concentrate" and the verb "concentrate" refer to protein concentration and encompass both protein concentration on a total solids basis and protein concentration on a total weight basis. This means, for example, that concentration does not necessarily require that the absolute protein concentration w / w of the composition be increased, as long as the protein content is increased relative to the total solids.

[0064] In the context of the present invention, the term "weight ratio" between component X and component Y refers to the calculated m X / m Y (In the formula, m X is the amount (weight) of component X, and m Y is the amount (by weight) of component Y).

[0065] In the context of the present invention, the term "at least pasteurized" relates to a heat treatment that has a microorganism-killing effect equal to or greater than that of a heat treatment at 70° C. for 10 seconds. The standard for determining the killer effect is Escherichia coli (E. coli) O157:H7.

[0066] In the context of the present invention, the term "whey protein feedstock" relates to a whey protein source from which a liquid BLG isolate is derived. A whey protein feedstock has a lower BLG content relative to total protein than a liquid BLG isolate and is typically a WPC, WPI, SPC or SPI.

[0067] In the context of the present invention, the term "BLG-enriched composition" refers to a BLG-enriched composition obtained by isolating BLG from a whey protein feedstock. The BLG-enriched composition typically contains the same whey protein as the whey protein feedstock, but BLG is present at a significantly higher concentration relative to total protein than in the whey protein feedstock. The BLG-enriched composition can be prepared from the whey protein feedstock by, for example, chromatography, protein crystallization, and / or membrane-based protein fractionation. The BLG-enriched composition contains BLG in an amount of at least 85% w / w, preferably at least 90% w / w, relative to total protein. In some cases, the BLG-enriched composition can be used directly as a liquid BLG isolate. However, additional processing is often required to convert the BLG-enriched composition into a liquid BLG isolate.

[0068] In the context of the present invention, the term "whey protein solution" is used to describe a particular aqueous whey protein composition that is supersaturated with respect to BLG in salt-soluble form and is useful for preparing BLG crystals.

[0069] In the context of the present invention, the term "sterile" means that the sterile composition or product in question does not contain viable microorganisms and is therefore free of microbial growth during storage at room temperature. A sterilized composition is sterile.

[0070] When a liquid, such as a beverage preparation, is sterilized and aseptically packaged in a sterile container, it typically has a shelf life of at least six months at room temperature. The sterilization process kills spores and microorganisms that can cause spoilage of the liquid.

[0071] In the context of the present invention, the term "energy content" refers to the total energy content contained in a food product. Energy content can be measured in kilojoules (kJ) or kilocalories (kcal) and is referred to as calories per amount of food, e.g., kcal per 100 grams of food. An example is a drink with an energy content of 350 kcal / 100 grams of drink.

[0072] The total energy content of a food includes the energy contributions from all macronutrients present in the food, such as energy from proteins, lipids, and carbohydrates. The distribution of energy from macronutrients in a food can be calculated based on the amount of macronutrients in the food and the contribution of the macronutrients to the total energy content of the food. Energy distribution can be expressed as the energy percentage (E%) of the total energy content of the food. For example, for a beverage containing 20E% protein, 50E% carbohydrates, and 30E% lipids, this means that 20% of the total energy comes from proteins, 50% of the total energy comes from carbohydrates, and 30% of the total energy comes from fats (lipids).

[0073] In the context of the present invention, the term "nutritionally complete dietary supplement" is understood as a food product comprising proteins, lipids and carbohydrates, and further comprising vitamins, minerals and trace elements, such that the beverage has a nutritional profile that corresponds to a complete and healthy diet.

[0074] In the context of the present invention, the term "nutritionally incomplete food supplement" means a food product that contains one or more key nutrients and optionally further contains vitamins, minerals and trace elements. Nutritionally incomplete beverages may contain protein as the only nutrient, or may contain, for example, protein and carbohydrates.

[0075] The term "food for special medical purposes (FSMP)" or "medical food" refers to a food for oral intake or tube feeding, used for specific medical disorders, diseases, or conditions with unique nutritional requirements, and used under medical supervision. Medical foods can be nutritionally complete or nutritionally incomplete supplemental foods / drinks.

[0076] The term "nutrient" refers to substances that an organism uses to survive, grow, and reproduce. Nutrients can be either macronutrients or micronutrients. Macronutrients are nutrients that provide energy when consumed, such as proteins, lipids, and carbohydrates. Micronutrients are nutrients such as vitamins, minerals, and trace elements.

[0077] The term "nutrients" refers to substances that living organisms use to survive, grow, and reproduce. Nutrients can be either macronutrients or micronutrients. Macronutrients are nutrients that provide energy when consumed, such as proteins, lipids, and carbohydrates. Micronutrients are nutrients that are vitamins, minerals, and trace elements.

[0078] The term "instant beverage powder" or "instant beverage powder product" means a powder that can be converted into a liquid beverage by the addition of a liquid, such as water.

[0079] In the context of the present invention, the terms "drink preparation" and "preparation" as used as entities relate to any aqueous liquid that can be consumed as a drink, for example by pouring, sipping or tube feeding.

[0080] In the context of the present invention, the term "protein fraction" relates to the proteins of the composition in question, for example the proteins of a powder or drink preparation.

[0081] In the context of the present invention, the term "astringent" relates to texture. Astringent taste is felt as a contraction of the buccal muscles, resulting in increased saliva production. Astringent taste is therefore not a taste per se, but rather a physical texture and time-dependent sensation in the mouth.

[0082] In the context of the present invention, the term "dry texture" relates to the sensation in the mouth, which feels like dryness of the mouth and teeth, resulting in minimal saliva production.

[0083] Thus, dry texture is not a taste per se, but rather a physical texture and time-dependent sensation in the mouth.

[0084] In the context of the present invention, the term "minerals" as used herein refers to any one of major minerals, trace or minor minerals, other minerals, and combinations thereof, unless otherwise specified. Major minerals include calcium, phosphorus, potassium, sulfur, sodium, chlorine, and magnesium. Trace or minor minerals include iron, cobalt, copper, zinc, molybdenum, iodine, selenium, and manganese, and other minerals include chromium, fluorine, boron, lithium, and strontium.

[0085] In the context of the present invention, the terms "lipid," "fat," and "oil" as used herein are used interchangeably to refer to lipid materials derived from or processed from plants or animals, unless otherwise specified. These terms also include synthetic lipid materials, provided that such synthetic lipid materials are suitable for human consumption.

[0086] In the context of the present invention, the term "transparent" encompasses beverage preparations that have a visually clear appearance and allow light to pass through them, through which a clear image can be seen. A transparent beverage has a turbidity of up to 200 NTU.

[0087] In the context of the present invention, the term "opaque" encompasses beverage preparations that have a visually unclear appearance, which have a turbidity of more than 200 NTU.

[0088] One aspect of the present invention is a packaged heat-treated beverage preparation having a pH in the range of 2.0 to 4.7, wherein the beverage a total amount of protein of 2 to 45% w / w by weight of the beverage, of which at least 85% w / w is BLG; optionally with sweeteners, sugar polymers and / or flavors The present invention relates to a preparation comprising:

[0089] Packaged heat-treated beverage preparations containing at least 85% w / w protein are highly beneficial for several reasons. High BLG content in acidic beverages allows for increased pH ranges and reduced heating temperatures, even when high protein concentrations are applied, while still maintaining clarity and lack of color. Surprisingly, BLG beverages have been found to have reduced astringency, dry texture, sourness, whey aroma, and citric flavor compared to WPI beverages containing lower amounts of BLG.

[0090] Another advantage of the present invention and the expanded pH range is that it allows for the production of dairy beverages that have high turbidity, low viscosity, while still being white, not yellowing, and still being stable.

[0091] In some preferred embodiments of the packaged thermally treated beverage preparations of the present invention, at least 85% w / w of the protein is BLG, preferably at least 88% w / w of the protein is BLG, more preferably at least 90% w / w, even more preferably at least 91% w / w, and most preferably at least 92% w / w of the protein is BLG.

[0092] Because even higher relative amounts of BLG are both achievable and desirable, in some preferred embodiments of the present invention, at least 94% w / w of the protein in the packaged heat-treated beverage preparation is BLG, more preferably at least 96% w / w of the protein is BLG, even more preferably at least 98% w / w of the protein is BLG, and most preferably approximately 100% w / w of the protein is BLG.

[0093] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation is at least pasteurized.

[0094] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation is sterilized.

[0095] In some preferred embodiments of the present invention, the native conformation of the protein is maintained.

[0096] The degree of nativeness of a protein depends on several factors, such as the protein concentration, pH, temperature and time of heat treatment.

[0097] The intrinsic tryptophan fluorescence emission ratio R = I330 / I350 is a measure of the nativity of a protein. When R is at least 1.11, the native conformation predominates, whereas when R is less than 1.11, at least partial unfolding and aggregation predominates. A method for analyzing intrinsic tryptophan fluorescence is described in Example 1.1.

[0098] The inventors have found that an intrinsic tryptophan fluorescence ratio R=I330 / I350 of at least 1.11 can be obtained for a heat-treated high-protein beverage, while still having a low viscosity and being clear, even when the protein fraction and / or beverage preparation is subjected to a heat treatment equivalent to pasteurization (e.g., a temperature below 90°C).

[0099] Thus, in some preferred embodiments of the present invention, the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11, thus indicating that the protein is in its native state.

[0100] In some preferred embodiments of the present invention, the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.12, preferably at least 1.13, more preferably at least 1.15, even more preferably at least 1.17, and most preferably at least 1.19.

[0101] In some preferred embodiments of the present invention, a packaged thermally treated beverage preparation comprising a protein fraction and, optionally, other ingredients such as lipids, carbohydrates, vitamins, minerals, food acids or emulsifiers has a tryptophan fluorescence ratio of at least 1.11.

[0102] Thus, in some preferred embodiments of the present invention, the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11.

[0103] In some preferred embodiments of the present invention, the thermally treated beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.12, preferably at least 1.13, more preferably at least 1.15, even more preferably at least 1.17, and most preferably at least 1.19.

[0104] In some preferred embodiments of the present invention, the protein is denatured or at least partially denatured.

[0105] Thus, in some preferred embodiments of the present invention, the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of less than 1.11, thus indicating that the protein is at least partially unfolded and aggregation predominates.

[0106] In some embodiments of the present invention, the thermally treated beverage preparation has an intrinsic tryptophan fluorescence ratio (I330nm / I350nm) of less than 1.10, more preferably less than 1.08, even more preferably less than 1.05, and most preferably less than 1.00.

[0107] In addition to the protein fraction, the beverage preparation may also optionally contain other food additives such as lipids, carbohydrates, vitamins, minerals, food acids or emulsifiers, etc. In some preferred embodiments of the present invention, the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of less than 1.11, thus indicating that the protein is at least partially unfolded and aggregation predominates.

[0108] In some preferred embodiments of the present invention, the thermally treated beverage preparation has an intrinsic tryptophan fluorescence ratio (I330nm / I350nm) of less than 1.10, more preferably less than 1.08, even more preferably less than 1.05, and most preferably less than 1.00.

[0109] Protein denaturation can also be demonstrated by other analytical methods besides tryptophan fluorescence, which are described in Example 1.3.

[0110] In some preferred embodiments of the present invention, the protein fraction of the packaged heat-treated beverage preparation has a degree of protein denaturation of at most 10%, preferably at most 8%, more preferably at most 5%, even more preferably at most 3%, even more preferably at most 1%, and most preferably at most 0.5%.

[0111] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation has a degree of protein denaturation of at most 10%, preferably at most 8%, more preferably at most 5%, even more preferably at most 3%, even more preferably at most 1%, and most preferably at most 0.5%.

[0112] In some embodiments of the present invention, when the protein fraction and / or beverage preparation is subjected to, for example, high temperature heat treatment, the degree of protein denaturation is greater than 10%, preferably greater than 20%, preferably greater than 30%, preferably greater than 40%, or preferably greater than 50%, or preferably greater than 70%, or preferably greater than 80%, or preferably greater than 90%, or preferably greater than 95%, or preferably greater than 99%.

[0113] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 3.0 to 4.3, which is particularly preferred for producing clear beverages with low viscosity and improved taste.

[0114] With respect to appearance, it has surprisingly been found that using a whey protein beverage in which at least 85% w / w of the protein is BLG, the pH can be increased during heat treatment, resulting in improved visual perception (color and turbidity) and viscosity when compared to a heat-treated WPI beverage.

[0115] Surprisingly, it has been found that there are significant differences in sensory parameters between beverages made with WPI compared to the BLG beverages of the present invention. Surprisingly and advantageously, it has been found that BLG beverages have lower levels of astringency, dry texture, sourness, whey aroma, and citric acid flavor compared to WPI beverages. It has also been found that by increasing the pH of an acidic beverage, less sweetener is needed to balance the acidity of the beverage, and therefore, a lower concentration of sweetener is needed in such beverages.

[0116] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation has a pH in the range of 3.0 to 4.1, or preferably 3.1 to 4.0, or preferably 3.2 to 3.9, or preferably 3.7 to 3.9, more preferably 3.4 to 3.9, and even more preferably 3.5 to 3.9.

[0117] These pH ranges are particularly suitable when the beverage preparation is pasteurized.

[0118] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation preferably has a pH in the range of 3.0 to 3.9, or preferably 3.2 to 3.7, or preferably 3.4 to 3.6, or preferably 3.5 to 3.7, or preferably 3.4 to 3.6.

[0119] These pH ranges in combination with high temperature processes such as sterilization are particularly suitable for producing clear beverages with low viscosity and improved taste.

[0120] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 4.1 to 4.7, which is particularly suitable for producing a stable beverage with a milky appearance and high turbidity while still having a low viscosity. In some embodiments of the present invention, the pH range is 4.2 to 4.6. In other embodiments of the present invention, the pH range is 4.2 to 4.5.

[0121] The visual appearance of a beverage preparation is important to consumers for both clear and opaque beverages. Particularly in the case of clear watery or white milky beverages, the inventors have found that it is advantageous to be able to control the color, or rather the lack of color, of the beverage.

[0122] However, even when dedicated colorants are added during beverage production, the inventors have found that it is advantageous to be able to avoid additional color sources to avoid undesirable variations or changes in the visual appearance of the beverage. The inventors have found that the high BLG protein profile described herein is more color-neutral / colorless than conventional WPIs and contributes to less color variation than conventional WPIs. Conventional WPIs have a yellowish appearance that can be alleviated to some extent by adding an oxidizing agent, such as bleach. However, the addition of an oxidizing agent is usually undesirable and is no longer necessary in the present invention.

[0123] The CIELAB color scale described in Example 1.9 is used to determine the color of the beverage. For example, a positive delta b * A negative delta b indicates a color that is more yellow than demineralized water. * A value of 0.01 indicates a beverage that is bluer than demineralized water. Therefore, to obtain a beverage that is neither yellow nor blue, the color delta b * A value close to 0 is usually preferred by consumers.

[0124] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation has a color value delta b in the range of -0.10 to +0.51 on the CIELAB color scale, especially when the preparation has a turbidity of at most 200 NTU, more preferably at most 40 NTU. * It has.

[0125] In another preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a color value delta b in the range of 0.0 to 0.40, preferably in the range of 0.10 to 0.25 on the CIELAB color scale. * It has.

[0126] For opaque beverage preparations, e.g. having a turbidity of more than 200 NTU, preferably more than 1000 NTU, the packaged heat-treated beverage preparation preferably has a color value delta b on the CIELAB color scale in the range of -6 to -1.7, preferably in the range of -5.0 to -2.0. * It has.

[0127] In some preferred embodiments of the present invention, the protein fraction of the packaged heat-treated beverage preparation has a color value delta b in the range of -0.10 to +0.51, especially when the preparation has a turbidity of at most 200 NTU, more preferably at most 40 NTU. * It has.

[0128] These drinks have a higher delta b * The beverages have a lower yellow color compared to beverages containing WPIs, which have a higher yellow color.

[0129] In another preferred embodiment of the present invention, the protein fraction of the packaged heat-treated beverage preparation has a color value delta b in the range of 0.0 to 0.40, preferably in the range of 0.10 to 0.25 on the CIELAB color scale. * It has.

[0130] a * The value represents the green-red component, with green going negative and red going positive. To obtain a beverage that is neither red nor green, the color delta a * A value near zero is usually preferred.

[0131] The protein fraction of the packaged heat-treated beverage preparation has a delta a in the range of -0.2 to 0.2 on the CIELAB color scale, especially when the preparation has a turbidity of at most 200 NTU, more preferably at most 40 NTU. * Preferably, the packaged heat-treated beverage preparation has a color value delta a in the range of -0.15 to 0.15, preferably in the range of -0.10 to 0.10 on the CIELAB color scale. * It has.

[0132] The inventors have found that it can be advantageous to control the mineral content in order to achieve some of the desired properties of a packaged heat-treated beverage preparation.

[0133] In some embodiments of the present invention, the packaged thermally processed beverage preparation comprises a plurality of minerals. In one exemplary embodiment, the packaged thermally processed beverage preparation comprises at least four minerals. In one embodiment, the four minerals are sodium, potassium, magnesium, and calcium.

[0134] The inventors have surprisingly found that the use of BLG isolate as defined herein and in Example 2 allows for the production of heat-treated beverage preparations with high mineral concentrations without compromising viscosity, thereby enabling the production of packaged heat-treated beverage preparations with high mineral content, allowing for the production of beverages that are nutritionally complete or nutritionally incomplete supplements.

[0135] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg and Ca in the packaged heat-treated beverage preparation is in the range of 0 to 750 mM, preferably in the range of 100 to 600 mM, or preferably in the range of 200 to 500 mM.

[0136] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg and Ca is at most 750 mM in the packaged heat-treated beverage preparation.

[0137] In another preferred embodiment of the present invention, the sum of the amounts of Na, K, Mg and Ca in the packaged heat-treated beverage preparation is at most 600 mM, preferably at most 500 mM, or preferably at most 400 mM, or preferably at most 300 mM, or preferably at most 200 mM, preferably at most 170 mM, most preferably at most 150 mM, or preferably at most 130 mM, or preferably at most 100 mM, or preferably at most 80 mM, or preferably at most 60 mM, or preferably at most 40 mM, or preferably at most 30 mM, or preferably at most 20 mM, or preferably at most 10 mM, or preferably at most 5 mM, or preferably at most 1 mM.

[0138] In another exemplary embodiment, the packaged thermally processed beverage preparation comprises a plurality of minerals selected from the group consisting of calcium, iodine, zinc, copper, chromium, iron, phosphorus, magnesium, selenium, manganese, molybdenum, sodium, potassium, and combinations thereof.

[0139] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises at most 150 mM KCl and at most 150 mM CaCl, or at most 130 mM KCl and at most 130 mM CaCl, or at most 110 mM KCl and at most 110 mM CaCl, or at most 100 mM KCl and at most 100 mM CaCl, or preferably at most 80 mM KCl and at most 80 mM CaCl, or preferably at most 50 mM KCl and at most 50 mM CaCl, or preferably at most 40 mM KCl and at most 40 mM CaCl.

[0140] In another preferred embodiment of the present invention, the thermally treated beverage preparation is a low-mineral beverage.

[0141] In the context of the present invention, the term "low in minerals" relates to a composition, such as a liquid, drink, powder or another food product, which has at least one, preferably two, and even more preferably all of the following: - ash content of maximum 1.2% w / w based on total solids; - a total content of calcium and magnesium of maximum 0.3% w / w based on total solids; - maximum total sodium and potassium content of 0.10% w / w based on total solids; - Total phosphorus content of up to 100mg phosphorus per 100g of protein.

[0142] Preferably, the low mineral composition has at least one, preferably two or more, and even more preferably all of the following: - ash content of maximum 0.7% w / w based on total solids; - a total content of calcium and magnesium of maximum 0.2% w / w based on total solids; - a total content of sodium and potassium of maximum 0.08% w / w based on total solids; - Total phosphorus content of up to 80 mg phosphorus per 100 g of protein.

[0143] Even more preferably, the low mineral composition has at least one, preferably two or more, and even more preferably all of the following: - ash content of maximum 0.5% w / w based on total solids; - a total calcium and magnesium content of maximum 0.15% w / w based on total solids; - maximum total sodium and potassium content of 0.06% w / w based on total solids; - Total phosphorus content of up to 50 mg phosphorus per 100 g of protein.

[0144] It is particularly preferred that the low mineral composition has: - ash content of maximum 0.5% w / w based on total solids; - a total calcium and magnesium content of maximum 0.15% w / w based on total solids; - maximum total sodium and potassium content of 0.06% w / w based on total solids; - Total phosphorus content of up to 50 mg phosphorus per 100 g of protein.

[0145] The inventors have found that the present invention makes it possible to prepare packaged heat-treated beverage preparations that have a very low content of phosphorus and other minerals such as potassium, which is advantageous for patients suffering from kidney disease or with reduced kidney function.

[0146] The packaged thermally treated beverage preparation is preferably a low phosphorus beverage preparation.

[0147] The packaged thermally treated beverage preparation is preferably a low potassium beverage preparation.

[0148] The packaged thermally treated beverage preparation is preferably a low phosphorus and low potassium beverage preparation.

