Ph neutral β-lactoglobulin beverage preparation

Abstract

To provide a packaged pH neutral heat-treated beverage preparation which is more achromatic than conventional whey-containing beverages.SOLUTION: A packaged, heat-treated beverage preparation having a pH in the range of 5.5-8.0 comprises: a total amount of protein of 1 to 20% w / w based on the weight of the heat-treated beverage preparation, where at least 85% w / w of the protein is β-lactoglobulin (BLG); and optionally, sweetener and / or flavor. The heat-treated beverage preparation is at least pasteurized and is at least sterile.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a novel packaged heat treated beverage preparation having a pH in the range of 5.5 to 8.0.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] Beverages developed for sports nutrition are particularly likely to contain whey protein, which is incorporated for its unique nutritional benefits to athletes. Some medical and therapeutic nutritional beverages also contain whey protein, due to its rich content of amino acids essential for protein synthesis, digestibility, and health benefits.

[0003] Whey proteins can be isolated from whey or whey. Whey typically contains a mixture of beta-lactoglobulin (BLG), alpha-lactalbumin (ALA), serum albumin, and immunoglobulins, with BLG being the most prevalent. Thus, whey protein concentrate (WPC) contains a mixture of these proteins. Whey protein isolate (WPI) is lower in fat and lactose than WPC. Whey products can have a yellow color, and therefore many attempts have been made in the past to eliminate or reduce the yellow color of whey products.

[0004] The traditional method of whitening / bleaching whey is to chemically bleach it using hydrogen peroxide (HP, H2O2), which can have a negative impact on taste and promote whey protein unfolding and aggregation (Kramer et al, 2017. "Effect of Oxidation and Protein Unfolding on Cross-Linking of beta-Lactoglobulin and alfa-Lactalbumin", J. Agric. Food Chem. 2017, 65, 10258-10269.

[0005] WO 2005 / 004616 A1 describes a method for bleaching or whitening dairy products, which comprises adding lipoxygenase (LOX) to the dairy product. This method can be used to whiten whey and dairy products.

[0006] Other methods to whiten whey include the addition of chlorophyll to dairy products or the use of titanium dioxide (TiO2), an inorganic inert white pigment used in cheese milk, candy, chewing gum, toothpaste, etc., and approved by the FDA as food grade.

[0007] WO 2018 / 115520 A1 discloses a method for producing an edible isolated β-lactoglobulin composition and / or a composition comprising crystallized β-lactoglobulin based on the crystallization of BLG in a salt-soluble form, after which the crystallized BLG can be separated from the remaining mother liquor.

[0008] WO 2011 / 112695 A1 discloses a nutritional composition and methods of making and using the same. The nutritional composition comprises whey protein micelles and leucine, providing a sufficient amount of leucine to improve protein synthesis in humans while maintaining a low viscosity fluid matrix and acceptable organoleptic properties.

[0009] WO 2011 / 051436 A1 discloses at least partially transparent compositions intended for human or animal consumption and relates to packaging of such compositions. One embodiment of the present invention relates to an at least partially transparent container comprising an at least partially transparent aqueous non-alcoholic composition. The container comprises at least one polarizer that renders the liquid crystals present in the composition visible.

[0010] WO 2004 / 049819 A2 discloses a method for improving the functional properties of globular proteins, comprising the steps of providing a solution of one or more globular proteins, in which the protein(s) are at least partially aggregated into fibrils, and performing one or more of the following steps in random order: increasing the pH; increasing the salt concentration; concentrating the solution; modifying the solvent quality of the solution. Preferably, the solution of one or more globular proteins is provided by heating at low pH or by adding a denaturing agent. The protein additive thus obtained, its use in food and non-food products, as well as food and non-food products containing said protein additive are also disclosed.

[0011] WO 2010 / 037736 A1 discloses the isolation of whey proteins and the preparation of whey products and whey isolates. In particular, the invention relates to the isolation of β-lactoglobulin products and the isolation of α-rich whey protein isolates from whey obtained from animals. The α-rich whey protein isolates provided by the invention are high in alpha-lactalbumin and immunoglobulin G in addition to being low in β-lactoglobulin.

[0012] French Patent No. 2 296 428 discloses protein compositions for nutritional and therapeutic use based on whey proteins obtained by any known separation process. The compositions can be used to treat or prevent digestive disorders (e.g. diarrhea) in infants and adults, to increase resistance to intestinal infections, and to treat certain metabolic disorders (e.g. hyperphenylalaninemia). They can also be used dermatologically or cosmetically and can form part of a low-protein diet. Summary of the Invention

[0013] The inventors have observed that the degree to which a whey protein-containing beverage is neutral or white in color affects consumer perception of the whey protein-containing beverage: a yellowish clear beverage or a yellowish milky beverage is not appealing to consumers.

[0014] It is an object of the present invention to provide a packaged, pH neutral, heat treated beverage preparation that is more neutral in color than conventional whey-containing beverages.

[0015] Another objective is to utilize a gentler method for reducing the yellow color. A further objective is that it must not adversely affect the stability of the beverage.

[0016] The inventors have discovered that such packaged thermally treated beverages can be provided within a broad neutral pH range of 5.5-8.0, and within a broad protein concentration range of 1-20% by weight, while still having a low viscosity, being stable, and being more neutral in color. The present invention provides both beverages that are clear, and in other embodiments, beverages that are opaque.

[0017] Thus, one aspect of the invention is a packaged thermally treated beverage preparation having a pH in the range of 5.5 to 8.0, the beverage comprising: - a protein having a total amount of 1-20% w / w by weight of the beverage, wherein at least 85% w / w of the protein is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, The present invention relates to a beverage comprising:

[0018] Another aspect of the invention is a method for producing a packaged thermally treated beverage preparation having a pH in the range of 5.5 to 8.0, comprising the steps of: a) - 1 to 20% by weight of a total amount of protein, wherein at least 85% of the protein is beta-lactoglobulin (BLG); - optionally providing a liquid solution comprising a sweetener and / or a flavoring agent, b) packaging the liquid solution; Including, wherein the liquid solution of step a) and / or the packaged liquid solution of step b) are subjected to a heat treatment comprising at least pasteurization.

[0019] Yet another aspect of the invention relates to the use of a protein solution comprising a total amount of protein of 1-20% w / w by weight of the solution, at least 85 w / w% of the protein being beta-lactoglobulin (BLG), for controlling whiteness of a sterile beverage preparation having a pH in the range of 5.5-8.0.

[0020] Yet another aspect of the present invention relates to a packaged thermally treated beverage preparation as defined herein for use in a method for the treatment of a disease associated with protein deficiency.

[0021] A further aspect of the present invention relates to the use of the packaged thermally treated beverage preparation, defined herein as a dietary supplement. [Brief description of the drawings]

[0022] [Figure 1] Shown is an image of a milky white BLG sample heated at 94° C. for 14 minutes at pH 6.0. [Diagram 2] Shown is an image of a gelled WPI sample heated at 94° C. for 14 minutes at pH 6.0. [Diagram 3] FIG. 1 shows the amount of insoluble protein material, native whey protein, soluble whey protein aggregates and protein nanogels in exemplary whey protein beverages A (BLG 98.2 w / w%), B (BLG 95.9 w / w%) and C (WPI). [Figure 4] FIG. 1 shows semi-dynamic in vitro digestion of exemplary beverages A (BLG 98.2 w / w%), B (BLG 95.9 w / w%), and C (WPI). [Diagram 5]Shown are: Top: SDS-PAGE analysis of protein aliquots collected at selected time points (17.5–105 min) during semi-dynamic in vitro digestion of the samples. Bottom: pH trace during digestion of the samples. [Figure 6] Figure 1 shows a simulation of gastric acidification. Gel strength was measured during acidification of three beverages containing either predominantly soluble whey protein aggregates (Drinks A and WPI) or protein nanogel (Drink B). [Figure 7] Liquid beverages with 10-16 w / w% protein are shown after heat treatment at 90°C for 5 minutes. [Figure 8] Viscosity measurements are shown for 10-16 w / w% protein after heat treatment at 90°C for 5 minutes. [Figure 9] The particle size of heat-treated beverages containing 10 to 16 w / w% protein is shown. [Figure 10] On the left are 10 w / w% beverages at pH 6.0 and pH 7.0 using BLG powder A. On the right is a 12 w / w% beverage at pH 6.0 using BLG powder A (from Table 9). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] definition In the context of the present invention, the term "β-lactoglobulin" or "BLG" relates to β-lactoglobulin derived from a mammalian species, e.g. in native, unfolded and / or glycosylated form, including naturally occurring genetic variants. The 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 by hydrophobic interactions and / or covalent bonds.

[0024] BLG is the most predominant protein in bovine whey and milk serum and exists in several genetic variants, the major ones in bovine milk 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 fighting pathogens. No homologue of BLG is found in human breast milk.

[0025] 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, bovine BLG is primarily a dimer, but dissociates into monomers below approximately pH 3 while retaining its native state as determined using nuclear magnetic resonance spectroscopy. Conversely, BLG also occurs in tetrameric, octameric, and other multimeric aggregated forms under a variety of natural conditions.

[0026] In the context of the present invention, the term "non-aggregated β-lactoglobulin" or "non-aggregated BLG" also relates to β-lactoglobulin derived from a mammalian species, e.g. in native, unfolded and / or glycosylated form, including naturally occurring genetic variants. 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.

[0027] The percentage of non-aggregated BLG to total BLG was calculated (m 総BLG -m 非凝集性BLG ) / m 総BLG * 100% determined by m 総BLG is the concentration or amount of BLG determined according to Example 1.31, m 非凝集性BLG is the concentration or amount of non-aggregated BLG determined according to Example 1.6.

[0028] In the context of the present invention, the term "crystal" relates to a solid material whose components (such as atoms, molecules or ions) are arranged in a highly ordered microscopic structure forming a crystal lattice that extends in all directions.

[0029] In the context of the present invention, the term "BLG crystal" refers to a protein crystal comprising primarily non-aggregated, preferably native, BLG arranged in a highly ordered microscopic structure forming a crystal lattice extending in all directions. BLG crystals may be, for example, monolithic or polycrystalline, and may 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 edges or corners of intact crystals. See, for example, FIG. 18 of PCT Application No. PCT / EP2017 / 084553 for examples of many intact BLG crystals, and FIG. 13 of PCT Application No. PCT / EP2017 / 084553 for examples of BLG crystal fragments. In either case, the BLG crystals or crystal fragments can be visually identified using an optical microscope as a well-defined, compact, coherent structure. BLG crystals or crystal fragments are often at least partially transparent. Protein crystals are further known to be birefringent, and this optical property can be used to identify unknown particles with crystalline structure. On the other hand, non-crystalline BLG aggregates are poorly defined, opaque, and often appear as open or porous masses of irregular size.

[0030] In the context of the present invention, the term "crystallization" relates to the formation of protein crystals. Crystallization can, for example, occur spontaneously or can be initiated by the addition of crystallization seeds.

[0031] In the context of the present invention, the term "edible composition" relates to a composition that is safe for human consumption and for use as a food ingredient and does not contain problematic amounts of toxic components such as toluene or other undesirable organic solvents.

[0032] In the context of the present invention, the term "ALA" or "alpha-lactalbumin" refers to alpha-lactalbumin derived from a mammalian species, e.g. in native and / or glycosylated form, including naturally occurring genetic variants. The term further includes aggregated ALA and precipitated BLG. When referring to the amount of ALA, for example, the total amount of ALA, including aggregated ALA, is referred to. The total amount of ALA is determined according to Example 1.31. The term "aggregated ALA" refers to ALA that is typically at least partially unfolded and further typically aggregated with other denatured ALA molecules and / or other denatured whey proteins by hydrophobic interactions and / or covalent bonds.

[0033] 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, while β-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 the main structural differences with β-lactoglobulin is that ALA does not have any free thiol groups that could serve as the initiation point for the covalent aggregation reaction.

[0034] In the context of the present invention, the term "non-aggregated ALA" also relates to ALA derived from a mammalian species, e.g. in native, unfolded and / or glycosylated form, including naturally occurring genetic variants. However, the term does not include aggregated or precipitated ALA. The amount or concentration of non-aggregated BLG is determined according to Example 1.6.

[0035] The percentage of nonaggregated ALA to total ALA was calculated (m 総ALA -m 非凝集性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 非凝集性ALA is the concentration or amount of non-aggregated ALA determined according to Example 1.6.

[0036] 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, including naturally occurring genetic variants such as chymosin, by aspartic proteinases, e.g., in native and / or glycosylated form.

[0037] In the context of the present invention, the term "BLG isolate" refers to a composition comprising 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.

[0038] In the context of the present invention, the term "BLG isolate powder" relates to a BLG isolate in powder form, preferably a free-flowing powder.

[0039] In the context of the present invention, the term "BLG isolate" relates to a BLG isolate in liquid form, preferably in aqueous solution.

[0040] The term "whey" refers to the liquid phase remaining after the casein of milk has been precipitated and removed. Precipitation of casein can be achieved, for example, by acidification of the milk and / or by the use of rennet enzymes. There are several types of whey, 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.

[0041] The term "whey" refers to the liquid remaining when casein and milk fat globules are removed from milk, for example by microfiltration or large pore ultrafiltration. Whey is sometimes also referred to as "ideal whey".

[0042] The term "whey protein" or "serum protein" relates to proteins present in whey.

[0043] 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 it may be the complete set of protein species found in whey or / and whey.

[0044] In the context of the present invention, the major non-BLG proteins of a standard whey protein concentrate derived from sweet whey are ALA, CMP, bovine serum albumin, immunoglobulins, osteopontin, lactoferrin, and lactoperoxidase. In the context of the present invention, the major non-BLG whey proteins by weight percentage of a standard whey protein concentrate derived 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 the total protein; BSA in an amount of 4% w / w relative to total protein; Casein species in an amount of 5% w / w based on total protein, Immunoglobulin in an amount of 6% w / w relative to the total protein, osteopontin in an amount of 0.5% w / w based on total protein; Lactoferrin in an amount of 0.1% w / w based on total protein, and Lactoperoxidase in an amount of 0.1% w / w of total protein.

[0045] The term casein refers to the casein proteins found in milk and includes both the native micellar casein and the caseins found in raw milk, which are individual casein species.

[0046] In the context of the present invention, the term "mother liquor" refers to the whey protein solution remaining after BLG has been crystallized and the BLG crystals have been at least partially removed. The mother liquor may still contain some BLG crystals, but usually contains only small BLG crystals that have escaped separation.

[0047] In the context of the present invention, a liquid that is "supersaturated" or "supersaturated with respect to BLG" comprises a concentration of dissolved non-aggregated BLG that exceeds the saturation point of non-aggregated BLG in that liquid at 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, p. 2286-2300, Issue 7, 2014), and supersaturation can be determined by various measurement techniques (e.g., spectroscopy or particle size analysis). In the context of the present invention, supersaturation with respect to BLG is determined by the following procedure.

[0048] Procedure to test if a liquid is supersaturated with respect to BLG at a specific set of conditions: a) Transfer a 50 mL sample of the liquid to be tested into a centrifuge tube (VWR catalogue 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 keep the sample and its subsequent fractions in the original physical and chemical conditions of the liquid: b) The samples are 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 subsample of the supernatant (subsample A). e) Add 10 mg of BLG crystals with particle size up to 200 microns (at least 98% pure non-agglomerated BLG based on total solids) to the second centrifuge tube and vortex the mixture. f) Leave the second tube at the original temperature for 60 minutes. ) Immediately after step f), centrifuge the second centrifuge tube at 500 g for 10 min and take another subsample of the supernatant (subsample B), 0.05 mL. h) Collect the centrifugation pellet of step g) and, if present, resuspend in milliQ water and immediately examine the suspension for the presence of crystals visible under a microscope. i) Determine the concentration of unaggregated BLG in sub-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 sub-sample. The concentration of unaggregated BLG in sub-sample A is C BLG,A and the concentration of non-aggregated BLG in subsample B is C BLG,B It is called. j) The liquid from which the sample was taken in step a) is C BLG,B C BLG,A and crystals are observed in step i), there was supersaturation (for the particular conditions).

[0049] In the context of the present invention, the terms "liquid" and "solution" encompass both compositions that do not contain particulate matter, e.g., protein crystals or other protein particles, and compositions that contain a combination of liquids and solid and / or semi-solid particles. Thus, a "liquid" or "solution" may be a suspension or even a slurry. However, a "liquid" and a "solution" are preferably pumpable.

[0050] In the context of the present invention, the terms "whey protein concentrate" (WPC) and "serum protein concentrate" (SPC) relate to dry or aqueous compositions comprising a total amount of protein of 20 to 89% w / w relative to the total solids content.

[0051] The WPC or SPC preferably comprises: 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 0-40% w / w relative to protein.

[0052] Alternatively, but also preferably, the WPC or SPC may comprise: 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 0-40% w / w relative to protein.

[0053] Preferably, the WPC or SPC comprises: 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 0-40% w / w relative to protein.

[0054] More preferably, the WPC or SPC comprises: 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 CMP 0-25% w / w on protein.

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

[0056] 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.

[0057] The WPI or SPI preferably comprises: 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 0–40% w / w of total protein.

[0058] Alternatively, but also preferably, the WPI or SPI may include: 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 0-25% w / w of total protein.

[0059] More preferably, the WPI or SPI may include: 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 0-25% w / w of total protein.

[0060] SPI typically contains no or only trace amounts of CMP.

[0061] In the context of the present invention, the term "additional protein" refers to a protein that is not BLG. Additional proteins present in the 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 protein.

[0062] The terms "consisting essentially of" and "consisting essentially of" mean that the claim or feature in question includes particular materials or steps, and that do not materially affect the basic and novel characteristic(s) of the claimed invention.

[0063] In the context of the present invention, the phrase "Y and / or X" means "Y" or "X" or "Y and X". Following the same logic, "n1, n2, ..., n i-1 , and / or n i " means "n1" or "n2" or ... or "n i-1 " or "n i " or components: n1, n2, ... n i-1 , and n i means any combination of

[0064] 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 water.

[0065] In the context of the present invention, the term "physical microbial reduction" refers to a physical interaction with the composition that results in a reduction in the total amount of viable microorganisms in the composition. The term does not include the addition of chemicals that result in the death of microorganisms. The term further does not include the heat exposure to which the sprayed droplets are exposed during spray drying, but does include possible pre-heating before spray drying.

[0066] In the context of the present invention, the pH of a powder refers to the pH of 10 g of powder mixed in 90 g of demineralized water, measured according to Example 1.16.

[0067] In the context of the present invention, the weight percent (% w / w) of a particular composition, product, or ingredient means the weight percent of that ingredient relative to the weight of the particular composition, product, or ingredient, unless a different measure (e.g., total solids or total protein) is specifically mentioned.

[0068] In the context of the present invention, the process step "concentrate" and the verb "concentrate" refer to protein concentration and include both protein concentration on a total solids basis and protein concentration on a total weight basis, which does not necessarily require, for example, that the absolute protein concentration w / w of the composition is increased, as long as the protein content is increased relative to the total solids.

[0069] In the context of the present invention, the term "weight ratio" between component X and component Y means 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.

[0070] In the context of the present invention, the term "at least pasteurized" relates to a heat treatment having a microbial killing effect equal to or greater than that of a heat treatment for 10 seconds at 70° C. The standard for determining the killing effect is E. coli O157:H7.

[0071] In the context of the present invention, the term "whey protein feed" relates to a whey protein source from which a liquid BLG isolate is derived. A whey protein feed has a lower BLG content relative to total protein than a liquid BLG isolate and is typically a WPC, WPI, SPC or SPI.

[0072] In the context of the present invention, the term "BLG-enriched composition" refers to a BLG-enriched composition resulting from isolating BLG from a whey protein feed. A BLG-enriched composition typically contains the same whey protein as a whey protein feed, but BLG is present in a significantly higher concentration relative to total protein than in a whey protein feed. A BLG-enriched composition can be prepared from a whey protein feed, for example, by chromatography, protein crystallization and / or membrane-based protein fractionation. A 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, a BLG-enriched composition can be used directly as a liquid BLG isolate. However, additional processing is often required to convert a BLG-enriched composition into a liquid BLG isolate.

[0073] In the context of the present invention, the term "whey protein solution" is used to describe a special aqueous whey protein composition that is supersaturated with respect to BLG in a salt-solution manner and is useful for preparing BLG crystals.

[0074] In the context of the present invention, the term "sterile" means that the sterile composition, or product in question, does not contain any viable microorganisms and is therefore free of microbial growth during storage at room temperature. A sterilized composition is aseptic.

[0075] When liquids such as beverage preparations are sterilized and aseptically packaged in sterile containers, they typically have a shelf life of at least six months at room temperature. The sterilization process kills spores and microorganisms that may cause spoilage of the liquid.

[0076] In the context of the present invention, the term "energy content" refers to the total content of energy 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 beverage with an energy content of 350 kcal / 100 grams of beverage. The total energy content of a food includes the energy contribution from all macronutrients present in the food, such as energy from protein, fat, and carbohydrate. The distribution of energy from macronutrients in a food can be calculated based on the amount of the macronutrients in the food and the contribution of the macronutrients to the total energy content of the food. The 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% carbohydrate, and 30E% lipid, this means that 20% of the total energy comes from protein, 50% of the total energy comes from carbohydrate, and 30% of the total energy comes from fat (lipid).

[0077] The term "nutritionally complete dietary supplement" means a food that contains proteins, lipids, and carbohydrates, and further contains vitamins, minerals, and trace elements, which food has a nutritional profile that corresponds to a complete and healthy diet.

[0078] In the context of the present invention, the term "nutritionally incomplete dietary supplement" refers to a food product that contains one or more macronutrients and, optionally, further contains vitamins, minerals, and trace elements. A nutritionally incomplete beverage may contain protein as the only nutrient or may contain, for example, protein and carbohydrates.

[0079] The term "food for special medical purposes" or "medical food" refers to a food for oral intake or tube feeding, used for a specific medical disorder, disease, or condition with unique nutritional requirements and for use under medical supervision. Medical foods may be nutritionally complete or nutritionally incomplete dietary supplements / drinks.

[0080] The term "nutrients" refers to substances that 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 such as vitamins, minerals, and trace elements.

[0081] The term "nutrients" refers to substances that 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 vitamins, minerals, and trace element nutrients.

[0082] 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.

[0083] In the context of the present invention, the terms "beverage preparation" and "preparation" as used substantively relate to any water-based liquid that can be consumed as a drink, for example by pouring, sipping or tube feeding.

[0084] In the context of the present invention, the term "protein fraction" relates to the proteins of the composition in question, e.g. the proteins of a powder or beverage preparation.

[0085] In the context of the present invention, the term "astringent taste" relates to the mouthfeel. Astringent taste feels like a contraction of the cheek muscles, increasing saliva production. Astringent taste is therefore not a taste per se, but a physical mouthfeel and time-dependent sensation in the mouth.

[0086] In the context of the present invention, the term "dry mouthfeel" relates to the sensation in the mouth, where the mouth and teeth feel dry and saliva production is minimized. So dry mouthfeel is not a taste in itself, but a physical mouthfeel and a time-dependent sensation in the mouth.

[0087] In the context of the present invention, the term "mineral" 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.

[0088] In the context of the present invention, the terms "lipid," "fats," and "oils" 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, so long as such materials are suitable for human consumption.

[0089] 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 and through which a clear image appears. A transparent beverage has a turbidity of up to 200 NTU.

[0090] In the context of the present invention, the term "opaque" encompasses beverage preparations that have a visually unclear appearance, which have a turbidity of greater than 200 NTU.

