Acidic beta-lactoglobulin beverage preparation

Abstract

The present invention relates to a novel packaged heat-treated beverage formulation having a pH in the range of 2.0 to 4.7. The present invention also relates to a method for manufacturing a packaged heat-treated beverage formulation and different uses of the packaged heat-treated beverage formulation.

Description

Technology Field

[0001] The present invention relates to a novel packaged heat-treated beverage formulation having a pH in the range of 2.0 to 4.7. The present invention also relates to a method for manufacturing a packaged heat-treated beverage formulation and other uses of the packaged heat-treated beverage formulation. Background Technology

[0002] Nutritional supplements containing whey protein are generally used for muscle synthesis, weight control, and maintaining muscle and body weight. These supplements are targeted at different types of consumers, e.g., sportsmen / women, athletes, children, the elderly, and patients who are malnourished or at risk of malnutrition, and / or patients with increased protein requirements. Whey protein can be isolated from milk serum or whey. Whey typically contains a mixture of beta-lactoglobulin (BLG), alpha-lactalbumin (ALA), serum albumin, and immunoglobulin, of which BLG is the most dominant. Therefore, whey protein concentrate (WPC) contains a mixture of proteins. Whey protein isolate (WPI) contains less fat and lactose than WPC. Beverages containing whey protein, such as acidic heat-treated beverages, are well known.

[0003] Etzel 2004 (Etzel, MR, 2004, Manufacture and use of dairy protein fraction. American Society for Nutritional Science, pp. 996-1002) describes a beverage containing 2.5 wt% WPI with a pH of 2 to 7. They found that a heat-treated beverage can only be obtained by adding an anti-coagulant.

[0004] WO 2018 / 115,520 A1 discloses a method for preparing an isolated beta-lactoglobulin composition for food and / or a composition containing crystallized beta-lactoglobulin based on the crystallization of BLG in a salting-in mode. The crystallized BLG can then be separated from the residual mother liquor.

[0005] WO 2004 / 049,819 A2 discloses a method for improving the functional properties of globular proteins, the method comprising: providing a solution of one or more globular proteins in which the protein(s) are at least partially aggregated in fibrils; and performing one or more of the following steps in a random order: increasing the pH; increasing the salt concentration; concentrating the solution; and changing the solvent quality of the solution. Preferably, the solution of one or more globular proteins is provided by heating at a low pH or by adding a denaturing agent. Also disclosed are the protein additive thus obtained, its use for food and non-food applications, and food and non-food products containing the protein additive.

[0006] WO 2014 / 055,830 A1 discloses a shelf-stable and clear liquid nutritional composition containing water with a pH in the range of 2.5 to 4.6; at least one source of EGCg in an amount sufficient to provide 200 to 1700 mg / L of EGCg; and at least one source of protein in an amount sufficient to provide 25 to 45 g / L of total protein. The shelf-stable and clear liquid nutritional composition loses 20 weight% or less of the solid EGCg content present in the initial formulation of the composition due to epimerization, degradation, or both epimerization and degradation during heat sterilization. In certain embodiments, the loss of EGCg is indicated by the amount of epimerization product EGCg present in the shelf-stable and clear liquid nutritional composition after heat sterilization. A method for preparing the shelf-stable and clear liquid nutritional composition is also disclosed.

[0007] WO 2011 / 112,695 A1 discloses a nutritional composition and a method for preparing and using the nutritional composition. The nutritional composition comprises whey protein micelles and leucine and provides a sufficient amount of leucine to improve human protein synthesis while also maintaining a low-viscosity fluid matrix and acceptable organoleptic properties.

[0008] WO 2011 / 051,436 A1 discloses at least partially transparent compositions for human or animal consumption and packaging of such compositions. One embodiment of the present invention relates to at least partially transparent containers containing at least partially transparent aqueous non-alcoholic compositions. The containers comprise at least one polarizer for visualizing liquid crystals present in the compositions.

[0009] WO 2010 / 037,736 A1 discloses the isolation of whey protein and the manufacture of whey products and whey isolates. In particular, the present 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 present invention are high in α-lactalbumin and immunoglobulin G in addition to being low in β-lactoglobulin.

[0010] FR 2 296 428 discloses dietary and therapeutic protein compositions based on lactoserum protein obtained by any known separation process. The compositions may be used for the treatment or prevention of digestive disorders (e.g., diarrhea) in infants and adults, for increasing resistance to intestinal infections, and for the treatment of certain metabolic disorders (e.g., hyperphenylalaninemia). They may also be used dermatologically or cosmetically and may form part of a low-protein diet.

[0011] The inventors have observed that sensory characteristics, such as astringency and texture, play an important role in the selection of liquid nutritional beverages by consumers. Some of the challenges that arise when incorporating whey protein into acidic heat-treated beverages are the formation of unstable precipitates settling in the beverage, high viscosity or even gel formation, and unpleasant tastes resulting from a high degree of astringency and / or dry texture.

[0012] The object of the present invention is to provide an acidic packaged heat-treated beverage formulation containing whey protein and having improved sensory reception and / or visual properties.

[0013] Another objective of the present invention is to provide a high-protein beverage that has low viscosity, a pleasant taste, optionally low astringency, and can be transparent or opaque.

[0014] In the present invention, the inventors have discovered that such packaged heat-treated beverages can be provided within a wide acidic pH range of pH 4.7 or lower, while still having low viscosity and optionally also low levels of astringency and dry texture. The present invention provides both clear beverages and opaque but stable beverages.

[0015] Accordingly, an aspect of the present invention relates to a packaged heat-treated beverage formulation having a pH in the range of 2.0 to 4.7, said beverage comprising:

[0016] - Total amount of protein of 2 to 45% w / w relative to the weight of the beverage - where at least 85% w / w of the protein is BLG -

[0017] - Optionally, sweeteners, sugar polymers and / or flavorings.