[0149] In the context of the present invention, the term "low phosphorus" relates to a composition, e.g., a liquid, powder or another food product, having a total phosphorus content of at most 100 mg phosphorus per 100 g protein. Preferably, the low phosphorus composition has a total phosphorus content of at most 80 mg phosphorus per 100 g protein. More preferably, the low phosphorus composition may have a total phosphorus content of at most 50 mg phosphorus per 100 g protein. Even more preferably, the low phosphorus composition may have a total phosphorus content of at most 20 mg phosphorus per 100 g protein. Even more preferably, the low phosphorus composition may have a total phosphorus content of at most 5 mg phosphorus per 100 g protein. The low phosphorus composition according to the present invention may be used as a food ingredient for producing a food product for patient groups with reduced renal function.

[0150] Thus, in some particularly preferred embodiments of the present invention, the packaged heat-treated beverage preparation contains at most 80 mg of phosphorus per 100 g of protein. Preferably, the packaged heat-treated beverage preparation contains at most 30 mg of phosphorus per 100 g of protein. More preferably, the packaged heat-treated beverage preparation contains at most 20 mg of phosphorus per 100 g of protein. Even more preferably, the packaged heat-treated beverage preparation contains at most 10 mg of phosphorus per 100 g of protein. Most preferably, the packaged heat-treated beverage preparation contains at most 5 mg of phosphorus per 100 g of protein.

[0151] The phosphorus content is determined in accordance with Example 1.19 with respect to the total amount of elemental phosphorus in the composition in question.

[0152] In the context of the present invention, the term "low potassium" relates to a composition, e.g., a liquid, powder, or another food product, having a total potassium content of at most 700 mg potassium per 100 g protein. Preferably, the low phosphorus composition has a total potassium content of at most 600 mg potassium per 100 g protein. More preferably, the low potassium composition may have a total potassium content of at most 500 mg potassium per 100 g protein. More preferably, the low potassium composition may have a total potassium content of at most 400 mg potassium per 100 g protein. More preferably, the low potassium composition may have a total potassium content of at most 300 mg potassium per 100 g protein. Even more preferably, the low potassium composition may have a total potassium content of at most 200 mg potassium per 100 g protein. Even more preferably, the low potassium composition may have a total potassium content of at most 100 mg potassium per 100 g protein. Even more preferably, the low potassium composition may have a total potassium content of up to 50 mg potassium per 100 g protein, and even more preferably, the low potassium composition may have a total potassium content of up to 10 mg potassium per 100 g protein.

[0153] The low potassium composition according to the present invention can be used as a food ingredient to prepare food for patient populations with reduced renal function.

[0154] Thus, in some particularly preferred embodiments of the present invention, the packaged heat-treated beverage preparation comprises at most 600 mg of potassium per 100 g of protein. More preferably, the packaged heat-treated beverage preparation comprises at most 500 mg of potassium per 100 g of protein. More preferably, the packaged heat-treated beverage preparation comprises at most 400 mg of potassium per 100 g of protein. More preferably, the packaged heat-treated beverage preparation comprises at most 300 mg of potassium per 100 g of protein. Even more preferably, the packaged heat-treated beverage preparation comprises at most 200 mg of potassium per 100 g of protein. Even more preferably, the packaged heat-treated beverage preparation comprises at most 100 mg of potassium per 100 g of protein. Even more preferably, the packaged heat-treated beverage preparation comprises at most 50 mg of potassium per 100 g of protein, and even more preferably, the packaged heat-treated beverage preparation comprises at most 10 mg of potassium per 100 g of protein.

[0155] The potassium content is determined in accordance with Example 1.19 relative to the total amount of elemental phosphorus in the composition in question.

[0156] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation comprises at most 100 mg phosphorus / 100 g protein and at most 700 mg potassium / 100 g protein, preferably at most 80 mg phosphorus / 100 g protein and at most 600 mg potassium / 100 g protein, more preferably at most 60 mg phosphorus / 100 g protein and at most 500 mg potassium / 100 g protein, more preferably at most 50 mg phosphorus / 100 g protein and at most 400 mg potassium / 100 g protein, or more preferably at most 20 mg phosphorus / 100 g protein and at most 200 mg potassium / 100 g protein, or even more preferably at most 10 mg phosphorus / 100 g protein and at most 50 mg potassium / 100 g protein. In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation comprises at most 100 mg phosphorus / 100 g protein and at most 340 mg potassium / 100 g protein.

[0157] The heat-treated beverage preparation containing small amounts of phosphorus and potassium can be advantageously supplemented with carbohydrates and lipids, and preferably further comprises a total amount of carbohydrates in the range of 30-60%, preferably in the range of 35-50E%, of the total energy content of the beverage, and a total amount of lipids in the range of 20-60%, preferably in the range of 30-50E%, of the total energy content of the beverage.

[0158] In one embodiment of the present invention, the packaged thermally processed beverage preparation comprises a plurality of vitamins. In one exemplary embodiment, the packaged thermally processed beverage preparation comprises at least 10 vitamins. In one exemplary embodiment, the packaged thermally processed beverage preparation comprises a plurality of vitamins selected from the group consisting of vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12, vitamin C, vitamin D, vitamin K, riboflavin, pantothenic acid, vitamin E, thiamine, niacin, folic acid, biotin, and combinations thereof.

[0159] In one embodiment of the present invention, a packaged thermally treated beverage comprises a plurality of vitamins and a plurality of minerals.

[0160] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation contains one or more food acids selected from the group consisting of citric acid, malic acid, tartaric acid, acetic acid, benzoic acid, butyric acid, lactic acid, fumaric acid, succinic acid, ascorbic acid, adipic acid, phosphoric acid, and mixtures thereof.

[0161] In one embodiment of the present invention, the packaged thermally treated beverage preparation further comprises a flavor selected from the group consisting of salt, flavorings, seasonings and / or spices. In a preferred embodiment of the present invention, the flavor comprises chocolate, cocoa, lemon, orange, lime, strawberry, banana, forest fruit flavors or combinations thereof. The choice of flavor may depend on the beverage to be produced.

[0162] Clarity is a parameter used by consumers to evaluate products. One way to determine the clarity of a liquid food product is by measuring the turbidity of the product as described in Example 1.7.

[0163] In some embodiments of packaged thermally treated beverage preparations, it is beneficial for the beverage preparation to be clear. This can be advantageous, for example, when the beverage is used in sports drinks or "protein waters," where it is beneficial for the beverage to resemble water in appearance.

[0164] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a turbidity of at most 200 NTU, such beverage being clear.

[0165] Surprisingly, the inventors have found that the heat-treated beverage preparation according to the invention allows for the production of clear heat-treated beverage preparations having a turbidity of up to 200 NTU.

[0166] This was seen when the heat treatment applied was both sterilization and pasteurization.

[0167] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation has a turbidity of at most 150 NTU, or preferably at most 100 NTU, or preferably at most 80 NTU, or preferably at most 60 NTU, or more preferably at most 40 NTU, or preferably at most 30 NTU, preferably at most 20 NTU, more preferably at most 10 NTU, more preferably at most 5 NTU, and even more preferably at most 2 NTU.

[0168] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a turbidity of more than 200 NTU, such beverage being opaque.

[0169] In some embodiments of the packaged heat-treated beverage preparation, it is beneficial for the beverage preparation to be opaque. This is advantageous, for example, when the beverage is milk-like and should have a milky appearance. The appearance of nutritionally complete dietary supplements is typically opaque.

[0170] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation has a turbidity of more than 250 NTU. Preferably, the packaged heat-treated beverage preparation has a turbidity of more than 300 NTU, more preferably more than 500 NTU, more preferably more than 1000, preferably more than 1500 NTU, and even more preferably more than 2000 NTU.

[0171] The amount of insoluble matter in a heat-treated beverage preparation is a measure of the instability of the beverage and the extent to which sediment settling occurs over time. Beverages with high amounts of insoluble matter are typically considered unstable.

[0172] In the context of the present invention, a whey protein beverage preparation is considered "stable" if centrifugation at 3000 x g for 5 minutes precipitates a maximum of 15% of the total protein in a heated sample. See analytical methods in Example 1.10.

[0173] Surprisingly, when BLG is used as a protein source in an amount of at least 85 w / w%, the protein fraction is found to contain up to 15% insoluble material after centrifugation at 3000 g for 5 minutes, compared to when a WPI with a low BLG content is used as a protein source, demonstrating that the beverage preparation is stable.

[0174] Thus, in some preferred embodiments of the present invention, the protein fraction of the heat-treated beverage preparation contains at most 15% insoluble material.

[0175] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation contains a maximum of 15% insoluble matter.

[0176] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation preferably contains at most 12% insoluble matter, more preferably at most 10% insoluble matter, even more preferably at most 8% insoluble matter, and most preferably at most 6% insoluble matter.

[0177] Even lower levels of insoluble matter are generally preferred, and in some preferred embodiments the packaged thermally treated beverage preparation contains at most 4% insoluble matter, preferably at most 2% insoluble matter, more preferably at most 1% insoluble matter, and most preferably no detectable insoluble matter.

[0178] Consumers prefer that heat-treated beverages are liquid, easy to drink, and do not gel.

[0179] One method of determining the viscosity of a beverage preparation is by measuring the viscosity of the beverage as described in Example 1.8.

[0180] In some embodiments of packaged thermally treated beverage preparations, it is beneficial for the beverage preparation to have a very low viscosity, which is advantageous when the beverage is used as a sports drink or in some embodiments as a nutritionally complete or nutritionally incomplete supplement.

[0181] Surprisingly, the inventors have found that beverage preparations having an acidic pH and subjected to a heat treatment such as pasteurization, and further subjected to sterilization, have a viscosity of up to 200 centipoise (cP) measured at a shear rate of 100 / sec and 22 degrees Celsius.

[0182] Thus, in some preferred embodiments of the present invention, the packaged heat-treated beverage preparation has a viscosity of at most 200 cP.

[0183] Preferably, the viscosity of the packaged heat treated beverage preparation is at most 150 cP, preferably at most 100 cP, more preferably at most 80 cP, even more preferably at most 50 cP, and most preferably at most 40 cP.

[0184] Even lower viscosities are typically preferred, and therefore in some preferred embodiments of the present invention, the viscosity of the packaged heat-treated beverage preparation is at most 20 cP, preferably at most 10 cP, more preferably at most 5 cP, even more preferably at most 3 cP, even more preferably at most 2 cP, and most preferably at most 1 cP.

[0185] It has previously been found that the addition of an anti-flocculation agent to a WPI-containing acidic, clear, heat-treated beverage having a pH above pH 3.0 is essential. See, e.g., Etzel 2004 (Etzel, MR, 2004, Manufacture and use of dairy protein fraction. American Society for Nutritional Science, pp. 996-1002).

[0186] Surprisingly, it has been found by the inventors that clear heat-treated beverages containing at least 85% w / w BLG can be produced even at pH above pH 3.0 without the addition of an aggregation inhibitor.

[0187] Thus, in some preferred embodiments of the present invention, the packaged heat-treated beverage preparation does not contain an agglomeration inhibitor, or instead contains only trace amounts of an agglomeration inhibitor.

[0188] In the context of the present invention, the term "flocculation inhibitor" relates to food-grade non-protein surfactants such as, for example, lauryl sulfate, polysorbates, and mono- and / or diglycerides.

[0189] In some embodiments of the present invention, the packaged thermally treated beverage preparation comprises up to 0.1% w / w of an agglomeration inhibitor, preferably up to 0.03% w / w of an agglomeration inhibitor, and most preferably no agglomeration inhibitor. These embodiments are particularly preferred for clear, low-fat beverages.

[0190] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation comprises a total amount of protein of 4.0 to 30% w / w based on the weight of the beverage.

[0191] In some embodiments of the present invention, the packaged heat-treated beverage preparation advantageously has a protein content of 2.0 to 10.0% w / w based on the weight of the beverage.

[0192] Thus, in some embodiments of the present invention, the packaged heat-treated beverage preparation preferably comprises a total amount of protein of 2.0 to 10% w / w by weight of the beverage, preferably a total amount of protein of 3.0 to 10% w / w by weight of the beverage, preferably a total amount of protein of 5.0 to 9.0% w / w by weight of the beverage, preferably a total amount of protein of 6.0 to 8.0% w / w by weight of the beverage.

[0193] In some embodiments of the invention, the protein content of the beverage is advantageously high, such as 10.0-45.0% w / w by weight of the beverage.

[0194] Thus, in some embodiments of the present invention, the packaged heat-treated beverage preparation preferably comprises a total amount of protein of 10.0 to 45.0% w / w by weight of the beverage, preferably a total amount of protein of 10.0 to 20% w / w by weight of the beverage, preferably a total amount of protein of 12 to 30% w / w by weight of the beverage, preferably a total amount of protein of 15 to 25% w / w by weight of the beverage, preferably a total amount of protein of 18 to 20% w / w by weight of the beverage.

[0195] The packaged thermally treated beverage preparations of the present invention are particularly useful as sports drinks, in which case they preferably also contain optionally only limited amounts of lipids and / or optionally limited amounts of carbohydrates.

[0196] In some preferred embodiments of the present invention, the preparations are particularly useful as sports drinks, for example comprising a total amount of protein in the range of 2 to 45% w / w by weight of the drink, preferably 2 to 20% w / w by weight of the drink, or preferably 2 to 10% w / w by weight of the drink, most preferably 2 to 6% w / w by weight of the drink.

[0197] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparations are particularly useful as nutritionally incomplete dietary supplements, for example comprising a total amount of protein in the range of 2-45% w / w by weight of the beverage, preferably 2-20% w / w by weight of the beverage, or preferably 3-10% w / w by weight of the beverage.

[0198] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparations are particularly useful as nutritionally complete dietary supplements, for example comprising a total amount of protein in the range of 4-45% w / w by weight of the beverage, or preferably 5-20% w / w by weight of the beverage.

[0199] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation is particularly advantageous for patients suffering from kidney disease or reduced kidney function.

[0200] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation comprises a total amount of protein in the range, for example, of 2 to 45% w / w by weight of the beverage, preferably 2 to 20% w / w by weight of the beverage, or preferably 3 to 12% w / w by weight of the beverage, or preferably 3 to 10% w / w by weight of the beverage.

[0201] It is particularly preferred that the packaged heat-treated beverage preparation comprises BLG isolate in combination with other protein sources, preferably as the main protein source, and possibly as the only protein source.

[0202] The packaged heat-treated beverage preparation of the present invention may contain major nutrients other than protein. In some embodiments of the present invention, the packaged heat-treated beverage preparation further contains carbohydrates. The total carbohydrate content in the heat-treated beverage preparation of the present invention depends on the intended use of the heat-treated beverage preparation.

[0203] In some preferred embodiments of the present invention, the packaged thermally processed beverage preparation further comprises at least one carbohydrate source. In one exemplary embodiment, the at least one carbohydrate source is selected from the group consisting of sucrose, maltodextrin, corn syrup solids, saccharose, maltose, sucromalt, maltitol powder, glycerin, glucose polymers, corn syrup, modified starch, resistant starch, rice-derived carbohydrates, isomaltulose, white sugar, glucose, fructose, lactose, high fructose corn syrup, honey, sugar alcohols, fructooligosaccharides, soy fiber, corn fiber, guar gum, konjac flour, polydextrose, Fibersol, and combinations thereof.

[0204] In some preferred embodiments, the packaged thermally treated beverage preparation further comprises carbohydrates in the range of 0-95% of the total energy content of the preparation, preferably in the range of 10-85% of the total energy content of the preparation, preferably in the range of 20-75% of the total energy content of the preparation, or preferably in the range of 30-60% of the total energy content of the preparation.

[0205] An even lower carbohydrate content is usually preferred, and thus in some preferred embodiments of the present invention, preferably in the range of 0-30% of the total energy content of the preparation, more preferably in the range of 0-20% of the total energy content of the preparation, and even more preferably in the range of 0-10% of the total energy content of the preparation.

[0206] In some preferred embodiments of the present invention, the preparation is particularly useful as a sports drink and comprises a total amount of carbohydrates of up to 75%, preferably up to 40E%, preferably up to 10E%, or preferably up to 5E% of the total energy content (E) of the drink.

[0207] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparations are particularly useful as nutritionally incomplete dietary supplements and contain a total amount of carbohydrates in the range of 70-95%, preferably 80-90E%, of the total energy content (E) of the beverage.

[0208] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparations are particularly useful as nutritionally complete dietary supplements and contain a total amount of carbohydrates in the range of 30-60%, preferably 35-50E%, of the total energy content of the beverage.

[0209] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation is particularly advantageous for patients suffering from kidney disease or reduced kidney function.

[0210] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises a total amount of carbohydrates in the range of 30-60%, preferably in the range of 35-50E% of the total energy content of the beverage.

[0211] In one embodiment of the present invention, the packaged thermally treated beverage preparation further comprises at least one additional ingredient selected from the group consisting of vitamins, flavors, minerals, sweeteners, antioxidants, food acids, lipids, carbohydrates, prebiotics, probiotics, and non-whey proteins.

[0212] In one embodiment of the present invention, the liquid solution further comprises at least one high-intensity sweetener. In one embodiment, the at least one high-intensity sweetener is selected from the group consisting of aspartame, cyclamate, sucralose, acesulfame salts, neotame, saccharin, stevia extract, steviol glycosides such as rebaudioside A, or combinations thereof. In some embodiments of the present invention, it is particularly preferred that the sweetener comprises or consists of one or more high-intensity sweeteners (HIS).

[0213] HIS are found in both natural and artificial sweeteners and typically have a sweetness intensity at least 10 times that of sucrose.

[0214] When used, the total amount of HIS is typically in the range of 0.01 to 2% w / w. For example, the total amount of HIS can be in the range of 0.05 to 1.5% w / w. Alternatively, the total amount of HIS can be in the range of 0.1 to 1.0% w / w.

[0215] The choice of sweetener may depend on the beverage being produced, for example, high-intensity sugar sweeteners (e.g., aspartame, acesulfame K, or sucralose) may be used in beverages where an energy contribution from the sweetener is not desired, while for beverages with a natural sweetener profile, natural sweeteners (e.g., steviol glycosides, sorbitol, or sucrose) may be used.

[0216] Furthermore, it may be more preferable for the sweetener to comprise or consist of one or more polyol sweeteners. Non-limiting examples of useful polyol sweeteners include maltitol, mannitol, lactitol, sorbitol, inositol, xylitol, threitol, galactitol, or combinations thereof. When used, the total amount of polyol sweeteners is typically in the range of 1 to 20% w / w. For example, the total amount of polyol sweeteners may be in the range of 2 to 15% w / w. Alternatively, the total amount of polyol sweeteners may be in the range of 4 to 10% w / w.

[0217] The packaged heat-treated beverage preparation of the present invention may contain major nutrients other than protein. In some embodiments of the present invention, the packaged heat-treated beverage preparation further contains lipids. The total lipid content in the heat-treated beverage preparation of the present invention depends on the intended use of the heat-treated beverage preparation.

[0218] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation has a lipid content of 0-50% of the total energy content of the preparation, or preferably in the range of 0-45% of the total energy content of the preparation, or preferably in the range of 0-30% of the total energy content of the preparation, or preferably in the range of 0-20% of the total energy content of the preparation, or preferably in the range of 0-10% of the total energy content of the preparation, or preferably in the range of 0-5% of the total energy content of the preparation.

[0219] The amount of lipids is determined according to ISO 1211:2010 (Determination of fat content - Rose-Gottlieb gravimetric method).

[0220] In some preferred embodiments of the present invention, the preparation is particularly useful as a sports drink, for example, containing a total amount of lipids of up to 10E%, preferably up to 1E%.

[0221] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparations are particularly useful as nutritionally incomplete dietary supplements, for example, containing a total amount of lipids of up to 10%, preferably up to 1E%, of the total energy content of the beverage.

[0222] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparations are particularly useful as nutritionally complete dietary supplements, for example, containing a total amount of lipids in the range of 20-50% of the total energy content, preferably in the range of 30-40E%.

[0223] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation is particularly advantageous for patients suffering from kidney disease or reduced kidney function.

[0224] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation comprises a total amount of lipids, for example in the range of 20-60% of the total energy content, preferably in the range of 30-50E%.

[0225] In some preferred embodiments of the invention, the sum of alpha lactalbumin (ALA) and caseinomacropeptide (CMP) constitute at least 40% w / w of the non-BLG proteins of the powder, preferably at least 60% w / w, even more preferably at least 70% w / w, and most preferably at least 90% w / w of the non-BLG proteins of the powder.

[0226] In other preferred embodiments of the present invention, each major non-BLG whey protein is present in a weight percentage of total protein that is at most 25%, preferably at most 20%, more preferably at most 15%, even more preferably at most 10%, and most preferably at most 6% of its weight percentage of total protein in the standard whey protein concentrate from sweet whey.

[0227] Even lower concentrations of the major non-BLG whey proteins may be desirable. Thus, in a further preferred embodiment of the present invention, each major non-BLG whey protein is present in a weight percent of total protein that is at most 4%, preferably at most 3%, more preferably at most 2%, and even more preferably at most 1% of its weight percent of total protein in the standard whey protein concentrate from sweet whey.

[0228] The present inventors have seen indications that reduction of lactoferrin and / or lactoperoxidase is particularly advantageous for obtaining a colour-neutral whey protein product.