[0091] One aspect of the invention is a packaged thermally treated beverage preparation having a pH in the range of 5.5 to 8.0, wherein the beverage - a protein having a total amount of 1-20% w / w by weight of the beverage, wherein at least 85% w / w of the protein is beta-lactoglobulin (BLG); - optionally with sweeteners and / or flavourings.

[0092] An advantage of the present invention is that it allows for the production of a drinkable beverage having a neutral pH and low viscosity.

[0093] It is highly beneficial for the packaged thermally treated beverage preparation to have at least 85% w / w of the protein be BLG for a number of reasons. The advantage is that the packaged thermally treated beverage preparations according to the invention are more stable and have less color compared to similar WPI beverages.

[0094] This is obtained by the packaged heat treated beverage preparation of the present invention, so that it has been surprisingly found that the packaged heat treated beverage has less color, even when high protein concentrations are applied, compared to the heat treated pH neutral WPI beverage, which has a more yellowish color.

[0095] Therefore, it is an advantage that due to the inventive composition of the packaged heat treated beverage of the present invention, no bleaching or additional whitening is required to remove or reduce the yellow color.

[0096] 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.

[0097] 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 thermally 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.

[0098] For example, the packaged thermally treated beverage preparation preferably comprises BLG in an amount of at least 97.5% w / w of total protein, preferably at least 98.0% w / w, more preferably at least 98.5% w / w, even more preferably at least 99.0%, most preferably at least 99.5% w / w of total protein, approximately 100.0% w / w of total protein, etc.

[0099] The proteins of the packaged heat-treated beverage preparation are preferably prepared from mammalian milk, preferably from ruminant milk, such as milk from cows, sheep, goats, buffaloes, camels, llamas, horses and / or deer. Proteins from bovine milk are particularly preferred. Thus, the proteins of the packaged heat-treated beverage preparation are preferably bovine milk proteins.

[0100] The protein of the packaged heat treated beverage preparation is preferably a whey protein or milk serum protein, even more preferably a bovine whey protein or milk serum protein.

[0101] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation is at least pasteurized.

[0102] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation is sterile.

[0103] The appearance of a beverage preparation is important to consumers for both clear and opaque beverages. Particularly for clear, watery beverages, 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.

[0104] However, even if a dedicated coloring agent is added during the production of the beverage, the inventors have found that it is advantageous to be able to avoid additional color sources in order to avoid undesirable variations or changes in the appearance of the beverage. The inventors have found that the high BLG protein profile described herein is more neutral / colorless in color than conventional WPIs and contributes less color change than conventional WPIs. Conventional WPIs have a yellowish appearance, which can be reduced to some extent by adding an oxidizing agent such as bleach. However, the addition of an oxidizing agent is often undesirable and in the present invention it is even no longer necessary.

[0105] The CIELAB color scale described in Example 1.9 is used to determine the color of beverages. For example, a positive delta b * values ​​indicate a yellower color than desalinated water, while a negative delta b * A value of 0.01 indicates a beverage that is more bluish than demineralized water. Thus, consumers are advised to choose a color delta b to obtain a beverage that is neither yellow nor blue. * A value close to 0 is often preferred.

[0106] 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 up to 200 NTU, more preferably up to 40 NTU. * has.

[0107] 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 on the CIELAB color scale, preferably in the range of +0.10 to +0.25. * has.

[0108] For example, for the preparation of opaque beverages 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 in the range of -6 to -1.7 on the CIELAB color scale, preferably in the range of -5.0 to -2.0. * has.

[0109] 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 on the CIELAB color scale, particularly when the preparation has a turbidity of up to 200 NTU, more preferably up to 40 NTU. * has.

[0110] These drinks contain delta b * There is less yellow color compared to beverages containing WPIs with higher values ​​and more yellow color.

[0111] In some other 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 0.0 to 0.40 on the CIELAB color scale, preferably in the range of +0.10 to +0.25. * has.

[0112] a * The values ​​represent the green and red components, green in the negative direction and red in the positive direction. To obtain a beverage that is neither red nor green, the color delta a * A value near zero is often preferred.

[0113] Typically, the protein fraction of a packaged heat treated beverage preparation will have 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 up to 200 NTU, more preferably up to 40 NTU. * has. Preferably, the packaged heat treated beverage preparation has a colour 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 colour scale. * has.

[0114] The inventors have found that it can be advantageous to control the mineral content to achieve some of the desired properties of a packaged thermally processed beverage preparation.

[0115] The inventors have surprisingly found that the use of BLG isolate as defined herein allows for the production of beverage preparations with high mineral concentrations without compromising viscosity and avoiding gelling (see Example 2). This potentially allows for the production of packaged heat-treated beverage preparations with high mineral content, potentially allowing for the production of beverages that are nutritionally complete or nutritionally incomplete dietary supplements.

[0116] 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.

[0117] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg and Ca is in the packaged thermally treated beverage preparation in the range of 0-400 mM, preferably in the range of 10-200 mM, or preferably in the range of 20-100 mM.

[0118] 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 in the range of 0-100 mM, more preferably in the range of 5-50 mM, even more preferably in the range of 10-35 mM.

[0119] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg and Ca is up to 400 mM in the packaged thermally treated beverage preparation.

[0120] In another preferred embodiment of the invention, the sum of the amounts of Na, K, Mg and Ca is in the packaged heat treated beverage preparation at most 300 mM, preferably at most 200 mM, 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 20 mM, or preferably at most 10 mM, or preferably at most 5 mM, or preferably at most 1 mM. In some preferred embodiments of the present invention, the sum of the amounts of Mg and Ca is up to 75 mM in the packaged, heat-treated beverage preparation, more preferably up to 40 mM in the packaged, heat-treated beverage preparation, more preferably up to 20 mM in the packaged, heat-treated beverage preparation.

[0121] In some preferred embodiments of the present invention, the sum of the amounts of Mg and Ca is up to 10 mM in the packaged heat-treated beverage preparation, more preferably up to 8.0 mM in the packaged heat-treated beverage preparation, more preferably up to 6.0 mM in the packaged heat-treated beverage preparation, even more preferably up to 4.0 mM in the packaged heat-treated beverage preparation, and most preferably up to 2.0 mM in the packaged heat-treated beverage preparation.

[0122] In another exemplary embodiment of the present invention, the packaged thermally treated 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.

[0123] In another preferred embodiment of the present invention, the thermally treated beverage preparation is a low-mineral beverage.

[0124] In the context of the present invention, the term "low-mineral" refers to: - Maximum 1.2% w / w transparent ash based on total solids, - total calcium and magnesium content of max. 0.3% w / w based on total solids, - total sodium and potassium content of maximum 0.10% w / w based on total solids; - a total phosphorus content of up to 100 mg per 100 g protein,

[0125] Preferably, the low mineral composition comprises: - maximum ash content of 0.7% w / w based on total solids; - total calcium and magnesium content of maximum 0.2% w / w based on total solids, - total sodium and potassium content of maximum 0.08% w / w based on total solids; a total phosphorus content of up to 80 mg per 100 g protein,

[0126] Even more preferably, the low mineral composition comprises: - maximum ash content of 0.5% w / w based on total solids, - total calcium and magnesium content of maximum 0.15% w / w based on total solids; - total sodium and potassium content of maximum 0.06% w / w based on total solids; a total phosphorus content of up to 50 mg per 100 g protein,

[0127] The low mineral composition is as follows: - maximum ash content of 0.5% w / w based on total solids, - total calcium and magnesium content of maximum 0.15% w / w based on total solids; - total sodium and potassium content of maximum 0.06% w / w based on total solids; It is particularly preferred that the protein has a total phosphorus content of up to 50 mg per 100 g of protein.

[0128] In another exemplary embodiment of the present invention, the packaged thermally treated 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.

[0129] The inventors have found that the present invention allows the preparation of packaged heat-treated beverage preparations that have very low contents of phosphorus and other minerals, such as potassium, which is advantageous for patients suffering from renal disease or who otherwise have reduced renal function.

[0130] The packaged thermally treated beverage preparation is preferably a low phosphorus beverage preparation.

[0131] The packaged thermally treated beverage preparation is preferably a low potassium beverage preparation.

[0132] The packaged thermally treated beverage preparation is preferably a low phosphorus and low potassium beverage preparation.

[0133] In the context of the present invention, the term "low phosphorus" relates to a composition, e.g. a liquid, powder or other food product, having a total phosphorus content of up to 100 mg per 100 g protein. Preferably, the low phosphorus composition has a total phosphorus content of up to 80 mg per 100 g protein. More preferably, the low phosphorus composition may have a total phosphorus content of up to 50 mg per 100 g protein. Even more preferably, the low phosphorus composition may have a total phosphorus content of up to 20 mg per 100 g protein. Even more preferably, the low phosphorus composition may have a total phosphorus content of up to 5 mg per 100 g protein. The low phosphorus composition according to the present invention can be used as a food ingredient for producing a food product for a patient group with reduced renal function.

[0134] Therefore, in some particularly preferred embodiments of the present invention, the packaged heat treated beverage preparation comprises up to 80 mg phosphorus per 100 g protein. Preferably, the packaged heat treated beverage preparation comprises up to 30 mg phosphorus per 100 g protein. More preferably, the packaged heat treated beverage preparation comprises up to 20 mg phosphorus per 100 g protein. Even more preferably, the packaged heat treated beverage preparation comprises up to 10 mg phosphorus per 100 g protein. Most preferably, the packaged heat treated beverage preparation comprises up to 5 mg phosphorus per 100 g protein.

[0135] The phosphorus content relates to the total amount of elemental phosphorus in the composition in question and is determined according to Example 1.19.

[0136] In the context of the present invention, the term "low potassium" relates to a composition, e.g. a liquid, powder or other food product, having a total potassium content of up to 700 mg per 100 g protein. Preferably, the low potassium composition has a total potassium content of up to 600 mg per 100 g protein. More preferably, the low potassium composition may have a total potassium content of up to 500 mg per 100 g protein. More preferably, the low potassium composition may have a total potassium content of up to 400 mg per 100 g protein. More preferably, the low potassium composition may have a total potassium content of up to 300 mg per 100 g protein. Even more preferably, the low potassium composition may have a total potassium content of up to 200 mg per 100 g protein. Even more preferably, the low potassium composition may have a total potassium content of up to 100 mg 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.

[0137] The low potassium composition according to the present invention can be used as a food ingredient for producing food for patient groups with reduced renal function.

[0138] Thus, in some particularly preferred embodiments of the present invention, the packaged heat treated beverage preparation comprises up to 600 mg potassium per 100 g protein. More preferably, the packaged heat treated beverage preparation comprises up to 500 mg potassium per 100 g protein. More preferably, the packaged heat treated beverage preparation comprises up to 400 mg potassium per 100 g protein. More preferably, the packaged heat treated beverage preparation comprises up to 300 mg potassium per 100 g protein. More preferably, the packaged heat treated beverage preparation comprises up to 200 mg potassium per 100 g protein. More preferably, the packaged heat treated beverage preparation comprises up to 100 mg potassium per 100 g protein. Even more preferably, the packaged heat treated beverage preparation comprises up to 50 mg potassium per 100 g protein, and even more preferably, the packaged heat treated beverage preparation comprises up to 10 mg potassium per 100 g protein.

[0139] The potassium content relates to the total amount of elemental potassium in the composition in question and is determined according to Example 1.19.

[0140] In some preferred embodiments of the present invention, the packaged heat treated beverage preparation comprises up to 100mg phosphorus / 100g protein and up to 700mg potassium / 100g protein, preferably up to 80mg phosphorus / 100g protein and up to 600mg potassium / 100g protein, more preferably up to 60mg phosphorus / 100g protein and up to 500mg potassium / 100g protein, more preferably up to 50mg phosphorus / 100g protein and up to 400mg potassium / 100g protein, or more preferably up to 20mg phosphorus / 100g protein and up to 200mg potassium / 100g protein, or even more preferably up to 10mg phosphorus / 100g protein and up to 50mg potassium / 100g protein. In some preferred embodiments of the present invention, the packaged heat treated beverage preparation comprises up to 100mg phosphorus / 100g protein and up to 340mg potassium / 100g protein.

[0141] Thermally processed beverage preparations containing small amounts of phosphorus and potassium can advantageously be supplemented with carbohydrates and lipids, the thermally processed beverage preparations preferably further comprising 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 a total amount of lipids in the range of 20-60% of the total energy content, preferably in the range of 30-50E%.

[0142] In one embodiment of the present invention, the packaged heat-treated beverage preparation comprises a plurality of vitamins. In one exemplary embodiment, the packaged heat-treated beverage preparation comprises at least 10 vitamins. In one exemplary embodiment, the substantially clear liquid nutritional composition 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.

[0143] In one embodiment of the present invention, the packaged thermally treated beverage comprises a plurality of vitamins and a plurality of minerals. In some embodiments of the present invention, the packaged thermally treated beverage preparation comprises one or more edible acids selected from the group consisting of citric acid, malic acid, tartaric acid, acetic acid, benzoic acid, butyric acid, lactic acid, lactobionic acid, fumaric acid, succinic acid, ascorbic acid, adipic acid, phosphoric acid, and mixtures thereof.

[0144] In some preferred embodiments, the packaged thermally treated beverage preparation optionally comprises a sweetener, a sugar polymer, and / or a flavoring.

[0145] In one embodiment of the present invention, the packaged thermally treated beverage preparation comprises a flavoring selected from the group consisting of salt, seasoning, umami seasoning and / or spices. In a preferred embodiment of the present invention, the flavoring comprises chocolate, cocoa, lemon, orange, lime, strawberry, banana, forest fruit flavors or combinations thereof. The choice of flavoring may vary depending on the beverage to be produced.

[0146] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation has a pH in the range of 6.5 to 7.5. Most preferably, the pH employed is a pH of 6.5 to 7.0 or a pH of 6.8 to 7.2.

[0147] With regard to appearance, it has surprisingly been found that the use of a whey protein beverage in which at least 85% w / w of the protein is BLG may improve both visual (color and turbidity) and viscosity when compared to a heat treated WPI beverage.

[0148] The packaged heat treated beverage preparation preferably has a pH in the range of 5.5 to 6.2, alternatively the packaged heat treated beverage preparation has a pH in the range of 6.2 to 8.0. Alternatively, the packaged thermally treated beverage preparation has a pH in the range of 6.8 to 8.0, more preferably the packaged thermally treated beverage preparation has a pH in the range of 6.2 to 8.0.

[0149] The packaged thermally treated beverage preparations of the present invention are preferably clear, transparent and have been found to have a low viscosity at a pH in the range of 6.2 to 8.0, preferably at a pH of 6.3 to 7.6, more preferably at a pH of 6.5 to 7.2.

[0150] The packaged heat treated beverage preparations of the present invention have been found to have low viscosity and milky appearance, preferably at a pH in the range of 5.5 to 8.0, preferably at a pH of 5.7 to 6.8, more preferably at a pH of 5.8 to 6.0.

[0151] In some preferred embodiments of the present invention, it has been found that the beverage preparation of the present invention is preferably heat treated at a pH in the range of 5.6-6.2, preferably at a pH of 5.6-8.0, optionally mixed with sources of carbohydrates, fats, minerals and vitamins, adjusted to a preferred pH of 6.2-8.0 and subjected to a second heat treatment (UHT).

[0152] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation has a turbidity of up to 200 NTU.

[0153] The appearance of a packaged heat-treated beverage preparation is of interest to the consumer. Clarity is a parameter that consumers use to evaluate the product. One way to determine the clarity of a beverage preparation is by measuring the turbidity of the beverage as described in Example 1.7.

[0154] In some embodiments of packaged thermally processed beverage preparations, it is beneficial for the beverage preparation to be transparent. This can be advantageous, for example, when the beverage is used as a sports drink or as a "protein water", in which case it is beneficial for the beverage to resemble water in appearance.

[0155] In a preferred embodiment of the present invention, the packaged thermally treated beverage preparation has a turbidity of up to 200 NTU, such beverage being clear and / or transparent.

[0156] Surprisingly, it has been found by the inventors that the heat treated beverage preparation according to the invention results in a clear heat treated beverage preparation having a turbidity of up to 200 NTU.

[0157] This was observed whether the heat treatment applied was sterilization or pasteurization.

[0158] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation has a turbidity of up to 150 NTU, or preferably a turbidity of up to 100 NTU, or preferably a turbidity of up to 80 NTU, or preferably a turbidity of up to 60 NTU, or more preferably a turbidity of up to 40 NTU, or preferably a turbidity of up to 30 NTU, preferably a turbidity of up to 20 NTU, more preferably a turbidity of up to 10 NTU, more preferably a turbidity of up to 5 NTU, and even more preferably a turbidity of up to 2 NTU.

[0159] In a preferred embodiment of the present invention, the packaged thermally treated beverage preparation has a turbidity of greater than 200 NTU, such beverages being opaque.

[0160] In some embodiments of packaged heat-treated beverage preparations, it is beneficial for the beverage preparation to be opaque. This is advantageous, for example, when the beverage is to resemble milk and have a milky appearance. Nutritionally complete dietary supplements typically have an opaque appearance.

[0161] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation has a turbidity of greater than 250 NTU. Preferably, the packaged thermally treated beverage preparation has a turbidity of greater than 300 NTU, more preferably, greater than 500 NTU, more preferably, greater than 1000, preferably greater than 1500 NTU, and even more preferably, greater than 2000 NTU.

[0162] 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 settling of sediment occurs over time. Beverages that contain a high amount of insoluble matter are usually considered to be unstable.

[0163] In the context of the present invention, a whey protein beverage preparation is considered to be "stable" if a maximum of 15% of the total protein in a heated sample is precipitated upon centrifugation at 3000g for 5 minutes. See analytical method in Example 1.10. Surprisingly, when BLG is used as a protein source in an amount of at least 85 w / w%, the protein fraction was found to contain up to 15% insoluble material after centrifugation at 3000 g for 5 minutes, compared to when a WPI with a lower BLG content is used as a protein source, demonstrating that the beverage preparation is stable.

[0164] Thus, in some preferred embodiments of the present invention, the protein fraction of the thermally treated beverage preparation comprises up to 15% insoluble material.

[0165] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation contains up to 15% insoluble matter.

[0166] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation preferably comprises a maximum of 12% insoluble matter, more preferably a maximum of 10% insoluble matter, even more preferably a maximum of 8% insoluble matter, and most preferably a maximum of 6% insoluble matter.

[0167] Even lower levels of insoluble matter are often preferred, and in some preferred embodiments the packaged thermally treated beverage preparation contains up to 4% insoluble matter, preferably up to 2% insoluble matter, more preferably up to 1% insoluble matter, and most preferably no detectable insoluble matter.

[0168] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation has a viscosity of up to 200 cP centipoise when measured at 22° C. at a shear rate of 100 / sec.

[0169] Consumers prefer their heat treated beverages to be liquids rather than gels.

[0170] One method of determining the viscosity of a beverage formulation is by measuring the viscosity of the beverage as described in Example 1.8.

[0171] In some embodiments of the packaged thermally treated beverage preparation, it is beneficial for the beverage preparation to have a low viscosity, which is advantageous when the beverage is used as a sports drink, or in some embodiments, as a nutritionally complete beverage.

[0172] Surprisingly, the inventors have found that beverage preparations having a neutral pH and that have been subjected to a thermal treatment such as pasteurization and further sterilized have a viscosity of up to 200 centipoise when measured at a shear rate of 100 / sec at 22°C. Thus, in some preferred embodiments of the present invention, the packaged thermally treated beverage preparation has a viscosity of up to 200 cP.

[0173] Preferably, the viscosity of the packaged thermally 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, most preferably at most 40 cP.

[0174] Since even lower viscosities are often preferred, in some preferred embodiments of the present invention, the viscosity of the packaged thermally 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 even more preferably at most 1 cP.

[0175] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises a total amount of protein of 2-18% w / w based on the weight of the beverage preparation.

[0176] In another preferred embodiment of the invention, the packaged thermally treated beverage preparation comprises a total amount of protein of 3-20% w / w, more preferably 3-18% w / w, even more preferably 3-15% w / w, and most preferably 3-10% w / w, based on the weight of the beverage.

[0177] In some embodiments of the present invention, the packaged thermally treated beverage preparation advantageously has a protein content of 1.0-10.0% w / w based on the weight of the beverage. In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises a total amount of protein of 1-10% w / w based on the weight of the beverage preparation.

[0178] The packaged heat-treated beverage preparation preferably comprises a total amount of protein of 2.0-9.0% w / w by weight of the beverage, or the packaged heat-treated beverage preparation preferably comprises a total amount of protein of 3.0-8.0% w / w by weight of the beverage, or the packaged heat-treated beverage preparation preferably comprises a total amount of protein of 5.0-7.5% w / w by weight of the beverage, or the packaged heat-treated beverage preparation preferably comprises a total amount of protein of 4.0-6.0% w / w by weight of the beverage.

[0179] Most preferably, the packaged heat treated beverage preparation comprises a total amount of protein of 4.0-6.0% w / w by weight of the beverage. This protein range is particularly relevant when the heat treated beverage preparation is a sports drink. However, this range is also relevant for some medical uses of the beverage.

[0180] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises a total amount of protein of 10-20% w / w based on the weight of the beverage preparation.

[0181] In some embodiments of the invention, the packaged thermally treated beverage preparation preferably comprises a total amount of protein of 10-18% w / w by weight of the beverage, or preferably comprises a total amount of protein of 12.0-16.0% w / w by weight of the beverage, or preferably comprises a total amount of protein of 13.0-15.0% w / w by weight of the beverage.

[0182] In some preferred embodiments, the packaged thermally treated beverage preparation comprises a total amount of protein of 1.0-6.0% w / w by weight of the beverage, and in other preferred embodiments, the packaged thermally treated beverage preparation comprises a total amount of protein of 6.0-12.0% w / w by weight of the beverage.

[0183] Or in another preferred embodiment, the packaged thermally treated beverage preparation comprises a total amount of protein of 12.0-20.0% w / w based on the weight of the beverage.

[0184] All proteins in the beverage are preferably whey proteins and / or whey proteins.

[0185] The packaged thermally treated beverage preparations of the present invention are particularly useful as sports drinks, in which case they preferably contain only lipids, optionally in limited amounts, and / or also optionally in limited amounts of carbohydrates.

[0186] In some preferred embodiments of the invention, the preparations are particularly useful as sports drinks, for example comprising a total amount of protein in the range of 1-20% w / w by weight of the drink, preferably 2-15% w / w by weight of the drink, or preferably 2-10% w / w by weight of the drink, most preferably 2-6% w / w by weight of the drink.

[0187] In some preferred embodiments of the invention, the packaged thermally treated beverage preparations are particularly useful as nutritionally incomplete dietary supplements, for example comprising a total amount of protein in the range of 2-20% w / w by weight of the beverage, or preferably 3-10% w / w by weight of the beverage.

[0188] In some preferred embodiments of the invention, the packaged thermally treated beverage preparations are particularly useful as nutritionally complete dietary supplements, for example comprising a total amount of protein in the range of 4-20% w / w by weight of the beverage, or preferably 5-18% w / w by weight of the beverage.

[0189] In certain preferred embodiments of the present invention, the packaged thermally treated beverage preparations are particularly advantageous for patients suffering from renal disease or otherwise experiencing reduced renal function.

[0190] In some preferred embodiments of the invention, the packaged thermally treated beverage preparation comprises a total amount of protein in the range, for example, of 2-20% w / w by weight of the beverage, or preferably of 3-12% w / w by weight of the beverage, or preferably of 3-10% w / w by weight of the beverage.