[0018] Another aspect of the present invention relates to a method for producing a packaged heat-treated beverage formulation having a pH in the range of 2.0 to 4.7, the method comprising the following steps:

[0019] a) Step of providing a liquid solution containing the following:

[0020] - Total amount of protein of 2 to 45 weight% - where, at least 85% of the protein is BLG -

[0021] - Optionally, sweeteners, sugar polymers and / or flavorings

[0022] b) Step of packaging the liquid solution,

[0023] Here, the liquid solution of step a) and / or the packaged liquid solution of step b) are heat-treated, including at least pasteurization.

[0024] Further aspects of the present invention relate to the use of a protein solution comprising a total amount of protein of 2 to 45% w / w relative to the weight of the solution, wherein at least 85% w / w of the protein is BLG to control the turbidity of a heat-treated acidic beverage formulation having a pH in the range of 2.0 to 4.7.

[0025] Another further aspect of the present invention relates to the use of a protein solution comprising a total amount of protein of 2 to 45% w / w relative to the weight of the solution, wherein at least 85% w / w of the protein is BLG to control the astringent taste of a heat-treated acidic beverage formulation having a pH in the range of 2.0 to 4.7.

[0026] A further aspect of the present invention relates to a packaged heat-treated beverage formulation according to the present invention for use in a method for treating a disease associated with poor protein absorption.

[0027] Further aspects of the present invention relate to the use of a packaged heat-treated beverage formulation according to the present invention as a dietary supplement. Brief explanation of the drawing

[0028] Figure 1 shows images of BLG and WPI beverages with a pH of 3.7 and a protein content of 6% w / w, heat-treated at 120°C for 20 seconds or at 75°C for 15 seconds. Figure 2 shows images of WPI-B pH 3.0 to 3.7 at 120℃ and BLG pH 3.7 at 120℃ / 20 seconds. Figure 3 shows images of WPI-B pH 3.0 to 3.7 at 75°C and BLG pH 3.7 at 75°C / 15 seconds. Figure 4 shows images of WPI-B pH 3.7 and BLG pH 3.9, 75℃ / 15 sec. Figure 5 shows the turbidity of a BLG beverage formulation treated with 6% UHT (120℃ / 20 sec). Figure 6 shows the turbidity of a 6% pasteurized (75℃ / 15 sec) BLG beverage formulation. Figure 7 shows the viscosity of a BLG beverage formulation treated with 6% UHT (120℃ / 20 sec). Figure 8 shows the yellowness (b*) of a beverage composition treated with 6% UHT (120°C / 20 seconds). Figure 9 shows the yellowness (b*) of a 6% pasteurized (75°C / 15 seconds) beverage composition. Figure 10 shows images of 15% BLG beverage pH 3.7 (left) and 6% WPI pH 3.7 (right) at 75℃ / 15 seconds. Figure 11 shows the results of the sensory evaluation of the high-protein BLG beverage formulation and images of 6 w / w% and 15 w / w% BLG samples at pH 3.7. Figure 12 shows high-protein beverage preparations prepared by heating 30, 27.5, 25, and 20% BLG beverage preparations (from left to right) at 75°C for 5 minutes. The viscosity remained low even after heating. Figure 13 shows images of different WPI and BLG beverage samples. Figure 14 shows the sensory evaluation of the beverages (scale from 0 to 15). WPI pH 3.0 120℃ / 20 sec and BLG pH 3.7 75℃ / 15 sec. Figure 15 demonstrates the effect of pH and temperature on the taste of protein drinks. Figure 16 shows sensory data for the astringent taste of BLG beverages at pH 3.0 (120℃ / 20 sec) and pH 3.7 (75℃ / 15 sec). Figure 17 shows sensory data on the dry texture of BLG beverages at pH 3.7 heat-treated at 120℃ / 20 seconds or 75℃ / 15 seconds. Figure 18 shows sensory data recording how heat-treatment temperature affects whey flavor. Figure 19 shows an image of a mineral-rich 6% BLG beverage with a pH of 3.7 and heat-treated at 95°C for 5 minutes. Figure 20 shows an image of a 6% BLG beverage with a pH of 3.7, which is rich in minerals and heat-treated at 75°C for 5 minutes. Figure 21 shows the stability of milky white BLG beverages with and without sucrose added at pH 4.3 when heat-treated at 93°C for 4 minutes. Figure 22 shows an image of an opaque 6% protein BLG beverage prepared by heat treatment at 75°C for 5 minutes with a pH of 4.2 or 4 to 5. Figure 23 shows images of BLG and SPI beverages with a pH of 3.7 and heat-treated at 75°C for 5 minutes. Figure 24 shows images of BLG and SPI beverages with a pH of 3.7. Figure 25 shows the yellowness (b*) of 6 w / w % BLG UHT-treated (120°C / 20 sec) beverage compositions (pH 3.0 and 3.7) stored in a dark place (20°C) for up to 6 months. Figure 26 shows the yellowness (b*) of 6 w / w % BLG pasteurized (75°C / 15 sec) beverages (pH 3.0 and 3.7) stored in a dark place (20°C) for up to 6 months. Specific details for implementing the invention

[0029] definition

[0030] In the context of the present invention, the term “beta-lactoglobulin” or “BLG” relates to beta-lactoglobulin from mammalian species in, for example, natural, unfolded, and / or glycosylated forms, and includes naturally occurring genetic variants. The term also includes aggregated BLG, precipitated BLG, and crystalline BLG. When referring to the amount of BLG, the total amount of BLG including aggregated BLG is referenced. 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 also aggregated with other modified BLG molecules and / or other modified whey proteins by hydrophobic interactions and / or covalent bonding.

[0031] BLG is the most dominant protein in bovine whey and milk serum, exists as several genetic variants, and the major proteins in cow's milk are designated as A and B. BLG is a lipocalin protein and can bind to many hydrophobic molecules, suggesting a role in their transport. BLG has also been shown to be able to bind to iron via siderophores and may play a role in fighting pathogens. Homologues of BLG are not found in human breast milk.

[0032] Bovine BLG is a relatively small protein consisting of approximately 162 amino acid residues with a molecular weight of about 18.3 to 18.4 kDa. Under physiological conditions, it is primarily a dimer but dissociates into monomers at pH below about 3, which are preserved in its natural state as determined using nuclear magnetic resonance spectroscopy. Conversely, BLG also occurs as tetramers, octamers, and other multiple aggregated forms under various natural conditions.