[0229] Thus, in some preferred embodiments of the present invention, lactoferrin is present in a weight percent of total protein that is at most 25%, preferably at most 20%, more preferably at most 15%, even more preferably at most 10%, and most preferably at most 6% of its weight percent of total protein in a standard whey protein concentrate from sweet whey. Even lower concentrations of lactoferrin may be desirable. Thus, in further preferred embodiments of the present invention, lactoferrin is present in a weight percent of total protein that is at most 4%, preferably at most 3%, more preferably at most 2%, and even more preferably at most 1% of its weight percent of total protein in a standard whey protein concentrate from sweet whey.

[0230] Similarly, in some preferred embodiments of the present invention, lactoperoxidase is present in a weight percent of total protein of up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, and most preferably up to 6% of its weight percent of total protein in the standard whey protein concentrate from sweet whey. Even lower concentrations of lactoperoxidase may be desirable. Thus, in further preferred embodiments of the present invention, lactoperoxidase is present in a weight percent of total protein of up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of its weight percent of total protein in the standard whey protein concentrate from sweet whey.

[0231] Lactoferrin and lactoperoxidase are quantified according to Example 1.29.

[0232] In one embodiment of the present invention, the packaged thermally treated beverage preparation is a sports drink.

[0233] In one embodiment of the present invention, the packaged thermally processed beverage preparation is a nutritionally complete dietary supplement.

[0234] In one embodiment of the present invention, the packaged thermally processed beverage preparation is a nutritionally incomplete dietary supplement.

[0235] In one embodiment of the present invention, the packaged thermally treated beverage preparation is a low phosphorus and low potassium beverage suitable for patients suffering from kidney disease or reduced kidney function.

[0236] The packaged thermally treated beverage preparations of the present invention are particularly useful as sports drinks, in which case they preferably also contain optionally only limited amounts of lipids and / or optionally limited amounts of carbohydrates.

[0237] In some preferred embodiments of the present invention, the preparation is particularly useful as a sports drink, for example: a total amount of protein in the range of 2 to 45% w / w by weight of the beverage, preferably 2 to 20% w / w by weight of the beverage, or preferably 2 to 10% w / w by weight of the beverage, most preferably 2 to 6% w / w by weight of the beverage, a total amount of carbohydrates of at most 75%, preferably at most 40E%, preferably at most 10E%, or preferably at most 5E% of the total energy content (E) of the beverage, and - maximum of 10E% total lipids, preferably at most 1E% Includes.

[0238] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparations are particularly useful as nutritionally incomplete dietary supplements, for example: a total amount of protein ranging from 2 to 45% w / w relative to the weight of the beverage, preferably from 2 to 20% w / w or preferably from 3 to 10% w / w relative to the weight of the beverage, a total amount of carbohydrates ranging from 70 to 95%, preferably from 80 to 90E%, of the total energy content (E) of the beverage, and - A total amount of fat of up to 10% of the total energy content of the beverage, preferably up to 1E% Includes.

[0239] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparations are particularly useful as nutritionally complete dietary supplements, for example: a total amount of protein ranging from 4 to 45% w / w relative to the weight of the beverage, preferably ranging from 5 to 20% w / w relative to the weight of the beverage, a total amount of carbohydrates in the range of 30-60% of the total energy content of the beverage, preferably in the range of 35-50E%; and - total amount of fat in the range of 20-50% of the total energy content, preferably in the range of 30-40E% Includes.

[0240] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation is particularly advantageous for patients suffering from kidney disease or reduced kidney function, as the beverage preparation has a very low content of phosphorus and other minerals such as potassium.

[0241] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises, for example: a total amount of protein in the range of 2 to 45% w / w relative to the weight of the beverage, preferably 2 to 20% w / w relative to the weight of the beverage, or preferably 3 to 12% w / w relative to the weight of the beverage, preferably 3 to 10% w / w, a total amount of carbohydrates in the range of 30-60% of the total energy content of the beverage, preferably in the range of 35-50E%; and - total amount of fat in the range of 20-60% of the total energy content, preferably in the range of 30-50E% Includes.

[0242] One aspect of the present invention is a method for producing a packaged thermally treated beverage preparation having a pH in the range of 2 to 4.7, comprising the steps of: a) - 2 to 45% by weight of a total amount of proteins, at least 85% of which are BLG; optionally with sweeteners, sugar polymers and / or flavors providing a liquid solution comprising: b) packaging the liquid solution; Including, The liquid solution of step a) and / or the packaged liquid solution of step b) are subjected to a heat treatment including at least pasteurization, Regarding the method.

[0243] In some preferred embodiments, the liquid solutions of the present invention have at least 85% w / w BLG protein, preferably at least 88% w / w BLG protein, more preferably at least 90% w / w BLG protein, even more preferably at least 91% w / w BLG protein, and most preferably at least 92% w / w BLG protein.

[0244] Since even higher relative amounts of BLG are both achievable and desirable, in some preferred embodiments of the invention, at least 94% w / w of the protein of the liquid solution is BLG, more preferably at least 96% w / w of the protein is BLG, even more preferably at least 98% w / w of the protein is BLG, and most preferably approximately 100% w / w of the protein is BLG.

[0245] The packaging in step b) can be any suitable packaging technique and any suitable container can be used to package the liquid solution.

[0246] However, in a preferred embodiment of the present invention, the packaging in step b) is aseptic packaging, i.e., the liquid solution is packaged under sterile conditions. For example, aseptic packaging can be performed by using an aseptic filling system, which preferably includes filling the liquid solution into one or more sterile containers.

[0247] Aseptic filling and sealing is particularly preferred where the liquid solution is already sterile or very low in microorganisms before filling.

[0248] Examples of useful containers are, for example, bottles, cartons, bricks and / or bags.

[0249] In some preferred embodiments of this method, the packaged liquid solution of step b) is subjected to a heat treatment that includes at least pasteurization. This embodiment, typically referred to as in-vessel heat treatment or retort treatment, involves heating the entire container and its contents to achieve pasteurization or even sterility. When in-vessel heat treatment is used, it is particularly preferred to maintain the temperature in the range of 70-82°C, more preferably in the range of 70-80°C, and most preferably in the range of 70-78°C. In this way, the level of protein unfolding is minimized.

[0250] In another preferred embodiment of the method of the present invention, the liquid solution of step a) is subjected to a heat treatment, including at least pasteurization, before being packaged in step b).

[0251] In a particularly preferred embodiment, the heat treatment comprises heating the beverage preparation to a temperature in the range of 70-82°C.

[0252] In some preferred embodiments of the present invention, the temperature of the heat treatment is in the range of 70 to 80°C, preferably in the range of 70 to 79°C, more preferably in the range of 71 to 78°C, even more preferably in the range of 72 to 77°C, and most preferably in the range of 73 to 76°C, for example around 75°C.

[0253] Preferably, the duration of the heat treatment is between 1 second and 30 minutes when carried out at a temperature range of 70-82°C. The maximum exposure time is optimal for the lowest temperature in the temperature range, and vice versa. The lower the pH of the liquid solution, the higher the temperature it can withstand without unfolding.

[0254] In a particularly preferred embodiment of the present invention, the heat treatment is provided at 70 to 80°C for 1 second to 30 minutes, more preferably at 71 to 77°C for 1 minute to 25 minutes, and even more preferably at 72 to 76°C for 2 minutes to 20 minutes.

[0255] In some preferred embodiments of the present invention, the heat treatment comprises heating to a temperature of 85° C. to 95° C. for 1 to 3 minutes.

[0256] In some embodiments, particularly when unfolding and optionally also aggregation of BLG is required, higher temperatures may be preferred, for example, the temperature of heat treatment may be at least 81°C, preferably at least 91°C, preferably at least 95°C, more preferably at least 100°C, even more preferably at least 120°C, and most preferably at least 140°C.

[0257] In some preferred embodiments of the present invention, sterilization comprises a temperature in the range of 120-150° C. for 4-30 seconds.

[0258] The heat treatment may involve, for example, a temperature in the range of 90-130°C and a duration in the range of 5 seconds to 10 minutes. The heat treatment may involve, for example, heating to a temperature in the range of 90-95°C for 1-10 minutes, e.g., approximately 120°C for approximately 20 seconds. Alternatively, the heat treatment may involve heating to a temperature in the range of 115-125°C for 5-30 seconds, e.g., approximately 120°C for approximately 20 seconds.

[0259] Alternatively, the heat treatment may be a UHT type treatment, for example, typically involving a temperature in the range of 135-144° C. and a duration in the range of 2-10 seconds.

[0260] Alternatively, but also preferably, the heat treatment may involve a temperature in the range of 145-180° C. and a duration in the range of 0.01-2 seconds, more preferably a temperature in the range of 150-180° C. and a duration in the range of 0.01-0.3 seconds.

[0261] The implementation of heat treatment may involve the use of equipment such as a plate or shell-and-tube heat exchanger, a scraped-surface heat exchanger, or a retort system. Alternatively, and particularly preferred for heat treatment above 95°C, direct steam-type heating may be used, for example, using direct steam injection, direct steam injection, or spray cooking. Furthermore, such direct steam-type heating is preferably used in combination with flash cooling. Suitable examples of spray cooking implementations can be found in WO2009113858, which is incorporated herein by reference for all purposes. Suitable examples of direct steam injection and direct steam injection implementations can be found in WO2009113858 and WO2010 / 085957, which are incorporated herein by reference for all purposes. General aspects of high-temperature processing can be found, for example, in "Thermal technologies in food processing," ISBN 185573558 X, which is incorporated herein by reference for all purposes.

[0262] In some preferred embodiments of the present invention, pasteurization is combined with another physical microbial reduction.

[0263] Useful examples of physical microbial reduction include one or more of: sterilization filtration, UV irradiation, high pressure treatment, pulsed electric field treatment, and ultrasound.

[0264] In some particularly preferred embodiments of the present invention, the heat treatment is selected to provide a degree of protein denaturation of at most 50%, preferably at most 20%, even more preferably at most 10%, and most preferably at most 5%.

[0265] It is further preferred that the heat treatment is selected to provide an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.11, preferably at least 1.13, more preferably at least 1.15, and even more preferably at least 1.17.

[0266] In some preferred embodiments of the present invention, the heat treatment is a sterilization that results in a sterile liquid beverage preparation. Such sterilization can be obtained, for example, by combining a sterile filtration with a heat treatment, such as pasteurization. Sterilization can include, for example, a heat treatment followed by a sterile filtration, or even more preferably, a sterile filtration followed by a heat treatment.

[0267] Depending on the heat treatment temperature used, it may be beneficial for the beverage preparation to be cooled. According to a preferred embodiment of the method of the present invention, after heat treatment the heat treated beverage preparation is optionally cooled to preferably 0-50°C, preferably 0-25°C, or preferably 0-20°C, or preferably 0-15°C, preferably 0-10°C, or preferably 4-8°C, or preferably 2-5°C, or preferably 1-5°C.

[0268] If the beverage preparation is pasteurized, it is preferably cooled to 0-15°C, preferably 1-10°C, more preferably 1-6°C after heat treatment.

[0269] According to one embodiment of this method, generally, any acid or base can be used to adjust the pH. Those skilled in the art will recognize suitable means for adjusting the pH. Suitable acids include, for example, citric acid, hydrochloric acid, malic acid, or tartaric acid, or phosphoric acid, most preferably citric acid and / or phosphoric acid.

[0270] Useful examples of bases include hydroxide salts, such as sodium hydroxide or potassium hydroxide, carbonate or bicarbonate salts, carboxylate salts, such as citrate or lactate salts, and combinations thereof. Preferably, the pH is adjusted using a base such as KOH or NaOH.

[0271] In some preferred embodiments of the present invention, the liquid solution has a pH in the range of 3.0 to 4.3. These pH ranges are particularly preferred for producing clear beverages with low viscosity and improved taste.

[0272] With respect to appearance, it has surprisingly been found that using a whey protein beverage in which at least 85% w / w of the protein is BLG, the pH can be increased during heat treatment, resulting in improved visual perception (color and turbidity) and viscosity when compared to a heat-treated WPI beverage.

[0273] Surprisingly, it has been found that there are significant differences in sensory parameters between beverages made with WPI compared to the BLG beverages of the present invention. Surprisingly and advantageously, it has been found that BLG beverages have lower levels of astringency, dry texture, sourness, whey aroma, and citric acid flavor compared to WPI beverages. It has also been found that by increasing the pH of an acidic beverage, less sweetener is needed to balance the acidity of the beverage, and therefore, a lower concentration of sweetener is needed in such beverages.

[0274] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation has a pH in the range of 3.0 to 4.1, or preferably 3.1 to 4.0, or preferably 3.2 to 3.9, or preferably 3.7 to 3.9, more preferably 3.4 to 3.9, and even more preferably 3.5 to 3.9.

[0275] These pH ranges are particularly suitable when the beverage preparation is pasteurized.

[0276] In some preferred embodiments of the present invention, the liquid solution preferably has a pH in the range of 3.0 to 3.9, or preferably 3.2 to 3.7, or preferably 3.4 to 3.6, or preferably 3.5 to 3.7, or preferably 3.4 to 3.6.

[0277] These pH ranges in combination with high temperature processes such as sterilization are particularly suitable for producing clear beverages with low viscosity and improved taste.

[0278] In some preferred embodiments of the present invention, the liquid solution has a pH in the range of 4.1 to 4.7, which is particularly suitable for producing a stable beverage with a milky appearance and high turbidity while still having a low viscosity. In some embodiments of the present invention, the pH range is 4.2 to 4.6. In other embodiments of the present invention, the pH range is 4.2 to 4.5.

[0279] In some preferred embodiments of the present invention, the liquid solution comprises a total amount of protein of 4.0 to 30% w / w based on the weight of the beverage.

[0280] In some embodiments of the present invention, the liquid solution advantageously has a protein content of 2.0 to 10.0% w / w based on the weight of the solution.

[0281] Thus, in some embodiments of the present invention, the liquid solution preferably comprises a total amount of protein of 2.0 to 10% w / w by weight of the liquid solution, preferably a total amount of protein of 3.0 to 10% w / w by weight of the liquid solution, preferably a total amount of protein of 5.0 to 9.0% w / w by weight of the liquid solution, preferably a total amount of protein of 6.0 to 8.0% w / w by weight of the liquid solution.

[0282] In some embodiments of the present invention, the protein content of the liquid solution is advantageously high, such as 10.0-45.0% w / w based on the weight of the liquid solution.

[0283] Thus, in some embodiments of the present invention, the liquid solution preferably comprises a total amount of protein of 10.0 to 45.0% w / w by weight of the liquid solution, preferably a total amount of protein of 10.0 to 20% w / w by weight of the liquid solution, preferably a total amount of protein of 12 to 30% w / w by weight of the liquid solution, preferably a total amount of protein of 15 to 25% w / w by weight of the liquid solution, preferably a total amount of protein of 18 to 20% w / w by weight of the liquid solution.

[0284] It is particularly preferred that the liquid solution comprises BLG isolate in combination with other protein sources, preferably as the main protein source, and possibly as the only protein source.

[0285] The BLG isolate is preferably a BLG isolate powder, or a liquid BLG isolate containing water and an amount of BLG isolate powder solids in the range of 1-50% w / w.

[0286] The beta-lactoglobulin (BLG) isolate powder is preferably prepared by spray drying and has a pH in the range of i) 2 to 4.9, ii) 6.1 to 8.5, or iii) 5.0 to 6.0; - total protein in an amount of at least 30% w / w, BLG in an amount of at least 85% w / w relative to the total protein, and - Maximum 10% w / w of water Includes.

[0287] The BLG isolate powder preferably has one or more of the following: -At least 0.2g / cm 3 Bulk density of an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.11; - protein denaturation up to 10%; - Heat stability at pH 3.9 up to 200 NTU, and -Up to 1000 colony forming units / g.

[0288] The BLG isolate powder is preferably an edible composition.

[0289] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of 2 to 4.9, making such powders particularly useful in acidic foods, especially acidic beverages.

[0290] In another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5.

[0291] In some preferred embodiments of the present invention, the BLG isolate powder comprises total protein in an amount of at least 40% w / w, preferably at least 50% w / w, at least 60% w / w, more preferably at least 70% w / w, and even more preferably at least 80% w / w.

[0292] Even higher protein contents may be required, and in some preferred embodiments of the present invention, the BLG isolate powder comprises total protein in an amount of at least 85% w / w, preferably at least 90% w / w, at least 92% w / w, more preferably at least 94% w / w, and even more preferably at least 95% w / w.

[0293] Total protein is determined according to Example 1.5.

[0294] In some preferred embodiments of the present invention, the BLG isolate powder comprises BLG in an amount of at least 92% w / w of total protein, preferably at least 95% w / w, more preferably at least 97% w / w, even more preferably at least 98%, and most preferably at least 99.5% w / w of total protein.

[0295] In some preferred embodiments of the invention, the sum of alpha lactalbumin (ALA) and caseinomacropeptide (CMP) constitute at least 40% w / w of the non-BLG proteins of the powder, preferably at least 60% w / w, even more preferably at least 70% w / w, and most preferably at least 90% w / w of the non-BLG proteins of the powder.

[0296] In other preferred embodiments of the present invention, each major non-BLG whey protein is present in a weight percentage of total protein that is at most 25%, preferably at most 20%, more preferably at most 15%, even more preferably at most 10%, and most preferably at most 6% of its weight percentage of total protein in the standard whey protein concentrate from sweet whey.

[0297] Even lower concentrations of the major non-BLG whey proteins may be desirable. Thus, in a further preferred embodiment of the present invention, each major non-BLG whey protein is present in a weight percent of total protein that is at most 4%, preferably at most 3%, more preferably at most 2%, and even more preferably at most 1% of its weight percent of total protein in the standard whey protein concentrate from sweet whey.

[0298] The present inventors have seen indications that reduction of lactoferrin and / or lactoperoxidase is particularly advantageous for obtaining a colour-neutral whey protein product.

[0299] Thus, in some preferred embodiments of the present invention, lactoferrin is present in a weight percent of total protein that is at most 25%, preferably at most 20%, more preferably at most 15%, even more preferably at most 10%, and most preferably at most 6% of its weight percent of total protein in a standard whey protein concentrate from sweet whey. Even lower concentrations of lactoferrin may be desirable. Thus, in further preferred embodiments of the present invention, lactoferrin is present in a weight percent of total protein that is at most 4%, preferably at most 3%, more preferably at most 2%, and even more preferably at most 1% of its weight percent of total protein in a standard whey protein concentrate from sweet whey.

[0300] Similarly, in some preferred embodiments of the present invention, lactoperoxidase is present in a weight percent of total protein of up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, and most preferably up to 6% of its weight percent of total protein in the standard whey protein concentrate from sweet whey. Even lower concentrations of lactoperoxidase may be desirable. Thus, in further preferred embodiments of the present invention, lactoperoxidase is present in a weight percent of total protein of up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of its weight percent of total protein in the standard whey protein concentrate from sweet whey.

[0301] Lactoferrin and lactoperoxidase are quantified according to Example 1.29.

[0302] In some preferred embodiments of the present invention, the BLG isolate powder has a moisture content in an amount of at most 10% w / w, preferably at most 7% w / w, more preferably at most 6% w / w, even more preferably at most 4% w / w, and most preferably at most 2% w / w.

[0303] In some preferred embodiments of the present invention, the BLG isolate powder comprises carbohydrates in an amount of up to 60% w / w, preferably up to 50% w / w, more preferably up to 20% w / w, even more preferably up to 10% w / w, even more preferably up to 1% w / w, and most preferably up to 0.1% w / w. The BLG isolate powder may contain carbohydrates such as, for example, lactose, oligosaccharides, and / or hydrolysates of lactose (i.e., glucose and galactose), sucrose, and / or maltodextrins.

[0304] In some preferred embodiments of the present invention, the BLG isolate powder comprises lipids in an amount of at most 10% w / w, preferably at most 5% w / w, more preferably at most 2% w / w, and even more preferably at most 0.1% w / w.

[0305] The inventors have found that it can be advantageous to control the mineral content in order to achieve some of the desired properties of the BLG isolate powder.

[0306] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolate powder is at most 10 mmol / g protein. Preferably, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolate powder is at most 6 mmol / g protein, more preferably at most 4 mmol / g protein, and even more preferably at most 2 mmol / g protein.

[0307] In another preferred embodiment of the present invention, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolate powder is at most 1 mmol / g protein. Preferably, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolate powder is at most 0.6 mmol / g protein, more preferably at most 0.4 mmol / g protein, even more preferably at most 0.2 mmol / g protein, and most preferably at most 0.1 mmol / g protein.

[0308] In another preferred embodiment of the present invention, the sum of the amounts of Mg and Ca in the BLG isolate powder is at most 5 mmol / g protein. Preferably, the sum of the amounts of Mg and Ca in the BLG isolate powder is at most 3 mmol / g protein, more preferably at most 1.0 mmol / g protein, and even more preferably at most 0.5 mmol / g protein.

[0309] In another preferred embodiment of the present invention, the sum of the amounts of Mg and Ca in the BLG isolate powder is at most 0.3 mmol / g protein. Preferably, the sum of the amounts of Mg and Ca in the BLG isolate powder is at most 0.2 mmol / g protein, more preferably at most 0.1 mmol / g protein, even more preferably at most 0.03 mmol / g protein, and most preferably at most 0.01 mmol / g protein.

[0310] The present inventors have discovered that it is possible to use a low phosphorus / low potassium variant of BLG isolate powder that is particularly useful for patients with kidney disease. To produce such a product, the BLG isolate powder must have an equally low content of phosphorus and potassium.