[0191] It is particularly preferred that the packaged thermally processed beverage preparation comprises a BLG isolate in combination with other protein sources, for example, preferably as the major protein source, and even possibly as the sole protein source.

[0192] The degree of nativeness of a protein depends on several factors, such as the protein concentration, pH, temperature, and duration of heat treatment.

[0193] The intrinsic tryptophan fluorescence ratio (I330nm / I350nm) is a measure of the extent of BLG unfolding, and the inventors have determined that a high BLG tryptophan fluorescence ratio correlates with low or no BLG unfolding. The intrinsic tryptophan fluorescence ratio (I330nm / I350nm) is measured according to Example 1.1.

[0194] In some preferred embodiments of the present invention, the BLG isolated powder has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11.

[0195] In some preferred embodiments of the invention, the BLG isolated powder 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.

[0196] If the BLG isolated 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 invention, the protein fraction of the BLG isolated powder has an intrinsic tryptophan fluorescence ratio of at least 1.11.

[0197] In some preferred embodiments of the invention, the protein fraction of the BLG isolated powder 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.

[0198] For example, a protein fraction can be separated from the BLG isolated powder by dissolving the BLG isolated powder in demineralized water and subjecting the solution to dialysis or ultrafiltration-based diafiltration using a protein-retaining filter.

[0199] Protein denaturation can also be described by analytical methods other than Trp fluorescence, which are described in Example 1.3. The principle of this method is that 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 in the solution is stable. Therefore, 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 ) / PpH7.0又は3.0 ) * 100% In the formula, (P pH7.0又は3.0 ) is the total protein content at pH 7.0 or 3.0, and (S pH4.6 ) is the total protein content in the supernatant at pH 4.6. See Example 1.3.

[0200] In some preferred embodiments of the present invention, the BLG isolated powder has a degree of protein denaturation of up to 10%, preferably up to 8%, more preferably up to 6%, even more preferably up to 3%, even more preferably up to 1%, and most preferably up to 0.2%.

[0201] In some embodiments of the 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%.

[0202] The packaged heat-treated beverage preparation of the present invention may contain macronutrients other than protein. In some embodiments of the present invention, the packaged heat-treated beverage preparation further comprises 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 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, saccharose, maltose, dextrose, galactose, maltodextrin, corn syrup solids, sucromalt, glucose polymers, corn syrup, modified starch, resistant starch, carbohydrates derived from rice, isomaltulose, white sugar, glucose, fructose, lactose, high fructose com syrup, honey, sugar alcohols, fructooligosaccharides, soybean fiber, corn fiber, guar gum, konjac flour, polydextrose, Fibersol, and combinations thereof. In some embodiments of the present invention, the packaged thermally processed beverage preparation comprises a non-digestible sugar such as a fructan, the fructan comprising inulin or fructooligosaccharide.

[0204] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparations and liquid solutions contain sugar polymers, i.e. oligosaccharides and / or polysaccharides.

[0205] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation 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.

[0206] An even lower carbohydrate content is often preferred, and in some preferred embodiments of the present invention, it is 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, even more preferably in the range of 0-10% of the total energy content of the preparation.

[0207] In some preferred embodiments of the present invention, the carbohydrate content of the packaged thermally treated beverage preparation is at most 3% of the total energy content of the preparation, more preferably at most 1% of the total energy content of the preparation, even more preferably at most 0.1% of the total energy content of the preparation.

[0208] In some preferred embodiments of the invention, the preparations are particularly useful as sports drinks, for example comprising 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.

[0209] In some preferred embodiments of the invention, the packaged thermally treated beverage preparations are particularly useful as nutritionally incomplete dietary supplements, for example comprising a total amount of carbohydrates in the range of 70-95% of the total energy content (E) of the beverage, preferably 80-90E%.

[0210] In some preferred embodiments of the invention, the packaged thermally treated beverage preparations are particularly useful as nutritionally complete dietary supplements, for example comprising 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 certain preferred embodiments of the present invention, the packaged thermally treated beverage preparations are particularly advantageous for patients suffering from renal disease or otherwise experiencing reduced renal function.

[0212] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises, for example, 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.

[0213] 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, flavorings, minerals, sweeteners, antioxidants, food acids, lipids, carbohydrates, prebiotics, probiotics, and non-whey proteins.

[0214] Further ingredients ensure that the packaged heat treated beverage preparation contains the desired nutrients, ie nutrients that are specifically adapted for patients suffering from protein deficiencies or athletes wanting to build muscle.

[0215] 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, stevia glycosides such as rebaudioside A, or combinations thereof. In some embodiments of the present invention, it is particularly preferred that the sweetener comprises or even consists of one or more high-intensity sweeteners (HIS).

[0216] HIS are found in both natural and artificial sweeteners and typically have a sweetening intensity at least 10 times that of sucrose.

[0217] When used, the total amount of HIS is typically in the range of 0.01-2% w / w. For example, the total amount of HIS can be in the range of 0.05-1.5% w / w. Alternatively, the total amount of HIS can be in the range of 0.1-1.0% w / w.

[0218] The choice of sweetener may depend on the beverage being produced, for example, high intensity sweeteners (e.g., aspartame, acesulfame K, sucralose, etc.) may be used in beverages where an energy contribution from the sweetener is not desired, whereas natural sweeteners (e.g., steviol glycosides, sorbitol, or sucrose) may be used in beverages with a natural profile.

[0219] Furthermore, it may be preferred that the sweetener comprises or even consists of one or more polyol sweetener(s). Non-limiting examples of useful polyol sweeteners are maltitol, mannitol, lactitol, sorbitol, inositol, xylitol, threitol, galactitol or combinations thereof. When used, the total amount of polyol sweetener is typically in the range of 1-20% w / w. For example, the total amount of polyol sweetener may be in the range of 2-15% w / w. Alternatively, the total amount of polyol sweetener may be in the range of 4-10% w / w.

[0220] The packaged heat-treated beverage preparation of the present invention may contain macronutrients other than protein. In some embodiments of the present invention, the packaged heat-treated beverage preparation further comprises 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.

[0221] 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 0-40% 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.

[0222] The amount of lipids is determined according to ISO 1211:2010 (Determination of fat content - Roese-Gottlieb gravimetric method).

[0223] In some preferred embodiments of the present invention, the lipid content of the packaged thermally treated beverage preparation is at most 3% of the total energy content of the preparation, more preferably at most 1% of the total energy content of the preparation, even more preferably at most 0.1% of the total energy content of the preparation.

[0224] In some preferred embodiments of the invention, the preparation is particularly useful as a sports drink, for example comprising a total amount of lipids of up to 10E%, preferably up to a maximum of 1E%.

[0225] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparations are particularly useful as nutritionally incomplete dietary supplements, for example comprising a total amount of lipids of up to 10%, preferably up to a maximum of 1E%, of the total energy content of the beverage.

[0226] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparations are particularly useful as nutritionally complete dietary supplements, for example comprising a total amount of lipids in the range of 20-50%, preferably in the range of 30-40E%, or more preferably 25-40E% of the total energy content.

[0227] In certain preferred embodiments of the present invention, the packaged thermally treated beverage preparations are particularly advantageous for patients suffering from renal disease or otherwise experiencing reduced renal function.

[0228] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises, for example, a total amount of lipids in the range of 20-60%, preferably in the range of 30-50E% of the total energy content.

[0229] In one embodiment of the invention, the packaged thermally processed beverage preparation comprises food grade fats, such as canola oil and / or MCT (medium chain triglycerides), preferably in an amount of 2-10% by weight. Preferably, these fats contain a significant proportion, such as at least 40%, preferably at least 60%, of unsaturated fatty acids, most preferably polyunsaturated fatty acids. Most preferably, the beverage is in an emulsified form, the lipids being preferably present as emulsified droplets in the aqueous phase of the beverage preparation.

[0230] The beverage preparation typically comprises a total amount of water in the range of 50-99% w / w, preferably in the range of 45-97% w / w, more preferably in the range of 40-95% w / w, even more preferably in the range of 35-90% w / w and most preferably in the range of 30-85% w / w.

[0231] In some preferred embodiments of the present invention, the beverage preparation comprises a total amount of water in the range of 55-90% w / w, preferably in the range of 57-85% w / w, more preferably in the range of 60-80% w / w, even more preferably in the range of 62-75% w / w, and most preferably in the range of 65-70% w / w.

[0232] In some preferred embodiments of the invention, the beverage preparation comprises a total amount of water in the range of 90-99% w / w, preferably in the range of 92-98.5% w / w, more preferably in the range of 94-98% w / w, even more preferably in the range of 95-98% w / w, and most preferably in the range of 96-98% w / w. These embodiments are useful, for example, for clear, watery beverages.

[0233] In some preferred embodiments of the present invention, the beverage preparation comprises up to 1.0% w / w ethanol, more preferably up to 0.5% w / w, even more preferably up to 0.1% w / w, and most preferably is non-alcoholic, meaning there is no detectable ethanol.

[0234] The beverage preparation typically comprises a total solids content in the range of 1-45% w / w, preferably in the range of 5-40% w / w, more preferably in the range of 10-35% w / w, even more preferably in the range of 12-30% w / w, and most preferably in the range of 16-25% w / w.

[0235] In some preferred embodiments of the present invention, the beverage preparation comprises a total solids content in the range of 10-45% w / w, preferably in the range of 15-43% w / w, more preferably in the range of 20-40% w / w, even more preferably in the range of 25-38% w / w, and most preferably in the range of 30-35% w / w.

[0236] In some preferred embodiments of the present invention, the beverage preparation comprises a total solids content in the range of 1-10% w / w, preferably in the range of 1.5-8% w / w, more preferably in the range of 2-6% w / w, even more preferably in the range of 2-5% w / w, and most preferably in the range of 2-4% w / w. These embodiments are useful, for example, for clear, watery beverages.

[0237] The part of the beverage preparation that is not a solid is preferably water.

[0238] In some preferred embodiments of the invention, the combined total of alpha-lactalbumin (ALA) and caseinomacropeptide (CMP) constitutes at least 40% w / w of the non-BLG proteins in the beverage, 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 in the beverage.

[0239] In some preferred embodiments of the present invention, ALA constitutes up to 80% w / w of the non-BLG proteins of the beverage preparation, preferably up to 60% w / w, even more preferably up to 40% w / w, and most preferably up to 30% w / w of the non-BLG proteins of the beverage preparation.

[0240] Even lower ALA contents may be preferred, and thus in some preferred embodiments of the present invention, ALA constitutes up to 20% w / w of the non-BLG proteins of the beverage preparation, preferably up to 15% w / w, even more preferably up to 10% w / w, and most preferably up to 5% w / w of the non-BLG proteins of the beverage preparation.

[0241] In other preferred embodiments of the 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 the total protein in the standard whey protein concentrate derived from sweet whey.

[0242] Even lower concentrations of the major non-BLG whey proteins may be desirable, so in additional 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 4%, preferably at most 3%, more preferably at most 2%, even more preferably at most 1% by weight of total protein in a standard whey protein concentrate derived from sweet whey.

[0243] The inventors have seen indications that reduction of lactoferrin and / or lactoperoxidase is particularly advantageous in obtaining neutral coloured whey protein products.

[0244] Therefore, in some preferred embodiments of the present invention, lactoferrin is present in a weight percentage of total protein, which is 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 the weight percentage of total protein in standard whey protein concentrate derived from sweet whey. Even lower concentrations of lactoferrin may be desirable. Therefore, in additional preferred embodiments of the present invention, lactoferrin is present in a weight percentage of total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of the weight percentage of total protein in standard whey protein concentrate derived from sweet whey.

[0245] Similarly, in some preferred embodiments of the present invention, lactoperoxidase is present in a weight percentage of total protein, which is 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 the weight percentage of total protein in standard whey protein concentrate derived from sweet whey.Even lower concentrations of lactoperoxidase may be desirable.Therefore, in a further preferred embodiment of the present invention, lactoperoxidase is present in a weight percentage of total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of the weight percentage of total protein in standard whey protein concentrate derived from sweet whey.

[0246] Lactoferrin and lactoperoxidase are quantified according to Example 1.29.

[0247] In one embodiment of the present invention, the packaged thermally processed beverage preparation is a nutritionally complete food supplement. In one embodiment of the present invention, the packaged thermally treated beverage preparation is a nutritionally incomplete dietary supplement. In one embodiment of the present invention, the packaged thermally treated beverage preparation is a sports drink. 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 renal disease or otherwise suffering from reduced renal function.

[0248] The packaged thermally treated beverage preparations of the present invention are particularly useful as sports drinks, in which case they preferably contain only lipids, optionally in limited amounts, and / or also optionally in limited amounts of carbohydrates.

[0249] In some preferred embodiments of the present invention, the preparations are particularly useful as sports drinks, for example those having the following properties: a total amount of protein in the range of 1 to 20% w / w by weight of the beverage, preferably 2 to 15% 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 75E%, preferably at most 40E%, preferably at most 10E%, or preferably at most 5E% of the total energy content (E) of the beverage, and - A total amount of lipids of up to 10E%, preferably up to 1E%.

[0250] 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 20% w / w by weight of the beverage, or preferably from 3 to 10% w / w by 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 lipids of up to 10% of the total energy content of the beverage, preferably up to 1E%.

[0251] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparations are particularly useful as nutritionally complete dietary supplements, e.g., a total amount of protein ranging from 4 to 20% w / w by weight of the beverage, or preferably from 5 to 18% w / w by weight of the beverage, a total amount of carbohydrates ranging from 30 to 60% of the total energy content of the beverage, preferably ranging from 35 to 50E%; and a total amount of lipids in the range of 20-50%, preferably in the range of 30-40E%, or preferably 25-45E% of the total energy content of the beverage.

[0252] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparations are particularly advantageous for patients suffering from renal disease or otherwise suffering from reduced renal function, as the beverage preparations are very low in phosphorus and other minerals such as potassium.

[0253] 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 20% w / w by weight of the beverage, preferably 3 to 12% w / w by weight of the beverage, or preferably 3 to 10% w / w by weight of the beverage, a total amount of carbohydrates ranging from 30 to 60% of the total energy content of the beverage, preferably ranging from 35 to 50E%; and - A total amount of lipids in the range of 20-60% of the total energy content, preferably in the range of 30-50E%.

[0254] The inventors have seen indications that beverages with a high content of protein nanogel relative to total protein have a lower viscosity when they reach the stomach than beverages containing significant amounts of soluble whey protein aggregates (see Example 9). Soluble whey protein aggregates are often formed when beverages containing conventional whey protein isolates are sterilized. The ability of foods to develop viscosity and / or structure in the stomach has previously been linked to satiety (Halford et al; Satiety-enhancing products for appetite control: science and regulation of functional foods for weight management; Proceedings of the Nutrition Society (2012), 71, 350-362), and the inventors have seen indications that beverages with a high content of protein nanogel are less satiating on ingestion than comparable beverages containing soluble whey protein aggregates. This is highly advantageous for individuals who have a poor or anorexic appetite, but require high energy nutrition to restore and / or maintain muscle mass or other bodily functions.

[0255] In the context of the present invention, the term "protein nanogel" or "protein nanogel" refers to submicron-sized particles of denatured whey proteins, typically spherical or nearly spherical. Protein nanogels are also referred to as whey protein micelles and are discussed, for example, in WO 2007 / 110421 A2, although their micellar nature is questionable. The amount of soluble whey protein aggregates is quantified according to Example 1.32. Protein nanogels, when suspended, have an opaque, milky appearance and are therefore highly suitable for opaque beverages.

[0256] In the context of the present invention, the term "soluble whey protein aggregates" refers to small aggregates of denatured whey proteins, which aggregates can form strong gels (much stronger than native whey proteins) during acidification to pH 4.6, which aggregates are typically linear, worm-like, branched or chain-like in shape and typically submicron in size. Soluble whey protein aggregates are well known to those skilled in the art and are, for example, referred to as linear aggregates and described in WO 2007 / 110421 A2. The amount of soluble whey protein aggregates is quantified according to Example 1.32. Soluble whey protein aggregates typically form clear solutions when dissolved in water, making them very suitable for clear beverages.

[0257] pH, mineral content (especially Ca 2+By controlling the pH of the whey protein solution, the amount of the protein that is added to the whey protein solution, and the protein concentration of the protein solution, it is possible to control whether a gel, a large gel fragment, a protein nanogel, or a soluble whey protein aggregate is formed. This is well known to those skilled in the art. Protein nanogels are typically formed by heating a whey protein solution having a pH in the range of about 5.5 to about 6.5, preferably about 5.8 to 6.2, and are supported by a whey protein solution with a lower mineral content compared to conventional whey protein concentrates. Soluble whey protein aggregates are typically formed by heating a whey protein solution having a pH in the range of about 6.5 to 8.5, preferably about 6.6 to 7.5, and are supported by a higher content of monovalent cations such as sodium. Such monovalent cations are often added when raising the pH.

[0258] Therefore, in some particularly preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises at least 50% w / w protein nanogel relative to total protein, preferably at least 60% w / w, more preferably at least 70% w / w, even more preferably at least 80% w / w protein nanogel, and most preferably at least 90% w / w protein nanogel relative to total protein.

[0259] For example, the packaged thermally treated beverage preparation may contain: a total amount of protein of between 5 and 20% w / w, preferably between 8 and 19% w / w, more preferably between 9 and 18% w / w, even more preferably between 10 and 17% w / w, most preferably between 11 and 16% by weight of the beverage, a total amount of BLG of at least 85% w / w, preferably at least 88% w / w, more preferably at least 90% w / w, even more preferably at least 90% w / w, and most preferably at least 92% w / w relative to the total protein; - a total amount of protein nanogel of at least 50% w / w relative to the total protein, preferably at least 60% w / w, more preferably at least 70% w / w, even more preferably at least 80% w / w relative to the total protein.

[0260] The packaged thermally treated beverage preparation comprises: a total amount of protein of 10-20% w / w, preferably 11-19% w / w, more preferably 12-18% w / w, even more preferably 13-17% w / w, most preferably 14-16% by weight of the beverage, a total amount of BLG of at least 90% w / w, preferably at least 92% w / w, more preferably at least 94% w / w, most preferably at least 96% w / w relative to the total protein; It is particularly preferred that the total amount of protein nanogel is at least 50% w / w relative to the total protein, preferably at least 60% w / w, more preferably at least 70% w / w, even more preferably at least 80% w / w relative to the total protein.

[0261] The inventors have further found that protein nanogels are less prone to viscosity generation than native whey proteins, making it possible to produce sterile, pH-neutral beverages that are high in protein but have a sufficiently low viscosity to make them easy to drink.

[0262] Protein nanogels made by heat denaturation of whey proteins containing at least 85% w / w BLG appear to be particularly advantageous for use in high protein beverages and, without being bound by theory, it is believed that high BLG protein nanogels provide a more compact protein nanogel structure than protein nanogels based on regular WPI and this difference allows more protein to be included in the beverage without affecting drinkability.

[0263] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises: - a protein nanogel of at least 50% w / w based on the total protein, - up to 30% w / w soluble whey protein aggregates based on total protein; - Contains up to 5% w / w insoluble protein material.

[0264] In some more preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises: - a protein nanogel of at least 60% w / w based on the total protein, - up to 20% w / w soluble whey protein aggregates based on total protein; - Contains up to 5% w / w insoluble protein material.

[0265] In some even more preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises: - a protein nanogel with at least 70% w / w of total protein, - up to 15% w / w soluble whey protein aggregates based on total protein; - Contains up to 5% w / w insoluble protein material.

[0266] The inventors have observed that beverages with an increased content of soluble whey protein aggregates relative to total protein have a higher viscosity when reaching a stomach-like environment than beverages containing less soluble whey protein aggregates (see Example 9). The ability of a food product to develop viscosity and / or structure in the stomach has previously been linked to satiety, and thus beverages with a high content of soluble whey protein aggregates induce an increased feeling of satiety upon ingestion. This is highly advantageous for those who wish to lose weight, particularly those suffering from obesity.

[0267] Therefore, in another particularly preferred embodiment of the present invention, the packaged thermally treated beverage preparation comprises at least 60% w / w soluble whey protein aggregates relative to total protein, preferably at least 70% w / w, more preferably at least 80% w / w, even more preferably at least 90% w / w soluble whey protein aggregates relative to total protein.

[0268] Alternatively, but also preferably, the packaged thermally treated beverage preparation comprises: a total amount of protein of 5-12% w / w, preferably 6-11% w / w, more preferably 7-10% w / w, even more preferably 8-10% w / w, most preferably 11-16% by weight of the beverage, a total amount of BLG of at least 94% w / w relative to the total protein, preferably at least 96% w / w relative to the total protein, even more preferably at least 98% w / w relative to the total protein, and - at least 60% w / w of soluble whey protein aggregates relative to the total protein, preferably at least 70% w / w, more preferably at least 80% w / w, even more preferably at least 90% w / w of soluble whey protein aggregates relative to the total protein, The packaged thermally treated beverage preparation preferably comprises: a turbidity of at most 100 NTU, preferably at most 40 NTU, even more preferably at most 10 NTU, and - 100s at 22 °C of maximum 100 cP, preferably maximum 50 cP, more preferably maximum 20 cP, more preferably maximum 10 cP -1 , and has a viscosity at a shear rate of

[0269] For example, the packaged thermally treated beverage preparation may contain: a total amount of protein of between 5 and 20% w / w, preferably between 8 and 19% w / w, more preferably between 9 and 18% w / w, even more preferably between 10 and 17% w / w, most preferably between 11 and 16% by weight of the beverage, a total amount of BLG of at least 85% w / w relative to the total protein, preferably at least 90% w / w relative to the total protein, even more preferably at least 94% w / w relative to the total protein, most preferably at least 6% w / w relative to the total protein, and - preferably at least 60% w / w soluble whey protein aggregates based on total protein, preferably at least 70% w / w, more preferably at least 80% w / w, even more preferably at least 90% w / w soluble whey protein aggregates based on total protein.

[0270] Soluble whey protein aggregates made from heat denaturation of whey proteins containing at least 85% w / w BLG appear to be particularly advantageous for use in protein beverages, and without being bound by theory, it is believed that high BLG soluble aggregates provide stronger gels upon acidification than soluble aggregates based on regular WPI. This difference makes it possible to produce beverages that form more gel / higher viscosity in the stomach upon digestion, thereby promoting satiety.

[0271] In some preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises: - soluble whey protein aggregates up to at least 60% w / w of total protein; - protein nanogels up to 20% w / w of total protein, -Contains a maximum of 2% w / w insoluble protein material.

[0272] In some more preferred embodiments of the present invention, the packaged thermally treated beverage preparation comprises: - at least 80% w / w of soluble whey protein aggregates relative to total protein; - protein nanogels up to 5% w / w of total protein, - Contains a maximum of 2% w / w insoluble protein material.

[0273] One aspect of the invention is a method for producing a packaged thermally treated beverage preparation having a pH in the range of 5.5 to 8.0, comprising the steps of: a) - A protein having a total amount of 1 to 20% by weight, wherein at least 85 w / w% of the protein is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, providing a liquid solution comprising: b) packaging the liquid solution; Including, wherein the liquid solution of step a) and / or the packaged liquid solution of step b) are subjected to a heat treatment comprising at least pasteurization.

[0274] Preferably, the method for producing a packaged heat treated beverage preparation having a pH in the range of 5.5 to 8.0 comprises: a) Below: - A total amount of protein of 1 to 20% by weight, wherein at least 85 w / w% of the protein is beta-lactoglobulin (BLG); - optionally providing a liquid solution comprising a sweetener, a sugar polymer and / or a flavouring, b) packaging the liquid solution, wherein 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.