[0033] In the context of the present invention, the term “non-aggregated beta-lactoglobulin” or “non-aggregated BLG” also relates to beta-lactoglobulin from mammalian species in a naturally unfolded and / or glycosylated form, for example, and includes naturally occurring genetic variants. However, the term does not include aggregated BLG, precipitated BLG, or crystallized BLG. The amount or concentration of non-aggregated BLG is determined according to Example 1.6.

[0034] The percentage of non-aggregated BLG relative to total BLG is (m 총 BLG - m 비응집체 BLG ) / m 총 BLG It is determined by calculating *100%. m 총 BLG is the concentration or amount of BLG determined according to Example 1.31, and m 비-응집된 BLG is the concentration or amount of non-aggregated BLG determined according to Example 1.6.

[0035] In the context of the present invention, the term “crystal” relates to a solid material in which components (e.g., atoms, molecules, or ions) are arranged in a highly ordered microscopic structure to form a crystal lattice that extends in all directions.

[0036] In the context of the present invention, the term “BLG crystal” relates to protein crystals that form a crystal lattice extending in all directions, containing natural BLG arranged in a non-aggregated and preferably highly ordered fine structure. The BLG crystals may be, for example, monolithic or polycrystalline, and may be, for example, whole crystals, fragments of crystals, or a combination thereof. Fragments of crystals are formed, for example, when whole crystals are subjected to mechanical shearing during processing. Fragments of crystals also have a highly ordered fine structure but may lack the uniform surface and / or uniform edges or corners of the whole crystals. For example, for examples of multiple whole BLG crystals, refer to Fig. 18 of PCT application no. PCT / EP2017 / 084553, and for examples of fragments of BLG crystals, refer to Fig. 13 of PCT application no. PCT / EP2017 / 084553. In both cases, BLG crystals or crystal fragments can be visually identified as having a well-defined, dense, and coherent structure using an optical microscope. BLG crystals or crystal fragments are often at least partially transparent. Protein crystals are also known to be birefringent, and this optical property can be used to identify unknown particles with a crystalline structure. On the other hand, amorphous BLG aggregates often appear as insufficiently defined, opaque, and irregularly sized open or porous masses.

[0037] In the context of the present invention, the term "crystallize" relates to the formation of protein crystals. Crystallization may occur spontaneously, for example, or be initiated by the addition of a crystallization seed.

[0038] In the context of the present invention, the term “edible composition” relates to a composition that is safe for human consumption and use as a food ingredient and does not contain problematic amounts of toxic components, such as toluene or other unwanted organic solvents.

[0039] In the context of the present invention, the term “ALA” or “alpha-lactalbumin” relates to alpha-lactalbumin from mammalian species in, for example, natural and / or glycosylated forms, and includes naturally occurring genetic variants. The term also includes aggregated ALA and precipitated BLG. When referring to the amount of ALA, the total amount of ALA including aggregated ALA is referred to, for example. The total amount of ALA is determined according to Example 1.31. The term “aggregated ALA” relates to ALA that is typically at least partially unfolded and aggregated with other modified ALA molecules and / or other modified whey proteins by hydrophobic interactions and / or covalent bonding.

[0040] Alpha-lactalbumin (ALA) is a protein present in the milk of almost all mammalian species. ALA forms the regulatory subunit of the lactose synthase (LS) heterodimer, and β-1,4-galactosyltransferase (β4Gal-T1) forms the catalytic component. Together, these proteins enable LS to produce lactose by transferring the galactose moiety to glucose. One of the major structural differences from beta-lactoglobulin is that ALA does not possess any free thiol groups that can act as initiators for covalent aggregation reactions.

[0041] In the context of the present invention, the term “non-aggregated ALA” also relates to ALA from mammalian species in a naturally unfolded and / or glycosylated form, for example, and includes naturally occurring genetic variants. However, the term does not include aggregated ALA or precipitated ALA. The amount or concentration of non-aggregated BLG is determined according to Example 1.6.

[0042] The percentage of non-aggregated ALA relative to total ALA is (m 총 ALA - m 비응집체 ALA ) / m 총 ALA It is determined by calculating *100%. m 총 ALAis the concentration or amount of ALA determined according to Example 1.31, and m 비-응집된 ALA is the concentration or amount of non-aggregated ALA determined according to Example 1.6.

[0043] In the context of the present invention, the term “caseinomacropeptide” or “CMP” comprises a naturally occurring genetic variant of an aspartate proteinase, for example, chymosin, derived from the hydrolysis of “κ-CN” or “kappa-casein” from mammalian species in a natural and / or glycosylated form, relating to residues 106 to 169.

[0044] In the context of the present invention, the term "BLG isolate" means a composition containing BLG in an amount of at least 85% w / w relative to total protein. The BLG isolate preferably has a total protein content of at least 30% w / w relative to total solids, and preferably at least 80% w / w.

[0045] In the context of the present invention, the term "BLG isolate powder" relates to BLG isolate in powder form and preferably to free-flowing powder.

[0046] In the context of the present invention, the term "BLG isolate liquid" relates to BLG isolate in liquid form and preferably an aqueous liquid.

[0047] The term "whey" refers to the liquid phase remaining after casein in milk has been precipitated and removed. Casein precipitation can be achieved, for example, by acidifying the milk and / or using rennet enzymes. There are various types of whey, such as "sweet whey," a whey product produced by rennet-based precipitation of casein, and "acid whey" or "sour whey," a whey product produced by acid-based precipitation of casein. Acid-based precipitation of casein can be achieved, for example, by adding food acids or bacterial culture.

[0048] The term "milk serum" refers to the liquid remaining when casein and milk fat globules are removed from milk, for example, by microfiltration or large-pore ultrafiltration. Milk serum may also be referred to as "ideal whey."

[0049] The term "milk serum protein" or "serum protein" relates to proteins present in milk serum.

[0050] In the context of the present invention, the term “whey protein” relates to a protein found in whey or milk serum. A whey protein may be a subset of protein species found in whey or milk serum, may even be a single whey protein species, or may be a complete set of protein species found in whey or / and milk serum.