[0311] Therefore, in some preferred embodiments of the present invention, the BLG isolate powder has a total phosphorus content of up to 100 mg phosphorus per 100 g protein. Preferably, the BLG isolate powder has a total phosphorus content of up to 80 mg phosphorus per 100 g protein. More preferably, the BLG isolate powder has a total phosphorus content of up to 50 mg phosphorus per 100 g protein. Even more preferably, the BLG isolate powder has a total phosphorus content of up to 20 mg phosphorus per 100 g protein. The BLG isolate powder has a total phosphorus content of up to 5 mg phosphorus per 100 g protein.

[0312] In some preferred embodiments of the present invention, the BLG isolate powder contains up to 600 mg of potassium per 100 g of protein. More preferably, the BLG isolate powder contains up to 500 mg of potassium per 100 g of protein. More preferably, the BLG isolate powder contains up to 400 mg of potassium per 100 g of protein. More preferably, the BLG isolate powder contains up to 300 mg of potassium per 100 g of protein. Even more preferably, the BLG isolate powder contains up to 200 mg of potassium per 100 g of protein. Even more preferably, the BLG isolate powder contains up to 100 mg of potassium per 100 g of protein. Even more preferably, the BLG isolate powder contains up to 50 mg of potassium per 100 g of protein, and even more preferably, the BLG isolate powder contains up to 10 mg of potassium per 100 g of protein.

[0313] The phosphorus content is determined in accordance with Example 1.19 relative to the total amount of elemental phosphorus in the composition in question. Similarly, the potassium content is determined in accordance with Example 1.19 relative to the total amount of elemental potassium in the composition in question.

[0314] In some preferred embodiments of the present invention, the BLG isolate powder comprises at most 100 mg phosphorus / 100 g protein and at most 700 mg potassium / 100 g protein, preferably at most 80 mg phosphorus / 100 g protein and at most 600 mg potassium / 100 g protein, more preferably at most 60 mg phosphorus / 100 g protein and at most 500 mg potassium / 100 g protein, more preferably at most 50 mg phosphorus / 100 g protein and at most 400 mg potassium / 100 g protein, or more preferably at most 20 mg phosphorus / 100 g protein and at most 200 mg potassium / 100 g protein, or even more preferably at most 10 mg phosphorus / 100 g protein and at most 50 mg potassium / 100 g protein. In some preferred embodiments of the present invention, the BLG isolate powder comprises at most 100 mg phosphorus / 100 g protein and at most 340 mg potassium / 100 g protein.

[0315] The low phosphorus and / or low potassium compositions according to the present invention may be used as food ingredients for the preparation of food for patient populations with reduced renal function.

[0316] The inventors have found that for some applications, such as acidic foods, and particularly acidic beverages, it is particularly advantageous to have an acidic BLG isolate powder with a pH of at most 4.9, and even more preferably at most 4.3. This is particularly true for high protein, clear, acidic beverages.

[0317] In the context of the present invention, a clear liquid has a turbidity of at most 200 NTU, measured according to Example 1.7.

[0318] Thus, in some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of 2 to 4.9. Preferably, the BLG isolate powder has a pH in the range of 2.5 to 4.7, more preferably 2.8 to 4.3, even more preferably 3.2 to 4.0, and most preferably 3.4 to 3.9. Alternatively, but also preferably, the BLG isolate powder can have a pH in the range of 3.6 to 4.3.

[0319] The inventors have found that for some applications, such as pH-neutral foods and especially pH-neutral beverages, it is particularly advantageous to have a pH-neutral BLG isolate powder, which is especially true for high protein, clear or opaque pH-neutral beverages.

[0320] Thus, in some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5. Preferably, the powder has a pH in the range of 6.1 to 8.5, more preferably 6.2 to 8.0, even more preferably 6.3 to 7.7, and most preferably 6.5 to 7.5.

[0321] In another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 5.0 to 6.0. Preferably, the powder has a pH in the range of 5.1 to 5.9, more preferably 5.2 to 5.8, even more preferably 5.3 to 5.7, and most preferably 5.4 to 5.6.

[0322] Advantageously, the BLG isolate powder used in the present invention has a density of at least 0.20 g / cm 3 , preferably at least 0.30 g / cm 3 , more preferably at least 0.40 g / cm 3 , and even more preferably at least 0.45 g / cm 3 , and even more preferably at least 0.50 g / cm 3 , most preferably at least 0.6 g / cm 3 The bulk density may be

[0323] Low-density powders, such as freeze-dried BLG isolate, are fluffy and easily drawn into the air in manufacturing environments during use. This is problematic because it increases the risk of cross-contamination of freeze-dried powders with other foods and dusty environments are known to cause sanitation problems. In extreme cases, dusty environments also increase the risk of dust explosions.

[0324] The high density variants of the present invention are easier to handle and less likely to bleed into the ambient air.

[0325] An additional advantage of the high density varieties of the present invention is that they take up less space during shipping, thereby increasing the weight of BLG isolate powder that can be shipped per volume unit.

[0326] A further advantage of the high density variants of the present invention is that they are less expensive than, for example, powdered sugar (bulk density approximately 0.56 g / cm 3 ), granulated sugar (bulk density approximately 0.71 g / cm 3 ), powdered citric acid (bulk density approximately 0.77 g / cm 3 ) and is less likely to separate when used in powder mixtures with other powdered food ingredients.

[0327] The BLG isolate powder of the present invention has a density of 0.2 to 1.0 g / cm 3 in the range of 0.30 to 0.9 g / cm 3 more preferably in the range of 0.40 to 0.8 g / cm 3 in the range of 0.45 to 0.75 g / cm 3 and even more preferably in the range of 0.50 to 0.75 g / cm 3 in the range of 0.6 to 0.75 g / cm 3 The bulk density may range from 0.1 to 0.5.

[0328] The bulk density of the powder is determined according to Example 1.17.

[0329] The inventors have found that maintaining the native conformation of BLG is advantageous and have seen indications that increased unfolding of BLG results in increased levels of dry texture when BLG is used in acidic beverages.

[0330] The intrinsic tryptophan fluorescence ratio (I330 / I350) is a measure of the degree of unfolding of BLG, and the inventors have found that a high intrinsic tryptophan fluorescence ratio correlates with low or no unfolding of BLG, resulting in less observed dry texture. The intrinsic tryptophan fluorescence ratio (I330 / I350) is measured according to Example 1.1.

[0331] In some preferred embodiments of the present invention, the BLG isolate powder has an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.11.

[0332] In some preferred embodiments of the present invention, the BLG isolate powder has an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.12, preferably at least 1.13, more preferably at least 1.15, even more preferably at least 1.17, and most preferably at least 1.19.

[0333] If the BLG isolate powder contains a significant amount of non-protein material, it is preferable to isolate the protein fraction before measuring the intrinsic tryptophan fluorescence ratio. Thus, in some preferred embodiments of the present invention, the protein fraction of the BLG isolate powder has an intrinsic tryptophan fluorescence ratio of at least 1.11.

[0334] In some preferred embodiments of the present invention, the protein fraction of the BLG isolate powder has an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.12, preferably at least 1.13, more preferably at least 1.15, even more preferably at least 1.17, and most preferably at least 1.19.

[0335] The protein fraction can be separated from the BLG isolate powder, for example, by dissolving the BLG isolate powder in demineralized water and subjecting the solution to dialysis using a protein-retaining filter or ultrafiltration-based dialysis. If the BLG isolate powder contains interfering levels of lipids, such lipids can be removed, for example, by microfiltration. The microfiltration and ultrafiltration / diafiltration steps can be combined to remove both lipids and small molecules from the protein fraction.

[0336] It is generally preferred that a substantial amount of the BLG in the BLG isolate powder is non-aggregated. Preferably, at least 50% of the BLG is non-aggregated. More preferably, at least 80% of the BLG is non-aggregated. Even more preferably, at least 90% of the BLG is non-aggregated. Most preferably, at least 95% of the BLG is non-aggregated. Even more preferably, approximately 100% of the BLG in the BLG isolate powder is non-aggregated.

[0337] In some preferred embodiments of the present invention, the BLG isolate powder has a degree of protein denaturation of at most 10%, preferably at most 8%, more preferably at most 6%, even more preferably at most 3%, even more preferably at most 1%, and most preferably at most 0.2%.

[0338] However, it may also be preferred that the BLG isolate powder have a significant level of protein denaturation, for example, if an opaque beverage is desired. Accordingly, in another preferred embodiment of the present invention, the BLG isolate powder has a degree of protein denaturation of at least 11%, preferably at least 20%, more preferably at least 40%, even more preferably at least 50%, even more preferably at least 75%, and most preferably at least 90%.

[0339] While the BLG isolate powder has a significant level of protein denaturation, it is generally preferable to maintain a low level of insoluble protein material, i.e., precipitated protein material that settles in the beverage during storage. The level of insoluble material is measured according to Example 1.10.

[0340] In some preferred embodiments of the present invention, the BLG isolate powder comprises at most 20% w / w insoluble protein material, preferably at most 10% w / w insoluble protein material, more preferably at most 5% w / w insoluble protein material, even more preferably at most 3% w / w insoluble protein material, and most preferably at most 1% w / w insoluble protein material. It may even be preferred that the BLG isolate powder contains no insoluble protein material at all.

[0341] The inventors have found that the heat stability of BLG isolate powder at pH 3.9 is a good indicator of its usefulness for clear, high-protein beverages. Heat stability at pH 3.9 is measured according to Example 1.2.

[0342] It is particularly preferred that the BLG isolate powder has a thermostability at pH 3.9 of at most 200 NTU, preferably at most 100 NTU, more preferably at most 60 NTU, even more preferably at most 40 NTU, and most preferably at most 20 NTU. Even better thermostability is possible, with the BLG isolate powder preferably having a thermostability at pH 3.9 of at most 10 NTU, preferably at most 8 NTU, more preferably at most 4 NTU, and even more preferably at most 2 NTU.

[0343] The microbial content of BLG isolate powder is preferably kept to a minimum. However, it is difficult to obtain both a high level of protein nativeness and a low content of microorganisms because microbial reduction methods tend to lead to protein unfolding and denaturation. The present invention makes it possible to obtain a very low content of microorganisms while maintaining a high level of BLG nativeness.

[0344] Thus, in some preferred embodiments of the present invention, the BLG isolate powder contains at most 15,000 colony forming units (CFU) / g. Preferably, the BLG isolate powder contains at most 10,000 CFU / g. More preferably, the BLG isolate powder contains at most 5,000 CFU / g. Even more preferably, the BLG isolate powder contains at most 1,000 CFU / g. Even more preferably, the BLG isolate powder contains at most 300 CFU / g. Most preferably, the BLG isolate powder contains at most 100 CFU / g, such as at most 10 CFU / g. In particularly preferred embodiments, the powder is sterile. Sterile BLG isolate powder can be prepared, for example, by combining several physical microbial reduction methods, such as microfiltration and heat treatment at acidic pH, during the manufacture of the BLG isolate powder.

[0345] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of: i) 2 to 4.9; ii) 6.1 to 8.5; or iii) 5.0 to 6.0; - total protein in an amount of at least 30% w / w, preferably at least 80% w / w, even more preferably at least 90% w / w; - beta-lactoglobulin (BLG) in an amount of at least 85% w / w, preferably at least 90% w / w, relative to the total protein; - water in an amount of up to 6% w / w, lipids in an amount of up to 2% w / w, preferably up to 0.5% w / w Including, The BLG isolate powder is an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.11; - protein denaturation of up to 10%, and -Heat stability up to 200 NTU at pH 3.9 It has.

[0346] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of i) 2 to 4.9, or ii) 6.1 to 8.5; - total protein in an amount of at least 30% w / w, preferably at least 80% w / w, even more preferably at least 90% w / w; - beta-lactoglobulin (BLG) in an amount of at least 85% w / w relative to the total protein, preferably at least 90% w / w, more preferably at least 94% w / w relative to the total protein, - water in an amount of up to 6% w / w, lipids in an amount of up to 2% w / w, preferably up to 0.5% w / w Including, The BLG isolate powder is an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.11; a degree of protein denaturation of at most 10%, preferably at most 5%, and - a thermal stability at pH 3.9 of at most 70 NTU, preferably at most 50 NTU, even more preferably at most 40 NTU It has.

[0347] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of i) 2 to 4.9 or ii) 6.1 to 8.5; - total protein in an amount of at least 30% w / w, - beta-lactoglobulin (BLG) in an amount of at least 85% w / w, preferably at least 90% w / w, relative to the total protein; -Water in an amount of up to 6% w / w Including, The BLG isolate powder is -At least 0.2g / cm 3 Bulk density of an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.11; - protein denaturation of up to 10%, and -Heat stability up to 200 NTU at pH 3.9 It has.

[0348] In another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 2 to 4.9, - total protein in an amount of at least 80% w / w, preferably at least 90% w / w, even more preferably at least 94% w / w; - beta-lactoglobulin (BLG) in an amount of at least 85% w / w relative to the total protein, preferably at least 90% w / w, even more preferably at least 94% w / w relative to the total protein, - water in an amount of up to 6% w / w, lipids in an amount of up to 2% w / w, preferably up to 0.5% w / w Including, The BLG isolate powder is -At least 0.2g / cm 3 , preferably at least 0.3 g / cm 3 , more preferably at least 0.4 g / cm 3 Bulk density of an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.11; a degree of protein denaturation of at most 10%, preferably at most 5%, more preferably at most 2%, and - a thermal stability at pH 3.9 of at most 50 NTU, preferably at most 30 NTU, even more preferably at most 10 NTU It has.

[0349] In yet another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5, - total protein in an amount of at least 80% w / w, preferably at least 90% w / w, even more preferably at least 94% w / w; - beta-lactoglobulin (BLG) in an amount of at least 85% w / w relative to the total protein, preferably at least 90% w / w, even more preferably at least 94% w / w relative to the total protein, - water in an amount of up to 6% w / w, lipids in an amount of up to 2% w / w, preferably up to 0.5% w / w Including, The BLG isolate powder is -At least 0.2g / cm 3 , preferably at least 0.3 g / cm 3 , more preferably at least 0.4 g / cm 3 Bulk density of a degree of protein denaturation of at most 10%, preferably at most 5%, more preferably at most 2%, and - a thermal stability at pH 3.9 of at most 50 NTU, preferably at most 30 NTU, even more preferably at most 10 NTU It has.

[0350] In a further preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5, - total protein in an amount of at least 80% w / w, preferably at least 90% w / w, even more preferably at least 94% w / w; - beta-lactoglobulin (BLG) in an amount of at least 85% w / w relative to the total protein, preferably at least 90% w / w, even more preferably at least 94% w / w relative to the total protein, - water in an amount of up to 6% w / w, lipids in an amount of up to 2% w / w, preferably up to 0.5% w / w Including, The BLG isolate powder is -At least 0.2g / cm 3 , preferably at least 0.3 g / cm 3 , more preferably at least 0.4 g / cm 3 Bulk density of a degree of protein denaturation of at most 10%, preferably at most 5%, more preferably at most 2%, and - a thermal stability at pH 3.9 of at most 50 NTU, preferably at most 30 NTU, even more preferably at most 10 NTU It has.

[0351] In a further preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 5.0 to 6.0, - total protein in an amount of at least 80% w / w, preferably at least 90% w / w, even more preferably at least 94% w / w; - beta-lactoglobulin (BLG) in an amount of at least 85% w / w relative to the total protein, preferably at least 90% w / w, even more preferably at least 94% w / w relative to the total protein, - water in an amount of up to 6% w / w, lipids in an amount of up to 2% w / w, preferably up to 0.5% w / w Including, The BLG isolate powder is -At least 0.2g / cm 3 , preferably at least 0.3 g / cm 3 , more preferably at least 0.4 g / cm 3 Bulk density of a degree of protein denaturation of at most 10%, preferably at most 5%, more preferably at most 2%, a thermostability at pH 3.9 of at most 50 NTU, preferably at most 30 NTU, even more preferably at most 10 NTU, and - BLG crystallinity preferably less than 10% It has.

[0352] BLG isolate powder containing BLG in an amount of at least 85% w / w relative to total protein is typically prepared by the following steps: a) i) a pH in the range of 2 to 4.9; ii) a pH in the range of 6.1 to 8.5, or iii) pH in the range of 5.0 to 6.0 providing a liquid BLG isolate having the formula: b) optionally subjecting the liquid BLG isolate to physical microbial reduction; c) drying the liquid BLG isolate, preferably by spray drying; The method is provided by a method comprising:

[0353] The BLG isolate is preferably prepared from mammalian milk, preferably from ruminant milk, such as cow, sheep, goat, buffalo, camel, llama, mare, and / or deer milk. Proteins from bovine milk are particularly preferred. Thus, the BLG is preferably bovine BLG.

[0354] The liquid BLG isolate can be provided in several different ways.

[0355] Typically, providing a liquid BLG isolate comprises or consists of isolating BLG from a whey protein feedstock to obtain a BLG concentrated composition by one or more of the following methods: - Crystallization or precipitation of BLG by salting out, - crystallization or precipitation of BLG by salting out, -ion ​​exchange chromatography, and -Fractionation of whey proteins by ultrafiltration.

[0356] A particularly preferred method of obtaining a BLG enriched composition is by crystallization of BLG, preferably by salting out or alternatively by salting out.

[0357] The whey protein source is preferably WPC, WPI, SPC, SPI or a combination thereof.

[0358] The term "whey protein feedstock" refers to the composition from which the BLG concentrate composition and subsequent liquid BLG isolate are derived.

[0359] In some embodiments of the present invention, the preparation of the BLG enriched composition comprises or consists of high salt BLG crystallization in the pH range of 3.6-4.0 according to US Pat. No. 2,790,790.

[0360] In another embodiment of the present invention, the preparation of the BLG-enriched composition comprises or consists of the method described by de Jongh et al. (Mild Isolation Procedure Discloses New Protein Structural Properties of β-Lactoglobulin, J. Dairy Sci., Vol. 84(3), 2001, 562-571) or Vyas et al. (Scale-Up of Native β-Lactoglobulin Affinity Separation Process, J. Dairy Sci. 85:1639-1645, 2002).

[0361] However, in a particularly preferred embodiment of the present invention, the BLG enriched composition is prepared by crystallization under salting conditions at a pH of 5-6, as described in PCT application PCT / EP2017 / 084553, which is incorporated herein by reference for all purposes.

[0362] In some preferred embodiments of the present invention, the BLG enriched composition is an edible BLG composition according to PCT / EP2017 / 084553, which contains at least 90% BLG relative to total protein, and preferably contains BLG crystals.

[0363] If it does not already have the necessary properties for use as a liquid BLG isolate, the BLG concentrate composition isolated from the whey protein feedstock may be subjected to one or more steps selected from the following group as part of providing a liquid BLG isolate: - desalination, -Addition of minerals, -dilution, -concentrated, -Physical microorganism reduction; -pH adjustment.

[0364] Non-limiting examples of desalting include, for example, dialysis, gel filtration, UF / diafiltration, NF / diafiltration, and ion exchange chromatography.

[0365] Non-limiting examples of mineral additions include the addition of soluble food-acceptable salts, such as salts of Na, K, Ca, and / or Mg. Such salts can be, for example, phosphate salts, chloride salts, or salts of food acids, such as citrate or lactate salts. Minerals can be added in solid, suspended, or dissolved form.

[0366] Non-limiting examples of dilution include the addition of a liquid diluent such as water, demineralized water, or an aqueous solution of a mineral, acid, or base.

[0367] Non-limiting examples of concentration include, for example, evaporation, reverse osmosis, nanofiltration, ultrafiltration, and combinations thereof.

[0368] If the concentration must increase the concentration of protein relative to the total solids, it is preferable to use a concentration step such as ultrafiltration or alternatively dialysis. If the concentration does not need to increase the concentration of protein relative to the total solids, methods such as evaporation, nanofiltration and / or reverse osmosis may be useful.

[0369] Non-limiting examples of physical microbial reduction include heat treatment, bacterial filtration, UV irradiation, high pressure treatment, pulsed electric field treatment, and ultrasound, which are well known to those skilled in the art.

[0370] Non-limiting examples of pH adjustment include, for example, the addition of a base and / or acid, preferably a food-acceptable base and / or acid. It is particularly preferred to use an acid and / or base that can chelate divalent metal cations. Examples of such acids and / or bases include citric acid, citrate salts, EDTA, lactic acid, lactate salts, phosphoric acid, phosphate salts, and combinations thereof.

[0371] In some preferred embodiments of the present invention, the liquid solution has a color value delta b in the range of -0.10 to +0.51 on the CIELAB color scale, especially when the preparation has a turbidity of at most 200 NTU, more preferably at most 40 NTU. * It has.

[0372] In another preferred embodiment of the present invention, the liquid solution has a color value delta b in the range of 0.0 to 0.40 on the CIELAB color scale, preferably in the range of +0.10 to +0.25. * It has.

[0373] The liquid solution of the present invention may contain macronutrients other than protein. In some embodiments of the present invention, the liquid solution further contains carbohydrates. The total carbohydrate content of the liquid solution of the present invention depends on the intended use of the final heat-treated beverage preparation.