[0275] The liquid solution of step a) preferably has the same composition as the heat-treated beverage preparation, except for the changes caused by the heat treatment. Therefore, the characteristics mentioned in the context of the heat-treated beverage preparation apply equally to the liquid solution, except that the liquid solution typically has a lower degree of protein denaturation than the heat-treated beverage preparation.

[0276] In some preferred embodiments of the liquid solution of the 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.

[0277] Even higher relative amounts of BLG are both achievable and desirable, and in some preferred embodiments of the invention, at least 94% w / w of the protein in 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.

[0278] For example, the liquid solution preferably comprises BLG in an amount of at least 97.5% w / w relative to total protein, preferably at least 98.0% w / w, more preferably at least 98.5% w / w, even more preferably at least 99.0%, most preferably at least 99.5% w / w relative to total protein, such as approximately 100.0%.

[0279] In some preferred embodiments of the invention, the sum of alpha-lactalbumin (ALA) and caseinomacropeptide (CMP) constitutes at least 40% w / w of the non-BLG proteins of the liquid solution, preferably at least 60% w / w, even more preferably 70% w / w, and most preferably at least 90% w / w of the non-BLG proteins of the liquid solution.

[0280] In some preferred embodiments of the invention, ALA constitutes up to 80% w / w of the non-BLG proteins of the liquid solution, preferably up to 60% w / w, even more preferably up to 40% w / w, and most preferably up to 30% w / w of the non-BLG proteins of the liquid solution.

[0281] Even lower ALA contents may be preferred, so that in some preferred embodiments of the invention, ALA constitutes up to 20% w / w of the non-BLG proteins of the liquid solution, preferably up to 15% w / w, even more preferably up to 10% w / w, and most preferably up to 5% w / w of the non-BLG proteins of the liquid solution.

[0282] In other preferred embodiments of the 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 the total protein in the standard whey protein concentrate derived from sweet whey.

[0283] Even lower concentrations of the major non-BLG whey proteins may be desirable, so in additional 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 4%, preferably at most 3%, more preferably at most 2%, even more preferably at most 1% by weight of total protein in a standard whey protein concentrate derived from sweet whey.

[0284] The inventors have seen indications that reduction of lactoferrin and / or lactoperoxidase is particularly advantageous in obtaining neutral coloured whey protein products.

[0285] Therefore, in some preferred embodiments of the present invention, lactoferrin is present in a weight percentage of total protein, which is 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 the weight percentage of total protein in standard whey protein concentrate derived from sweet whey. Even lower concentrations of lactoferrin may be desirable. Therefore, in additional preferred embodiments of the present invention, lactoferrin is present in a weight percentage of total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of the weight percentage of total protein in standard whey protein concentrate derived from sweet whey.

[0286] Similarly, in some preferred embodiments of the present invention, lactoperoxidase is present in a weight percentage of total protein, which is 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 the weight percentage of total protein in standard whey protein concentrate derived from sweet whey.Even lower concentrations of lactoperoxidase may be desirable.Therefore, in a further preferred embodiment of the present invention, lactoperoxidase is present in a weight percentage of total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of the weight percentage of total protein in standard whey protein concentrate derived from sweet whey.

[0287] The protein of the liquid solution is preferably prepared from mammalian milk, preferably from ruminant milk, such as milk from cows, sheep, goats, buffaloes, camels, llamas, horses and / or deer. Proteins from cow's milk are particularly preferred. Thus, the protein of the liquid solution is preferably a cow's milk protein.

[0288] The protein of the liquid solution is preferably whey protein and / or milk serum protein, even more preferably bovine whey protein and / or milk serum protein.

[0289] In some preferred embodiments of the invention, the liquid solution has an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.11, more preferably at least 1.13, even more preferably at least 1.15, and most preferably at least 1.17.

[0290] In some preferred embodiments of the present invention, the liquid solution has a degree of protein denaturation of at most 20%, more preferably at most 10%, even more preferably at most 5%, and most preferably at most 1%.

[0291] Both the degree of protein denaturation and the low fluorescence emission ratio are characteristic of liquid solutions in which the protein is predominantly in the native conformation, which is particularly preferred for producing clear beverages.

[0292] The packaging in step b) can be any suitable packaging technique and any suitable container can be used to package the liquid solution.

[0293] 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, and preferably includes filling the liquid solution into one or more sterile containers.

[0294] Aseptic filling and sealing is particularly preferred where the liquid solution is already sterile or has very low levels of microorganisms prior to filling.

[0295] Examples of useful containers are, for example, bottles, cartons, bricks, and / or bags.

[0296] In some preferred embodiments of the invention, the container wall has a light transmission of at most 10%, preferably at most 1%, more preferably at most 0.1%, even more preferably at most 0.01%, and most preferably at most 0.001% at any wavelength in the range of 250-500 nm.

[0297] In another preferred embodiment of the invention, the container wall has a light transmission in the range of 250-500 nm of at most 10%, preferably at most 1%, more preferably at most 0.1%, even more preferably at most 0.01% and most preferably at most 0.001%.

[0298] The optical transmission of a container wall is measured by providing a plane portion of the container wall and measuring the optical transmission through the container wall at any relevant wavelength. Measurements are made using a standard spectrophotometer by inserting the portion of the container wall into the optical path (e.g., using a cuvette or similar arrangement) such that the plane of the portion of the container wall is positioned perpendicular to the optical path. The transmission at wavelength i is given by T i =I i,後 / I i,前 * Calculated as 100%, where I i,前 is the light intensity at wavelength i before reaching the container wall, and I i,後 is the intensity of the light beam at wavelength i after it has passed through a portion of the container wall.

[0299] The average optical transmission is the sum of all transmission measurements T taken within a given wavelength range. i and dividing the sum by the number of transmission measurements in a given wavelength range.

[0300] In some preferred embodiments of the invention, the container wall has a light transmission of at most 10%, preferably at most 1%, more preferably at most 0.1%, even more preferably at most 0.01%, and most preferably at most 0.001% at any wavelength in the range of 250-800 nm.

[0301] In another preferred embodiment of the invention, the container wall has a light transmission in the range of 250-800 nm of at most 10%, preferably at most 1%, more preferably at most 0.1%, even more preferably at most 0.01% and most preferably at most 0.001%.

[0302] No-light or low-light transmissive containers can be manufactured, for example, using pigmented, absorbent or coated polymers, or pigmented or coated glass, or alternatively, by incorporating a metal layer into the container wall, for example in the form of aluminum foil. Such no-light or low-light transmissive containers are known in the food and pharmaceutical industries.

[0303] Non-limiting examples of suitable polymeric materials include, for example, polyethylene terephthalate (PET) or PET-like polymers.

[0304] In other preferred embodiments of the invention, at least a portion of the container wall is transparent, preferably the entire container is transparent. In some preferred embodiments of the invention, at least a portion of the container wall, preferably the entire container wall, has an average light transmittance in the range of 400-700 nm of at least 11%, preferably at least 20%, more preferably at least 50%, even more preferably at least 60%, and most preferably at least 80%.

[0305] In some preferred embodiments of the method of the present invention, the liquid solution of step a) is subjected to a heat treatment, including at least pasteurization, and then packaged in step b).

[0306] In another embodiment of the method of the present invention, the packaged liquid solution of step b) is subjected to a heat treatment including at least pasteurization.

[0307] In a particular embodiment, the heat treatment comprises heating the beverage preparation to a temperature in the range of 70-80°C.

[0308] In some preferred embodiments of the present invention, the temperature of the heat treatment is in the range of 70-80°C, preferably in the range of 70-79°C, more preferably in the range of 71-78°C, even more preferably in the range of 72-77°C, and most preferably in the range of 73-76°C, such as about 75°C.

[0309] Preferably, the duration of the heat treatment is between 1 second and 60 minutes when carried out in the temperature range of 70 to 80. The longest exposure time is optimal for the lowest temperature of the temperature range and vice versa. In other preferred embodiments, the temperature of the heat treatment is 70° C. for at least 60 minutes, or preferably 75° C. for at least 45 minutes, or preferably 80° C. for at least 30 minutes, or preferably 85° C. for at least 22 minutes, or preferably 90° C. for at least 10 minutes.

[0310] In a particularly preferred embodiment of the present invention, the heat treatment is provided at 70-78°C for 1 second to 30 minutes, more preferably at 71-77°C for 1 minute to 25 minutes, and even more preferably at 72-76°C for 2 minutes to 20 minutes.

[0311] In some preferred embodiments of the invention, the method comprises heat treating at a temperature of 85° C. to 95° C. for 1 to 3 minutes.

[0312] In some embodiments, higher temperatures may also be preferred, especially if unfolding, and optionally aggregation of BLG is required. 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.

[0313] In some preferred embodiments of the present invention, sterilization involves a temperature in the range of 120-150° C. for 4-30 seconds.

[0314] The heat treatment may, for example, include 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, for example, include heating at a temperature in the range of 90-95° C. for 1-10 minutes, such as approximately 120° C. for approximately 20 seconds. Alternatively, the heat treatment may include heating at a temperature in the range of 115-125° C. for 5-30 seconds, such as approximately 120° C. for approximately 20 seconds.

[0315] The heat treatment may for example be a UHT type treatment, which typically involves a temperature in the range of 135-144° C. and a duration in the range of 2-10 seconds.

[0316] 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.

[0317] The implementation of the heat treatment may include the use of equipment such as plate or tubular heat exchangers, scraped surface heat exchangers or retort systems. Alternatively, and particularly preferably for heat treatments above 95°C, direct steam-based heating can be used, for example using direct steam injection, direct steam infusion or spray-cooking. Furthermore, such direct steam-based heating is preferably used in combination with flash cooling. Suitable examples of the implementation of spray cooking can be found in WO2009113858 A1, which are incorporated herein for all purposes. Suitable examples of the implementation of direct steam injection and direct steam infusion can be found in WO2009113858 A1 and WO2010 / 085957 A3, which are incorporated herein for all purposes. General aspects of high temperature processing are described, for example, in "Thermal technologies in food processing" ISBN 185573558 X, which are incorporated herein by reference for all purposes.

[0318] In some preferred embodiments of the present invention, pasteurization is combined with physical microbial reduction.

[0319] Useful examples of physical microbial reduction include one or more of heating, bacterial filtration, UV irradiation, high pressure treatment, pulsed electric field treatment, and ultrasound.

[0320] In some preferred embodiments of the present invention, the heat treatment is sterilization resulting in a sterile liquid beverage preparation. Such sterilization can preferably be obtained by combining bacterial filtration and pasteurization.

[0321] In the context of the present invention, the term "bacterial filtration" refers to filtration carried out with a pore size sufficient to retain microorganisms such as bacteria and spores, but not native BLG. Bacterial filtration is sometimes also called sterile filtration and includes microfiltration of the liquid in question. Bacterial filtration is typically carried out using membranes with a pore size of at most 1 micron, preferably at most 0.8 micron, more preferably at most 0.6 micron, even more preferably at most 0.4 micron, and most preferably at most 0.2 micron.

[0322] Bacterial filtration may involve, for example, membranes having pore sizes of 0.02 to 1 micron, preferably 0.03 to 0.8 micron, more preferably 0.04 to 0.6 micron, even more preferably 0.05 to 0.4 micron, and most preferably 0.1 to 0.2 micron.

[0323] In some preferred embodiments of the invention, the liquid solution is subjected to bacterial filtration followed by heat treatment using a temperature of up to 80° C., preferably up to 75° C. The combination of temperature and duration of this heat treatment is preferably selected to provide a sterile beverage preparation.

[0324] In another preferred embodiment of the invention, the liquid solution is subjected to a bacterial filtration followed by a heat treatment using a temperature of at least 150° C. for a duration of at most 0.2 seconds, preferably at most 0.1 seconds. The combination of temperature and duration of this heat treatment is preferably selected to provide a sterile beverage preparation.

[0325] Depending on the heat treatment temperature used, it may be beneficial to subject the beverage preparation to cooling. According to a preferred embodiment of the process of the present invention, following the heat treatment, the heat treated beverage preparation is optionally cooled, preferably to 0-50°C, preferably to 0-25°C, or preferably to 0-20°C, or preferably to 0-15°C, preferably to 0-10°C, or preferably to 4-8°C, or preferably to 2-5°C, or preferably to 1-5°C.

[0326] If the beverage preparation is pasteurized, it is preferably cooled to 0-15°C, more preferably 1-5°C after heat treatment.

[0327] According to one embodiment of the method, generally any acid or base is used to adjust the pH of the liquid solution. Those skilled in the art will recognize suitable means for adjusting the pH, such as sodium or potassium carbonate, sodium or potassium bicarbonate, or ammonium hydroxide. Preferably, a base such as KOH or NaOH is used to adjust the pH, although other bases including NaOH can also be used to adjust the pH. Those skilled in the art will recognize other 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.

[0328] In some preferred embodiments of the invention, the liquid solution has a pH in the range of 6.5 to 7.5. Most preferably, the pH employed is a pH of 6.5 to 7.0 or a pH of 6.8 to 7.2.

[0329] The liquid solution preferably has a pH in the range of 5.5 to 6.2, alternatively the liquid solution has a pH in the range of 6.2 to 8.0.

[0330] Alternatively, the liquid solution may have a pH in the range of 6.8 to 8.0, and more preferably, the liquid solution has a pH in the range of 6.2 to 8.0. It has been found that the liquid solution of the present invention has low viscosity, is clear and transparent, preferably in the pH range of 6.2 to 8.0, preferably 6.3 to 7.6, more preferably 6.5 to 7.2.

[0331] It has been found that the liquid solution of the present invention has low viscosity and a milky appearance, preferably at a pH in the range of 5.5 to 8.0, preferably 5.7 to 6.8, more preferably 5.8 to 6.0.

[0332] In some preferred embodiments of the present invention, it has been found that the liquid solution of the present invention is preferably heat treated at a pH in the range of 5.6-6.2, preferably at a pH of 5.6-8.0, optionally mixed with sources of carbohydrates, fats, minerals and vitamins, adjusted to a preferred pH of 6.2-8.0 and subjected to a second heat treatment (UHT).

[0333] In some preferred embodiments of the invention, the liquid solution comprises a total amount of protein of 4.0-20% w / w based on the weight of the beverage.

[0334] In some embodiments of the invention, the liquid solution advantageously has a protein content of 2.0-10.0% w / w based on the weight of the solution. Thus, in some embodiments of the present invention, the liquid solution preferably comprises a total amount of protein of 2.0-10% w / w by weight of the liquid solution, preferably a total amount of protein of 3.0-10% w / w by weight of the liquid solution, preferably a total amount of protein of 5.0-9.0% w / w by weight of the liquid solution, preferably a total amount of protein of 6.0-8.0% w / w by weight of the liquid solution.

[0335] In some embodiments of the invention, the protein content of the liquid solution is advantageously high, such as 10.0-20% w / w based on the weight of the liquid solution.

[0336] Thus, in some embodiments of the invention, the liquid solution preferably comprises a total amount of protein of 10.0-20% w / w by weight of the liquid solution, preferably a total amount of protein of 12-19% w / w by weight of the liquid solution, even more preferably a total amount of protein of 15-18% w / w by weight of the liquid solution, and most preferably a total amount of protein of 16-17% w / w by weight of the liquid solution.

[0337] It is particularly preferred that the liquid solution comprises a BLG isolate in combination with other protein sources, for example, preferably as the primary protein source, and even perhaps as the sole protein source.

[0338] The BLG isolate is preferably a BLG isolate powder or a liquid BLG isolate comprising water and the solids of the BLG isolate powder in an amount in the range of 1-50% w / w.

[0339] The beta-lactoglobulin (BLG) isolated 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, and is selected from the following: - 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 - containing water in an amount of up to 10% w / w.

[0340] The BLG isolated powder preferably comprises: - 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 up to 200 NTU at pH 3.9, and - Up to 1000 colony forming units / g, or more.

[0341] The BLG isolate powder is preferably an edible composition.

[0342] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of 2 to 4.9. Such powders are particularly useful in acidic foods, especially acidic beverages.

[0343] In another preferred embodiment of the present invention, the BLG isolated powder has a pH in the range of 6.1 to 8.5.

[0344] In some preferred embodiments of the present invention, the BLG isolated 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.

[0345] Even higher protein contents may be required, and in some preferred embodiments of the present invention, the BLG isolated 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. Total protein is determined according to Example 1.5.

[0346] In some preferred embodiments of the present invention, the BLG isolated powder comprises BLG in an amount of at least 92% w / w relative to total protein, preferably at least 95% w / w, more preferably at least 97% w / w, more preferably at least 98%, and most preferably at least 99.5% w / w relative to total protein.

[0347] In some preferred embodiments of the invention, the combined weight of alpha-lactalbumin (ALA) and caseinomacropeptide (CMP) constitutes at least 40% w / w of the non-BLG proteins of the powder, preferably at least 60% w / w, more preferably at least 70% w / w, and most preferably at least 90% w / w of the non-BLG proteins of the powder.

[0348] In other preferred embodiments of the 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 the total protein in the standard whey protein concentrate derived from sweet whey.

[0349] Even lower concentrations of the major non-BLG whey proteins may be desirable, so in additional 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 4%, preferably at most 3%, more preferably at most 2%, even more preferably at most 1% by weight of total protein in a standard whey protein concentrate derived from sweet whey.

[0350] The inventors have seen indications that reduction of lactoferrin and / or lactoperoxidase is particularly advantageous in obtaining neutral coloured whey protein products.

[0351] Therefore, in some preferred embodiments of the present invention, lactoferrin is present in a weight percentage of total protein, which is 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 the weight percentage of total protein in standard whey protein concentrate derived from sweet whey. Even lower concentrations of lactoferrin may be desirable. Therefore, in additional preferred embodiments of the present invention, lactoferrin is present in a weight percentage of total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of the weight percentage of total protein in standard whey protein concentrate derived from sweet whey.

[0352] Similarly, in some preferred embodiments of the present invention, lactoperoxidase is present in a weight percentage of total protein, which is 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 the weight percentage of total protein in standard whey protein concentrate derived from sweet whey.Even lower concentrations of lactoperoxidase may be desirable.Therefore, in a further preferred embodiment of the present invention, lactoperoxidase is present in a weight percentage of total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of the weight percentage of total protein in standard whey protein concentrate derived from sweet whey.

[0353] Lactoferrin and lactoperoxidase are quantified according to Example 1.29.

[0354] In some preferred embodiments of the invention, the BLG isolated powder has a moisture content in an amount of up to 10% w / w, preferably up to 7% w / w, more preferably up to 6% w / w, even more preferably up to 4% w / w, and most preferably up to 2% w / w.

[0355] In some preferred embodiments of the present invention, the BLG isolated 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%. For example, the BLG isolated powder may comprise carbohydrates such as, for example, lactose, oligosaccharides, and / or hydrolysis products of lactose (i.e., glucose and galactose), sucrose, and / or maltodextrins.

[0356] In some preferred embodiments of the invention, the BLG isolated powder comprises lipids in an amount of up to 10% w / w, preferably up to 5% w / w, more preferably up to 2% w / w, and even more preferably up to 0.1% w / w.

[0357] The present inventors have found that it can be advantageous to control the mineral content to achieve some of the desired properties of the BLG isolate powder.

[0358] In some preferred embodiments of the invention, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolation powder is up to 10 mmol / g protein. Preferably, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolation powder is up to 6 mmol / g protein, more preferably up to 4 mmol / g protein, and even more preferably up to 2 mmol / g protein.

[0359] In another preferred embodiment of the invention, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolation powder is at most 1 mmol / g protein. Preferably, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolation 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.

[0360] In another preferred embodiment of the present invention, the sum of the Mg and Ca amounts of the BLG isolated powder is at most 5 mmol / g protein. Preferably, the sum of the Mg and Ca amounts of the BLG isolated powder is at most 3 mmol / g protein, more preferably at most 1.0 mmol / g protein, even more preferably at most 0.5 mmol / g protein.

[0361] In another preferred embodiment of the present invention, the sum of the Mg and Ca amounts of the BLG isolated powder is at most 0.3 mmol / g protein. Preferably, the sum of the Mg and Ca amounts of the BLG isolated 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.

[0362] The inventors have found that it is possible to create a low phosphorus / low potassium version of BLG isolate powder that is particularly useful for patients with renal disease. To produce such a product, the phosphorus and potassium content of the BLG isolate powder must be equally low.

[0363] Thus, in some preferred embodiments of the present invention, the BLG isolated powder has a total phosphorus content of up to 100 mg phosphorus per 100 g protein. Preferably, the BLG isolated powder has a total phosphorus content of up to 80 mg phosphorus per 100 g protein. More preferably, the BLG isolated powder has a total phosphorus content of up to 50 mg phosphorus per 100 g protein. Even more preferably, the BLG isolated powder has a total phosphorus content of up to 20 mg phosphorus per 100 g protein. The total phosphorus content of the BLG isolated powder is up to 5 mg phosphorus per 100 g protein.

[0364] In some preferred embodiments of the present invention, the BLG isolated powder comprises up to 600 mg potassium per 100 g protein. More preferably, the BLG isolated powder comprises up to 500 mg potassium per 100 g protein. More preferably, the BLG isolated powder comprises up to 400 mg potassium per 100 g protein. More preferably, the BLG isolated powder comprises up to 300 mg potassium per 100 g protein. Even more preferably, the BLG isolated powder comprises up to 200 mg potassium per 100 g protein. Even more preferably, the BLG isolated powder comprises up to 100 mg potassium per 100 g protein. Even more preferably, the BLG isolated powder comprises up to 50 mg potassium per 100 g protein, and even more preferably, the BLG isolated powder comprises up to 10 mg potassium per 100 g protein.

[0365] The phosphorus content is related to the total amount of elemental phosphorus in the composition in question and is determined according to Example 1.19. Similarly, the potassium content is related to the total amount of elemental potassium in the composition in question and is determined according to Example 1.19.

[0366] In some preferred embodiments of the invention, the BLG isolated powder comprises up to 100mg phosphorus / 100g protein and up to 700mg potassium / 100g protein, preferably up to 80mg phosphorus / 100g protein and up to 600mg potassium / 100g protein, more preferably up to 60mg phosphorus / 100g protein and up to 500mg potassium / 100g protein, more preferably up to 50mg phosphorus / 100g protein and up to 400mg potassium / 100g protein, or more preferably up to 20mg phosphorus / 100g protein and up to 200mg potassium / 100g protein, or even more preferably up to 10mg phosphorus / 100g protein and up to 50mg potassium / 100g protein. In some preferred embodiments of the invention, the BLG isolated powder comprises up to 100mg phosphorus / 100g protein and up to 340mg potassium / 100g protein.

[0367] The low phosphorus and / or low potassium composition according to the present invention can be used as a food ingredient for producing food for patient groups with reduced renal function.

[0368] In the context of the present invention, a transparent liquid has a turbidity of at most 200 NTU, measured according to Example 1.7.

[0369] Thus, in some preferred embodiments of the present invention, the BLG isolated powder has a pH in the range of 2 to 4.9. Preferably, the BLG isolated 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 isolated powder can have a pH in the range of 3.6 to 4.3.

[0370] The inventors have found that for some applications, such as pH neutral foods, and especially pH neutral beverages, it is particularly advantageous to use a pH neutral BLG isolate powder, this being especially true for high protein, clear or opaque pH neutral beverages.

[0371] Thus, in some preferred embodiments of the present invention, the BLG isolated 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.

[0372] In another preferred embodiment of the invention, the BLG isolated 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.