[0051] In the context of the present invention, the major non-BLG proteins of the standard whey protein concentrate from sweet whey are ALA, CMP, bovine serum albumin, immunoglobulin, osteopontin, lactoferrin, and lactoperoxidase. In the context of the present invention, the weight percentage of the major non-BLG whey proteins of the standard whey protein concentrate from sweet whey is as follows:

[0052] 18% w / w of total protein ALA,

[0053] 18% w / w of CMP relative to total protein,

[0054] 4% w / w of total protein amount of BSA,

[0055] 5% w / w amount of casein species relative to total protein,

[0056] 6% w / w of total protein in immunoglobulin,

[0057] 0.5% w / w of total protein of osteopontin,

[0058] 0.1% w / w amount of lactoferrin relative to total protein, and

[0059] 0.1% w / w amount of lactoperoxidase relative to total protein.

[0060] In the context of the present invention, the term “mother liquor” relates to a whey protein solution remaining after BLG has been crystallized and at least partially removed. The mother liquor may still contain some BLG crystals, but generally may contain only small BLG crystals that have escaped separation.

[0061] The term casein relates to casein proteins found in milk and includes natural micellar casein found in raw milk, individual casein species, and caseates.

[0062] 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 higher than the saturation point of non-aggregated BLG in the liquid under given physical and chemical conditions. The term “supersaturation” is well known in the field of crystallization (see, for example, Gerard Coquerela, “Crystallization of molecular systems from solution: phase diagrams, supersaturation and other basic concepts”, Chemical Society Reviews, p. 2286–2300, Issue 7, 2014), and supersaturation can be determined by a number of different measurement techniques (e.g., by spectroscopy or particle size analysis). In the context of the present invention, supersaturation with respect to BLG is determined by the following procedure.

[0063] Procedure for testing whether a liquid becomes supersaturated with respect to BLG under a specific set of conditions:

[0064] a) Transfer a 50 ml sample of the liquid to be tested to a centrifuge tube (VWR Catalog No. 525-0402) having a height of 115 mm, an inner diameter of 25 mm, and a capacity of 50 mL. During steps a) through h), care must be taken to maintain the sample and its subsequent fractions under the original physical and chemical conditions of the liquid.

[0065] b) Immediately centrifuge the sample at 3000 g for 3.0 minutes with a maximum acceleration of 30 seconds and a maximum deceleration of 30 seconds.

[0066] c) Immediately after centrifugation, transfer as much of the supernatant as possible (without disturbing the pellet if one has formed) to a second centrifugation tube (of the same type as in step a).

[0067] d) Take a 0.05 mL subsample (subsample A) of the supernatant.

[0068] e) Add 10 mg of BLG crystals (at least 98% pure, non-aggregated BLG relative to total solids) having a particle size of up to 200 microns to the second centrifuge tube and shake the mixture.

[0069] f) Leave the second centrifuge tube at its original temperature for 60 minutes.

[0070] g) Immediately after step f), centrifuge the second centrifuge tube at 500 g for 10 minutes, then take another 0.05 mL subsample (subsample B) of the supernatant.

[0071] h) If the centrifugation pellet from step g) is present, recover it, resuspend it in milliQ water, and immediately inspect the suspension for the presence of microscopic crystals.

[0072] i) Determine the concentration of non-aggregated BLG in subsamples A and B using the method summarized in Example 1.6—the results are expressed as % BLG w / w relative to the total weight of the subsamples. The concentration of non-aggregated BLG in subsample A is C BLG, AIt is referred to as, and the concentration of non-aggregated BLG in subsample B is C BLG, B It is referred to as.

[0073] j) The liquid from which the sample of step a) was taken is c BLG, B a c BLG, A If a crystal is observed at a lower level and at step i) (under specific conditions), it is supersaturated.

[0074] In the context of the present invention, the terms “liquid” and “solution” include both compositions free of particulate matter and compositions containing a combination of liquid and solid and / or semi-solid particles, such as protein crystals or other protein particles. Accordingly, the “liquid” or “solution” may be a suspension or even a slurry. However, the “liquid” and “solution” are preferably pumpable.

[0075] In the context of the present invention, the terms "whey protein concentrate (WPC)" and "serum protein concentrate (SPC)" relate to dry or aqueous compositions containing a total amount of protein of 20 to 89% w / w relative to the total solids.

[0076] WPC or SPC preferably contains the following:

[0077] 20 to 89% w / w protein relative to total solids,

[0078] 15 to 70% w / w BLG relative to total protein,

[0079] 8 to 50% w / w ALA relative to total protein, and

[0080] 0 to 40% w / w CMP relative to protein.

[0081] Alternatively, but also preferably, WPC or SPC may contain the following:

[0082] 20 to 89% w / w protein relative to total solids,

[0083] 15 to 90% w / w BLG relative to total protein,

[0084] 4 to 50% w / w ALA relative to total protein, and

[0085] 0 to 40% w / w CMP relative to protein.

[0086] Preferably, WPC or SPC contains the following:

[0087] 20 to 89% w / w protein relative to total solids,

[0088] 15 to 80% w / w BLG relative to total protein,

[0089] 4 to 50% w / w ALA relative to total protein, and

[0090] 0 to 40% w / w CMP relative to protein.

[0091] More preferably, WPC or SPC contains the following:

[0092] 70 to 89% w / w protein relative to total solids,

[0093] 30 to 90% w / w BLG relative to total protein,

[0094] 4 to 35% w / w ALA relative to total protein, and

[0095] 0 to 25% w / w CMP relative to protein.

[0096] SPC typically contains no CMP or only trace amounts of CMP.

[0097] 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 to 100% w / w relative to total solids.

[0098] WPI or SPI preferably contains the following:

[0099] 90 to 100% w / w protein relative to total solids,

[0100] 15 to 70% w / w BLG relative to total protein,

[0101] 8 to 50% w / w ALA relative to total protein, and

[0102] 0 to 40% w / w CMP relative to total protein.

[0103] Alternatively, but also preferably, WPI or SPI may contain the following:

[0104] 90 to 100% w / w protein relative to total solids

[0105] 30 to 95% w / w BLG relative to total protein,

[0106] 4 to 35% w / w ALA relative to total protein, and

[0107] 0 to 25% w / w CMP relative to total protein.