[0374] In some preferred embodiments of the present invention, the liquid solution further comprises at least one source of carbohydrate. In one exemplary embodiment, the at least one source of carbohydrate is selected from the group consisting of sucrose, maltodextrin, corn syrup solids, sucromalt, glucose polymers, corn syrup, modified starch, resistant starch, rice-derived carbohydrates, isomaltulose, white sugar, glucose, fructose, lactose, galactose, maltose, dextrose, high fructose corn syrup, honey, sugar alcohols, fructooligosaccharides, soy fiber, corn fiber, guar gum, konjac flour, polydextrose, Fibersol, and combinations thereof.

[0375] In some preferred embodiments, the liquid solution further comprises carbohydrates in the range of 0-95% of the total energy content of the liquid solution, preferably in the range of 10-85% of the total energy content of the liquid solution, preferably in the range of 20-75% of the total energy content of the liquid solution, or preferably in the range of 30-60% of the total energy content of the liquid.

[0376] An even lower carbohydrate content is usually preferred, and thus in some preferred embodiments of the present invention, preferably in the range of 0-30% of the total energy content of the preparation, more preferably in the range of 0-20% of the total energy content of the preparation, and even more preferably in the range of 0-10% of the total energy content of the preparation.

[0377] In one embodiment of the present invention, the liquid solution further comprises at least one additional ingredient selected from the group consisting of vitamins, flavors, minerals, sweeteners, antioxidants, food acids, lipids, carbohydrates, prebiotics, probiotics, and non-whey proteins.

[0378] In one embodiment of the present invention, the liquid solution further comprises at least one high-intensity sweetener. In one embodiment, the at least one high-intensity sweetener is selected from the group consisting of aspartame, cyclamate, sucralose, acesulfame salts, neotame, saccharin, stevia extract, steviol glycosides such as rebaudioside A, or combinations thereof. In some embodiments of the present invention, it is particularly preferred that the sweetener comprises or consists of one or more high-intensity sweeteners (HIS).

[0379] HIS are found in both natural and artificial sweeteners and typically have a sweetness intensity at least 10 times that of sucrose.

[0380] When used, the total amount of HIS is typically in the range of 0.01 to 2% w / w. For example, the total amount of HIS can be in the range of 0.05 to 1.5% w / w. Alternatively, the total amount of HIS can be in the range of 0.1 to 1.0% w / w.

[0381] The choice of sweetener may depend on the beverage being produced, but high-intensity sugar sweeteners (e.g., aspartame, acesulfame K, or sucralose) may be used in beverages where an energy contribution from the sweetener is not desired, while for beverages with a natural profile, natural sweeteners (e.g., steviol glycosides, sorbitol, or sucrose) may be used.

[0382] Furthermore, it may be more preferable for the sweetener to comprise or consist of one or more polyol sweeteners. Non-limiting examples of useful polyol sweeteners include maltitol, mannitol, lactitol, sorbitol, inositol, xylitol, threitol, galactitol, or combinations thereof. When used, the total amount of polyol sweeteners is typically in the range of 1 to 20% w / w. For example, the total amount of polyol sweeteners may be in the range of 2 to 15% w / w. Alternatively, the total amount of polyol sweeteners may be in the range of 4 to 10% w / w.

[0383] The liquid solution of the present invention may contain macronutrients other than protein. In some embodiments of the present invention, the liquid solution further contains lipids. The total lipid content in the final heat-treated beverage preparation of the present invention depends on the intended use of the heat-treated beverage preparation.

[0384] In some preferred embodiments of the present invention, the liquid solution has a lipid content of 0-50% of the total energy content of the liquid solution, or preferably in the range of 0-45% of the total energy content of the liquid solution, or preferably in the range of 0-30% of the total energy content of the liquid solution, or preferably in the range of 0-20% of the total energy content of the liquid solution, or preferably in the range of 0-10% of the total energy content of the liquid solution, or preferably in the range of 0-5% of the total energy content of the liquid solution.

[0385] The amount of lipids is determined according to ISO 1211:2010 (Determination of fat content - Rose-Gottlieb gravimetric method).

[0386] The inventors have found that it can be advantageous to control the mineral content in order to achieve some of the desired properties of a packaged heat-treated beverage preparation.

[0387] In some embodiments of the present invention, the packaged thermally processed beverage preparation comprises a plurality of minerals. In one exemplary embodiment, the liquid solution comprises at least four minerals. In one embodiment, the four minerals are sodium, potassium, magnesium, and calcium.

[0388] The inventors have surprisingly found that the use of BLG isolate as defined herein and in Example 2 allows for the production of heat-treated beverage preparations with high mineral concentrations without compromising viscosity, thereby enabling the production of packaged heat-treated beverage preparations with high mineral content, allowing for the production of beverages that are nutritionally complete or nutritionally incomplete supplements.

[0389] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg and Ca in the liquid solution is in the range of 0 to 750 mM, preferably in the range of 100 to 600 mM, or preferably in the range of 200 to 500 mM.

[0390] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg and Ca is at most 750 mM in the liquid solution.

[0391] In another preferred embodiment of the invention, the sum of the amounts of Na, K, Mg and Ca in the liquid solution is at most 600 mM, preferably at most 500 mM, or preferably at most 400 mM, or preferably at most 300 mM, or preferably at most 200 mM, preferably at most 170 mM, most preferably at most 150 mM, or preferably at most 130 mM, or preferably at most 100 mM, or preferably at most 80 mM, or preferably at most 60 mM, or preferably at most 40 mM, or preferably at most 30 mM, or preferably at most 20 mM, or preferably at most 10 mM, or preferably at most 5 mM, or preferably at most 1 mM.

[0392] In another exemplary embodiment, the liquid solution comprises a plurality of minerals selected from the group consisting of calcium, iodine, zinc, copper, chromium, iron, phosphorus, magnesium, selenium, manganese, molybdenum, sodium, potassium, and combinations thereof.

[0393] In some preferred embodiments of the invention, the liquid solution comprises up to 150 mM KCl and up to 150 mM CaCl, or up to 130 mM KCl and up to 130 mM CaCl, or up to 110 mM KCl and up to 110 mM CaCl, or up to 100 mM KCl and up to 100 mM CaCl, or preferably up to 80 mM KCl and up to 80 mM CaCl, or preferably up to 50 mM KCl and up to 50 mM CaCl, or preferably up to 40 mM KCl and up to 40 mM CaCl.

[0394] In another preferred embodiment of the present invention, the liquid solution is a low-mineral drink.

[0395] In the context of the present invention, the term "low in minerals" relates to a composition, such as a liquid, drink, powder or another food product, which has at least one, preferably two, and even more preferably all of the following: - ash content of maximum 1.2% w / w based on total solids; - a total content of calcium and magnesium of maximum 0.3% w / w based on total solids; - maximum total sodium and potassium content of 0.10% w / w based on total solids; - Total phosphorus content of up to 100mg phosphorus per 100g of protein.

[0396] Preferably, the low mineral composition has at least one, preferably two or more, and even more preferably all of the following: - ash content of maximum 0.7% w / w based on total solids; - a total content of calcium and magnesium of maximum 0.2% w / w based on total solids; - a total content of sodium and potassium of maximum 0.08% w / w based on total solids; - Total phosphorus content of up to 80 mg phosphorus per 100 g of protein.

[0397] Even more preferably, the low mineral composition has at least one, preferably two or more, and even more preferably all of the following: - ash content of maximum 0.5% w / w based on total solids; - a total calcium and magnesium content of maximum 0.15% w / w based on total solids; - maximum total sodium and potassium content of 0.06% w / w based on total solids; - Total phosphorus content of up to 50 mg phosphorus per 100 g of protein.

[0398] It is particularly preferred that the low mineral composition has: - ash content of maximum 0.5% w / w based on total solids; - a total calcium and magnesium content of maximum 0.15% w / w based on total solids; - maximum total sodium and potassium content of 0.06% w / w based on total solids; - Total phosphorus content of up to 50 mg phosphorus per 100 g of protein.

[0399] The inventors have found that the present invention makes it possible to prepare packaged heat-treated beverage preparations that have a very low content of phosphorus and other minerals such as potassium, which is advantageous for patients suffering from kidney disease or with reduced kidney function.

[0400] The liquid solution is preferably a low phosphorus solution.

[0401] The liquid solution is preferably a low potassium solution.

[0402] The liquid solution is preferably a low phosphorus and low potassium solution.

[0403] In the context of the present invention, the term "low phosphorus" relates to a composition, e.g., a liquid, powder or another food product, having a total phosphorus content of at most 100 mg phosphorus per 100 g protein. Preferably, the low phosphorus composition has a total phosphorus content of at most 80 mg phosphorus per 100 g protein. More preferably, the low phosphorus composition may have a total phosphorus content of at most 50 mg phosphorus per 100 g protein. Even more preferably, the low phosphorus composition may have a total phosphorus content of at most 20 mg phosphorus per 100 g protein. Even more preferably, the low phosphorus composition may have a total phosphorus content of at most 5 mg phosphorus per 100 g protein. The low phosphorus composition according to the present invention may be used as a food ingredient for producing a food product for patient groups with reduced renal function.

[0404] Thus, in some particularly preferred embodiments of the present invention, the liquid solution contains at most 80 mg of phosphorus per 100 g of protein. Preferably, the liquid solution contains at most 30 mg of phosphorus per 100 g of protein. More preferably, the liquid solution contains at most 20 mg of phosphorus per 100 g of protein. Even more preferably, the liquid solution contains at most 10 mg of phosphorus per 100 g of protein. Most preferably, the liquid solution contains at most 5 mg of phosphorus per 100 g of protein.

[0405] The phosphorus content is determined in accordance with Example 1.19 with respect to the total amount of elemental phosphorus in the composition in question.

[0406] In the context of the present invention, the term "low potassium" relates to a composition, e.g., a liquid, powder, or another food product, having a total potassium content of at most 700 mg potassium per 100 g protein. Preferably, the low phosphorus composition has a total potassium content of at most 600 mg potassium per 100 g protein. More preferably, the low potassium composition may have a total potassium content of at most 500 mg potassium per 100 g protein. More preferably, the low potassium composition may have a total potassium content of at most 400 mg potassium per 100 g protein. More preferably, the low potassium composition may have a total potassium content of at most 300 mg potassium per 100 g protein. Even more preferably, the low potassium composition may have a total potassium content of at most 200 mg potassium per 100 g protein. Even more preferably, the low potassium composition may have a total potassium content of at most 100 mg potassium per 100 g protein. Even more preferably, the low potassium composition may have a total potassium content of up to 50 mg potassium per 100 g protein, and even more preferably, the low potassium composition may have a total potassium content of up to 10 mg potassium per 100 g protein.

[0407] The low potassium composition according to the present invention can be used as a food ingredient to prepare food for patient populations with reduced renal function.

[0408] Thus, in some particularly preferred embodiments of the present invention, the liquid solution comprises at most 600 mg of potassium per 100 g of protein. More preferably, the liquid solution comprises at most 500 mg of potassium per 100 g of protein. More preferably, the liquid solution comprises at most 400 mg of potassium per 100 g of protein. More preferably, the liquid solution comprises at most 300 mg of potassium per 100 g of protein. Even more preferably, the liquid solution comprises at most 200 mg of potassium per 100 g of protein. Even more preferably, the liquid solution comprises at most 100 mg of potassium per 100 g of protein. Even more preferably, the liquid solution comprises at most 50 mg of potassium per 100 g of protein, and even more preferably, the liquid solution comprises at most 10 mg of potassium per 100 g of protein.

[0409] The potassium content is determined in accordance with Example 1.19 relative to the total amount of elemental phosphorus in the composition in question.

[0410] In some preferred embodiments of the present invention, the liquid solution comprises at most 100 mg phosphorus / 100 g protein and at most 700 mg potassium / 100 g protein, preferably at most 80 mg phosphorus / 100 g protein and at most 600 mg potassium / 100 g protein, more preferably at most 60 mg phosphorus / 100 g protein and at most 500 mg potassium / 100 g protein, more preferably at most 50 mg phosphorus / 100 g protein and at most 400 mg potassium / 100 g protein, or more preferably at most 20 mg phosphorus / 100 g protein and at most 200 mg potassium / 100 g protein, or even more preferably at most 10 mg phosphorus / 100 g protein and at most 50 mg potassium / 100 g protein. In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation comprises at most 100 mg phosphorus / 100 g protein and at most 340 mg potassium / 100 g protein.

[0411] The liquid solution containing small amounts of phosphorus and potassium can be advantageously supplemented with carbohydrates and lipids, and the heat-treated beverage preparation preferably further comprises carbohydrates in a total amount in the range of 30-60%, preferably in the range of 35-50E%, of the total energy content of the liquid solution, and lipids in a total amount in the range of 20-60%, preferably in the range of 30-50E%, of the total energy content.

[0412] In one embodiment of the present invention, the liquid solution comprises a plurality of vitamins. In one exemplary embodiment, the liquid solution comprises at least 10 vitamins. In one exemplary embodiment, the liquid solution comprises a plurality of vitamins selected from the group consisting of vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12, vitamin C, vitamin D, vitamin K, riboflavin, pantothenic acid, vitamin E, thiamine, niacin, folic acid, biotin, and combinations thereof.

[0413] In one embodiment of the present invention, the liquid solution comprises multiple vitamins and multiple minerals.

[0414] In some preferred embodiments of the present invention, the liquid solution contains one or more food acids selected from the group consisting of citric acid, malic acid, tartaric acid, acetic acid, benzoic acid, butyric acid, lactic acid, fumaric acid, succinic acid, ascorbic acid, adipic acid, phosphoric acid, and mixtures thereof.

[0415] In one embodiment of the present invention, the liquid solution further comprises a flavor selected from the group consisting of salt, flavorings, seasonings and / or spices. In a preferred embodiment of the present invention, the flavor comprises chocolate, cocoa, lemon, orange, lime, strawberry, banana, forest fruit flavors or combinations thereof. The choice of flavor may depend on the beverage to be produced.

[0416] One aspect of the present invention relates to the use of a protein solution comprising a total amount of protein of 3 to 35% w / w by weight of the solution, at least 90 w / w% of which is BLG, for controlling the turbidity of a heat-treated acidic beverage preparation having a pH in the range of 2.0 to 4.7.

[0417] Another aspect of the present invention relates to the use of a protein solution comprising a total amount of protein of 2 to 45% w / w, based on the weight of the solution, of which at least 90 w / w% is BLG, for controlling astringency in heat-treated acidic beverage preparations having a pH in the range of 2.0 to 4.7.

[0418] Another aspect of the present invention relates to a packaged thermally treated beverage preparation as defined herein for use in a method for treating a disease associated with protein malabsorption.

[0419] Another aspect of the present invention relates to the use of the packaged thermally treated beverage preparations provided herein as a dietary supplement.

[0420] In a preferred embodiment of the present invention, the packaged thermally treated beverage preparation as defined herein is used as a dietary supplement and is taken before, during or after exercise.

[0421] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage a total amount of 2 to 45% w / w of protein relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; -optionally sweeteners and / or flavors Including, - the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11; - Fat content of up to 5% of the total energy content of the preparation Includes.

[0422] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage a total amount of 2 to 10% w / w of protein by weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; -optionally sweeteners and / or flavors Including, - the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11; - Fat content of up to 5% of the total energy content of the preparation Includes.

[0423] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage a total amount of 10 to 45% w / w, preferably 10 to 35% w / w, of the protein by weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w is BLG; -optionally sweeteners and / or flavors Including, - the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11; - Fat content of up to 5% of the total energy content of the preparation Includes.

[0424] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 32.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage a protein in a total amount of 2 to 45% w / w relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; and -optionally sweeteners and / or flavors Including, The packaged heat-treated beverage preparation has a turbidity of at most 200 NTU, preferably at most 40 NTU.

[0425] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage a total amount of 2 to 10% w / w of protein by weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; -optionally sweeteners and / or flavors Including, The packaged heat-treated beverage preparation has a turbidity of at most 200 NTU, preferably at most 40 NTU.

[0426] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage a total amount of 10 to 45% w / w, preferably 10 to 205 w / w, of protein relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; and -optionally sweeteners and / or flavors Including, The packaged heat-treated beverage preparation has a turbidity of at most 200 NTU, preferably at most 40 NTU.

[0427] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage a protein in a total amount of 2 to 45% w / w relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; and -optionally sweeteners and / or flavors Including, - the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11; - the protein fraction of the beverage preparation has a color value delta b ranging from -0.10 to +0.51 on the CIELAB color scale * and delta b * = measured at room temperature, b 6.0w / w%タンパク質に標準化された試料 * -b 脱塩水 * is.

[0428] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage a protein in a total amount of 2 to 10% w / w relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; and -optionally sweeteners and / or flavors Including, - the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11; - the protein fraction of the beverage preparation has a color value delta b ranging from -0.10 to +0.51 on the CIELAB color scale * and delta b * = measured at room temperature, b 6.0w / w%タンパク質に標準化された試料 * -b 脱塩水 * is.

[0429] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage a protein in a total amount of 10 to 45% w / w, preferably 10 to 20% w / w, based on the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; and -optionally sweeteners and / or flavors Including, - the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11; - the protein fraction of the beverage preparation has a color value delta b ranging from -0.10 to +0.51 on the CIELAB color scale * and delta b * = measured at room temperature, b 6.0w / w%タンパク質に標準化された試料 * -b 脱塩水 * is.

[0430] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage a total amount of 2 to 45% w / w of protein relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; -optionally sweeteners and / or flavors Including, - the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11; The sum of the amounts of Na, K, Mg and Ca is at most 750 mM, preferably at most 400 mM, preferably at most 200 mM.

[0431] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage a total amount of 2 to 10% w / w of protein by weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; -optionally sweeteners and / or flavors Including, - the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11; The sum of the amounts of Na, K, Mg and Ca is at most 750 mM, preferably at most 400 mM, preferably at most 200 mM.

[0432] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage a total amount of 10 to 45% w / w, preferably 10 to 20% w / w, of protein relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w is BLG; -optionally sweeteners and / or flavors Including, - the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11; The sum of the amounts of Na, K, Mg and Ca is at most 750 mM, preferably at most 400 mM, preferably at most 200 mM.

[0433] In a preferred embodiment of the present invention, the packaged heat-treated opaque beverage preparation has a pH in the range of 3.0 to 4.7, preferably 3.9 to 4.6, more preferably 4.0 to 4.5, and the beverage is a total amount of 2 to 45% w / w of proteins relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; and -optionally sweeteners and / or flavors Including, - the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11, and / or the protein fraction has a degree of protein denaturation of at most 5%, and / or - Fat content of more than 5% of the total energy content of the preparation Includes.

[0434] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 3.0 to 4.7, preferably 3.9 to 4.6, more preferably 4.0 to 4.5, and the beverage is a total amount of 2 to 45% w / w of protein relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; -optionally sweeteners and / or flavors Including, a turbidity greater than 200 NTU, preferably greater than 1000 NTU, and / or - Viscosity is up to 200 cP.

[0435] In a preferred embodiment of the present invention, the packaged heat-treated opaque beverage preparation has a pH in the range of 3.0 to 4.7, preferably 3.9 to 4.6, more preferably 4.0 to 4.5, and the beverage is a total amount of 2 to 10% w / w of proteins relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; and -optionally sweeteners and / or flavors Including, - the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11, and / or the protein fraction has a degree of protein denaturation of at most 5%, and / or - Fat content of more than 5% of the total energy content of the preparation Includes.

[0436] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 3.0 to 4.7, preferably 3.9 to 4.6, more preferably 4.0 to 4.5, and the beverage is a total amount of 2 to 10% w / w of proteins relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; -optionally sweeteners and / or flavors Including, a turbidity greater than 200 NTU, preferably greater than 1000 NTU, and / or - Viscosity is up to 200 cP.

[0437] In a preferred embodiment of the present invention, the packaged heat-treated opaque beverage preparation has a pH in the range of 3.0 to 4.7, preferably 3.9 to 4.6, more preferably 4.0 to 4.5, and the beverage is a total amount of 10 to 45% w / w, preferably 10 to 20% w / w, of proteins relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; and -optionally sweeteners and / or flavors Including, - the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11, and / or the protein fraction has a degree of protein denaturation of at most 5%, and / or - Fat content of more than 5% of the total energy content of the preparation Includes.

[0438] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 3.0 to 4.7, preferably 3.9 to 4.6, more preferably 4.0 to 4.5, and the beverage is a total amount of 10 to 45% w / w of protein relative to the weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, is BLG; -optionally sweeteners and / or flavors Including, a turbidity greater than 200 NTU, preferably greater than 1000 NTU, and / or - Viscosity is up to 200 cP.

[0439] In some embodiments of the present invention, the thermally treated beverage has a shelf life of at least 6 months at 25° C. an edible BLG composition as defined in PCT / EP2017 / 084553, which provides a total amount of BLG of at least 1% (w / w), preferably at least 5% (w / w), sweeteners, such as sugar sweeteners and / or non-sugar sweeteners, at least one food acid, such as citric acid or other suitable food acid; optionally, flavorings, and -Maximum 80mg phosphorus / 100g protein Including, It has a pH range of 2.5 to 4.0.

[0440] In a preferred embodiment of the invention, the invention relates to the use of a protein solution comprising a total amount of 3-30% w / w of protein by weight of the solution, of which at least 85 w / w%, preferably at least 90% w / w, is BLG, for controlling the turbidity of a heat-treated acidic beverage preparation having a pH in the range of 3.0-4.5.