[0373] Advantageously, the BLG isolated powder of 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 , and most preferably at least 0.6 g / cm 3 The bulk density may be

[0374] Low density powders such as freeze-dried BLG isolate are fluffy and easily drawn into the air in the manufacturing area during use. This is problematic as freeze-dried powders increase the risk of cross-contamination to other foods and dusty environments are known to cause hygiene problems. In extreme cases, dusty environments also increase the risk of dust explosions.

[0375] The high density variants of the present invention are easier to handle and less likely to bleed into the surrounding air.

[0376] An additional advantage of the high density variants of the present invention is that they take up less space during shipping, thereby increasing the weight of BLG isolated powder that can be shipped in one volumetric unit.

[0377] A further advantage of the high density variants of the present invention is that they are more efficient than other powdered food ingredients, such as 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 ) etc., separation is difficult.

[0378] The BLG isolated powder of the present invention has a density of, for example, 0.2 to 1.0 g / cm 3 in the range of 0.30 to 0.9 g / cm 3 More preferably, the range is 0.40 to 0.8 g / cm 3 and even more preferably in the range of 0.45 to 0.75 g / cm 3 , and even more preferably 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 0.01 to 0.01.

[0379] The bulk density of the powder is determined according to Example 1.17.

[0380] 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 mouthfeel when BLG is used in acidic beverages.

[0381] The intrinsic tryptophan fluorescence ratio (I330 / I350) is a measure of the degree of BLG unfolding, and we have found that a higher intrinsic tryptophan fluorescence ratio correlates with less or no unfolding of BLG, and thus a less dry mouthfeel is observed. The intrinsic tryptophan fluorescence ratio (I330 / I350) is measured according to Example 1.1.

[0382] In some preferred embodiments of the present invention, the BLG isolated powder has an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.11.

[0383] In some preferred embodiments of the present invention, the BLG isolated 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.

[0384] If the BLG isolated 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 invention, the protein fraction of the BLG isolated powder has an intrinsic tryptophan fluorescence ratio of at least 1.11.

[0385] In some preferred embodiments of the invention, the protein fraction of the BLG isolated 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.

[0386] For example, the BLG isolated powder can be dissolved in demineralized water and the solution can be subjected to dialysis or ultrafiltration-based diafiltration using a protein-retaining filter to separate the protein fraction from the BLG isolated powder. If the BLG isolated 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.

[0387] It is often preferred that a significant amount of the BLG in the BLG isolated powder is non-aggregated BLG. Preferably, at least 50% of the BLG is non-aggregated BLG. More preferably, at least 80% of the BLG is non-aggregated BLG. Even more preferably, at least 90% of the BLG is non-aggregated BLG. Most preferably, at least 95% of the BLG is non-aggregated BLG. Even more preferably, approximately 100% of the BLG in the BLG isolated powder is non-aggregated BLG.

[0388] In some preferred embodiments of the present invention, the BLG isolated powder has a degree of protein denaturation of up to 10%, preferably up to 8%, more preferably up to 6%, even more preferably up to 3%, even more preferably up to 1%, and most preferably up to 0.2%.

[0389] However, it may also be preferred for the BLG isolate powder to have a significant level of protein denaturation, for example when an opaque beverage is desired. Thus, 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%.

[0390] Where the BLG isolate powder has a significant level of protein denaturation, it is often preferable to maintain a low level of insoluble protein material, i.e., precipitated protein material that may precipitate in the beverage during storage. The level of insoluble material is measured according to Example 1.10.

[0391] In some preferred embodiments of the invention, the BLG isolated powder comprises up to 20% w / w insoluble protein material, preferably up to 10% w / w insoluble protein material, more preferably up to 5% w / w insoluble protein material, even more preferably up to 3% w / w insoluble protein material, and most preferably up to 1% w / w insoluble protein material. It may even be preferred that the BLG isolated powder is completely free of insoluble protein material.

[0392] 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.

[0393] The BLG isolated powder has a thermal stability at pH 3.9 of up to 200 NTU, preferably up to 100 NTU, more preferably up to 60 NTU, even more preferably up to 40 NTU, and most preferably up to 20 NTU. Even better thermal stability is possible, the BLG isolated powder preferably has a thermal stability at pH 3.9 of up to 10 NTU, preferably up to 8 NTU, more preferably up to 4 NTU, and even more preferably up to 2 NTU.

[0394] The microbial content of BLG isolated powder is preferably kept to a minimum.However, it is difficult to obtain both high protein nativeity and low content of microbial, because the microbial reduction process tends to lead to protein unfolding and denaturation.The present invention makes it possible to obtain very low microbial content while maintaining high level of nativeity of BLG.

[0395] Thus, in some preferred embodiments of the present invention, the BLG isolated powder comprises up to 15,000 colony forming units (CFU) / g. Preferably, the BLG isolated powder comprises up to 10,000 CFU / g. More preferably, the BLG isolated powder comprises up to 5,000 CFU / g. Even more preferably, the BLG isolated powder comprises up to 1,000 CFU / g. Even more preferably, the BLG isolated powder comprises up to 300 CFU / g. Most preferably, the BLG isolated powder comprises up to 100 CFU / g, such as up to 10 CFU / g. In particularly preferred embodiments, the powder is sterile. Sterile BLG isolated powder is prepared by combining several physical microbial reduction processes during the manufacture of the BLG isolated powder, such as, for example, microfiltration and heat treatment at acidic pH.

[0396] In some preferred embodiments of the present invention, the BLG isolated 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, and is selected from the following: - 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, The BLG isolated powder has the following properties: - an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.11; - protein denaturation up to 10%, and -Heat stability up to 200 NTU at pH 3.9.

[0397] In some preferred embodiments of the present invention, the BLG isolated powder has a pH in the range of i) 2 to 4.9, or ii) 6.1 to 8.5, and is as follows: - 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, The BLG isolated powder has the following properties: - an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.11; a protein denaturation degree of at most 10%, preferably at most 5%, and -Has a thermal stability of at most 70 NTU, preferably at most 50 NTU, even more preferably at most 40 NTU at pH 3.9.

[0398] In some preferred embodiments of the present invention, the BLG isolated powder has a pH in the range of i) 2 to 4.9, or ii) 6.1 to 8.5, and is as follows: -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 isolated powder has the following properties: - 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%, and -Heat stability up to 200 NTU at pH 3.9.

[0399] In another preferred embodiment of the present invention, the BLG isolated powder has a pH in the range of 2 to 4.9, and is as follows: - 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, The BLG isolated powder has the following properties: - 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 of at most 50 NTU, preferably at most 30 NTU, even more preferably at most 10 NTU at pH 3.9.

[0400] In yet another preferred embodiment of the present invention, the BLG isolated powder has a pH in the range of 6.1 to 8.5, and has the following: - 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, The BLG isolated powder has the following properties: - 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 of at most 50 NTU, preferably at most 30 NTU, even more preferably at most 10 NTU at pH 3.9.

[0401] In yet another preferred embodiment of the present invention, the BLG isolated powder has a pH in the range of 6.1 to 8.5, and has the following: - 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, The BLG isolated powder has the following properties: - 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 of at most 50 NTU, preferably at most 30 NTU, even more preferably at most 10 NTU at pH 3.9.

[0402] In yet another preferred embodiment of the present invention, the BLG isolated powder has a pH in the range of 5.0 to 6.0 and has the following: - 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, The BLG isolated powder has the following properties: - 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 thermal stability of at most 50 NTU, preferably at most 30 NTU, even more preferably at most 10 NTU at pH 3.9, and - preferably has a BLG crystallinity of less than 10%.

[0403] BLG isolate powders containing BLG in an amount of at least 85% w / w relative to total protein typically have the following: 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) a pH in the range of 5.0 to 6.0; providing a liquid BLG isolate having said liquid BLG isolate comprising BLG in an amount of at least 85 w / w relative to total protein. b) optionally subjecting the liquid BLG isolate to physical microbial reduction; c) drying the liquid BLG isolate, preferably by spray drying.

[0404] The liquid BLG isolate is preferably prepared from mammalian milk, preferably from ruminant milk, such as milk from cows, sheep, goats, buffaloes, camels, llamas, horses and / or deer. Proteins from bovine milk are particularly preferred. Thus, the BLG is preferably bovine BLG.

[0405] The liquid BLG isolate can be provided in a number of different ways.

[0406] Typically, providing a liquid BLG isolate comprises, or even consists of, separating BLG from a whey protein feed to provide a BLG-enriched 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.

[0407] A particularly preferred method for providing a BLG-rich composition is by crystallization of BLG, preferably by salting out or by salting out.

[0408] The whey protein feed is preferably WPC, WPI, SPC, SPI, or a combination thereof.

[0409] The term "whey protein feed" relates to the BLG-enriched composition and the subsequent composition from which the liquid BLG isolate is derived.

[0410] In some embodiments of the present invention, the preparation of the BLG-enriched composition comprises or even consists of high salt BLG crystallization in the pH range 3.6-4.0 according to US Pat. No. 2,790,790 A1.

[0411] In another embodiment of the invention, the preparation of the BLG-enriched composition comprises or even 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, pages 562-571) or Vyas et al. (Scale-Up of Native β-Lactoglobulin Affinity Separation Process, J. Dairy Sci. 85:1639-1645, 2002).

[0412] However, in a particularly preferred embodiment of the present invention, the BLG-enriched composition is prepared by crystallization at pH 5-6 under salting conditions as described in PCT application PCT / EP2017 / 084553, which is incorporated herein by reference for all purposes.

[0413] In some preferred embodiments of the invention, the BLG-rich composition is an edible BLG composition according to PCT / EP2017 / 084553, comprising at least 90% BLG relative to total protein, and preferably comprising BLG crystals.

[0414] If it does not already have the requisite properties for use as a liquid BLG isolate, the BLG-rich composition isolated from the whey protein feed may be prepared as part of providing a liquid BLG isolate by: - Desalination, -Addition of minerals, -Dilution, -concentrated, -Physical microorganism reduction; -pH adjustment, The process may be subjected to one or more steps selected from the group consisting of:

[0415] Non-limiting examples of desalting methods include, for example, dialysis, gel filtration, UF / diafiltration, NF / diafiltration, and ion exchange chromatography.

[0416] Non-limiting examples of mineral additions include the addition of soluble food acceptable salts such as, for example, Na, K, Ca, and / or Mg salts. Such salts can be, for example, phosphate salts, chloride salts, or salts of food acids such as, for example, citrate or lactate salts. Minerals can be added in solid, suspended, or dissolved form.

[0417] 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.

[0418] Non-limiting examples of concentration techniques include, for example, evaporation, reverse osmosis, nanofiltration, ultrafiltration, and combinations thereof.

[0419] If concentration is required to increase the concentration of protein relative to total solids, then it is preferred to use a concentration step such as ultrafiltration or alternatively dialysis. If concentration is not required to increase the concentration of protein relative to total solids, then methods such as evaporation, nanofiltration and / or reverse osmosis may be useful.

[0420] Non-limiting examples of physical microbial reduction include, for example, heat treatment, bacterial filtration, UV irradiation, high pressure treatment, pulsed electric field treatment, and ultrasound, which methods are well known to those skilled in the art.

[0421] Non-limiting examples of pH adjustment include, for example, the addition of bases and / or acids, preferably food-acceptable bases and / or acids. It is particularly preferred to use acids and / or bases capable of chelating divalent metal cations. Examples of such acids and / or bases are citric acid, citrate salts, EDTA, lactic acid, lactate salts, phosphoric acid, phosphate salts, and combinations thereof.

[0422] In some preferred embodiments of the present invention, particularly when the preparation has a turbidity of up to 200 NTU, more preferably up to 40 NTU, the liquid solution has a color value delta b in the range of -0.10 to +0.51 on the CIELAB color scale. * has.

[0423] In another preferred embodiment of the 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. * has.

[0424] The liquid solutions of the present invention may contain macronutrients other than protein.

[0425] In some embodiments of the present invention, the liquid solution further comprises a carbohydrate. The total carbohydrate content in the liquid solution of the present invention depends on the intended use of the final heat treated beverage preparation.

[0426] In some 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, saccharose, maltose, dextrose, galactose, maltodextrin, corn syrup solids, sucromalt, glucose polymers, corn syrup, modified starch, resistant starch, carbohydrates derived from rice, isomaltulose, white sugar, glucose, fructose, lactose, high fructose com syrup, honey, sugar alcohols, fructooligosaccharides, soybean fiber, corn fiber, guar gum, konjac flour, polydextrose, Fibersol, and combinations thereof. In some embodiments of the present invention, the packaged thermally processed beverage preparation comprises a non-digestible sugar such as a fructan, the fructan comprising inulin or fructooligosaccharide.

[0427] 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 solution.

[0428] An even lower carbohydrate content is often preferred, and in some preferred embodiments of the present invention, it is 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, even more preferably in the range of 0-10% of the total energy content of the preparation.

[0429] In some preferred embodiments of the invention, the carbohydrate content of the liquid solution is at most 3% of the total energy content of the liquid solution, more preferably at most 1% of the total energy content of the liquid solution, even more preferably at most 0.1% of the total energy content of the liquid solution.

[0430] In one embodiment of the invention, the liquid solution further comprises at least one additional ingredient selected from the group consisting of vitamins, flavorings, minerals, sweeteners, antioxidants, food acids, lipids, carbohydrates, prebiotics, probiotics, and non-whey proteins. Further ingredients ensure that the final packaged heat treated beverage preparation contains the desired nutrients, i.e. nutrients specifically adapted for patients suffering from protein deficiencies or athletes wanting to build muscle.

[0431] 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, stevia glycosides such as rebaudioside A, or combinations thereof. In some embodiments of the present invention, it is particularly preferred that the sweetener comprises or even consists of one or more high-intensity sweeteners (HIS).

[0432] HIS are found in both natural and artificial sweeteners and typically have a sweetening intensity at least 10 times that of sucrose.

[0433] When used, the total amount of HIS is typically in the range of 0.01-2% w / w. For example, the total amount of HIS can be in the range of 0.05-1.5% w / w. Alternatively, the total amount of HIS can be in the range of 0.1-1.0% w / w.

[0434] The choice of sweetener may depend on the beverage being produced, for example, high intensity sweeteners (e.g., aspartame, acesulfame K, sucralose, etc.) may be used in beverages where an energy contribution from the sweetener is not desired, whereas natural sweeteners (e.g., steviol glycosides, sorbitol, or sucrose) may be used in beverages with a natural profile.

[0435] Alternatively or additionally, carbohydrate sweeteners can be used.

[0436] Furthermore, it may be preferred that the sweetener comprises or even consists of one or more polyol sweetener(s). Non-limiting examples of useful polyol sweeteners are maltitol, mannitol, lactitol, sorbitol, inositol, xylitol, threitol, galactitol, or combinations thereof. When used, the total amount of polyol sweetener is typically in the range of 1-20% w / w. For example, the total amount of polyol sweetener may be in the range of 2-15% w / w. Alternatively, the total amount of polyol sweetener may be in the range of 4-10% w / w.

[0437] The liquid solution of the present invention may contain macronutrients other than protein. In some embodiments of the present invention, the liquid solution further comprises 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.

[0438] In some preferred embodiments of the invention, the liquid solution has a lipid content in the range 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.

[0439] The amount of lipids is determined according to ISO 1211:2010 (Determination of fat content - Roese-Gottlieb gravimetric method).

[0440] In some preferred embodiments of the invention, the lipid content of the liquid solution is at most 3% of the total energy content of the liquid solution, more preferably at most 1% of the total energy content of the liquid solution, even more preferably at most 0.1% of the total energy content of the liquid solution.

[0441] The liquid solution typically comprises a total amount of water in the range of 50-99% w / w, preferably in the range of 45-97% w / w, more preferably in the range of 40-95% w / w, even more preferably in the range of 35-90% w / w, and most preferably in the range of 30-85% w / w.

[0442] In some preferred embodiments of the invention, the liquid solution comprises a total amount of water in the range of 55-90% w / w, preferably in the range of 57-85% w / w, more preferably in the range of 60-80% w / w, even more preferably in the range of 62-75% w / w, and most preferably in the range of 65-70% w / w.

[0443] In some preferred embodiments of the invention, the liquid solution comprises a total amount of water in the range of 90-99% w / w, preferably in the range of 92-98.5% w / w, more preferably in the range of 94-98% w / w, even more preferably in the range of 95-98% w / w, and most preferably in the range of 96-98% w / w.

[0444] In some preferred embodiments of the invention, the liquid solution comprises up to 1.0% w / w ethanol, more preferably up to 0.5% w / w, even more preferably up to 0.1% w / w, and most preferably is non-alcoholic, meaning there is no detectable ethanol.

[0445] The liquid solution typically comprises a total solids content in the range of 1-45% w / w, preferably in the range of 5-40% w / w, more preferably in the range of 10-35% w / w, even more preferably in the range of 12-30% w / w, and most preferably in the range of 16-25% w / w.

[0446] In some preferred embodiments of the invention, the liquid solution comprises a total solids content in the range of 10-45% w / w, preferably in the range of 15-43% w / w, more preferably in the range of 20-40% w / w, even more preferably in the range of 25-38% w / w, and most preferably in the range of 30-35% w / w.

[0447] In some preferred embodiments of the invention, the liquid solution comprises a total solids content in the range of 1-10% w / w, preferably in the range of 1.5-8% w / w, more preferably in the range of 2-6% w / w, even more preferably in the range of 2-5% w / w, and most preferably in the range of 2-4% w / w.

[0448] The portion of the liquid solution that is not a solid is preferably water.

[0449] The inventors have found that it can be advantageous to control the mineral content to achieve some of the desired properties of a packaged thermally processed beverage preparation.

[0450] The inventors have surprisingly found that the use of BLG isolate as defined herein and in Examples 2 and 3 allows for the production of heat treated beverage preparations without compromising viscosity and avoiding gelling, potentially allowing the production of packaged heat treated beverage preparations with high mineral content, potentially allowing the production of beverages that are nutritionally complete or nutritionally incomplete dietary supplements.

[0451] In some embodiments of the present invention, the liquid solution 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.

[0452] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg and Ca is in the range of 0-400 mM, preferably in the range of 10-200 mM, or preferably in the range of 20-100 mM in the liquid solution.

[0453] In some preferred embodiments of the invention, the sum of the amounts of Na, K, Mg and Ca is up to 400 mM in the liquid solution.

[0454] In another preferred embodiment of the invention the sum of the amounts of Na, K, Mg and Ca is at most 300 mM, preferably at most 200 mM, 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 20 mM, or preferably at most 10 mM, or preferably at most 5 mM, or preferably at most 1 mM in the liquid solution.

[0455] In some preferred embodiments of the invention, the sum of the amounts of Mg and Ca is at most 75 mM in the liquid solution, more preferably at most 40 mM in the liquid solution, more preferably at most 20 mM in the liquid solution.

[0456] In another preferred embodiment of the invention, the sum of the amounts of Mg and Ca is at most 10 mM in the liquid solution, more preferably at most 8.0 mM in the liquid solution, more preferably at most 6.0 mM in the liquid solution, even more preferably at most 4.0 mM in the liquid solution, and most preferably at most 2.0 mM in the liquid solution.

[0457] In another exemplary embodiment of the invention, 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.

[0458] In another preferred embodiment of the present invention, the liquid solution is a low-mineral solution.

[0459] In the context of the present invention, the term "low-mineral" refers to: - Maximum 1.2% w / w transparent ash based on total solids, - total calcium and magnesium content of max. 0.3% w / w based on total solids, - maximum total sodium and potassium content of 0.10% w / w based on total solids; - a total phosphorus content of up to 100 mg per 100 g protein,

[0460] Preferably, the low mineral composition comprises: - maximum ash content of 0.7% w / w based on total solids; - total calcium and magnesium content of maximum 0.2% w / w based on total solids, - total sodium and potassium content of maximum 0.08% w / w based on total solids; a total phosphorus content of up to 80 mg per 100 g protein,

[0461] Even more preferably, the low mineral composition comprises: - maximum ash content of 0.5% w / w based on total solids, - total calcium and magnesium content of maximum 0.15% w / w based on total solids; - total sodium and potassium content of maximum 0.06% w / w based on total solids; a total phosphorus content of up to 50 mg per 100 g protein,

[0462] The low mineral composition is as follows: - maximum ash content of 0.5% w / w based on total solids, - total calcium and magnesium content of maximum 0.15% w / w based on total solids; - total sodium and potassium content of maximum 0.06% w / w based on total solids; It is particularly preferred that the protein has a total phosphorus content of up to 50 mg per 100 g of protein.

[0463] In another exemplary embodiment of the invention, 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.

[0464] The inventors have found that the present invention allows the preparation of packaged heat-treated beverage preparations that have very low contents of phosphorus and other minerals, such as potassium, which is advantageous for patients suffering from renal disease or who otherwise have reduced renal function.

[0465] The liquid solution is preferably a low phosphorus beverage preparation.

[0466] The liquid solution is preferably a low potassium beverage preparation.

[0467] The liquid solution is preferably a low phosphorus and low potassium beverage preparation.

[0468] In the context of the present invention, the term "low phosphorus" relates to a composition, e.g. a liquid, powder or other food product, having a total phosphorus content of up to 100 mg per 100 g protein. Preferably, the low phosphorus composition has a total phosphorus content of up to 80 mg per 100 g protein. More preferably, the low phosphorus composition may have a total phosphorus content of up to 50 mg per 100 g protein. Even more preferably, the low phosphorus composition may have a total phosphorus content of up to 20 mg per 100 g protein. Even more preferably, the low phosphorus composition may have a total phosphorus content of up to 5 mg per 100 g protein. The low phosphorus composition according to the present invention can be used as a food ingredient for producing a food product for a patient group with reduced renal function.

[0469] The phosphorus content relates to the total amount of elemental phosphorus in the composition in question and is determined according to Example 1.19.

[0470] The potassium content relates to the total amount of elemental potassium in the composition in question and is determined according to Example 1.19.

[0471] In some preferred embodiments of the present invention, the liquid solution comprises up to 100mg phosphorus / 100g protein and up to 700mg potassium / 100g protein, preferably up to 80mg phosphorus / 100g protein and up to 600mg potassium / 100g protein, more preferably up to 60mg phosphorus / 100g protein and up to 500mg potassium / 100g protein, more preferably up to 50mg phosphorus / 100g protein and up to 400mg potassium / 100g protein, or more preferably up to 20mg phosphorus / 100g protein and up to 200mg potassium / 100g protein, or even more preferably up to 10mg phosphorus / 100g protein and up to 50mg potassium / 100g protein. In some preferred embodiments of the present invention, the packaged heat treated beverage preparation comprises up to 100mg phosphorus / 100g protein and up to 340mg potassium / 100g protein.

[0472] The liquid solution containing small amounts of phosphorus and potassium can advantageously be supplemented with carbohydrates and lipids, and the heat-treated beverage preparation preferably further comprises a total amount of carbohydrates in the range of 30-60% of the total energy content of the liquid solution, preferably in the range of 35-50E%, and a total amount of lipids in the range of 20-60% of the total energy content, preferably in the range of 30-50E%.

[0473] 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.

[0474] In one embodiment of the invention, the liquid solution includes a plurality of vitamins and a plurality of minerals.

[0475] In some preferred embodiments of the present invention, the liquid solution comprises one or more edible 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.

[0476] In one embodiment of the present invention, the liquid solution further comprises a flavoring selected from the group consisting of salt, seasoning, umami seasoning and / or spices. In a preferred embodiment of the present invention, the flavoring comprises chocolate, cocoa, lemon, orange, lime, strawberry, banana, forest fruit flavors or combinations thereof. The choice of flavoring depends on the beverage to be produced.