[0108] More preferably, WPI or SPI may contain the following:

[0109] 90 to 100% w / w protein relative to total solids,

[0110] 30 to 90% w / w BLG relative to total protein,

[0111] 4 to 35% w / w ALA relative to total protein, and

[0112] 0 to 25% w / w CMP relative to total protein.

[0113] SPI typically contains no CMP or only trace amounts of CMP.

[0114] In the context of the present invention, the term “additional protein” means a protein that is not BLG. The additional protein present in the whey protein solution typically comprises one or more of the non-BLG proteins found in milk serum or whey. Non-limiting examples of such proteins are alpha-lactalbumin, bovine serum albumin, immunoglobulin, caseinomacropeptide (CMP), osteopontin, lactoferrin, and milk fat globule membrane protein.

[0115] The terms "essentially made" and "essentially made" mean that the claim or feature includes materials or steps specified and that do not substantially affect the basic and novel characteristic(s) of the claimed invention.

[0116] 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, the phrase "n1, n2, ..., n i-1 , and / or n i " is " n1" or " n2" or ... or "n i-1 " or "n i " or means any combination of components: n1, n2,...n i-1 , and n i .

[0117] In the context of the present invention, the term “dry” or “dried” means that the composition or product contains up to 10% w / w, preferably up to 6% w / w, more preferably less water.

[0118] In the context of the present invention, the term "physical microbial reduction" relates to physical interactions with the composition that reduce the total amount of viable microorganisms in the composition. The term does not include the addition of chemicals that kill microorganisms. The term also does not include heat exposure to which the sprayed droplets are exposed during spray-drying, but includes possible preheating prior to spray-drying.

[0119] In the context of the present invention, the pH of the powder is 10 g mixed in 90 g of deionized water. It refers to the pH of the powder and is measured according to Example 1.16.

[0120] In the context of the present invention, the weight percentage (% w / w) of a component of a particular composition, product, or material means the weight percentage of that component relative to the weight of the particular composition, product, or material, unless otherwise specifically mentioned (e.g., total solids or total protein).

[0121] In the context of the present invention, the process step "concentration" and the verb "to concentrate" relate to the concentration of protein and include both the concentration of protein based on total solids and the concentration of protein based on total weight. This means that, for example, as long as the protein content increases relative to total solids, the concentration does not necessarily require an increase in the absolute concentration w / w of the protein in the composition.

[0122] In the context of the present invention, the term "weight ratio" between component X and component Y is m X / m Y It refers to the value obtained by calculation, where m X is the amount (weight) of component X, and m Y is the amount (weight) of component Y.

[0123] In the context of the present invention, the terms “at least pasteurized” and “at least pasteurized” relate to a heat treatment having a microbial killing effect equal to or greater than that of a heat treatment at 70°C for 10 seconds. The criterion for determining the bactericidal effect is E. coli O157:H7.

[0124] In the context of the present invention, the term “whey protein feed” relates to a whey protein source from which liquid BLG isolates are derived. The whey protein feed has a lower content of BLG relative to total protein than the liquid BLG isolates and is typically WPC, WPI, SPC, or SPI.

[0125] In the context of the present invention, the term “BLG-rich composition” relates to a BLG-rich composition produced by isolating BLG from a whey protein feed. The BLG-rich composition typically contains the same whey protein as the whey protein feed, but BLG is present at a significantly higher concentration relative to total protein than in the whey protein feed. The BLG-rich composition may be prepared from a whey protein feed, for example, by chromatography, protein crystallization, and / or membrane-based protein fractionation. The BLG-rich composition contains BLG in an amount of at least 85% w / w, preferably at least 90% w / w, relative to total protein. In some cases, the BLG-rich composition may be used directly as a liquid BLG isolate. However, additional processing is often required to convert the BLG-rich composition into a liquid BLG isolate.

[0126] In the context of the present invention, the term "whey protein solution" is used to describe a specific aqueous whey protein composition that is supersaturated relative to BLG in a salting mode and is useful for producing BLG crystals.

[0127] In the context of the present invention, the term “sterile state” means that the sterile composition or product does not contain any arbitrarily viable microorganisms and therefore there is no microbial growth during storage at room temperature. The sterile composition is in a sterile state.

[0128] When a liquid, such as a beverage preparation, is sterilized and packaged in a sterile container under aseptic conditions, it typically has a shelf life of at least six months at room temperature. Sterilization treatment kills spores and microorganisms that can cause the liquid to spoil.

[0129] In the context of the present invention, the term “energy content” refers to the total energy content contained in a food product. Energy content may be measured in kilojoules (kJ) or kilocalories (kcal) and is referred to as calories per serving of the food product, for example, kcal per 100 grams of the food product. An example is a beverage having an energy content of 350 kcal per 100 grams of the beverage.

[0130] The total energy content of a food product includes the energy contribution from all macronutrients present in the food product, such as energy from protein, lipids, and carbohydrates. The energy distribution from macronutrients within the food product can be calculated based on the amount of macronutrients within the food product and the contribution of macronutrients relative to the total energy content of the food product. The energy distribution may be referred to as the energy percentage (E%) of the total energy content of the food product. For example, in the case of a beverage containing 20 E% protein, 50 E% carbohydrates, and 30 E% lipids, this means that 20% of the total energy comes from protein, 50% of the total energy comes from carbohydrates, and 30% of the total energy comes from fat (lipids).

[0131] In the context of the present invention, the term “nutritionally complete nutritional supplement” is understood as a food product containing proteins, lipids, and carbohydrates, and additionally containing vitamins, minerals, and trace elements, wherein the beverage has a nutritional profile consistent with a complete and healthy diet.

[0132] In the context of the present invention, the term "nutritionally incomplete supplement" refers to a food product containing one or more macronutrients and optionally additionally containing vitamins, minerals, and trace elements. A nutritionally incomplete beverage may contain protein as the sole nutrient, or, for example, may contain protein and carbohydrates.