[0441] In a preferred embodiment of the present invention, the present invention relates to the use of a protein solution comprising a total amount of 3-30% w / w of protein by weight of the solution, of which at least 85 w / w%, preferably at least 90% w / w, is BLG, for controlling astringency in heat-treated acidic beverage preparations having a pH in the range of 2.0-4.0.

[0442] A preferred embodiment of the present invention relates to a thermally treated beverage preparation obtained by one or more of the methods described herein.

[0443] It should be noted that embodiments and features described in the context of one aspect of the invention also apply to other aspects of the invention.

[0444] All patent and non-patent references cited in this application are incorporated herein by reference in their entirety.

[0445] The invention will now be described in further detail in the following non-limiting examples.

[0446] example Example 1: Analysis method Example 1.1: Determining protein nativeity by intrinsic tryptophan fluorescence Tryptophan (Trp) fluorescence spectroscopy is a well-known tool for monitoring protein folding and unfolding. Buried Trp residues in native proteins typically exhibit the highest fluorescence emission near 330 nm, rather than in more solvent-exposed positions such as unfolded proteins. In unfolded proteins, the wavelength of Trp fluorescence emission typically shifts to higher wavelengths and is usually measured near 350 nm. Here, we exploit this transition to monitor thermally induced unfolding by calculating the ratio of fluorescence emissions at 330 nm and 350 nm to investigate the effect of heating temperature.

[0447] The analysis includes the following steps: The beverage composition was diluted to 0.6 mg / ml with MQ water. 300 μl of sample was transferred to a white 96-well plate, avoiding air bubbles, or 3 mL was transferred to a 10 mm quartz cuvette. · Tryptophan fluorescence emission intensity was recorded from above between 310 and 400 nm by excitation at 295 using a 5 nm slit. Samples were measured using a Cary Eclipse fluorescence spectrophotometer equipped with a plate reader accessory (G9810A) or a single cuvette holder. The emission intensity ratio was calculated by dividing the fluorescence emission intensity measured at 330 nm by the emission intensity at 350 nm, R = I330 / I350, and was used as a measure of protein nativeness. An R of at least 1.11 represents the predominant native BLG conformation; An R less than 1.11 reports at least partial unfolding and aggregation.

[0448] Example 1.2: Thermal stability at pH 3.9 Heat stability at pH 3.9: Heat stability at pH 3.9 is a measure of the ability of a protein composition to remain clear upon extended pasteurization at pH 3.9.

[0449] Heat stability at pH 3.9 is determined by mixing a sample of the powder or liquid to be tested with water (or alternatively, if it is a dilute liquid, concentrating it by low-temperature evaporation) to form an aqueous solution having a pH of 3.9 and containing 6.0% w / w protein, and adjusting the pH to 3.9 with the minimum amount of 0.1 M NaOH or 0.1 M HCl necessary.

[0450] After allowing the pH-adjusted mixture to stand for 30 minutes, 25 mL of the mixture was transferred to a 30 mL thin-walled glass test tube. This was then heated to 75.0°C for 300 seconds by immersion in a water bath at 75.0°C. Immediately after heating, the glass test tube was transferred to an ice bath and cooled to 1-5°C. The turbidity of the heat-treated sample was measured according to Example 1.7.

[0451] Example 1.3: Determination of the degree of protein denaturation of whey protein compositions Since denatured whey proteins are known to be less soluble at pH 4.6 than at pH values ​​below or above pH 4.6, the degree of denaturation of a whey protein composition is determined by measuring the amount of soluble protein at pH 4.6 relative to the total amount of protein at the pH at which the protein is stable in solution.

[0452] More specifically, in the case of whey protein, the whey protein composition to be analyzed (e.g., powder or aqueous solution) is converted into: a first aqueous solution containing 5.0% (w / w) total protein and having a pH of 7.0 or 3.0; and - A second aqueous solution containing 5.0% (w / w) total protein and having a pH of 4.6. pH adjustment is performed using 3% (w / w) NaOH (aqueous) or 5% (w / w) HCl (aqueous).

[0453] The total protein content (P pH7.0又は3.0 ) is determined according to Example 1.5.

[0454] The second aqueous solution is stored at room temperature for 2 hours and then centrifuged at 3000 g for 5 minutes. A sample of the supernatant is collected and analyzed according to Example 1.5 to determine the protein concentration (S pH4.6 ) is obtained. The degree of protein denaturation, D, of the whey protein composition is calculated as follows: D=((P pH7.0又は3.0 -S pH4.6 ) / P pH7.0又は3.0 ) * 100%

[0455] Example 1.4 Determining Protein Denaturation (by pH 4.6 Acid Precipitation) Using Reversed-Phase UPLC Analysis BLG samples (including unheated reference and heated BLG beverage compositions) were diluted to 2% with MQ water. 5 mL of protein solution, 10 mL of Milli-Q water, 4 mL of 10% acetic acid, and 6 mL of 1.0 M NaOAc were mixed and stirred for 20 minutes to precipitate denatured proteins at a pH of approximately 4.6. The solution was filtered through a 0.22 μm filter to remove aggregates and non-native proteins.

[0456] All samples were subjected to the same degree of dilution by adding polishing water.

[0457] For each sample, the same volume was loaded onto a UPLC system equipped with a UPLC column (Protein BEH C4; 300 Å; 1.7 μm; 150 × 2.1 mm) and detected at 214 nm. Samples were run using the following conditions: Buffer A: Milli-Q water, 0.1%w / w TFA Buffer B: HPLC grade acetonitrile, 0.1% w / w TFA Flow rate: 0.4ml / min

[0458] Gradient: 0–6.00 min 24–45% B; 6.00–6.50 min 45–90% B; 6.50–7.00 min 90% B; 7.00–7.50 min 90–24% B and 7.50–10.00 min 24% B. The area of ​​the BLG peak relative to a protein standard (Sigma L0130) was used to determine the concentration of native bLG in the samples (5-level calibration curve).

[0459] Samples were further diluted and re-injected if outside the linear range.

[0460] Example 1.5: Determination of total protein The total protein content (true protein) of the sample is determined by: 1) Determine the total nitrogen of the sample according to ISO 8968-1 / 2 | IDF 020-1 / 2 - Milk - Determination of the nitrogen content - Part 1 / 2: Determination of the nitrogen content using the Kjeldahl method. 2) Determine the non-protein nitrogen of the sample according to ISO 8968-4 | IDF 020-4 - Milk - Determination of the nitrogen content - Part 4: Determination of the non-protein nitrogen content. 3) The total amount of protein (m 総窒素 -m 非タンパク質窒素 ) * Calculate as 6.38.

[0461] Example 1.6: Determination of non-aggregated BLG, ALA and CMP The contents of unaggregated α-lactalbumin (ALA), β-lactoglobulin (BLG), and caseinomacropeptide (CMP) were analyzed by HPLC at 0.4 mL / min. 25 μL of the filtered sample was injected onto two TSKgel3000PWxl (7.8 mm x 30 cm, Tosohass, Japan) columns connected in series with a coupled precolumn PWxl (6 mm x 4 cm, Tosohass, Japan) equilibrated with eluent (465 g Milli-Q water, 417.3 g acetonitrile, and 1 mL trifluoroacetic acid) and analyzed using a UV detector at 210 nm.

[0462] Natural α-lactalbumin (C α ), β-lactoglobulin (C β ) and caseinomacropeptide (C CMPQuantitative determination of the content of ) was performed by comparing the peak area obtained for the corresponding standard protein with that of the sample.

[0463] The total amount of additional proteins (non-BLG proteins) was determined by subtracting the amount of BLG from the amount of total protein (determined according to Example 1.5).

[0464] Example 1.7: Determining turbidity Turbidity is the cloudiness or blurring of a fluid caused by numerous particles that are generally invisible to the naked eye, similar to smoke in the air.

[0465] Turbidity is measured in Nephelometric Turbidity Units (NTU).

[0466] 20 mL of beverage / sample was added to an NTU glass and placed in a Turbiquant® 3000IR turbidity meter. NTU values ​​were measured after stabilization and were repeated twice.

[0467] Example 1.8: Determining viscosity The viscosity of the beverage preparations was measured using a rheometer (Anton Paar, Physica MCR301).

[0468] 3.8 mL of sample was added to cup DG26.7. The sample was equilibrated to 22°C and then cooled for 50 seconds. -1 Pre-shear at 40°C for 30 seconds, followed by a 30 second equilibration period and a 1 second -1 ~200 seconds -1 ~1 second -1 A shear rate sweep was performed between .

[0469] Unless otherwise stated, viscosity is 100 seconds -1 It is given in units of centipoise (cP) at shear rates of 1000 psi. The higher the measured cP value, the higher the viscosity.

[0470] Alternatively, viscosity is estimated using Viscoman by Gilson, approximately 300 s -1 The shear rate is reported as

[0471] Example 1.9: Color Determination Color was measured using a Chroma Meter (Konica Minolta, CR-400). 15 g of sample was added to a small Petri dish (55 x 14.2 mm, VWR catalog number 391-0895) to avoid air bubble formation. The protein content of the samples was standardized to ≤ 6.0 w / w% protein.

[0472] The Chroma Meter was calibrated to a white calibration plate (number 19033177). The illuminant was set to D65 and the observer was set at 2°. Color (CIELAB color space, a * -, b * -, L * value) was measured as the average of three individual readings at different locations on the Petri dish with the lid covering the suspension.

[0473] The demineralized water reference has the following values: L * 39.97±0.3 a * 0.00±0.06 b * -0.22±0.09

[0474] Measurements were converted to delta / difference values ​​based on demineralized water measurements. Delta L * = measured at room temperature, L 6.0w / w%タンパク質に標準化された試料 * -L 脱塩水 * Delta A * = measured at room temperature, a 6.0w / w%タンパク質に標準化された試料 * -a 脱塩水 * Delta b * = measured at room temperature, b 6.0w / w%タンパク質に標準化された試料 * -b 脱塩水 *

[0475] Samples are standardized to ≦6.0 w / w% protein.

[0476] L * a * b * The CIELAB color space (also known as the CIELAB space) is a uniform color space defined by the International Commission on Illumination (CIE) in 1976 and used to quantitatively report lightness and hue (ISO 11664-4:2008(E) / CIE S 014-4 / E:2007).

[0477] In this space, L * indicates brightness (value 0 to 100), and L * = 0 is the darkest black, L * =100 indicates the brightest white.

[0478] Color channel a * and b * is a * =0 and b * = 0 represents a true neutral gray value. * The axis represents the green-red component, with green going negative and red going positive. * The axis represents the blue-yellow component, with blue going negative and yellow going positive.

[0479] Example 1.10 Beverage Stability Test / Insoluble Protein Substances A whey protein beverage composition was considered stable if less than 15% of the total protein in the heated sample precipitated upon centrifugation at 3000 g for 5 minutes. Approximately 20 g of sample was added to a centrifuge tube and centrifuged at 3000 g for 5 minutes. Protein recovery was quantified using Kjeldahl analysis of the protein before centrifugation and the supernatant after centrifugation. See Example 1.5.

[0480] Calculate protein loss:

number

[0481] This parameter is sometimes referred to as the level of insoluble protein material and can be used to analyze both liquid and powder samples. If the sample is a powder, 10 g of powder is suspended in 90 g of demineralized water and allowed to hydrate for 1 hour at 22°C with gentle stirring. Approximately 20 g of sample (e.g., liquid sample or suspended powder sample) is placed in a centrifuge tube and centrifuged at 3000 g for 5 minutes. The protein (P) before centrifugation is 総 ) and the supernatant after centrifugation (P 3000xg ) Kjeldahl analysis was used to quantify protein recovery according to Example 1.5.

[0482] Calculate the amount of insoluble protein material:

number

[0483] Example 1.11: Sensory evaluation The heat treated beverage preparations were subjected to descriptive sensory evaluation. The beverage preparations were subjected to heat using a plate heat exchanger. One volume of sample is mixed with one volume of water and compared to unheated whey protein isolate, lactic acid and citric acid, which are also used to generate an attribute list prior to a final tasting session. [Table 1]

[0484] Crackers, white tea, melon, and water were used to rinse participants' mouths between each sample.

[0485] A 15 mL test sample at ambient temperature (20-25°C) was provided in a small cup.

[0486] The test samples were given in a randomized order, three times in three different blocks to 10 people each.

[0487] The attributes (see table above) were rated on a 15cm scale with 0 = low intensity and 15 = high intensity.

[0488] Statistical analysis was performed using a three-way ANOVA test with multiple replicates in the "Panelcheck" software. Samples were fixed and panels were randomly set up.

[0489] Significant differences between samples were assessed using the Bonferroni correction, which means the smallest significant difference value (pairwise comparison of groups associated with letters).

[0490] Example 1.12: Determining Transparency Through Imaging Photography of the beverage preparations was performed by placing the sample in a turbidity NTU measurement vial touching a piece of paper with the text "lorem ipsen" written on it. Using a smartphone, the vial was photographed and we assessed whether the text could be clearly observed through the vial.

[0491] Example 1.13: Determination of ash content The ash content of the food is determined in accordance with NMKL 173:2005 "Gravimetric determination of ash in food."

[0492] Example 1.14: Determining Conductivity The "conductivity" (sometimes called "specific conductivity") of an aqueous solution is a measure of the solution's ability to conduct electricity. Conductivity can be determined, for example, by measuring the AC resistance of the solution between two electrodes, with the results typically given in units of milliSiemens / cm (mS / cm). Conductivity can be measured, for example, according to EPA (U.S. Environmental Protection Agency) Method No. 120.1.

[0493] Conductivity values ​​referred to herein are normalized to 25°C unless otherwise specified.

[0494] The conductivity is measured with a conductivity meter (WTW Cond 3210 with tetracon 325 electrodes).

[0495] The system is calibrated before use as described in the manual. The electrode is thoroughly rinsed with the same type of medium in which the measurement will be made to avoid local dilution. The electrode is lowered into the medium so that the area in which the measurement will be made is completely submerged. The electrode is then agitated to remove any air trapped in the electrode. The electrode is then held stationary until a stable value can be obtained and recorded from the display.

[0496] Example 1.15: Determining the total solids content of a solution The total solids content of a solution may be determined according to NMKL 110 2nd Edition, 2005 (Total solids (water) - gravimetric determination for milk and dairy products). NMKL is the abbreviation for "European Standard Methods of Analysis and Nordic Committee for Food Analysis".

[0497] The water content of a solution can be calculated relative to 100% minus the total solids (% w / w).

[0498] Example 1.16: Determining pH All pH values ​​are measured using a pH glass electrode and normalized to 25°C.

[0499] pH glass electrodes (with temperature compensation) are carefully rinsed before use and calibrated before use.

[0500] If the sample is in liquid form, the pH is measured directly in the liquid solution at 25°C.

[0501] If the sample is a powder, 10 grams of powder is dissolved in 90 ml of demineralized water at room temperature with vigorous stirring. The pH of the solution is then measured at 25°C.

[0502] Example 1.17: Determination of loose and bulk density The density of a dry powder is defined as the relationship between the weight and volume of the powder analyzed using a special Stampf volumeter (i.e., a graduated cylinder) under specified conditions. Density is typically expressed in g / ml or kg / L.

[0503] In this method, a sample of dry powder is packed into a graduated cylinder. After a specified number of taps, the volume of the product is read and its density is calculated.

[0504] This method allows us to define three types of density: Packed density, which is the mass divided by the volume of the powder after it has been transferred to a specified graduated cylinder. Loose density, which is the mass divided by the volume of the powder after 100 taps according to the conditions specified in this standard. Bulk density, which is the mass divided by the volume of the powder after 625 taps according to the conditions specified in this standard.

[0505] These methods use a special measuring cylinder, 250 ml, graduated 0-250 ml, weighing 190±15 g (J. Engelsmann AG 67059 Ludwigshafen / Rh) and a Stampf volumeter (e.g., J. Engelsmann AG).

[0506] The loose density and bulk density of the dried product are determined by the following procedure.

[0507] Pretreatment: The sample to be measured is stored at room temperature.

[0508] The sample is then thoroughly mixed by repeatedly rotating and swirling the container (avoiding crushing of particles). The container is not filled more than 2 / 3 full.

[0509] procedure: Weigh out 100.0±0.1 g of powder and transfer it into a measuring cylinder. Read off the volume V0 in ml.

[0510] If 100 g of powder will not fit into the cylinder, the amount should be reduced to 50 or 25 g.

[0511] Fix the measuring cylinder to the Stampf volumeter and tap it 100 times. Flatten the surface with a spatula and measure the volume V. 100 Read in ml.

[0512] The number of tabs is changed to 625 (including 100 taps). After tapping, the surface is flattened and the volume V 625 Read in ml.

[0513] Density calculation: Calculate the loose density and bulk density in g / ml according to the following formula: Bulk density = M / V (where M represents the sample weight in grams and V represents the volume after 625 taps in ml).

[0514] Example 1.18: Determining the moisture content of a powder The moisture content of foods is determined according to ISO 5537:2004 (Milk powder - Determination of moisture content (reference method)). NMKL is the abbreviation for "Nordic Committee for European Standard Methods of Analysis and Food Analysis".

[0515] Example 1.19: Determination of calcium, magnesium, sodium, potassium, and phosphorus (ICP-MS method) The total amounts of calcium, magnesium, sodium, potassium and phosphorus are determined using a procedure in which the sample is first digested using microwave digestion and then the total amount of minerals is determined using an ICP instrument.

[0516] Device: The microwave is manufactured by Anton Paar and the ICP is an Optima 2000DV manufactured by PerkinElmer Inc.

[0517] material: 1 M HNO3 Yttrium in 2% HNO3 Suitable standards for calcium, magnesium, sodium, potassium and phosphorus in 5% HNO3

[0518] Pretreatment: Weigh out a certain amount of powder and transfer this powder to a microwave digestion tube. Add 35 mL of 1 M HNO. Microwave digest the sample according to the microwave instructions. Place the digested tube in a fume cupboard, remove the lid, and allow the volatile fumes to evaporate.

[0519] Measurement procedure : Transfer the pretreated sample to a DigiTUBE using a known amount of Milli-Q water. Add a 2% solution of yttrium in HNO3 to the digestion tube (approximately 0.25 mL per 50 mL of diluted sample) and dilute to a known volume using Milli-Q water. Analyze the sample by ICP using the procedure described by the manufacturer.

[0520] A blank sample is prepared by diluting a mixture of 10 mL of 1 M HNO3 and 0.5 mL of a 2% solution of yttrium in HNO3 to a final volume of 100 mL using Milli-Q water.

[0521] Prepare at least three standard samples with concentrations that bracket the expected sample concentration.

[0522] Example 1.20: Determining the Furosine Value: Furosine values ​​are determined as described in "Maillard Reaction Evaluation by Furosine Determination During Infant Cereal Processing," Guerra-Hernandez et al., Journal of Cereal Science 29 (1999) 171-176, and total protein is determined according to Example 1.5. Furosine values ​​are reported in mg furosine per 100 g protein.

[0523] Example 1.21: Determining the crystallinity of BLG in a liquid Use the following method to determine the crystallinity of BLG in liquids with a pH ranging from 5 to 6. a) A 10 mL sample of the liquid in question is transferred to a Maxi-Spin filter equipped with a 0.45 micron pore size CA membrane. b) Immediately spin the filter at 1500 g for 5 minutes and keep the centrifuge at 2°C. c) Add 2 mL of cold Milli-Q water (2°C) to the retentate side of the spin filter and immediately, with the centrifuge still cooled to 2°C, spin the filter at 1500 g for 5 minutes, collect the permeate (Permeate A), measure the volume, and determine the BLG concentration via HPLC using the method outlined in Example 1.31. d) Add 4 mL of 2 M NaCl to the retentate side of the filter, vortex rapidly, and allow the mixture to stand at 25° C. for 15 minutes. e) Immediately spin the filter at 1500 g for 5 minutes and collect the permeate (Permeate B). f) Using the method outlined in Example 1.31, determine the total weight of BLG in Permeate A and Permeate B and convert these results to total weight of BLG rather than weight percent. 透過液A The weight of BLG in permeate B is called m 透過液B It is called. g) Determine the liquid crystallinity for BLG as follows: Crystallinity=m 透過液B / (m 透過液A +m 透過液B ) * 100%

[0524] Example 1.22: Determination of Crystallinity of BLG in Dry Powder This method is used to determine the crystallinity of BLG in dry powders. a) Mix 5.0 grams of powder sample with 20.0 grams of cold Milli-Q water (2°C) and let stand at 2°C for 5 minutes. b) The liquid sample in question is transferred to a Maxi-Spin filter equipped with a 0.45 micron CA membrane. c) Immediately spin the filter at 1500g for 5 minutes and keep the centrifuge at 2°C. d) Add 2 mL of cold Milli-Q water (2°C) to the retentate side of the spin filter and immediately spin the filter at 1500 g for 5 minutes. Collect the permeate (Permeate A), measure the volume, and determine the BLG concentration via HPLC using the method outlined in Example 1.31, converting the results to total weight of BLG rather than weight %. The weight of BLG in Permeate A is m 透過液A It is called. f) The crystallinity of the BLG in the powder is then calculated using the following formula:

number

[0525] If the total amount of BLG in a powder sample is unknown, it can be determined by suspending another 5 g of the powder sample (from the same powder source) in 20.0 grams of Milli-Q water, adjusting the pH to 7.0 by adding aqueous NaOH solution, leaving the mixture at 25°C for 1 hour with stirring, and finally determining the total amount of BLG in the powder sample using Example 1.31.