[0477] The inventors have discovered that the present invention, and in particular the use of a protein fraction comprising at least 85% w / w BLG relative to the total protein, allows for the formation of protein nanogels at surprisingly high protein concentrations that were previously thought to lead to uncontrolled gel formation, which is advantageous as it allows for the direct production of high protein beverages without the need to subsequently concentrate the protein content after denaturing and concentrating the protein nanogel.

[0478] Therefore, in some preferred embodiments of the present invention, the liquid solution, or a previous solution used to prepare the liquid solution, comprises one or more of the following: - a total amount of protein in the range of 5 to 20% w / w, preferably in the range of 8 to 19% w / w, more preferably in the range of 10 to 18% w / w, most preferably in the range of 12 to 16% w / w; - 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 92% w / w relative to the total protein, most preferably at least 96% w / w relative to the total protein, below: a degree of protein denaturation of at most 30%, preferably at most 20%, even more preferably at most 10%, most preferably at most 5%, and It has a pH in the range of -5.6 to 6.5, preferably 5.8 to 6.2, and more preferably 5.9 to 6.1.

[0479] In the context of the present invention, the term "previous solution" or "previous solution used to prepare the liquid solution" relates to an aqueous solution used to prepare the liquid solution, typically having the same protein content as the liquid solution or a slightly higher protein content. The use of a previous solution is preferred when proteins, especially native BLG, must be modified, for example by heat denaturation, prior to forming the liquid solution.

[0480] In some preferred embodiments of the present invention, the liquid solution, or a previous solution used to prepare the liquid solution, comprises a total amount of protein in the range of 10-20% w / w, preferably in the range of 10-19% w / w, more preferably in the range of 10-18% w / w, and most preferably in the range of 10-16% w / w.

[0481] In another preferred embodiment of the invention, the liquid solution, or a previous solution used to prepare the liquid solution, comprises a total amount of protein in the range of 5-20% w / w, preferably in the range of 8-19% w / w, more preferably in the range of 10-18% w / w, most preferably in the range of 12-16% w / w. A pH in the range of -5.6 to 6.5, preferably 5.8 to 6.2, more preferably 5.9 to 6.1.

[0482] In some preferred embodiments of the invention, the liquid solution, or a previous solution used to prepare the liquid solution, has a degree of protein denaturation of at most 30%, preferably at most 20%, even more preferably at most 10%, and most preferably at most 5%.

[0483] An even lower degree of protein denaturation may be preferred, so in some preferred embodiments of the invention, the liquid solution, or a previous solution used to prepare the liquid solution, has a degree of protein denaturation of at most 4%, preferably at most 2%, even more preferably at most 1%, and most preferably at most 0.2%.

[0484] In some preferred embodiments of the invention, the liquid solution, or a previous solution used to prepare the liquid solution, has a lipid content of up to 5% w / w, more preferably up to 2% w / w, even more preferably up to 0.5% w / w, and most preferably up to 0.1% w / w.

[0485] In some preferred embodiments of the invention, the liquid solution, or a previous solution used to prepare the liquid solution, has a carbohydrate content of up to 12% w / w, more preferably up to 6% w / w, even more preferably up to 2% w / w, and most preferably up to 0.1% w / w.

[0486] However, in other preferred embodiments of the invention, the liquid solution, or the previous solution used to prepare the liquid solution, has a carbohydrate content of 2-25% w / w, more preferably 4-20% w / w, even more preferably 5-18% w / w, most preferably 6-15% w / w. These embodiments are useful, for example, when a significant content of carbohydrates is required in the final beverage. Alternatively, carbohydrates can be added to the heat-treated previous solution after the protein nanogel has been formed.

[0487] For example, in some preferred embodiments of the present invention that are useful for forming protein nanogels, the liquid solution, or a previous solution used to prepare the liquid solution, has a pH in the range of 5.6 to 6.5, preferably 5.8 to 6.2, and more preferably 5.9 to 6.1.

[0488] The liquid solution, or a previous solution used to prepare the liquid solution, is preferably subjected to a first heat treatment using a temperature in the range of 70-145°C, preferably 75-120°C, more preferably 80-98°C, even more preferably 82-96°C, and most preferably 85-95°C.

[0489] The first heat treatment, which causes protein nanogelation, can be carried out under high shear conditions, for example using a scraped surface heat exchanger or similar high shear device, and surprisingly it is also possible to form protein nanogels by heating the packaged liquid solution under low shear or even non-shear conditions, for example by immersing the packaged liquid solution in an oil bath or using a plate heat exchanger.

[0490] The first heat treatment preferably has a duration sufficient to provide a degree of protein denaturation of at least 40%, preferably at least 60%, more preferably at least 70%, and most preferably at least 80%. Even higher degrees of protein denaturation, preferably at least 90%, more preferably at least 95%, can be obtained by the first heat treatment.

[0491] The first heat treatment may have a duration in the range of, for example, 0.1 seconds to 2 hours, preferably 0.5 minutes to 1 hour, more preferably 2 minutes to 30 minutes, and most preferably 5 to 20 minutes.

[0492] In some preferred embodiments, the first heat treatment is the only heat treatment in the process for producing the packaged heat treated beverage preparation, in which case the first heat treatment is applied to the liquid solution before or after packaging.

[0493] In another preferred embodiment, a first heat treatment is used to form a protein nanogel, followed preferably by a second heat treatment, which serves the purpose of final pasteurization or sterilization of the liquid solution. In this case, the first heat treatment is preferably applied to the previous solution used to prepare the liquid solution. This previous solution can then be processed by mixing it with other ingredients mentioned herein, such as carbohydrates, fats, minerals, and / or vitamins, to form a liquid solution. Such processing may also include other steps, such as pH adjustment, homogenization, and / or emulsification. Preferably, the liquid solution comprises the previous solution and, optionally, additional ingredients mixed with the previous solution.

[0494] In some preferred embodiments of the present invention: - a first heat treatment is applied to the previous solution used to prepare the liquid solution, thereby forming a protein nanogel; - Optionally, the heated solution is combined with other ingredients, for example by mixing; - the liquid solution is packaged in a suitable container in the form of a pre-heated solution by itself or in the form of a combination of the pre-heated solution with other ingredients; and - The packaged liquid solution is subjected to a second heat treatment which includes at least pasteurization and is preferably sufficient to provide a sterile beverage preparation.

[0495] In some preferred embodiments of the invention, minerals such as, for example, Ca, Mg, K and / or K are added to the previous heat treated solution containing nanogel. The inventors have observed that whey protein is more resistant to pasteurization or even sterilization heat treatment in the presence of high mineral content if it is first converted to nanogel.

[0496] The term "beverage preparation" refers to a liquid solution that has been subjected to a heat treatment that includes at least pasteurization.

[0497] The total amount of Ca and Mg in the liquid solution, or in the previous solution used to prepare the liquid solution, is preferably in the range of 0.001-0.1% w / w, more preferably 0.005-0.06% w / w, and most preferably 0.02-0.04% w / w.

[0498] The total amount of Na and K in the liquid solution, or in the previous solution used to prepare the liquid solution, is preferably in the range of 0.001 to 0.2% w / w, more preferably 0.01 to 0.1% w / w, and most preferably 0.04 to 0.06% w / w.

[0499] In some preferred embodiments of the invention, minerals such as, for example, Ca, Mg, K and / or K are added to the previous heat treated solution containing nanogel. The inventors have observed that whey protein is more resistant to pasteurization or even sterilization heat treatment in the presence of high mineral content if it is first converted to nanogel.

[0500] Therefore, it is often preferred that the above heat treated solution containing nanogel is mixed with other ingredients containing minerals in sufficient amounts to provide a liquid solution containing a total amount of Ca and Mg of at least 0.1% w / w, preferably at least 0.3% w / w, more preferably at least 0.5% w / w.

[0501] The heat treated solution containing the nanogel is preferably mixed with other ingredients containing minerals in sufficient amounts to provide a liquid solution containing a total amount of Ca and Mg of 0.1-1.5% w / w, more preferably 0.3-1.2% w / w, and even more preferably 0.5-1.0% w / w.

[0502] In addition, it is often preferred that the above heat treated solution containing nanogel is mixed with other ingredients containing minerals in sufficient amounts to provide a liquid solution containing a total amount of Na and K of at least 0.2% w / w, preferably at least 0.5% w / w, more preferably at least 0.7% w / w.

[0503] The heat treated solution containing the nanogel is preferably mixed with other ingredients containing minerals in sufficient amounts to provide a liquid solution containing a total amount of Na and K of 0.2-1.5% w / w, more preferably 0.5-1.2% w / w, and even more preferably 0.7-1.0% w / w.

[0504] The liquid solution, or the previous solution used to prepare the liquid solution, is preferably prepared by mixing the BLG isolate described herein, preferably obtained according to WO 2018 / 115520 A1, with water, preferably desalted or pH adjusted, optionally with particular preference pH adjustment to obtain the desired protein content and a pH in the range of 5.6 to 6.4. It is preferred to stop the pH adjustment as soon as the liquid becomes clear.

[0505] Soluble whey protein aggregates are preferably formed by heat treatment at a pH in the range of 6.6-8.0, more preferably in the range of 6.7-7.5, even more preferably in the range of 6.9-7.3. The heat treatments described in the context of protein nanogels are similarly useful for the formation of soluble whey protein aggregates. However, the protein content of the liquid solution is preferably in the range of 1-12% w / w, more preferably in the range of 3-11% w / w, even more preferably in the range of 5-10% w / w, and most preferably in the range of 6-9% w / w.

[0506] For preparing soluble whey protein aggregates using liquid solutions having high protein concentrations, the total amount of Ca and Mg in the liquid solution is preferably at most 0.01% w / w, more preferably at most 0.005% w / w, even more preferably at most 0.001% w / w.

[0507] For preparing soluble whey protein aggregates using liquid solutions having high protein concentrations, the total amount of Na and K in the liquid solution is preferably at most 0.05% w / w, more preferably at most 0.01% w / w, most preferably at most 0.005% w / w.

[0508] One aspect of the invention relates to the use of a protein solution comprising a total amount of protein of 1-20% w / w by weight of the solution, at least 85 w / w% of the protein being beta-lactoglobulin (BLG), to control the whiteness of a sterile beverage preparation having a pH in the range of 5.5-8.0.

[0509] Another aspect of the present invention relates to a packaged thermally treated beverage preparation as defined herein for use in a method of treating a disease associated with protein deficiency.

[0510] Another aspect of the invention relates to the use of the packaged thermally treated beverage preparations, defined herein as dietary supplements.

[0511] In a preferred embodiment of the present invention, the packaged thermally treated beverage preparation defined herein is used as a dietary supplement and is consumed before, during or after exercise.

[0512] In some preferred embodiments of the present invention, a packaged thermally treated beverage preparation having a pH in the range of 5.8 to 8.0, the beverage comprising: - a total amount of protein of 1-20% w / w relative to the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); and optionally a sweetener and / or a flavoring, and the protein fraction of the beverage preparation has a color value delta b in the range of -0.10 to +0.51 on the CIELAB color scale. * having Delta b * =b 6.0w / w%タンパク質に標準化したサンプル * -b 脱塩水 * and is measured at room temperature.

[0513] In some preferred embodiments of the present invention, a packaged thermally treated beverage preparation having a pH in the range of 5.8 to 8.0, the beverage comprising: - a protein in a total amount of 1-20% w / w relative to the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); - optionally containing sweeteners and / or flavorings; 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. * having Delta b * =b 6.0w / w%タンパク質に標準化したサンプル * -b 脱塩水 * and is measured at room temperature. The lipid content is up to 5% of the total energy content of the preparation.

[0514] In some preferred embodiments of the present invention, a packaged thermally treated beverage preparation having a pH in the range of 5.8 to 8.0, the beverage comprising: - a total amount of protein of 1-20% w / w relative to the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); and optionally a sweetener and / or a flavoring, and the protein fraction of the beverage preparation has a color value delta b in the range of -0.10 to +0.51 on the CIELAB color scale. * having Delta b * =b 6.0w / w%タンパク質に標準化したサンプル* -b 脱塩水 * and is measured at room temperature. It also has a lipid content of more than 5% and preferably more than 20E% of the total energy content of the preparation.

[0515] In some preferred embodiments of the present invention, a packaged thermally treated beverage preparation having a pH in the range of 5.8 to 8.00, the beverage comprising: - a total amount of protein of 1-20% w / w relative to the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, It has a turbidity of more than 200 NTU, preferably more than 40 NTU.

[0516] In some preferred embodiments of the present invention, a packaged thermally treated beverage preparation having a pH in the range of 5.8 to 8.0, the beverage comprising: - a protein in a total amount of 1-20% w / w relative to the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, It has a turbidity of maximum 200 NTU, preferably maximum 40 NTU.

[0517] In some preferred embodiments of the present invention, a packaged thermally treated beverage preparation having a pH in the range of 5.8 to 8.0, the beverage comprising: a protein in a total amount of 3-20% w / w, more preferably 3-18% w / w, even more preferably 3-15% w / w, most preferably 3-10% w / w, based on the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, It has a turbidity of more than 200 NTU, preferably more than 40 NTU.

[0518] In some preferred embodiments of the present invention, a packaged thermally treated beverage preparation having a pH in the range of 5.8 to 8.0, the beverage comprising: - a protein in a total amount of 3-20% w / w, more preferably 3-18% w / w, even more preferably 3-15% w / w, most preferably 3-10% w / w based on the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, It has a turbidity of maximum 200 NTU, preferably maximum 40 NTU.

[0519] In some preferred embodiments of the present invention, the beverage is a packaged thermally treated beverage preparation having a pH in the range of 6.2 to 8.0, preferably 6.3 to 7.6, preferably 6.5 to 7.2, the beverage comprising: a protein in a total amount of 1-20% w / w relative to the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, It has a turbidity of maximum 200 NTU, preferably maximum 40 NTU.

[0520] More preferably, the beverage preparation is a packaged heat treated beverage preparation having a pH in the range of 6.5 to 8.0, preferably 6.7 to 7.6, preferably 6.9 to 7.2, the beverage preparation comprising: a protein in a total amount of 5-12% w / w relative to the weight of the beverage preparation, wherein at least 90 w / w% of the protein, preferably at least 94% w / w, is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, The beverage preparation has a turbidity of maximum 40 NTU, preferably maximum 20 NTU, and the beverage preparation is preferably sterile.

[0521] In some preferred embodiments of the present invention, the beverage is a packaged thermally treated beverage preparation having a pH in the range of 6.2 to 8.0, preferably 6.3 to 7.6, preferably 6.5 to 7.2, the beverage comprising: - a protein in a total amount of 3-20% w / w, more preferably 3-18% w / w, even more preferably 3-15% w / w, most preferably 3-10% w / w based on the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, It has a turbidity of maximum 200 NTU, preferably maximum 40 NTU.

[0522] In some preferred embodiments of the present invention, there is provided a packaged thermally treated beverage preparation having a pH in the range of 6.5 to 8.0, preferably 6.7 to 7.6, preferably 6.9 to 7.2, the beverage preparation comprising: - a protein in a total amount of 3-20% w / w, more preferably 3-18% w / w, even more preferably 3-15% w / w, most preferably 3-10% w / w based on the weight of the beverage preparation, wherein at least 90 w / w%, preferably at least 94% w / w of the protein is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, The beverage preparation has a turbidity of more than 200 NTU, preferably more than 400 NTU.

[0523] In some preferred embodiments of the present invention, the beverage is a packaged thermally treated beverage preparation having a pH in the range of 5.5 to 6.2, preferably 5.7 to 6.1, preferably 5.8 to 6.0, the beverage comprising: - a protein in a total amount of 1-20% w / w relative to the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, It has a turbidity of more than 200 NTU, preferably more than 400 NTU.

[0524] In some preferred embodiments of the present invention, the beverage is a packaged thermally treated beverage preparation having a pH in the range of 5.8 to 8.0, preferably 6.3 to 7.6, preferably 6.5 to 7.2, the beverage comprising: - a total amount of protein of 2-10.0% w / w based on the weight of the beverage preparation, preferably a total amount of protein of 3.0-8.0% w / w based on the weight of the beverage preparation, preferably a total amount of protein of 5.0-7.5% w / w based on the weight of the beverage preparation, more preferably a total amount of protein of 4.0-6.0% w / w based on the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); -optionally containing sweeteners and / or flavouring agents.

[0525] In some preferred embodiments of the present invention, the beverage is a packaged thermally treated beverage preparation having a pH in the range of 5.8 to 8.0, preferably 6.3 to 7.6, preferably 6.5 to 7.2, the beverage comprising: a protein in a total amount of 2-10.0% w / w, preferably 3.0-8.0, preferably 5.0-7.5, more preferably 4.0-6.0% w / w based on the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, The total amount of magnesium and calcium is up to 10 mM.

[0526] In some preferred embodiments of the present invention, the beverage is a packaged thermally treated beverage preparation having a pH in the range of 5.8 to 8.0, preferably 6.3 to 7.6, preferably 6.5 to 7.2, the beverage comprising: a protein in a total amount of 2-10.0% w / w, preferably 3.0-8.0, preferably 5.0-7.5, more preferably 4.0-6.0% w / w based on the weight of the beverage preparation, wherein at least 85 w / w%, preferably at least 90% w / w of the protein is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, It contains up to 100mg phosphorus / 100g protein and up to 700mg potassium / 100g protein, preferably up to 50mg phosphorus / 100g protein and up to 400mg potassium / 100g protein, or preferably up to 10mg phosphorus / 100g protein and up to 50mg potassium / 100g protein.

[0527] A preferred embodiment of the present invention relates to a thermally treated beverage preparation obtainable by one or more of the methods described herein.

[0528] 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.

[0529] All patent and non-patent references cited in this application are incorporated herein by reference in their entirety.

[0530] The invention will now be described in further detail in the following non-limiting examples.

[0531] Example 1: Method of analysis 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. Trp residues buried in native proteins typically exhibit the highest fluorescence emission around 330 nm, rather than when they are present in solvent-exposed positions such as in unfolded proteins. In unfolded proteins, the wavelength of Trp fluorescence emission typically shifts to higher wavelengths and is often measured around 350 nm. We here exploit this transition to monitor thermally induced unfolding by calculating the ratio of the fluorescence emissions at 330 nm and 350 nm and investigating the effect of heating temperature.

[0532] 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 from 310 to 400 nm was recorded from the top by exciting at 295 using a 5 nm slit. Samples were measured at 22°C 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, i.e., R=I330 / I350, and used as a measure of the nativity of the protein. o An R of at least 1.11 represents the predominant native BLG conformation; An R less than o1.11 reports at least partial unfolding and aggregation.

[0533] 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.

[0534] Heat stability at pH 3.9 is determined by forming an aqueous solution of pH 3.9, containing 6.0% w / w protein by mixing a sample of the powder or liquid to be tested with water (or, in the case of dilute liquids, alternatively, concentrating by low temperature evaporation) and adjusting the pH to 3.9 with a minimum of 0.1 M NaOH or 0.1 M HCl.

[0535] Allow the pH adjusted mixture to stand for 30 min, then transfer 25 mL of the mixture to a 30 mL thin-walled glass test tube. Heat to 75.0 °C for 300 s by immersion in a 75.0 °C water bath. Immediately after heating, transfer the glass test tube to an ice bath to cool to 1-5 °C, and measure the turbidity of the heat-treated sample according to Example 1.7.

[0536] Example 1.3: Determination of the degree of protein denaturation of a whey protein composition 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.

[0537] More specifically, in the case of whey protein, the whey protein composition to be analyzed (e.g., a powder or an aqueous solution) is converted to: 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.

[0538] pH adjustment is performed using 3% (w / w) NaOH (aq) or 5% (w / w) HCl (aq).

[0539] The total protein content (P pH7.0又は3.0 ) is determined according to Example 1.5. The second aqueous solution is stored at room temperature for 2 h, followed by centrifugation at 3000 g for 5 min. A sample of the supernatant is collected and analyzed according to Example 1.5 to determine the protein concentration in the supernatant (S pH4.6 ) is obtained. Calculate the degree of protein denaturation, D, of the whey protein composition as follows: D=((P pH7.0又は3.0 -S pH4.6 ) / P pH7.0又は3.0 ) * 100%

[0540] Example 1.4 Determination of protein denaturation using reversed-phase UPLC analysis (with pH 4.6 acid precipitation). BLG samples (such as unheated reference and heated BLG beverage compositions) were diluted to 2% with MQ water. 5 mL of protein solution, 10 mL of Milli-Q, 4 mL of 10% acetic acid, and 6 mL of 1.0 M NaOAc were mixed and stirred for 20 minutes to precipitate and aggregate denatured proteins at around pH 4.6. The solution was filtered through a 0.22 μm filter to remove aggregates and non-native proteins. All samples were subjected to the same degree of dilution by adding polished water. 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 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 (five-level calibration curve). Samples were further diluted and re-injected if outside the linear range.

[0541] Example 1.5: Determination of total protein The total protein content (true protein) of the samples is determined by: 1) Determination of total nitrogen in samples according to ISO 8968-1 / 2|IDF 020-1 / 2-Milk-Determination of nitrogen content-Part 1 / 2: Determination of nitrogen content using the Kjeldahl method. 2) Determination of non-protein nitrogen of samples in accordance with 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.

[0542] Example 1.6: Determination of non-aggregating BLG, ALA, and CMP The contents of non-aggregated alpha-lactalbumin (ALA), beta-lactoglobulin (BLG), and caseinomacropeptide (CMP), respectively, were analyzed by HPLC analysis at 0.4 mL / min. 25 microL of the filtered sample was injected into two TSKgel3000PWxl (7.8 mm 30 cm, Tosohass, Japan) columns connected in series, fitted with a pre-column PWxl (6 mm × 4 cm, Tosohass, Japan) equilibrated with eluent (consisting of 465 g Milli-Q water, 417.3 g acetonitrile, and 1 mL trifluoroacetic acid), and a UV detector at 210 nm was used.

[0543] Native alpha-lactalbumin (C アルファ ), β-lactoglobulin (C β ), and caseinomacropeptide (C CMP Quantitative determination of the content of ) was performed by comparing the peak areas obtained with the corresponding standard proteins with the peak areas of the samples.

[0544] 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).

[0545] Example 1.7: Determining turbidity Turbidity is the cloudiness or haze of a fluid caused by numerous particles that are generally invisible to the naked eye, similar to smoke in the air. Turbidity is measured in Nephelometric Turbidity Units (NTU).

[0546] 20 mL of beverage / sample was added to an NTU glass and placed in a Turbiquant® 3000 IR turbidity meter. NTU values ​​were measured after stabilization and were repeated twice.

[0547] Example 1.8: Determining viscosity The viscosity of the beverage formulations was measured using a rheometer (Anton Paar, Physica MCR301). 3.8 mL of sample was added to the cup DG26.7. The sample was equilibrated to 22 °C and then incubated for 50 s. -1 Preshear for 30 s at 400 C, followed by a 30 s equilibration period and a 1 s -1 ~200s -1 and 1s -1 A shear rate sweep was performed. Unless otherwise stated, viscosity is 100s -1 It is measured in centipoise (cP) at shear rate. The higher the measured cP value, the higher the viscosity.