[0133] The term "Food for Special Medical Purposes (FSMP)" or "medical food" refers to food products intended for oral intake or tube administration used for specific medical disorders, diseases, or conditions that have unique nutritional requirements and are used under medical supervision. Medical foods may be nutritionally complete supplements / beverages or nutritionally incomplete supplements / beverages.

[0134] The term "nutrient" refers to a substance used by an organism for survival, growth, and reproduction. Nutrients can be 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.

[0135] The term "nutrient" refers to a substance used by an organism for survival, growth, and reproduction. Nutrients can be 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.

[0136] The term "instant beverage powder" or "instant beverage powder product" refers to powder that can be converted into a liquid beverage by adding a liquid, such as water.

[0137] In the context of the present invention, the terms "beverage" and "beverage preparation" and "preparation" as substantially used relate to any water-based liquid that can be consumed as a beverage, for example, by pouring, sipping, or supplying via tube.

[0138] In the context of the present invention, the term "protein fraction" relates to proteins of the composition, for example, proteins of powder or beverage formulations.

[0139] In the context of the present invention, the term “astringent taste” relates to mouthfeeling. Astringent taste is felt like the contraction of cheek muscles and increases saliva production. Therefore, astringent taste is not a taste in itself, but a physical mouthfeel and a time-dependent sensation in the mouth.

[0140] In the context of the present invention, the term "dry texture" relates to a sensation in the mouth, which feels as if the mouth and teeth are drying out, thereby minimizing saliva production.

[0141] Therefore, dry texture is not a taste in itself, but a physical texture and a time-dependent sensation in the mouth.

[0142] 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.

[0143] In the context of the present invention, the terms “lipid,” “fat,” and “oil” as used herein are used interchangeably to refer to lipid materials derived from or processed from plants or animals, unless otherwise specified. These terms also include synthetic lipid materials insofar as such synthetic materials are suitable for human consumption.

[0144] In the context of the present invention, the term "transparent" includes beverage formulations that have a noticeably clear appearance, allow light to pass through, and display a distinct image. The transparent beverage has a turbidity of up to 200 NTU.

[0145] In the context of the present invention, the term "opaque" includes beverage formulations that are not noticeably clear in appearance and have a turbidity of more than 200 NTU.

[0146] An aspect of the present invention relates to a packaged heat-treated beverage formulation having a pH in the range of 2.0 to 4.7, said beverage comprising:

[0147] - Total amount of protein of 2 to 45% w / w relative to the weight of the beverage - where at least 85% w / w of the protein is BLG - and

[0148] - Optionally, sweeteners, sugar polymers and / or flavorings.

[0149] Packaged heat-treated beverage formulations containing at least 85% w / w of protein are highly beneficial for a number of reasons. In acidic beverages, high BLG content also increases the pH range and reduces the heating temperature while maintaining clarity and color, which is possible even when high protein concentrations are applied. This means that clear, colorless high-protein beverages can be produced at a less acidic pH level than is possible with traditional WPI.

[0150] Surprisingly, BLG beverages were found to have lower astringent taste, dry texture, sourness, whey flavor, and citric acid flavor compared to traditional acidic WPI beverages.

[0151] Another advantage of the present invention and its extended pH range is that a stable milky white beverage can be produced while having high turbidity and low viscosity, yet remaining white and not turning yellow.

[0152] In some preferred embodiments of the packaged heat-treated beverage formulation 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.

[0153] Since even higher relative amounts of BLG are feasible and desirable, in some preferred embodiments of the present invention, at least 94% w / w of the protein in the packaged heat-treated beverage formulation 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 about 100% w / w.

[0154] For example, a packaged heat-treated beverage formulation preferably contains 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% w / w relative to total protein, and most preferably contains BLG in an amount of at least 99.5% w / w relative to total protein, e.g., approximately 100.0% w / w relative to total protein.

[0155] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation is at least pasteurized.

[0156] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation is sterilized and is therefore in a sterile state.

[0157] In some preferred embodiments of the present invention, the natural form of the protein is maintained. The natural form of the protein is preferably maintained using heat treatment that does not cause irreversible changes to the protein form of at least BLG, preferably all proteins.

[0158] The degree of naturalness of a protein depends on a number of factors, including protein concentration, pH, temperature, and heat treatment time.

[0159] The intrinsic tryptophan fluorescence emission ratio R=I330 / I350 is a measure of protein nativity. When R is at least 1.11, the natural form is dominant, whereas when R is less than 1.11, at least partial unfolding and aggregation are dominant. A method for analyzing intrinsic tryptophan fluorescence is described in Example 1.1.

[0160] The inventors have discovered that an intrinsic tryptophan fluorescence emission ratio of at least 1.11 R=I330 / I350 can be obtained for a heat-treated high-protein beverage that is transparent and still has low viscosity. This is possible even when the protein fraction and / or beverage formulation is heat-treated to a temperature equivalent to pasteurization (e.g., a temperature below 90°C).

[0161] Accordingly, in some preferred embodiments of the present invention, the protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio of at least 1.11 (I330 nm / I350 nm), thus indicating that the protein is in its natural state.

[0162] In some preferred embodiments of the present invention, the protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) 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.

[0163] It may be particularly desirable for the beverage formulation or the protein fraction of the beverage formulation itself to have an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.15, more preferably at least 1.16, and even more preferably at least 1.17.

[0164] In some preferred embodiments of the present invention, a packaged heat-treated beverage formulation comprising a protein fraction and any other component, such as lipids, carbohydrates, vitamins, minerals, food acids, or emulsifiers, has a tryptophan fluorescence emission ratio of at least 1.11.

[0165] Accordingly, in some preferred embodiments of the present invention, the beverage formulation has an intrinsic tryptophan fluorescence emission ratio of at least 1.11 (I330 nm / I350 nm).

[0166] In some preferred embodiments of the present invention, the heat-treated beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.12, preferably at least 1.13, more preferably at least 1.15, even more preferably at least 1.17, most preferably at least 1.19.

[0167] In some preferred embodiments of the present invention, the protein is denatured or at least partially denatured.

[0168] Accordingly, in some preferred embodiments of the present invention, the protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of less than 1.11, which indicates that the protein is at least partially unfolded and aggregation is dominant.