[0526] Example 1.23: Determining UF permeate conductivity Transfer 15 mL of sample to an Amicon Ultra-15 centrifugal filter unit with a 3 kDa cutoff (3000 NMWL) and centrifuge at 4000 g for 20-30 minutes, or until a sufficient volume of UF permeate accumulates at the bottom of the filter unit to measure conductivity. Conductivity is measured immediately after centrifugation. Sample handling and centrifugation are performed at the temperature of the sample source.

[0527] Example 1.24: Detection of dry BLG crystals in powder The presence of dry BLG crystals in the powder can be identified in the following way.

[0528] The powder sample to be analyzed is resuspended in demineralized water having a temperature of 4°C in a weight ratio of 2 parts water to 1 part powder, mixed gently and allowed to rehydrate at 4°C for 1 hour.

[0529] The rehydrated sample is examined by microscopy, preferably using plane polarized light to detect birefringence, to identify the presence of crystals.

[0530] The crystalline material is isolated and subjected to X-ray crystallography to confirm the presence of crystalline structure and preferably also to confirm that the crystal lattice (space group and unit cell dimensions) corresponds to that of a BLG crystal.

[0531] The chemical composition of the isolated crystalline material is analyzed to confirm that the solid is primarily composed of BLG.

[0532] Example 1.25: Determining the Total Amount of Lactose The total lactose content is determined according to ISO 5765-2:2002 (IDF 79-2:2002) "Milk powders, dry ice mixtures and processed cheese - Determination of lactose content - Part 2: Enzymatic method utilizing the galactose moiety of lactose."

[0533] Example 1.26: Determining the total amount of carbohydrates: The amount of carbohydrate is determined using the Sigma Aldrich Total Carbohydrate Assay Kit (catalog MAK104-1KT), which hydrolyzes carbohydrates and converts them to furfural and hydroxyfurfural, which are converted to chromagens that are monitored spectrophotometrically at 490 nm.

[0534] Example 1.27: Determining the total amount of lipids The amount of lipids is determined according to ISO 1211:2010 (Determination of fat content - Rose-Gottlieb gravimetric method).

[0535] Example 1.28: Determining Brix Brix measurements were performed using a PAL-α digital handheld refractometer (Atago) calibrated against polished water (water filtered by reverse osmosis to obtain a maximum conductivity of 0.05 mS / cm).

[0536] Approximately 500 μl of sample was transferred to the prism face of the instrument and the measurement was initiated. The measurement value was read and recorded.

[0537] Example 1.29 Determination of lactoferrin and lactoperoxidase Lactoferrin concentrations are determined by ELISA immunoassay as outlined in Soyeurt 2012 (Soyeurt et al.; Mid-infrared prediction of lactoferrin content in bovine milk: potential indicator of mastitis; Animal (2012), 6:11, pp. 1830-1838).

[0538] The concentration of lactoperoxidase is determined using a commercially available bovine lactoperoxidase kit.

[0539] Example 1.30: Determining the number of colony-forming units The determination of the number of colony forming units per gram of sample is carried out according to ISO 4833-1:2013(E): Microbiology of food and animal feed - Horizontal method for enumerating microorganisms - Colony count technique at 30°C.

[0540] Example 1.31: Determination of the total amount of BLG, ALA, and CMP This procedure is a liquid chromatography (HPLC) method for quantitative analysis of proteins such as ALA, BLG, and CMP, and optionally other protein species, in a composition. In contrast to the method of Example 1.6, this method also measures proteins that exist in aggregates, and therefore provides a measure of the total amount of protein species in the composition in question.

[0541] The separation method is size exclusion chromatography (SEC), which uses 6 M guanidine HCl buffer as both the sample solvent and the HPLC mobile phase. Mercaptoethanol is used as a reducing agent to reduce disulfides (SS) in proteins or protein aggregates, creating unfolded monomeric structures.

[0542] Sample preparation is easily accomplished by dissolving the equivalent of 10 mg protein in the mobile phase.

[0543] Two TSK-GEL G3000SWXL (7.7 mm x 30.0 cm) columns (GPC columns) and a guard column are arranged in series to achieve sufficient separation of the main proteins in the raw material.

[0544] The eluted analytes are detected and quantified by UV detection (280 nm).

[0545] Equipment / Materials: 1. HPLC Pump 515 (Waters) with manual seal wash 2. HPLC Pump Controller Module II (Waters) 3. Autosampler 717 (Waters) 4. Dual Absorbance Detector 2487 (Waters) 5. Computer software capable of generating quantitative reports (Empower 3, Waters) 6. Analytical columns: two TSK-GEL G3000SWXL (7.8 x 300 mm, P / N: 08541) Guard column: TSK-Guard column SWxL (6.0 x 40 mm, P / N: 08543) 7. Ultrasonic bath (Branson 5200) 8. 25 mm syringe filter with 0.2 μm cellulose acetate membrane (514-0060, VWR)

[0546] procedure: Mobile phase: A. Stock buffer 1. Weigh 56.6g of NaHPO, 3.5g of NaHPO, and 2.9g of EDTA into a 1000mL beaker. Dissolve in 800 mL of water. 2. Measure the pH and adjust to 7.5±0.1 with HCl (to lower the pH) or NaOH (to raise the pH) as needed. 3. Transfer to a 1000 mL volumetric flask and dilute to volume with water.

[0547] B. 6M guanidine HCl mobile phase 1. Weigh 1146 g of guanidine HCl into a 2000 mL beaker and add 200 mL of stock buffer (A). 2. Dilute this solution to approximately 1600 mL with water while mixing with a magnetic stir bar (50°C). 3. Adjust the pH to 7.5 ± 0.1 with NaOH. 4. Transfer to a 2000 mL volumetric flask and dilute to volume with water. 5. Filter using a solvent filtration apparatus equipped with a 0.22 μm membrane filter.

[0548] Calibration Standards Calibration standards for each protein to be quantified are prepared in the following manner: 1. Approximately 25 mg of protein reference standard in 10 mL Accurately weigh (to the nearest 0.01 mg) into a volumetric flask and dissolve in 10 mL of water. This will be the protein stock standard solution (S1). 2. Pipette 200 μl of S1 into a 20 ml volumetric flask and dilute to volume with mobile phase. This becomes the low use standard solution WS1. 3. Pipette 500 μL of S1 into a 10 mL volumetric flask and dilute to volume with mobile phase. This is the standard solution WS2. 4. Pipette 500 μL of S1 into a 5 mL volumetric flask and dilute to volume with mobile phase. This is the standard solution WS3. 5. Pipette 750 μL of S1 into a 5 mL volumetric flask and dilute to volume with mobile phase. This is standard solution WS4. 6.Pipette 1.0 mL of S1 into a 5 mL volumetric flask and dilute to volume with mobile phase. This is the high use standard solution WS5. 7. Using a graduated disposable pipette, transfer 1.5 mL of WS1-5 into separate vials. Add 10 μL of 2-mercaptoethanol to each vial and cap. Vortex the solution for 10 seconds. The standards are allowed to stand at ambient temperature for approximately 1 hour. 8. Filter the standard using a 0.22 μm cellulose acetate syringe filter.

[0549] Protein purity is determined using Kjeldahl (N x 6.38) and area % from standard solution WS5 using HPLC. Protein (mg) = "Standard Protein Weight" (mg) x P1 x P2 P1=P% (Kjeldahl) P2 = Protein area % (HPLC)

[0550] Sample preparation 1. Weigh out the equivalent of 25 mg of protein from the original sample into a 25 mL volumetric flask. 2. Add approximately 20 mL of mobile phase and allow the sample to dissolve for approximately 30 minutes. 3. Add the mobile phase to volume and add 167 μL of 2-mercaptoethanol to 25 ml of sample solution. 4. Sonicate for approximately 30 minutes, then allow the sample to stand at ambient temperature for approximately 1.5 hours. 5. Mix the solution and filter using a 0.22 μl cellulose acetate syringe filter.

[0551] HPLC Systems / Columns Column equilibration 1. Connect a GPC guard column and two GPC analytical columns in series. A new column is typically placed in a phosphate buffer solution. 2. Slowly run water through the new column at 0.1-0.5 mL / min over 30-60 minutes. Continue to run for about an hour. 3. Gradually reduce the flow rate from 0.5 mL / min to 0.1 mL / min and replace with the mobile phase in the reservoir. 4. To avoid pressure shock, gradually increase the pump flow rate to 0.1-0.5 mL / min over 30-60 min and leave it at 0.5 mL / min. 5. Inject 10 samples to saturate the column and wait for the peaks to elute. This helps condition the column. This process is performed without the need to wait for each injection to be completed before the next injection. 6. Equilibrate with the mobile phase for at least 1 hour.

[0552] Calculating the results The quantitative determination of the content of the proteins to be quantified, such as α-lactalbumin, β-lactoglobulin and caseinomacropeptide, is carried out by comparing the peak area obtained for the corresponding standard protein with the peak area of ​​the sample. The results are reported as g of specific protein / 100 g of original sample or as the weight percentage of the specific protein relative to the weight of the original sample.

[0553] Example 2: Preparation of spray-dried acidic BLG isolate powder Whey Protein Source Lactose-depleted UF retentate derived from sweet whey from a standard cheese-making process was filtered through a 1.2 micron filter and fat-reduced via a Synder FR membrane before being used as feedstock for the BLG crystallization process. The chemical composition of the feedstock is shown in Table A. Note that all weight percentages of specific proteins, such as BLG, ALA, etc., referred to in this example refer to the weight percentage of non-aggregated protein relative to total protein.

[0554] adjustment The sweet whey feedstock was filtered through a Koch HFK-328 membrane (70 ml) at a 46-mil spacer feed pressure of 1.5-3.0 bar. 2The feed concentration was adjusted to 21% ± 5 total solids (TS) at an ultrafiltration setting of 20°C using a diafiltration membrane and polished water (water filtered by reverse osmosis to a maximum conductivity of 0.05 mS / cm) as the diafiltration medium. The pH was then adjusted by adding HCl to achieve a pH of approximately 5.5. Diafiltration was continued until the retentate conductivity declined below 0.1 mS / cm over 20 minutes. The permeate flow was then increased to 1.43 L / hr / m 2 The retentate was concentrated until the concentration was less than 0.05g. A first sample of the concentrated retentate was taken and centrifuged at 3000g for 5 minutes. The supernatant of the first sample was used to determine the BLG yield.

[0555] Crystallization The concentrated retentate was transferred to a 300 L crystallization tank and seeded with pure BLG crystal material made from rehydrated and spray-dried BLG crystals. The seeded whey protein solution was then cooled from 20° C. to approximately 6° C. over approximately 10 hours to allow the BLG crystals to form and grow.

[0556] After cooling, a sample of the crystal-containing whey protein solution (the second sample) was taken, and the BLG crystals were separated by centrifugation at 3000 g for 5 minutes. The supernatant and crystal pellet from the second sample were subjected to HPLC analysis as described below. The crystallization yield was calculated as outlined below and determined to be 57%.

[0557] [Table A]

[0558] Determination of BLG yield using HPLC: The supernatants of the first and second samples were diluted to the same extent by adding polishing water, and the diluted supernatants were filtered through a 0.22 μm filter. The same volume of each filtered and diluted supernatant was loaded onto an HPLC system equipped with a Phenomenex Jupiter® 5 μm C4 300 Å LC column 250 × 4.6 mm, Ea., and detected at 214 nm.

[0559] Samples were run using the following conditions: Buffer A: MilliQ water, 0.1%w / w TFA Buffer B: HPLC grade acetonitrile, 0.085% w / w TFA Flow rate: 1mL / min Column temperature: 40℃ Gradient: 0–30 min 82–55% A and 18–45% B; 30–32 min 55–10% A and 45–90% B; 32.5–37.5 min 10% A and 90% B; 38–48 min 10–82% A and 90–18% B.

[0560] Data Processing: Because both supernatants were processed in the same way, the areas of the BLG peaks can be directly compared to calculate the relative yield. Because the crystals contain only BLG and the samples were all processed in the same way, the concentration of alpha-lactalbumin (ALA), and therefore the area of ​​ALA, should be the same for all samples. Therefore, the area of ​​ALA before and after crystallization is used as a correction factor (cf) when calculating the relative yield.

number

[0561] The relative yield is calculated by the following formula:

number

[0562] Acid dissolution of BLC crystals Prior to separation, the feed was mixed 1:2 with polishing water and the remainder of the material from the crystallization tank was separated using a decanter at 350g, 2750 RPM, 150 RPM Diff., using a 64 spacer and a feed flow of 75 L / hr. The BLG crystals / solid phase from the decanter was then mixed with polishing water to form a thinner slurry, after which phosphoric acid was added to lower the pH to approximately 3.0 to rapidly dissolve the crystals.

[0563] After dissolving the BLG crystals, the pure BLG protein liquid was concentrated to 15 Brix using the same UF settings used to prepare the feedstock for crystallization, and the pH was adjusted to a final pH of approximately 3.8. The liquid BLG isolate was then heated to 75°C for 5 minutes and then cooled to 10°C. Heat treatment was found to reduce the microbial load from 137,000 CFU / g before heat treatment to less than 1,000 CFU / g after heat treatment. Heat treatment did not cause protein denaturation, and the intrinsic tryptophan fluorescence ratio (I330nm / I350nm) was determined to be 1.20, indicating the native conformation of the BLG molecule.

[0564] The BLG was dried in a pilot plant spray dryer with an inlet temperature of 180°C and an outlet temperature of 75°C. The resulting powder, sampled at the outlet, had a moisture content of approximately 4% w / w, and the chemical composition of the powder is shown in Table 1. A sample of the dried powder was dissolved and the degree of protein denaturation was determined to be 1.5%, and the intrinsic tryptophan fluorescence ratio (I330 / I350) was measured to be 1.20.

[0565] [Table B]

[0566] The bulk density (625 tap) of the spray-dried powder is 0.2-0.3 g / cm 3 It was estimated that:

[0567] Example 3: Preparation of a typical whey protein drink Disperse dried BLG isolate protein powder containing 85% or more BLG on a protein basis in approximately 75% demineralized water required to reach the desired final protein concentration.

[0568] Acidic BLG isolate powder is prepared as outlined in Example 2 and pH 5.5 BLG isolate powder is prepared as outlined in Example 7 of PCT / EP2017 / 084553.

[0569] As described in PCT / EP2017 / 084553, dissolution of the BLG material can be facilitated by the addition of an acid (selected from one or more food-grade acids such as phosphoric acid, hydrochloric acid, citric acid, malic acid, or dissolved or powdered salts thereof). If the addition of an acid reduces the pH during dissolution, the pH should preferably not exceed the desired target pH (i.e., avoiding unnecessary titration with acid and / or base).

[0570] Optionally, other ingredients may be added, including minerals, sweeteners, flavors, stabilizers, emulsifiers, or sources of fat and carbohydrates.

[0571] Adjust to final pH using 10% phosphoric acid (or other food grade acid) or 10% NaOH.

[0572] The remaining water is added to reach the desired protein concentration and the composition is optionally homogenized.

[0573] For comparison, whey protein isolate replaces 85% or more of the BLG product in the preparation of the reference sample, while maintaining the remaining steps.

[0574] The samples were stored at 20°C in a dark environment.

[0575] Example 4: Heat Treatment of Whey Protein Compositions The beverages were heat treated using a plate heat exchanger (manufacturer: OMVE HTST / UHT Pilot Plant HT320-20) equipped with a 10 μm bonded microfiber filter element, code 12-57-60k (Headline filter), by heating to 120°C for 20 seconds (high temperature, short time (HTST), resulting in denaturation of BLG) or at 75°C for a holding time of 15 seconds to 5 minutes (BLG remains native). Other heat treatment conditions may also be applied.

[0576] The heat-treated beverage composition was tapped in a 100 mL sterilized bottle at 75 to 85°C, then immediately sealed and placed on ice.

[0577] In other experiments, the whey protein source was heat-treated by transferring it to a thin-walled glass vial containing 15-30 mL of sample. The vial was immersed in a water bath pre-equilibrated at a target temperature ranging from 75°C to 95°C for 1-5 minutes, followed by cooling on ice.

[0578] Example 5: Production of a heat-treated beverage preparation In this example, a BLG beverage and a WPI beverage were prepared containing 6% protein and having a pH of 3.7.

[0579] The BLG beverage was prepared by dissolving pH 5.5 BLG isolate powder (described in Example 7 of PCT / EP2017 / 084553) in demineralized water at 10°C. 10% H3PO4 was slowly added to the solution. The final pH was adjusted to pH 3.7.

[0580] The solution was heat-treated at 120°C for 20 seconds using a plate heat exchanger as described in Example 4, or at 75°C with a holding time of 15 seconds to 5 minutes. The beverage was tapped to obtain a heat-sterilized whey protein beverage composition.

[0581] The same procedure was used to prepare a WPI beverage from the WPI powder.

[0582] Table 1 below shows the composition of the BLG powder used in preparing the beverage formulations, and also lists the composition of the WPI for comparison.

[0583] [Table 1]

[0584] A beverage preparation containing BLG and WPI with a pH of 3.7 and a protein content of 6% w / w was heat treated at 120°C for 20 seconds and 75°C for 15 seconds, with 95.9 w / w% of the protein being BLG. In the WPI beverage (WPI-B), 57 w / w% of the protein was BLG. The different samples were analyzed for turbidity (Example 1.7), viscosity (Example 1.8), and color (Example 1.9).

[0585] The results are presented in Table 2 below and in FIG. [Table 2]

[0586] Conclusion: The turbidity of the BLG sample remained low at 75° C., while the turbidity of the WPI sample was high. The WPI sample was also opaque (see Figure 1).

[0587] The sterile BLG sample had a turbidity of 7.0 NTU compared to the WPI which had a turbidity of 263 NTU.

[0588] The viscosity also remained low.

[0589] Thus, it is possible to produce a clear beverage with a BLG content of approximately 96 w / w% of the protein content at pH 3.7, which is not possible with the WPI sample that became opaque under the same conditions.

[0590] Example 6: Demonstration that the usable pH range of a clear whey protein beverage can be extended A BLG sample was prepared in which approximately 92 w / w% of the 6 w / w% protein was BLG, and for comparison, two different WPI samples were prepared containing approximately 60 w / w% (WPI-A) and 57 w / w% (WPI-B) BLG, respectively.

[0591] A 6 w / w% whey protein composition was prepared as described in Example 3 (BLG isolate powder was produced according to Example 2) and the final pH was adjusted using 10% phosphoric acid to obtain selected pH values ​​of 3.0 to 3.9, respectively. In one aspect of the experiment, samples adjusted to pH levels of 3.0 to 3.9 were UHT treated at 120°C for 20 seconds, tapped, sealed, and cooled. In another aspect of the experiment, samples at pH 3.0 and 3.9 were pasteurized at 75°C for 15 seconds, as described in Example 4.

[0592] The different samples were analyzed for turbidity (Example 1.7), viscosity (Example 1.8), color (Example 1.9) and appearance (Example 1.12).

[0593] The results are presented in Figures 2 to 10.

[0594] result: Figure 2 shows images of a pH 3.0-3.7 WPI-B and a pH 3.7 BLG beverage that were heat-treated at 120°C for 20 seconds. Figure 3 shows images of a pH 3.0-3.7 WPI-B and a pH 3.7 BLG beverage that were heat-treated at 75°C for 15 seconds. Figure 4 shows images of a pH 3.7 WPI-B and a pH 3.9 BLG beverage that were heat-treated at 75°C for 15 seconds.

[0595] Surprisingly, the inventors found that BLG beverage preparations remain clear even at pH 3.7 when UHT sterilized (FIG. 2), and can exceed pH 3.7 (pH 3.9-4.1) when pasteurized (FIGS. 3 and 4), under circumstances where the WPI is opaque. These findings are further supported by turbidity measurements shown in FIG. 5 (UHT) and FIG. 6 (pasteurized), which remained below 40 NTU even at pH 3.7 and 3.9, respectively, where the WPI significantly exceeds 40 NTU.

[0596] Upon UHT treatment of the BLG beverage preparation, the viscosity remains low. The low viscosity demonstrates that the beverage sample is easily drinkable. The viscosity increases dramatically with the use of WPI, especially at higher pH values ​​(Figure 7).

[0597] The authors further investigated the yellowness (b * values) significantly exceeded BLG up to at least pH 3.7. See Figure 8 (UHT) and Figure 9 (pasteurized).

[0598] Conclusion: The use of whey protein beverages in which at least 85% w / w of the protein is BLG enables at least two significant opportunities to provide whey protein beverages with desirable attributes to consumers: 1. Increasing the pH during heat treatment improves visual perception (color, turbidity) and viscosity compared to WPI. 2. Enables pasteurization that maintains the benefits of 1) while further expanding the available pH range.

[0599] Example 7: Preparation of heat-sterilized high-protein beverage using BLG A BLG sample was prepared in which approximately 92 w / w% of the protein was BLG (0.42 w / w% was ALA), and for comparison, a WPI sample was prepared using WPI-A, in which approximately 60 w / w% of the protein was BLG (8 w / w% was ALA) and the pH of the WPI powder was 3.3.

[0600] BLG isolate powder product (from Example 2, powder pH 3.9) was dispersed in tap water to produce beverages with protein concentrations ranging from 6.0 to 30.0 w / w% and adjusted to pH 3.7 using 10% phosphoric acid.