[0548] Alternatively, the viscosity can be estimated using Viscoman by Gilson, at approximately 300s -1 The shear rates were reported as:

[0549] Example 1.9: Color Determination Color was measured using a colorimeter (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) while avoiding the formation of air bubbles. Protein content of the samples was standardized to ≦6.0 w / w% protein. The colorimeter was calibrated to a white calibration plate (No. 19033177). The illuminant was set to D65 and the observer was set to 2 degrees. Color (CIELAB color space, a * Value, b * Value, L * The concentration of β-aminobutyric acid in the suspension was measured as the average of three separate readings at different locations on the Petri dish with the lid covering the suspension. The values ​​for desalinated water standards are as follows: L * 39.97±0.3 a * 0.00±0.06 b * -0.22±0.09 Measurements were converted to delta / difference values ​​based on demineralized water measurements. Delta L * =L 6.0w / w%タンパク質に標準化したサンプル * -L 脱塩水 * , measured at room temperature. Delta A * =a 6.0w / w%タンパク質に標準化したサンプル * -a 脱塩水 * , measured at room temperature. Delta b * =b 6.0w / w%タンパク質に標準化したサンプル * -b 脱塩水 * , measured at room temperature.

[0550] Samples were standardized to ≦6.0% w / w protein. L * a * b *The CIELAB color space (also called 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). In this space, L * indicates lightness (value from 0 to 100), and L * =0 is the darkest black, L * =100 indicates the brightest white. Color channel a * and b * is a * =0 and b * = 0 represents the true neutral gray value. * The axes represent the green and red components, with green going in the negative direction and red going in the positive direction. * The axes represent the blue and yellow components, with the negative direction being blue and the positive direction being yellow.

[0551] Example 1.10 Beverage Stability Testing / Insoluble Protein Substances A whey protein beverage was considered to have a stable composition if less than 15% of the total protein in the heated sample precipitated upon centrifugation at 3000g for 5 minutes: Approximately 20g of sample was added to a centrifuge tube and centrifuged at 3000g for 5 minutes. Kjeldahl analysis of the protein before centrifugation and the supernatant after centrifugation was used to quantitate protein recovery, see Example 1.5.

[0552] Calculate protein loss:

number

[0553] This parameter is sometimes referred to as the level of insoluble proteinaceous 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. Centrifuge approximately 20 g of sample (e.g., liquid sample or suspended powder sample) into a centrifuge tube at 3000 g for 5 minutes. Protein (P) before centrifugation is total ) and the supernatant after centrifugation (P 3000×g ) Kjeldahl analysis was used to quantitate protein recovery according to Example 1.5.

[0554] Calculate the amount of insoluble protein material:

number

[0555] Example 1.11: Measurement of gel strength upon acidification A rheometer (Anton Paar, Physica MCR301) was used to simulate the structure evolution of the beverage in the stomach during acidification. The beverage was diluted to 2 w / w% protein and warmed to 42 °C for 30 min. Then, 1 w / w% GDL (D-gluconic acid, Sigma Aldrich, 49-53% by weight) was added to the solution and stirred for 5 min. The solution (19.6 mL) was added to the rheometer cup CC27-SS. The rheometer was equilibrated to 42 °C and G' (storage modulus, Pa) was measured at 0.1 Hz and 0.5% strain for 60 min. The pH during acidification was followed using a pH logger (WTW, Multi 3410).

[0556] 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 "lorem ipsum" sentence written on it. Using a smartphone, the vial was photographed and we assessed whether the sentence was clearly observable through the vial.

[0557] Example 1.13: Determination of ash content Determine the ash content of foods in accordance with NMKL 173:2005 "Gravimetric Determination of Ash in Foods".

[0558] Example 1.14: Determining electrical conductivity The "conductivity" of an aqueous solution (sometimes referred to as "specific conductivity") is a measure of the solution's ability to conduct electricity. Conductivity is determined, for example, by measuring the AC resistance of the solution between two electrodes, with the results typically given in units of millisiemens per centimeter (mS / cm). Conductivity can be measured, for example, according to EPA (United States Environmental Protection Agency) Method No. 120.1. Conductivity values ​​reported herein are normalized to 25° C. unless otherwise specified. The conductivity is measured with a conductivity meter (WTW Cond 3210 with Tetracon 325 electrodes). The system is calibrated before use as described in the manual. The electrodes are rinsed well with the same type of medium in which the measurements are to be made to avoid local dilution. The electrodes are lowered into the medium so that the area in which the measurements are to be made is completely submerged. The electrodes are then agitated to remove any air trapped in the electrodes. The electrodes are then kept stationary until a stable value is obtained from the display and recorded.

[0559] Example 1.15: Determining the total solids content of a solution The total solids content of a solution can be determined according to NMKL 110 2nd Edition, 2005 (Total Solids (Water) - Gravimetric Analysis of Milk and Dairy Products). NMKL is the abbreviation for "Nordic Committee for Standard Methods of Analysis and Food Analysis (Nordisk Metodikkomite for Näeringsmidler)".

[0560] The water content of a solution can be calculated as 100% minus the relative total solids content (% w / w).

[0561] Example 1.16: Determining pH All pH values ​​were measured using a pH glass electrode and normalized to 25°C. pH glass electrodes (with temperature compensation) are rinsed carefully before use and calibrated before use. If the sample is a liquid solution, the pH is measured directly in the solution at 25°C. 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.

[0562] 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 (graduated cylinder) under specified conditions. Density is typically expressed in g / ml or kg / L.

[0563] 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.

[0564] This method allows the definition of three different densities: Packing 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 tappings according to the conditions specified in this standard. Bulk density, which is the mass divided by the volume of a powder after 625 tappings according to the conditions specified in this standard.

[0565] This method uses a special measuring cylinder of 250 ml, graduated from 0 to 250 ml and weighing 190±15 g (J. Engelsmann AG 67059 Ludwigshafen / Rh) and, for example, a Stampf volumeter from J. Engelsmann AG.

[0566] The loose density and bulk density of the dried product are determined by the following procedure.

[0567] Pretreatment: The samples to be measured are stored at room temperature.

[0568] The container is then rotated and inverted repeatedly to thoroughly mix the sample (avoid crushing of particles). The container is not filled more than 2 / 3 full.

[0569] procedure: Weigh out 100.0±0.1 grams of powder into a graduated cylinder. Read off the volume, V0, in ml.

[0570] If 100 g of powder will not fit into the cylinder, the amount will need to be reduced to 50 or 25 grams.

[0571] Fix the graduated cylinder to a Stampf volumeter and tap it 100 times. Smooth the surface with a spatula and measure the volume V 100 Read in ml.

[0572] Change the number of tabs to 625 (including 100 taps). After tapping, level the surface and measure the capacity V 625 Read in ml.

[0573] Density calculation: Calculate the loose density and bulk density, expressed in g / ml, according to the following formula: Bulk density = M / V where M is the weight of the sample in grams and V is the volume in ml after 625 taps.

[0574] 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 Standard Methods of Analysis and Food Analysis (Nordic Metodikkomite for Naeringsmidler)". Example 1.19: Determination of the amounts of calcium, magnesium, sodium, potassium and phosphorus (ICP-MS method)

[0575] 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 the mineral(s) is determined using an ICP instrument.

[0576] Device: The microwave is manufactured by Anton Paar and the ICP is an Optima 2000DV manufactured by PerkinElmer Inc.

[0577] material: 1M HNO3 Yttrium in 2% HNO3 Appropriate standards for calcium, magnesium, sodium, potassium, and phosphorus in 5% HNO3

[0578] Pretreatment: Weigh out a certain amount of powder and transfer the powder to a microwave digestion tube. Add 5 mL of 1 M HNO3. Digest the sample in a microwave according to the microwave instructions. Place the digested tube in a fume hood and remove the lid to allow volatile fumes to evaporate.

[0579] Measurement steps: Transfer the pretreated sample to the DigiTUBE using a known volume of Milli-Q water. Add a solution of yttrium in 2% 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.

[0580] Blind samples are prepared by diluting a mixture of 10 mL of 1 M HNO3 and 0.5 mL of yttrium 2% HNO3 solution to a final volume of 100 mL using Milli-Q water.

[0581] Prepare at least three standard samples with concentrations bracketing the expected sample concentrations. The detection limits for liquid samples are 0.005g / 100g sample for Ca, Na, K and Phosphor, and 0.0005g / 100g sample for Mg. The detection limits for powder samples are 0.025g / 100g sample for Ca, Na, K and Pho, and 0.0005g / 100g sample for Mg. If below the detection limit for phosphorus, the detection limit value is used in the example to indicate the maximum amount of Pho present as a worst case scenario.

[0582] Example 1.20: Determining the Furosine Value: Furosine value is 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 value is reported in mg furosine per 100 g protein.

[0583] Example 1.21: Determining the crystallinity of BLG in liquid Use the following method to determine the crystallinity of BLG in liquids with a pH range of 5-6.

[0584] 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) Keep the centrifuge at 2° C. and immediately spin the filters at 1500 g for 5 minutes. c) Add 2mL of cold Milli-Q water (2°C) to the retentate side of the spin filter and immediately, with the centrifuge cooled to 2°C, spin the filter at 1500g for 5 minutes and 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 2M 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 the result to total weight of BLG instead of weight percent. The weight of BLG in Permeate A is m 透過液A The weight of the BLG in the permeate B is m 透過液B It is called. g) The liquid crystallinity for BLG is determined as follows: Crystallinity=m 透過液B / (m 透過液A +m 透過液B ) * 100%

[0585] Example 1.22: Determination of crystallinity of BLG in dry powder This method is used to determine the crystallinity of BLG in dry powders.

[0586] a) Mix 5.0 grams of powder sample with 20.0 grams of cold Milli-Q water (2°C) and allow to stand at 2°C for 5 minutes. b) A sample of the liquid in question is transferred to a Maxi-Spin filter equipped with a 0.45 micron CA membrane. c) Keep the centrifuge at 2° C. and immediately spin the filters at 1500 g for 5 minutes. 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 via HPLC using the method outlined in Example 1.31, converting the results to total weight of BLG rather than weight percent. The weight of BLG in Permeate A is m 透過液A It is called. e) The crystallinity of the BLG in the powder is then calculated using the following formula:

number

[0587] If the total amount of BLG in the powder sample is unknown, it may be determined by suspending another 5 g 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, leaving the mixture under stirring at 25°C for 1 hour, and finally determining the total amount of BLG in the powder sample using Example 1.31.

[0588] Example 1.23: Determination of UF permeation conductivity Transfer 15 mL of sample to an Amicon Ultra-15 centrifugal filter unit with 3 kDa cutoff (3000 NMWL) and centrifuge at 4000 g for 20-30 min or until a sufficient volume of UF permeate accumulates at the bottom of the filter unit to measure conductivity. Measure conductivity immediately after centrifugation. Sample handling and centrifugation are performed at the temperature of the sample source.

[0589] 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:

[0590] A sample of the powder to be analysed is resuspended and gently mixed in demineralised water at a temperature of 4°C in a weight ratio of 1 part powder to 2 parts water and left to rehydrate at 4°C for 1 hour.

[0591] The rehydrated sample is examined by microscopy to identify the presence of crystals, preferably using plane polarized light to detect birefringence.

[0592] The crystalline material is isolated and subjected to X-ray crystallography to confirm the presence of crystalline structure, and preferably also that the crystal lattice (space group and unit cell dimensions) corresponds to that of BLG crystals.

[0593] The chemical composition of the isolated crystalline material is analyzed to confirm that the solid is composed primarily of BLG.

[0594] Example 1.25: Determining the total amount of lactose The total lactose content is determined in accordance with ISO 5765-2:2002 (IDF 79-2:2002) "Milk powder, dry ice mix and processed cheese - Determination of lactose content - Part 2: Enzymatic method utilizing the galactose moiety of lactose".

[0595] Example 1.26: Determining the total amount of carbohydrates: The amount of carbohydrates was determined using the Sigma Aldrich Total Carbohydrate Assay Kit (Cat MAK104-1KT), which hydrolyzes carbohydrates and converts them to furfural and hydroxyfurfural, which are converted to chromagens that are monitored spectrophotometrically at 490 nm.

[0596] Example 1.27: Determining the total amount of lipids The amount of lipids is determined according to ISO 1211:2010 (Determination of fat content - Roese-Gottlieb gravimetric method).

[0597] 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 give a conductivity of up to 0.05 mS / cm).

[0598] Approximately 500 μl of sample was transferred to the prism face of the device and the measurement was started. The measurement value was read and recorded.

[0599] The Brix of a whey protein solution is proportional to its total solids (TS) content, and TS (%w / w) is approximately the Brix. * It is 0.85.

[0600] Example 1.29 Determination of Lactoferrin and Lactoperoxidase Lactoferrin concentrations are determined by ELISA immunoassay as reviewed 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). The concentration of lactoperoxidase is determined using a commercially available bovine lactoperoxidase kit.

[0601] 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 ingredients - Horizontal method for enumeration of microorganisms - Colony counting technique at 30 °C.

[0602] Example 1.31: Determination of the total amount of BLG, ALA, and CMP This procedure is a liquid chromatography (HPLC) method for the 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 present in aggregates, and therefore provides a measure of the total amount of a protein species in the composition in question.

[0603] The isolation mode 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) of proteins or protein aggregates to produce unfolded monomeric structures.

[0604] Sample preparation is easily accomplished by dissolving the equivalent of 10 mg of protein in the mobile phase.

[0605] Two TSK-GEL G3000SWXL (7.7 mm x 30.0 cm) columns (GPC columns) and a guard column are arranged in series to achieve adequate isolation of the major proteins in the raw material.

[0606] The eluted analytes are detected and quantified by UV detection (280 nm).

[0607] Equipment / Materials: 1. HPLC Pump 515 (Waters) with manual seal washer 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 (7.8×300 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)

[0608] procedure: Mobile phase: A. Stock buffer. 1. Weigh 56.6 g Na2HPO4, 3.5 g NaH2PO4, and 2.9 g EDTA into a 1,000 mL beaker. Dissolve in 800mL of water. 2. Measure pH and adjust to 7.5 ± 0.1 as necessary with HCl (to lower pH) or NaOH (to raise pH). 3. Transfer to a 1000 mL volumetric flask and dilute to volume with water.

[0609] B. 6M guanidine HCl mobile phase. 1. Weigh out 1,146 g of guanidine HCl into a 2,000 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 stirrer (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.

[0610] Calibration standards. Calibration standards for each protein to be quantified are prepared in the following manner: 1. Accurately weigh out (to the nearest 0.01 mg) approximately 25 mg of protein reference standard. Place in a 10 mL volumetric flask and dissolve in 10 mL of water. This is the protein stock standard solution (S1) of protein. 2. Pipette 200 μl of S1 into a 20 ml volumetric flask and dilute to volume with mobile phase. This is the low dilution working 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 the 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 dilution working solution WS5. 7. Using graduated disposable pipettes, 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 standards using 0.22 μm cellulose acetate syringe filters.

[0611] Protein purity is measured using Kjeldahl (N x 6.38) and HPLC to measure area % from standard solution WS5. Protein (mg) = "Standard protein weight" (mg) x P1 x P2 P1=P% (Kjeldahl) P2 = Protein area % (HPLC)

[0612] Sample preparation 1. Weigh out an amount of the original sample equivalent to 25 mg of protein 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. Adjust the volume with mobile phase and add 167 μL of 2-mercaptoethanol to 25 ml of sample solution. 4. Sonicate for approximately 30 minutes, then leave the sample at ambient temperature for approximately 1.5 hours. 5. Mix the solution and filter using a 0.22 μl cellulose acetate syringe filter.

[0613] HPLC Systems / Columns Column Equilibration 1. Connect a GPC guard column and two GPC analytical columns in series. New columns are usually shipped in phosphate buffer. 2. Slowly run water through the new column at 0.1-0.5 mL / min for 30-60 minutes. Continue flushing for approximately 1 hour. 3. Gradually reduce the flow rate from 0.5 mL / min to 0.1 mL / min. Exchange with mobile phase in reservoir. 4. Gradually increase the pump flow rate from 0.1 to 0.5 mL / min over 30-60 minutes and leave it at 0.5 mL / min to avoid pressure shock. 5. Inject 10 samples to saturate the column and wait for the peaks to elute. This helps in conditioning the column. This process is accomplished without having to wait for each injection to be completed before injecting the next. 6. Equilibrate with mobile phase for at least 1 hour.

[0614] Calculating the results Quantitative determination of the content of the quantified proteins, e.g., alpha-lactalbumin, beta-lactoglobulin, and caseinomacropeptide, was performed by comparing the peak areas of the samples with those obtained with the corresponding standard proteins. Results are reported as specific protein / 100 g of original sample or as weight percentage of specific protein relative to the weight of the original sample.

[0615] Example 1.32: Quantification of the amount of microgels, protein nanogels, soluble whey protein aggregates and native proteins Quantification of the amount of insoluble protein material, protein nanogel, soluble whey protein aggregates, and native protein in a beverage or powder sample is carried out using the following steps: a) converting the sample to be tested (by mixing with demineralized water or by concentrating) into a solution containing 20 g total protein / L (e.g. by diluting with demineralized water or by concentrating) and confirming the concentration of total protein by measuring the total protein in an aliquot of the solution using Example 1.5; a Report as. b) A first aliquot of the solution from a) is centrifuged at 3.000×g for 5 minutes to precipitate insoluble protein material, followed by measuring the protein concentration in the supernatant as described in step a) and averaging the total protein content of the supernatant at c b The content of insoluble protein aggregates (percent of total protein) is reported as (c a -cb ) / c a * It is calculated as 100. c) A second aliquot of the solution of a) is centrifuged at 50.000×g for 1 hour to precipitate both the insoluble protein material and the protein nanogel, and the concentration of protein in the supernatant is measured as described in step a), c The protein nanogel fraction is reported as (c a -c c ) / c a * 100-(c a -c b ) / c a * It is calculated as 100. d) The third aliquot is adjusted to pH 4.6 to precipitate any denatured and / or aggregated proteins in the sample. The sample is left at room temperature for 15 min and then centrifuged at 50.000 × g for 1 h to separate the precipitate. The concentration of total protein (mainly native protein) in the resulting supernatant is measured as described in step a) and c d Report as. The fraction of soluble whey protein aggregates in solution is calculated as follows: (c a -c d ) / c a * 100-(c a -c c ) / c a * 100. The percentage of native protein in the solution is c d / c a * Calculate as 100.

[0616] If absolute concentrations of insoluble protein material, protein nanogel, soluble whey protein aggregates and / or native protein of the original sample are required, these are easily calculated using information about the amount of original sample used to prepare the solution of step a).

[0617] All centrifugation steps are performed at 25° C. using a Beckmann Coulter Avanti JXN-30 centrifuge equipped with a JA-30.50 rotor, using 50 mL samples in 50 mL Beckmann centrifuge tubes (29×103 mm).

[0618] Example 1.33: Hydrodynamic diameter The hydrodynamic diameter (mean intensity size (d.nm)) of protein particles was determined by dynamic light scattering using a Nanosizer (Malvern). 800 μL of demineralized water and 5 μL of heat-treated protein drink were mixed and added to a UV cuvette. Size measurements were performed at room temperature (22°C).

[0619] Example 2. Preparation of spray-dried acidic BLG isolate powder Whey Protein Feed 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 with a Synder FR membrane before being used as feed for the BLG crystallization process. The chemical composition of the feed is shown in Table 1. The inventors recognize 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.

[0620] conditioning Sweet whey feed was centrifuged through a Koch HFK-328 type membrane (70m) at a feed concentration of 21% total solids (TS) ±5, with a 46 mil spacer and a feed pressure of 1.5-3.0 bar. 2The diafiltration medium was conditioned on an ultrafiltration setup at 20° C. using a 1000 mL filtration membrane and polished water (water filtered by reverse osmosis to obtain a conductivity of up to 0.05 mS / cm) as the diafiltration medium. The pH was then adjusted by adding HCl to a pH of about 5.5. Diafiltration was continued until the drop in retentate conductivity was below 0.1 mS / cm for 20 minutes. The permeate flow was then increased to 1.43 L / hr / m 2 The retentate was concentrated until the Cd content was below 0.05%. A first sample of the concentrated retentate was taken and centrifuged at 3000 g for 5 min. The supernatant of the first sample was used to measure the BLG yield.

[0621] Crystallization The concentrated retentate was transferred to a 300 L crystallization tank where it was 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.

[0622] After cooling, a sample of the crystal-containing whey protein solution (second sample) was taken and the BLG crystals were isolated 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%.

[0623] [Table 1]

[0624] 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.

[0625] 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.

[0626] 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 in all samples. Therefore, the area of ​​ALA before and after crystallization is used as a correction factor (cf) when calculating the relative yield.

number

[0627] The relative yield is calculated according to the following formula:

number

[0628] Acid dissolution of BLC crystals The remaining material from the crystallization tank was separated using a decanter at 350g, 2750RPM, 150RPM difference with 64 spacer, and 75L / hr feed flow, after which the feed was mixed 1:2 with polishing water.The BLG crystals / solid phase from the decanter was then mixed with polishing water to make a thinner slurry, after which phosphoric acid was added to lower the pH to approximately 3.0 to quickly dissolve the crystals.

[0629] After dissolving the BLG crystals, the pure BLG protein liquid was concentrated to 15 Brix in the same UF setup used to prepare the feed for crystallization and the pH was adjusted to a final pH of approximately 3.8. The liquid BLG isolate was then heated to 75 degrees for 5 minutes and subsequently cooled to 10°C. It was found that the heat treatment reduced the microbial load from 137.000 CFU / g before heat treatment to less than 1000 CFU / g after heat treatment. The heat treatment did not cause any protein denaturation and the intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) was determined to be 1.20, indicating the native conformation of the BLG molecule.

[0630] 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 about 4% w / w and the chemical composition of the powder is shown in Table 2. 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.

[0631] [Table 2]

[0632] Example 3: Preparation of spray-dried PH neutral BLG isolated powder Using the same protocol and experimental setup as in Example 2, the lactose-reduced whey protein isolate shown in Table 3 was conditioned and used as feed for crystallization. The crystallization yield was calculated to be 68%. The inventors recognize that all weight percentages of specific proteins, such as BLG and ALA, referenced in this example relate to the weight percentage of non-aggregated protein relative to total protein.

[0633] [Table 3]

[0634] The remaining material from the crystallization tank is decanted into a decanter. After separation at 350g, 2750RPM, 150RPM difference with 64 spacer, and 75L / hr feed flow, the feed was mixed 1:2 with polishing water. The BLG crystals / solid phase from the decanter was then mixed with polishing water to make a thinner slurry, after which the pH was adjusted to approximately 7 by adding 0.1M potassium hydroxide to quickly dissolve the crystals.

[0635] After dissolving the crystals, the pure BLG protein liquid was concentrated to 15 Brix in the same UF setup used to prepare the whey protein solution for crystallization and the pH was adjusted to a final pH of approximately 7.0. 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 about 4% w / w. The composition of the powder is shown in Table 4. After drying, a portion of the powder was dissolved in demineralized water and the degree of protein denaturation was determined to be 9.0% and the intrinsic tryptophan fluorescence emission ratio (330 nm / 350 nm) was 1.16.

[0636] [Table 4]

[0637] The bulk density (625 tap) of the spray-dried powder is 0.2-0.3 g / cm 3 It was estimated that.

[0638] Example 4: Preparation of a typical whey protein drink Dry BLG isolate protein powder containing 85% or more BLG on a protein basis is dispersed in up to about 95% of the demineralized water needed to reach the desired final protein concentration. A neutral pH BLG isolate powder is prepared as outlined in Example 3, while a pH 5.5 BLG isolate powder is prepared as outlined in Example 7 of PCT / EP 2017 / 084553.

[0639] Optionally, other ingredients including minerals, sweeteners, flavorings, stabilizers, emulsifiers, or sources of fat and carbohydrate may be added.

[0640] Adjust pH to final pH using 10% NaOH or 10% phosphoric acid (or other food grade acid). The remaining water is added to reach the desired protein concentration and the composition is optionally homogenized. For comparison, 85% or more of the BLG product is replaced with whey protein isolate in the preparation of a reference sample while maintaining the remaining steps. The samples were stored in the dark at 20°C.