[0169] In some embodiments of the present invention, the heat-treated beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of less than 1.10, more preferably less than 1.08, even more preferably less than 1.05, and most preferably less than 1.00.

[0170] However, in another preferred embodiment of the present invention, the protein fraction of the heat-treated beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of less than 1.15, more preferably less than 1.14, even more preferably less than 1.13, and most preferably less than 1.12.

[0171] In some preferred embodiments of the present invention, the heat-treated beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of less than 1.15, more preferably less than 1.14, even more preferably less than 1.13, and most preferably less than 1.12.

[0172] The beverage formulation may optionally include other food additives, such as lipids, carbohydrates, vitamins, minerals, food acids, or emulsifiers, in addition to the protein fraction. In some preferred embodiments of the present invention, the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of less than 1.11, which indicates that the protein is at least partially unfolded and aggregation is dominant.

[0173] In some preferred embodiments of the present invention, the heat-treated beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of less than 1.10, more preferably less than 1.08, even more preferably less than 1.05, and most preferably less than 1.00.

[0174] Protein denaturation can also be described by another analytical method other than tryptophan fluorescence. This method is described in Example 1.3.

[0175] In some preferred embodiments of the invention, the protein fraction of the packaged heat-treated beverage formulation has a protein denaturation degree of up to 10%, preferably up to 8%, more preferably up to 5%, even more preferably up to 3%, even more preferably up to 1%, and most preferably up to 0.5%.

[0176] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation has a protein denaturation degree of up to 10%, preferably up to 8%, more preferably up to 5%, even more preferably up to 3%, even more preferably up to 1%, and most preferably up to 0.5%.

[0177] In some embodiments of the present invention, when the protein fraction and / or beverage formulation is, for example, heat-treated at high temperature, 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%.

[0178] For example, the protein fraction of the beverage formulation may have a protein denaturation degree of more than 10%, preferably more than 20%, more preferably more than 30%, even more preferably more than 40%, and most preferably more than 50%. A higher degree of denaturation may be desirable, and thus the protein fraction of the beverage formulation may have a protein denaturation degree of more than 70%, preferably more than 80%, more preferably more than 90%, even more preferably more than 95%, and most preferably more than 99%.

[0179] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 3.0 to 4.3. This pH range is particularly desirable for the production of clear beverages with low viscosity and improved taste.

[0180] Regarding appearance, it has been found that using a whey protein drink in which at least 85% w / w of the protein is BLG has the potential to increase the pH during heat treatment, thereby improving both visual perception (color and turbidity) and viscosity compared to a heat-treated WPI drink. Accordingly, the present invention heat-treats an acidic whey protein drink and increases the pH range and, in particular, the pH upper limit, which can produce a heat-treated low-viscosity and, preferably, also clear acidic drink.

[0181] Surprisingly, it was found that there are significant differences in sensory parameters between the BLG beverage of the present invention and the beverage produced with WPI. Surprisingly and favorably, compared to the WPI beverage, the BLG beverage It was found that the levels of astringency, dry texture, sourness, whey flavor, and citric acid flavor were low. Additionally, it was found that increasing the pH of acidic beverages requires less sweetener to balance the acidity of the beverage, and therefore, such beverages require lower concentrations of sweetener.

[0182] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 3.0 to 4.1, or preferably in the range of 3.1 to 4.0 or preferably in the range of 3.2 to 3.9, or preferably in the range of 3.7 to 3.9, more preferably in the range of 3.4 to 3.9, and even more preferably in the range of 3.5 to 3.9.

[0183] The pH of the packaged heat-treated beverage formulation may preferably be in the range of 3.7 to 4.3, more preferably in the range of 3.9 to 4.3, and even more preferably in the range of 4.1 to 4.3.

[0184] Alternatively, but also preferably, the pH of the packaged heat-treated beverage formulation may be in the range of 3.7 to 4.1, more preferably in the range of 3.9 to 4.1.

[0185] This pH range is particularly relevant when pasteurizing beverage formulations.

[0186] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation preferably has a pH in the range of 3.0 to 3.9, or preferably 3.2 to 3.7, or preferably 3.4 to 3.6, or preferably 3.5 to 3.7, or preferably 3.4 to 3.6.

[0187] This pH range combined with high-temperature processing, such as sterilization, is particularly relevant for the production of clear beverages with low viscosity and improved taste.

[0188] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 4.1 to 4.7, which is particularly relevant for the production of a stable beverage having a milky white appearance and high turbidity while still having low viscosity. In some embodiments of the present invention, the pH range is 4.2 to 4.6. In some other embodiments of the present invention, the pH range is 4.2 to 4.5.

[0189] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 3.0 to 3.9 and a total amount of protein of 10 to 34% w / w, more preferably 12 to 30% w / w, and even more preferably 15 to 25% w / w relative to the weight of the beverage formulation.

[0190] In another preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 3.7 to 3.9 and a total amount of protein of 10 to 34% w / w, more preferably 12 to 30% w / w, and even more preferably 15 to 25% w / w relative to the weight of the beverage formulation.

[0191] In a further preferred embodiment of the present invention, the packaged heat-treated beverage formulation has the following:

[0192] - pH in the range of 3.0 to 3.9, preferably 3.7 to 3.9,

[0193] - A total amount of protein of 10 to 34% w / w, more preferably 12 to 30% w / w, and even more preferably 15 to 25% w / w based on the weight of the beverage formulation, and

[0194] - An intrinsic tryptophan fluorescence emission ratio of at least 1.13, more preferably at least 1.15, even more preferably at least 1.17, and most preferably at least 1.19 (I330 nm / I350 nm).

[0195] In a further preferred embodiment of the present invention, the packaged heat-treated beverage formulation has the following:

[0196] - pH in the range of 3.0 to 3.9, preferably 3.7 to 3.9,

[0197] - A total amount of protein of 10 to 34% w / w, more preferably 12 to 30% w / w, and even more preferably 15 to 25% w / w based on the weight of the beverage formulation, and

[0198] - Protein denaturation of up to 10%, preferably up to 5%, more preferably up to 1%.