[0601] The solutions were thermally treated at 75-120°C for durations of 15 seconds to 5 minutes as described in Table 3 and immediately cooled on ice.

[0602] The different samples were analyzed for viscosity (Example 1.8), protein nativeity determined as the intrinsic tryptophan fluorescence emission ratio R=I330 / I350 (Example 1.1), appearance (Example 1.12) and turbidity (Example 1.7).

[0603] [Table 3]

[0604] result: The results are shown in Table 3 above and in FIGS.

[0605] Figure 10 shows images of a 15 w / w% BLG beverage at pH 3.7 heated at 75°C / 15 seconds that was clear and transparent (left), while a 6% WPI-A beverage at pH 3.7 heated at 75°C / 15 seconds (right) was opaque.

[0606] FIG. 11 shows the sensory evaluation of the high protein BLG beverage compositions and images of the 6 w / w% and 15 w / w% BLG samples at pH 3.7, both of which are clear.

[0607] Figure 12 shows high-protein beverage preparations prepared by heating BLG beverages with protein contents of 30 w / w%, 27.5 w / w%, 25 w / w%, and 20 w / w% (from left to right) at 75°C for 5 minutes; all samples had low viscosity and were liquid.

[0608] We surprisingly found that all solutions remained low viscosity even when heated at 75°C for up to 5 minutes, suggesting little or no denaturation.

[0609] The observed viscosity at high protein concentrations, typical of non-aggregated native proteins (flow behavior described by Inthavong, Kharlamova, Nicolai, Chassenieux, & Nicolai, 2016), is approximately 10 cP at 200 g / l.

[0610] Tryptophan fluorescence spectroscopy confirmed that BLG remains in the native conformation when gently heated (75 °C), as evidenced by an intrinsic tryptophan emission ratio (I330 / I350) of at least 1.11, whereas more severe heating causes denaturation as indicated by an intrinsic tryptophan emission ratio (I330 / I350) of less than 1.11.

[0611] RP-HPLC analysis confirmed the tryptophan fluorescence results revealing 3.6% denaturation of the 6% BLG beverage heated at 75°C for 5 min and 41% denaturation when heated at 95°C for 5 min.

[0612] It was shown that the viscosity remained low even after heating.

[0613] It was found that BLG beverage preparations can be heated above their denaturation temperature. However, when heated at 95°C for 5 min, gelation occurred in BLG beverages containing more than 16 w / w% protein, while 10% remained liquid at 90°C for 5 min and 6% remained liquid at 120°C for 15 s. At least partial denaturation / aggregation occurred under these heating conditions, as evidenced by a decrease in the intrinsic tryptophan emission ratio (I330 / I350).

[0614] Much to the inventors' surprise, a sensory panel (see Example 1.11 and Figure 11 for analysis) did not identify any significant differences in the dry texture of 6% and 15% BLG beverage preparations heated at 75°C, clearly suggesting the use of high protein beverages for consumers who have difficulty swallowing, for example.

[0615] Example 8: Improved Taste Whey Protein Beverage Preparation BLG and WPI samples were prepared with the following compositions:

[0616] The BLG isolate powder used is prepared according to Example 2. [Table 3-2]

[0617] The samples were analyzed by a 10-person sensory panel (see Example 1.11). The WPI samples were yellower and had a higher b than the BLG beverage, especially at higher pH values. * The analytical data are presented in Table 3.

[0618] [Table 4]

[0619] The visual appearance of the samples in Table 4 is shown in FIG.

[0620] The sensory evaluation data are shown in FIGS.

[0621] Delta b * To calculate, use the following formula: Delta b * = measured at room temperature, b 6.0w / w%タンパク質に標準化された試料 * -b 脱塩水 * Delta A * To calculate, use the following formula: Delta A * = measured at room temperature, a 6.0w / w%タンパク質に標準化された試料 * -a 脱塩水 * Delta L * To calculate, use the following formula: Delta L * = measured at room temperature, L 6.0w / w%タンパク質に標準化された試料 * -L 脱塩水 * The color values ​​of demineralized water are: L * =39.97, a * =0 and b * =-0.22.

[0622] result: By taking advantage of the opportunity to increase the pH and decrease the heating temperature while maintaining clarity and colorlessness, significant differences were observed in the taste of beverages made with WPI-A and BLG. The BLG beverage had lower astringency, dry texture, sourness, whey aroma, and citric acid flavor compared to the WPI beverage, as shown in Figure 14.

[0623] Figure 15 shows that increasing the pH to 3.7 before heat treatment reduces the acidity of the BLG beverage at both 120°C and 75°C while maintaining the clarity and low color of the product. As seen in Table 2 and Figure 1, this was not possible with WPI, as a pH of 3.7 does not produce a clear, crisp beverage.

[0624] Figure 16 demonstrates that changing both temperature and pH from pH 3.0, 120°C / 20 sec to pH 3.7, 75°C / 15 sec significantly reduces astringency.

[0625] Figure 17 demonstrates that reducing the heating temperature from 120°C / 20 seconds to 75°C / 15 seconds significantly reduces dry texture (native at 75°C vs. denatured protein at 120°C).

[0626] FIG. 18 demonstrates that maintaining BLG in its native state by using 75°C / 15 seconds heating at pH 3.7, which fails to produce a clear, crisp, colorless WPI beverage, reduces whey aroma.

[0627] WPI heat treated at 75°C / 15 seconds at pH 3.7 failed to produce a clear beverage. See also Figure 3.

[0628] Example 9: Low Color Sweet BLG Beverage Preparation A 6% w / w BLG beverage was prepared. See BLG powder composition below. The beverage was prepared as described in Example 3. [Table 4-2]

[0629] The prepared BLG drinks contained 6% protein and had pH 3.7 and pH 4.3.

[0630] 8% w / w sucrose was used as the carbohydrate sucrose. Tests were also conducted using the high-intensity sweetener sucralose. Samples were subjected to heat treatment at 93°C in a water bath for 4 minutes, then cooled in an ice bath.

[0631] The different samples were analyzed for clarity (Example 1.12), color (Example 1.9), turbidity (Example 1.7) and viscosity (Example 1.8).

[0632] The results are presented in Table 4 below.

[0633] [Table 5]

[0634] result: It was found that sweetened BLG beverages could be produced by using 8% sucrose as the sweetener and subjecting them to a heat treatment at 93° C. for 4 minutes. The addition of sucrose had only a small effect on viscosity, turbidity, and clarity (see Table 5), and the color was not affected by the addition of sucrose.

[0635] A commercially available beverage, e.g., a BLG beverage containing additives typically found in sports nutrition, was prepared, heat treated at 75°C for 5 minutes, 6% w / w protein BLG beverage at pH 3.7°C. See Table 5 below.

[0636] [Table 6] [Table 7]

[0637] result: It can be seen in Table 7 that both the additive-containing and additive-free BLG beverages remained low viscosity, clear, and essentially colorless.

[0638] Example 10: Exemplary Method for Preparation of Clear BLG Beverage with Added Minerals The BLG powder used in this example had a pH of 5.5 and contained approximately 96% w / w of the protein as BLG (and 0.4% w / w of the protein as ALA).

[0639] An acidic BLG isolate powder was prepared according to Example 2 and a beverage preparation was prepared according to Example 5.

[0640] High temperature heat treatment of beverage preparations: A 6% BLG beverage preparation was prepared with a pH of 3.7. KCl and CaCl were added in liquid form from 1 M stock solutions. These were heat treated at less than 95°C for 5 minutes.

[0641] result: The results are summarized in Table 8 below and in FIG.

[0642] FIG. 19 shows an image of a 6% BLG beverage with pH 3.7, heat treated at 95° C. for 5 minutes, and with added minerals. A: 0mM added minerals B: Added CaCl2 15mM C: Added KCl 20 mM D: Added KCl 10 mM and added CaCl2 15 mM

[0643] The turbidity of BLG beverage preparations spiked with minerals (0-20 mM KCl, 0-15 mM CaCl2, or 10 mM CaCl2 and 10 mM) remained below 30 NTU when heated at 95°C for 5 min at pH 3.7.

[0644] Gelation was observed at 30 mM added KCl (cloudy gel).

[0645] Gelation was observed at 20 mM added CaCl2 (clear gel).

[0646] The results clearly suggest that protein composition is more important than mineral differences relative to WPI, as the mineral additions in Table 8 significantly exceed the differences between the BLG and WPI products. The sample remained clear (see Figure 19) and had a low viscosity within the limits of Table 8 below.

[0647] [Table 8]

[0648] Low-temperature heat treatment of beverage preparations A 6% BLG beverage formulation with a pH of 3.7 was prepared. KCl and CaCl were added in liquid form from 1 M stock solutions. These were heat treated at a pasteurization temperature of 75°C for 5 minutes.

[0649] result. The inventors have surprisingly found that using pasteurization temperatures (75°C, 5 minutes) allows for exceptionally high mineral concentrations, see Table 9 below.

[0650] FIG. 20 shows an image of a 6% BLG beverage with pH 3.7 that was heat treated at 75° C. for 5 minutes and had minerals added. A: Added minerals 0mM; B: Added KCl 100 mM; C: Added CaCl2 100 mM, D: Added KCl 100 mM and added CaCl2 100 mM

[0651] When 100 mM KCl or 100 mM CaCl was added to the beverage composition before heating, the beverage preparation remained clear. See Figure 20. Furthermore, the viscosity was surprisingly low when both 100 mM KCl and 100 mM CaCl were added.

[0652] [Table 9]

[0653] Example 11: Dairy whey protein beverage, high temperature heat treatment An exemplary method for producing an opaque dairy beverage containing BLG and optionally a carbohydrate source is shown in Figure 1. The BLG powder is dissolved in tap water, adjusted to a pH according to Example 3, and thermally treated at 93°C for 4 minutes. The BLG beverages contain approximately 92% w / w of the protein as BLG and approximately 0.42% w / w of the protein as ALA, and are produced based on an acidic BLG isolate powder having a pH of 3.9 (Example 2).

[0654] A 6% BLG beverage was prepared with a pH of 4.3. 8% sucrose was added as a carbohydrate source. Turbidity, viscosity, color, and clarity were measured according to the procedures described in Examples 1.7, 1.8, and 1.9, as well as beverage stability as in Example 1.10.

[0655] The results are presented in Tables 10 and 11 below and in FIG.

[0656] [Table 10]

[0657] [Table 11]

[0658] WPI samples containing 6% protein and having a pH of 4.3 were prepared. The WPI samples were thermally treated at 94°C for 5 minutes. 0% or 8% sucrose was added to WPI-A samples, and 0% or 6% sucrose was added to WPI-B samples. [Table 11-2]

[0659] result: Figure 21 shows the stability of a pH 4.3 dairy BLG beverage with and without added sucrose, heat-treated at 93°C for 4 minutes. A: 0% sucrose (before centrifugation), B: 8% sucrose (before centrifugation), C: 0% sucrose (after centrifugation), D: 8% sucrose (after centrifugation).

[0660] The results presented in Tables 10 and 11 and Figure 21 demonstrate that a high-end pH, such as pH 4.3, allows for the production of dairy-based beverages, which may be preferred in some embodiments of the present invention, for example, when consumers prefer whey protein beverages with a milky appearance. Also, at pH 4.3, low viscosity was found for both formulations with and without sucrose.

[0661] The color also remained neutral, which is especially appreciated by consumers who prefer their dairy drinks not to have a yellowish color. * At higher values, a yellowish color is seen.

[0662] After centrifugation at 3000 × g for 5 min, the beverage was found to be stable as evidenced by less than 15% protein loss and high turbidity.

[0663] Dairy 6 w / w% protein WPI beverages based on WPI-A or WPI-B at pH 4.3 could not be produced due to gelation and high viscosity, which was true for both WPI samples with and without added sucrose.

[0664] Example 12: Dairy whey protein beverage, long-term low-temperature heat treatment Exemplary Method for Producing Dairy Drinks Containing BLG at Different pHs: BLG powder is dissolved in tap water and adjusted to pH 4.2-4.5 using 10% phosphoric acid according to Example 3. The preparation was thermally treated at 75°C for 5 minutes and had a protein content of 6% w / w. The BLG drink contains approximately 92% w / w of the protein as BLG and approximately 0.42% w / w of the protein as ALA, and is produced based on BLG powder with a pH of 3.9.

[0665] Turbidity, viscosity, color and visual clarity were measured according to the procedures described in Examples 1.7, 1.8, 1.9 and 1.12.

[0666] The results are presented in Table 12 below and in FIG.

[0667] Figure 22 shows images of an opaque 6% protein BLG beverage prepared by heating at 75°C for 5 minutes at pH 4.2-4.5.

[0668] [Table 12]

[0669] result: The beverages at pH 4.2-4.5 were found to have a milky, opaque appearance and high turbidity, while still having low viscosity.

[0670] Example 13: Colorless whey protein beverage containing greater than 85% bLG A beverage preparation was prepared in which approximately 92% w / w of the protein was BLG and approximately 0.42% w / w of the protein was ALA (the pH of the BLG isolate powder was 3.9). See Example 3.

[0671] For comparison, a whey protein sample containing SPI (serum protein isolate) containing approximately 80% w / w BLG and approximately 4% w / w ALA was prepared (the pH of the SPI powder was 6.7).

[0672] The sample had a protein content of 6% w / w.

[0673] The pH of the beverage was adjusted to pH 3.7.

[0674] The turbidity, viscosity, color and clarity of the formulations were measured according to the procedures described in Examples 1.7, 1.8, 1.9, as well as the beverage stability as in Example 1.10.

[0675] The results are presented in Table 13 below and in Figures 23 and 24.

[0676] [Table 13]

[0677] result: The viscosity of SPI (approximately 80% BLG, approximately 4% ALA) was found to be further increased by heat treatment compared to the BLG preparation at pH 3.7.

[0678] Furthermore, the SPI beverage had a higher b *It had a high value and therefore a yellowish color.

[0679] Example 14: Nutritional whey protein beverage containing 85% or more BLG, a carbohydrate source, and a fat source Example 14 describes an exemplary method for preparing a heat-sterilized beverage preparation in which at least 85% w / w of the protein is BLG.

[0680] The inventors have surprisingly found that BLG beverages (85% or greater) tolerate surprisingly high mineral concentrations present during sterilization by pasteurization at 75°C with a maximum hold time of at least 5 minutes (Example 10), as a 6% nutritional composition containing 100 mM added KCl and 100 mM added CaCl remained liquid (viscosity approximately 1 cP) even after heating at 75°C for 5 minutes.

[0681] Because the heat stability of whey proteins is typically compromised at high mineral dosages, the inventors further investigated the opportunity to produce a nutritionally complete, acidified BLG beverage to produce a sterile nutritional beverage containing 85% or more BLG, a carbohydrate source, a fat source, and minerals in a combination that meets current FSMP (Food for Special Medical Purposes) requirements.

[0682] The protein is dissolved and mixed with lipids and carbohydrates in example ratios based on the energy distribution listed in Table 14.

[0683] Food-grade acids and minerals were selected to meet the requirements set out for Foods for Special Medical Purposes (FSMP).

[0684] Vitamins may also be provided in the beverage to meet FSMP requirements and produce a nutritionally complete dietary supplement.

[0685] [Table 14]

[0686] A 6 w / w% BLG nutritional drink, also containing 13.5 w / w% sucrose and 4.7 w / w% rapeseed oil, was mixed at 70° C. The protein, fat and carbohydrate composition was selected to correspond to medical nutrition recommendations.

[0687] In certain embodiments, (1) 40 mM KCl and 14 mM CaCl, or (2) 80 mM KCl and 28 mM CaCl, were added with additional ingredients or (3) without further mineral addition as shown in Table 14.

[0688] The solution was homogenized at 200 bar.

[0689] The solutions were thermally treated by immersion in a 75°C or 95°C water bath for 5 minutes and cooled on ice.

[0690] [Table 15]

[0691] result: It was found that BLG can be used in combination with a fat source and a carbohydrate source to produce an opaque beverage by heating at 75°C and 95°C.

[0692] At 75°C, it remains in its native state (despite the fat content, it had a Trp fluorescence ratio of 1.18), but at 95°C it undergoes denaturation (Trp fluorescence). The viscosity remains low. Maintaining the native conformation allows for the administration of minerals important for medical nutrition (FSMP requirement). Furthermore, the ability of the nutritional composition to remain liquid in the presence of selected minerals clearly suggests its feasibility for use in medical nutrition.

[0693] Example 15: Low-phosphorus protein drink The purified BLG product from Example 3 (crystal preparation obtained from Feedstock 3) is used to prepare four low phosphorus beverage samples. All dry ingredients are mixed with demineralized water to give 10 kg of each sample and allowed to hydrate at 10° C. for 1 hour. [Table 15-2]

[0694] The sample is subjected to 90°C for 180 seconds and aseptically filled into sterile containers.

[0695] The packaged beverage has a shelf life of at least one year at ambient temperature.

[0696] Because all of the ingredients used to prepare the five drinks are low in phosphorus, the resulting drinks have phosphorus contents well below 80 mg / 100 g protein, making them suitable for use as protein drinks for patients with kidney disease.

Claims

1. 1. A packaged thermally treated beverage preparation having a pH in the range of 2 to 4.7, comprising: - a total amount of 4-45% w / w of proteins relative to the weight of the heat-treated beverage preparation, at least 90% w / w of which is beta-lactoglobulin (BLG); - Optionally, sweeteners, sugar polymers and / or flavors.

2. 10. The packaged thermally treated beverage preparation of claim 1, which is at least pasteurized.

3. 10. The packaged thermally treated beverage preparation of claim 1, which is sterile.

4. A packaged heat-treated beverage preparation as described in claim 1 or 3, wherein the protein fraction of the heat-treated beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.

11.

5. A packaged heat-treated beverage preparation described in any of claims 1 to 3, wherein the protein fraction of the heat-treated beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of less than 1.

11.

6. 6. The packaged heat-treated beverage preparation according to any one of claims 1 to 5, having a degree of protein denaturation of up to 10%.

7. A packaged heat-treated beverage preparation according to any one of claims 1 to 6, having a pH in the range of 3.0 to 4.

3.

8. The protein fraction of the heat-treated beverage preparation has a color value delta b in the range of −0.10 to +0.51 on the CIELAB color scale. ＊ and delta b ＊ = measured at room temperature, b ６．０ｗ／ｗ％タンパク質に標準化された試料 ＊ -b 脱塩水 ＊ 8. The packaged heat-treated beverage preparation according to any one of claims 1 to 7, wherein

9. Color value delta b ranging from -0.10 to +0.51 on the CIELAB color scale ＊ and delta b ＊ = measured at room temperature, b ６．０ｗ／ｗ％タンパク質に標準化された試料 ＊ -b 脱塩水 ＊ 9. The packaged heat-treated beverage preparation according to any one of claims 1 to 8, wherein

10. 10. The packaged heat-treated beverage preparation according to any one of claims 1 to 9, wherein the sum of the amounts of Na, K, Mg and Ca is at most 750 mM.

11. A packaged heat-treated beverage preparation according to any one of claims 1 to 10, having a turbidity of at most 200 NTU.

12. A packaged heat-treated beverage preparation according to any one of claims 1 to 10, having a turbidity of more than 200 NTU.

13. A packaged heat-treated beverage preparation according to any one of claims 1 to 12, wherein the protein fraction contains at most 15% insoluble material after centrifugation at 3000g for 5 minutes.

14. 14. The packaged heat-treated beverage preparation of any one of claims 1 to 10, 12 and 13, having a viscosity of up to 200 cP centipoise measured at 22 degrees Celsius at a shear rate of 100 / sec.

15. A packaged heat-treated beverage preparation according to any one of claims 1 to 14, which does not contain any anti-agglomeration agent.

16. 16. A packaged heat-treated beverage preparation according to any preceding claim, comprising a total amount of protein of 4.0 to 30% w / w by weight of the beverage.

17. A packaged heat-treated beverage preparation as described in any one of claims 1 to 16, further comprising carbohydrates in the range of 0 to 95% of the total energy content of the heat-treated beverage preparation.

18. A packaged heat-treated beverage preparation as described in any one of claims 1 to 17, further having a lipid content of 0 to 60% of the total energy content of the heat-treated beverage preparation.

19. Having carbohydrates in the range of 30-60% of the total energy content of the heat-treated beverage preparation, A packaged thermally treated beverage preparation according to any one of claims 1 to 18, having a lipid content of 20 to 50% of the total energy content.

20. A packaged thermally treated beverage preparation according to any preceding claim, comprising a BLG isolate.

21. A packaged thermally treated beverage preparation according to any preceding claim, wherein at least 92% w / w is beta-lactoglobulin (BLG).

22. 1. A method for producing a packaged thermally treated beverage preparation having a pH in the range of 2 to 4.7, comprising the steps of: a) - 2 to 45% by weight of a total amount of proteins, of which at least 90% w / w is BLG; optionally sweeteners, sugar polymers and / or flavors, providing a liquid solution comprising: b) packaging the liquid solution; The liquid solution of step a) and / or the packaged liquid solution of step b) are subjected to a heat treatment including at least pasteurization.

23. 22. A packaged thermally treated beverage preparation according to any one of claims 1 to 21 for use in a method for treating a disease associated with protein malabsorption.

24. Use of a packaged heat-treated beverage preparation according to any one of claims 1 to 21 as a dietary supplement.

25. 25. Use of the packaged thermally treated beverage preparation of claim 24, taken before, during or after exercise.

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