[0641] Example 5: Heat Treatment of Whey Protein Compositions Heat treatment was carried out by heating at 143°C for 2-6 seconds (high temperature, short time (HTST)) using a plate or tubular heat exchanger (manufacturer: OMVE HTST / UHT pilot plant HT320-20). The heat-treated beverage composition was dispensed into 100 mL sterile bottles at 10° C. and immediately sealed. In other experiments, heat treatment was performed by transferring the whey protein source into thin-walled glass vials containing 15–30 mL of sample. The vials were immersed in a water bath pre-equilibrated at the target temperature, ranging from 86 °C to 95 °C, for 1–18 min and then cooled on ice.

[0642] Example 6: 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 7.0. The BLG beverage was prepared by dissolving diafiltered pH 7.0 BLG isolate powder in 10° C. demineralized water. For comparison, a WPI sample was prepared using WPI-A. WPI-A was dissolved in demineralized water at 10° C. 10% NaOH was slowly added to the solution. The final pH was adjusted to pH 7.0.

[0643] The solution was heat treated at 143°C / 4 seconds using a plate heat exchanger as described in Example 5 and tapped to provide a heat sterilized whey protein beverage composition.

[0644] Table 5 below shows the composition of the BLG powder used to prepare the beverage formulations, and for comparison, the composition of the WPI.

[0645] [Table 5]

[0646] Example 7: Clear and colorless whey protein beverage containing >85% BLG A beverage preparation was prepared in which approximately 92% w / w of the protein was BLG, see Example 4. For comparison, a whey protein sample based on WPI powder containing approximately 61% w / w BLG was prepared. The protein content of the sample was 6% w / w. The pH was adjusted to pH 7.0 using NaOH. The preparation was heat treated at 143° C. for 4 seconds. Turbidity, viscosity, color and clarity were measured according to the procedures described in Examples 1.7, 1.8 and 1.9.

[0647] The results are shown in Table 6 below.

[0648] [Table 6] Delta b * To calculate, use the following formula: Delta b * =b6.0w / w%タンパク質に標準化したサンプル * -b 脱塩水 * , measured at room temperature. Delta A * To calculate, use the following formula: Delta A * =a 6.0w / w%タンパク質に標準化したサンプル * -a 脱塩水 * , measured at room temperature. Delta L * To calculate, use the following formula: Delta L * =L 6.0w / w%タンパク質に標準化したサンプル * -L 脱塩水 * , measured at room temperature.

[0649] The colour values ​​of demineralised water are: L * =39.97, a * =0 and b * =-0.22.

[0650] result: The results, shown in Table 6, indicate that when protein fractions containing at least 85 w / w% BLG were used, clear, water-white beverages were produced at pH 7.0. The BLG beverages also had low viscosity. In contrast, the sample containing WPI, in which approximately 61 wt % of the protein was BLG, was yellowish and had a higher b * had value.

[0651] Example 8: Milky whey protein beverage, high temperature heat treatment An opaque, milky beverage containing BLG was produced by dissolving BLG powder (pH 5.5) in tap water, adjusting the pH to 6.0 using 3% NaOH, and heat treating at 94°C for 14 minutes. The BLG beverage contains approximately 96% w / w of protein as BLG. A 10 w / w % BLG beverage was prepared having a pH of 6.0.

[0652] Consider the composition of the BLG and WPI samples below: [Table 6-2]

[0653] The turbidity, viscosity, color and clarity were measured according to the procedures described in Examples 1.7, 1.8, 1.9, and the stability of the beverages was measured as described in Example 1.12.

[0654] The results are shown in Table 7 below and FIG.

[0655] [Table 7]

[0656] result: The results presented in Table 7 and Figure 1 show that a milky / opaque colorless beverage was produced at pH 6.0 when 10 w / w% protein containing at least 85 w / w% BLG was used and subjected to a heat treatment corresponding to sterilization. In contrast, the samples containing WPI (WPI-A and WPI-B) gelled and did not allow the production of a beverage (see FIG. 2).

[0657] Example 9: Digestion of an exemplary BLG beverage containing primarily protein nanogels or soluble whey protein aggregates. The aim of this example is to investigate the formation of structures during gastric digestion of different whey protein drinks by in vitro simulation of gastric digestion. Three thermally processed nutritional compositions were prepared, two of which contained 85% or more BLG (Drinks A and B) and a conventional WPI beverage (Drink C). The beverage was prepared according to Example 4 and heat treated according to Example 5. The composition of the protein powder used to prepare the exemplary beverages can be found in Table 8.

[0658] [Table 8]

[0659] Beverage A A 6 w / w% BLG solution was prepared by dissolving protein powder containing 98.2% BLG (Table 8). The pH was adjusted to pH 7.0 using 10% NaOH and sterilized by UHT treatment at 143°C for 6 seconds to produce BLG beverage solution A. Beverage B A 6 w / w% BLG solution was prepared by dissolving a powder containing 95.9% BLG (Table 8). The pH was adjusted to pH 6.0 using 10% NaOH and heat-treated at 86° C. for 18 minutes to produce BLG beverage solution B. Beverage C A 6 w / w% WPI beverage was prepared by dissolving protein powder "WPI" containing 61% BLG (Table 8). The pH was adjusted to pH 7.0 using 10% NaOH and heat treated at 143°C for 6 seconds. Drink C is used as a reference.

[0660] Drinks A and C are clear, while drink B is opaque and milky.

[0661] The types and amounts of insoluble protein material, soluble aggregates, protein nanogels, and native whey protein present in the beverage were determined as described in Example 1.32, and the results are shown in FIG.

[0662] Results - Soluble aggregates, protein nanogels, insoluble protein material, and native whey protein: In contrast to the milky appearance of the BLG beverage (B), the BLG beverage (A) and the WPI beverage (C) were found to result in clear beverages. The aggregate composition was evaluated as described in Example 1.32 and is shown in Figure 3. Figure 3 shows that beverages A (BLG pH 7.0) and C (WPI reference, pH 7.0) contain primarily soluble whey protein aggregates (67% and 44%, respectively), while beverage B (BLG pH 6.0) contains primarily protein nanogels (74%). The content of insoluble protein material was less than 1% in all three beverages.

[0663] Approximately 28–33% residual native protein was observed in both the BLG (A) and WPI (C) beverages, which may be due to an incomplete aggregation process, while 13% native protein remained in the BLG (B). Therefore, the beverage compositions contain different protein structures that may lead to differences in their digestibility.

[0664] A fourth beverage (Beverage D) was prepared by mixing 1 volume of heat-treated solution A with 1 volume of heat-treated solution B. Beverage D was found to contain 45% protein nanogel, 19% soluble whey protein aggregates, and 37% native protein.

[0665] Simulated gastric digestion method: To investigate the formation of structures during gastric digestion of different whey protein beverages, the inventors performed the following in vitro simulation of gastric digestion.

[0666] Samples were subjected to simulated oral and gastric digestion according to the protocol previously described in Mulet-Cabero, A.-I., Mackie, A.R., Wilde, P.J., Fenelon, M.A., & Brodkorb, A. (2019): Structural mechanisms and kinetics of in vitro gastric digestion influenced by process-induced changes in milk. Food Hydrocolloids, 86, 172-183, with minor modifications as described below.

[0667] Before carrying out the digestion, the samples (30 g) were mixed with a solution of buffer salts at 37° C. in the same amounts and concentrations as would be used later during the actual simulated digestion. This mixture was titrated to pH 2.0 with 0.1 M HCl using a pH stat (Metrohm 602 pH stat), which is necessary to determine the actual buffering capacity of the food and to program the titrator for the actual digestion. For the actual digestion, 30 g of sample was mixed with model human saliva (minus salivary amylase because no starch was present in the drink) at 37°C for 2 min, and the amount of model saliva (1.65 g) was determined by the solids content of the sample (6%). The samples were transferred to a thermostated reaction vessel for the gastric digestion process, which initially contained a total of 10% acid and salts of gastric juice, simulating the fasted state of the stomach. The remaining 90% of the buffer salts, 0.1 M HCl and water were added at a rate such that the addition was completed after 105 min, the calculated duration of the gastric digestion process. A solution of pepsin (0.5 ml, 254400 U / ml) was added at 4.762 μl / min using a syringe pump, so that all the pepsin solution was added by the end of the 105 min digestion. The solution was added against the wall of the vessel to mimic secretion by the gastric mucosal surface. The pH of the mixture was adjusted to 1.0 ± 5% from the horizontal using a rotating vibrating mixer (15 rpm, ± 5% from the horizontal) that allows gentle mixing of the samples. ° The temperature was monitored using a Metrohm Unitrode (6.0258.010) with integrated PT100 thermometer, mounted vertically and centrally within the digestion vessel, which was placed at the center of the digestion chamber. Six samples were taken from the bottom of the digestion vessel throughout the course of the digestion (17.5, 35, 52.5, 70, 87.5, and 105 min) using a sampling pipette with a 4 mm diameter opening. A mobile phone (Samsung Galaxy S8+) was used to photograph the samples in a glass Petri dish against a black background at a resolution of 4032 × 3024 pixels and a height of approximately 15 cm in ambient light. SDS PAGE samples were analyzed under reducing conditions on 4-12% gradient gels (Bolt Gel, Invitrogen) using the manufacturer's protocol (constant voltage, 200 V, 22 min). Sample lanes are framed with standards (Invitrogen Mark 12). After running the gel, the gel was fixed in acid solution (50% water, 40% methanol, 10% acetic acid) for 2 h, washed in water (3 times 5 min, 100 ml water per wash) and then stained overnight (50 ml Simply Blue, Invitrogen). Gels were imaged using a Chemi Doc XRS system (Bio-Rad).

[0668] The results are shown in Figures 4 and 5. Figure 4 shows the semi-dynamic in vitro digestion of beverages A, B, and C (WPI), while the top part of Figure 5 shows the SDS-PAGE analysis of protein aliquots collected at selected time points (17.5-105 min) during the semi-dynamic in vitro digestion of the samples. The pH at various time points of the digestion study is shown below.

[0669] Results – Simulated gastric digestion: Surprisingly, despite the higher content of ALA and CMP in the WPI beverages, very similar visual protein coagulation behavior was observed throughout the digestion of beverages A (BLG at pH 7.0) and C (WPI at pH 7.0). In both beverages (A and C), protein coagulation was already observed at 17.5 min, likely due to the initial mixing with acid and gastric salts, with the amount of coagulation increasing to form an opaque liquid at 35-52.5 min (see Figure 4). The latter time point corresponds to a pH range of approximately 5.6-4.2, with visible protein coagulation / aggregation observed up to 70 min (see Figure 5). Moreover, the coagulation / aggregation was found to be accompanied by an increase in viscosity. Further addition of pepsin and gastric salts effectively lowered the pH, resulting in a clearer digesta, lowered viscosity and a gradual disappearance of protein coagulation.

[0670] Surprisingly, however, beverage B (BLG at pH 6.0) remained opaque throughout digestion and was found to contain no visible protein coagulum from 35 to 52.5 min, and was also assessed as being slightly viscous at this stage of digestion (Figure 4). Figure 5 shows that at the early stage of digestion (17.5 min), the protein band of the 14–21 kDa marker (corresponding to BLG) is indeed predominant in drink A, whereas multiple bands are present in WPI (drink C). Beyond the major BLG band, drink B contains further bands around the 37 kDa marker band that may correspond to BLG dimers. With increasing digestion time (with continued addition of gastric salts and pepsin), protein bands of lower molecular weight than BLG appear in both drinks A and C (WPI).

[0671] Surprisingly, the intensity of these low molecular weight protein / peptide bands in beverages A and C (WPI) beverages was found to be significantly lower in beverage B. This may be due to the increased resistance to gastric proteolysis of BLG, which is predominantly present in protein nanogel form compared to soluble aggregates. This finding may allow beverage manufacturers to tailor which proteins in their beverages are delivered from the gastrointestinal tract to the intestine by using beverages containing predominantly soluble whey protein aggregates or protein nanogels, or by using a mixture.

[0672] Gel strength upon acidification: The gel strength of beverages A, B and C was measured during acidification as described in Example 1.11, and the results are shown in Figure 6.

[0673] Results - Measurement of gel strength upon acidification: Figure 6 shows a simulation of gastric acidification. Gel strength was measured during acidification of three beverages containing mainly soluble aggregates (drinks A and C) or protein nanogel (drink B). The concentration of soluble whey protein aggregates in WPI is expected to be low due to the low content of BLG and high content of other proteins such as CMP (Table 8).

[0674] Although the digestion patterns of drinks A and C are similar (see Figures 4 and 5), it was surprisingly found that the viscosity of BLG drink A dramatically increased when acidified compared to the WPI sample, which may be due to the higher purity of BLG being incorporated into the aggregates and being significantly more abundant in BLG compared to WPI, which is less prone to aggregation of CMP and ALA (see Table 8). Example 10: High protein drink containing BLG (protein nanogel)

[0675] Nutritional high protein beverages were prepared from BLG Powder A (shown in Table 9) containing 10 w / w%, 11 w / w%, 12 w / w%, 13 w / w%, 14 w / w%, 15 w / w%, and 16 w / w% whey protein, with 95.9% w / w or more BLG. The pH was adjusted to pH 6.0 with 3% w / w NaOH. The solutions were heat treated in a 90°C water bath for 5 minutes as described in Example 5. The samples were then cooled in ice water and warmed to room temperature according to Example 5.

[0676] [Table 9]

[0677] The various beverages were analyzed for viscosity (Example 1.8), pictorial appearance (Example 1.9), size (hydrodynamic diameter), and hydrodynamic diameter (Example 1.33).

[0678] result: The results are shown in Figures 7, 8 and 9. Table 10 below shows the chemical composition of the 16 w / w % BLG beverage heat treated at 90°C for 5 minutes.

[0679] The inventors found that the viscosity of the BLG samples surprisingly remained significantly lower even after heat treatment at 90° C. for 5 minutes at concentrations from 10% w / w protein and even more surprisingly at least 16% w / w protein (see Figures 7 and 8). This was completely unexpected, as the comparable WPI samples gelled at 10% w / w WPI pH 6.0 (see Example 8 and Figure 2). Despite such high protein concentrations during thermal aggregation, no sedimentation or particles were observed in the beverage, making the nutritional composition particularly suitable for high protein beverage applications.

[0680] The hydrodynamic diameter of the protein particles was measured (Example 1.33) and is shown in Figure 9. The hydrodynamic diameter of the protein particles was measured to be 185-323 nm, indicating that the high protein beverage contained protein nanogel particles, which have been described by Phan-Xuan et al., 2014 (Phan-Xuan, T., Durand, D., Nicolai, T., Donato, L., Schmitt, C., & Bovetto, L. (2014). Heat induced formation of beta-lactoglobulin microgels driven by addition of calcium ions. Food Hydrocolloids, 2012, 34, 227-235) as particles with a hydrodynamic diameter of 100-300 nm.

[0681] [Table 10]

[0682] Example 11: Neutral pH high protein beverage Nutritional high protein beverages containing 6 w / w%, 10 w / w%, and 12 w / w% whey protein with 95.9% w / w BLG powder A (shown in Table 9) were prepared and the stability and turbidity of the beverages were evaluated. The pH was adjusted to pH 6.0 and 7.0 with 3% w / w NaOH or HCL. The solutions were heat treated in a 90°C water bath for 5 minutes as described in Example 5. The samples were then cooled in ice water and warmed to room temperature according to Example 5. Various samples were analyzed for viscosity (Example 1.8), turbidity (Example 1.7), color (Example 1.9), and amount of insoluble protein material (Example 1.10).

[0683] The results are shown in Table 10 below and FIG.

[0684] JPEG2025013864000017.jpg71168

[0685] result: Clear beverages: Surprisingly, it was found that powder A containing 0.1455 w / w% calcium (Table 9) can be used to produce a stable, clear beverage at pH 7.0 even at a high protein concentration of 10 w / w% protein. The inventors have experienced that WPI-based beverages can form gels under similar conditions.

[0686] Milky drinks: It was found that a milky white beverage with a significantly lower viscosity (4.5 cP) could be produced without visible signs of grain or sedimentation, even at a protein concentration of 12 w / w% protein at pH 6.0, after heat treatment at 90° C. for 5 minutes. In contrast, we found that at 10 w / w% protein, the corresponding WPI samples contained only 57-61 w / w% BLG of the total protein gel, as shown in Example 8.

[0687] Example 12: High protein drinks of neutral pH with lipid content of 25% and 50% of the total energy content Nutritional high protein beverages were prepared from BLG powder A (see Table 9) containing 3 w / w%, 6 w / w%, 10 w / w% and 12 w / w% whey protein, with 95.9% w / w BLG. To evaluate the opportunity to prepare nutritional beverages in the presence of fat, lipids were added up to lipid contents of 25% and 50% of the total energy content. Water and lipids were equilibrated in a water bath at 70° C. 0.2% Grindsted Citrem LR10 was dissolved in heated oil and then slowly mixed with preheated water. The solution was cooled to 60° C., powder was added and stirred for 30-45 minutes to obtain the beverage composition. The pH was adjusted to pH 6.0 or pH 7.0 with 3% NaOH or HCl. The solutions were heat treated in a water bath at 90° C. for 5 minutes. The samples were then cooled in ice water and allowed to warm to room temperature according to Example 5.

[0688] If necessary to obtain a homogenous sample, a homogenization step may be further included in the above steps.

[0689] Various samples were analyzed for viscosity (Example 1.8), turbidity (Example 1.7), color (Example 1.9), and amount of insoluble protein material (Example 1.10).

[0690] The results are shown in Table 11 below.

[0691] [Table 11]

[0692] result: It was found that it was possible to produce stable high protein BLG beverages with lipid contents of 25% and 50% of the total energy content and still have significantly lower viscosities. At pH 7.0, a milky white beverage (containing at least 85 w / w% BLG) with 10 w / w% protein and very low viscosity (5.5 cP) could be produced when 50% of the total energy content was lipid. It was also found that 12 w / w% beverages at pH 6.0 could be produced with a lipid content of 25% and a total energy content of 50%. They had a white milky appearance and a very low viscosity (7.8 cP). The turbidity was at least 11000 NTU. The beverages were stable and no insoluble protein material was observed after 5 minutes at 3000 g.

[0693] Example 13: High PH beverage A nutritional beverage containing BLG was prepared at pH 8.0 to demonstrate stability and appearance at high pH. BLG powders A and B (shown in Table 12) were used to prepare nutritional high protein beverages containing 3 w / w% whey protein with at least 85% by weight being BLG.

[0694] [Table 12]

[0695] The pH was adjusted to pH 8.0 with 3% NaOH. The solution was heat treated in a water bath at 90° C. for 5 minutes. The sample was then cooled in ice water and allowed to warm to room temperature according to Example 5. The results are shown in Table 13.

[0696] [Table 13]

[0697] result: The results show that a clear and stable beverage can be produced at pH 8.0. The beverages were surprisingly clear and colorless with very low viscosity and turbidity at pH 8.0 using either Powder A or Powder B.

Claims

1. 1. A sterilized, packaged, heat-treated beverage preparation having a pH in the range of 5.5 to 8.0, comprising: a total amount of protein of 4-20% w / w relative to the weight of the beverage preparation, at least 92% w / w of said protein being beta-lactoglobulin (BLG); and at least 50% w / w protein nanogel relative to total protein, optionally sweeteners and / or flavorings, However, the beverage preparation has been subjected to a heat treatment, including sterilization.

2. 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 When measured at room temperature, delta b ＊ = b ６．０ｗ／ｗ％タンパク質に標準化したサンプル ＊ -b 脱塩水 ＊ 2. The packaged thermally processed beverage preparation of claim 1, wherein

3. The beverage preparation has a color value delta b ranging from −0.10 to +0.51 on the CIELAB color scale. ＊ and When measured at room temperature, delta b ＊ = b ６．０ｗ／ｗ％タンパク質に標準化したサンプル ＊ -b 脱塩水 ＊ 2. The packaged thermally processed beverage preparation of claim 1, wherein

4. 4. The packaged heat-treated beverage preparation according to any one of claims 1 to 3, wherein the sum of the amounts of Na, K, Mg and Ca is up to 400 mM.

5. A packaged heat-treated beverage preparation according to any one of claims 1 to 4, wherein the beverage preparation comprises up to 100 mg phosphor / 100 g protein and up to 700 mg potassium / 100 g protein.

6. A packaged heat-treated beverage preparation according to any one of claims 1 to 5, having a pH in the range of 6.5 to 7.5, preferably 6.5 to 7.

0.

7. A packaged heat-treated beverage preparation according to any one of claims 1 to 6, having a turbidity of up to 200 NTU.

8. A packaged heat-treated beverage preparation according to any one of claims 1 to 7, having a turbidity of more than 200 NTU.

9. 9. The packaged heat-treated beverage preparation of any one of claims 1 to 8, having a viscosity of up to 200 cP centipoise when measured at 22°C at a shear rate of 100 / sec.

10. A packaged heat-treated beverage preparation according to any one of claims 1 to 9, comprising a total amount of protein of 4 to 10% w / w based on the weight of the beverage preparation.

11. A packaged heat-treated beverage preparation according to any one of claims 1 to 10, comprising a total amount of protein of 10 to 20% w / w based on the weight of the beverage preparation.

12. A packaged thermally processed beverage preparation according to any one of claims 1 to 11, further comprising carbohydrates representing 0-95% of the total energy content of the preparation.

13. A packaged thermally processed beverage preparation according to any one of claims 1 to 12, further comprising a lipid content of 0 to 50% of the total energy content of the preparation.

14. A packaged, heat-treated beverage preparation according to any one of claims 1 to 13, comprising a BLG isolate.

15. 15. A packaged heat-treated beverage preparation according to any one of claims 1 to 14, wherein each major non-BLG whey protein is present in a weight percentage of total protein that is up to 10% by weight of total protein in a standard whey protein concentrate derived from sweet whey.

16. 16. A packaged heat-treated beverage preparation according to any one of claims 1 to 15, wherein each major non-BLG whey protein is present in a weight percentage of total protein that is up to 6% by weight of total protein in a standard whey protein concentrate derived from sweet whey.

17. 16. A packaged heat-treated beverage preparation according to any one of claims 1 to 15, wherein each major non-BLG whey protein is present in a weight percentage of total protein that is up to 4% by weight of total protein in a standard whey protein concentrate derived from sweet whey.

18. 1. A method for producing a sterile, packaged heat-treated beverage preparation having a pH in the range of 5.5 to 8.0, wherein said packaged heat-treated beverage preparation comprises at least 50% w / w protein nanogel relative to total protein, the method comprising the steps of: a) providing a liquid solution comprising: - 4 to 20 wt. % total amount of protein, of which at least 92 w / w% is beta-lactoglobulin (BLG); optionally sweeteners and / or flavorings, 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 which includes at least sterilization.

19. 19. The method of claim 18, wherein the sterilization comprises a temperature in the range of 120 to 150°C for 4 to 30 seconds.

20. A packaged heat-treated beverage preparation described in any one of claims 1 to 17, wherein at least 94% by weight of the protein is BLG.

21. A packaged heat-treated beverage preparation described in any one of claims 1 to 17, wherein at least 96% by weight of the protein is BLG.

22. 22. A packaged, thermally treated beverage preparation according to any one of claims 1 to 17, 20 or 21 for use in a method for the treatment of a disease associated with protein malabsorption.

23. A nutritional supplement comprising a packaged heat-treated beverage preparation according to any one of claims 1 to 17, 20 or 21.

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