[0199] The visual appearance of beverage formulations is important to consumers in relation to both clear and opaque beverages. In particular, for clear beverages such as water or white and milky white beverages, the inventors have found that it is advantageous to be able to control the color of the beverage—or rather, to control the lack of color in the beverage.

[0200] However, even if dedicated coloring agents are added during beverage production, the inventors have found that it is advantageous to avoid additional sources of color to avoid unwanted changes or alterations in the visual appearance of the beverage. The inventors have found that the high BLG protein profile described herein is more neutral / colorless than conventional WPI and contributes to less color change than conventional WPI. Conventional WPI has a yellow appearance that can be reduced to some extent by adding oxidizing agents, such as bleaching agents. However, the addition of oxidizing agents is often undesirable and is no longer necessary in the present invention.

[0201] The CIELAB color scale described in Example 1.9 is used to determine the color of a beverage. For example, a positive delta b* value indicates a color that is more yellow than demineralized water, while a negative delta b* value indicates a beverage that is more blue than demineralized water. Therefore, consumers often prefer that the color delta b* value be close to 0 in order to drink a beverage that is neither yellow nor blue.

[0202] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation has a color value delta b* in the range of -0.10 to +0.51 on the CIELAB color scale, particularly if the formulation has a turbidity of up to 200 NTU, more preferably up to 40 NTU.

[0203] In another preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a color value delta b* in the range of 0.0 to 0.40, preferably 0.10 to 0.25, on the CIELAB color scale.

[0204] For example, in the case of an opaque beverage formulation having a turbidity of greater than 200 NTU, preferably greater than 1000 NTU, the packaged heat-treated beverage formulation 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. In another preferred embodiment of the present invention, the beverage formulation having a turbidity of greater than 200 NTU, preferably greater than 1000 NTU, has a color value delta b* in the range of -10 to -0.5, more preferably -9 to -1.0 on the CIELAB color scale.

[0205] In some preferred embodiments of the present invention, the protein fraction of a packaged heat-treated beverage formulation has a color value delta b* in the range of -0.10 to +0.51, particularly if the formulation has a turbidity of up to 200 NTU, more preferably up to 40 NTU.

[0206] These beverages have a higher delta b* value and less yellow compared to beverages containing WPI that are more yellow.

[0207] In another preferred embodiment of the present invention, the protein fraction of the packaged heat-treated beverage formulation has a color value delta b* in the range of 0.0 to 0.40, preferably 0.10 to 0.25, on the CIELAB color scale.

[0208] The a*-value represents the green-red component, where green is in the negative direction and red is in the positive direction. It is often preferred that the color delta a* value be close to 0 to drink a beverage that is neither red nor green.

[0209] The protein fraction of a packaged heat-treated beverage formulation is typically preferred to have a delta a* in the range of -0.2 to 0.2 on the CIELAB color scale, particularly if the formulation has a turbidity of up to 200 NTU, more preferably up to 40 NTU. Preferably, the packaged heat-treated beverage formulation has a color value delta a* in the range of -0.15 to 0.15 on the CIELAB color scale, preferably in the range of -0.10 to 0.10.

[0210] The inventors have discovered that it may be advantageous to control the mineral content to achieve some of the desired properties of a packaged heat-treated beverage formulation.

[0211] In some embodiments of the present invention, the packaged heat-treated beverage formulation comprises a plurality of minerals. In one exemplary embodiment, the packaged heat-treated beverage formulation comprises at least four minerals. In one embodiment, the four minerals are sodium, potassium, magnesium, and calcium.

[0212] The inventors have surprisingly discovered that when the BLG isolate is used as defined herein, a heat-treated beverage formulation having a high mineral concentration without viscosity impairment can be produced in Example 2. This provides the possibility that a packaged heat-treated beverage formulation can be produced with a high mineral content, and that a beverage that is a nutritionally complete or nutritionally incomplete supplement can be produced.

[0213] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg and Ca in the packaged heat-treated beverage formulation is in the range of 0 to 750 mM, preferably in the range of 100 to 600 mM or preferably in the range of 200 to 500 mM.

[0214] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg and Ca in the packaged heat-treated beverage formulation is up to 750 mM.

[0215] 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 formulation is up to 600 mM, preferably up to 500 mM, or preferably up to 400 mM, or preferably up to 300 mM, or preferably up to 200 mM, preferably up to 170 mM, most preferably up to 150 mM, or preferably up to 130 mM, or preferably up to 100 mM, or preferably up to 80 mM, or preferably up to 60 mM, or preferably up to 40 mM, or preferably up to 30 mM, or preferably up to 20 mM, or preferably up to 10 mM, or preferably up to 5 mM, or preferably up to 1 mM.

[0216] In another exemplary embodiment, the packaged heat-treated beverage formulation 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.

[0217] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation comprises up to 150 mM KCl and up to 150 mM CaCl2, the packaged heat-treated beverage formulation comprises up to 130 mM KCl and up to 130 mM CaCl2, the packaged heat-treated beverage formulation comprises up to 110 mM KCl and up to 110 mM CaCl2, the packaged heat-treated beverage formulation comprises up to 100 mM KCl and up to 100 mM CaCl2, preferably the packaged heat-treated beverage formulation comprises up to 80 mM KCl and up to 80 mM CaCl2, preferably the packaged heat-treated beverage formulation comprises up to 50 mM KCl and up to 50 mM CaCl2, and preferably the packaged heat-treated beverage formulation comprises up to 40 mM KCl and up to 40 mM CaCl2.

[0218] In another preferred embodiment of the present invention, the heat-treated beverage formulation is a low-mineral beverage.

[0219] In the context of the present invention, the term “low mineral” relates to a composition, e.g., a liquid, beverage, powder, or other food product having at least one, preferably two, and more preferably all of the following:

[0220] - Up to 1.2% w / w ash content relative to total solids,

[0221] - Total content of calcium and magnesium up to 0.3% w / w relative to total solids,

[0222] - Total content of sodium and potassium up to 0.10% w / w relative to total solids,

[0223] - Total phosphorus content of up to 100 mg of phosphorus per 100 g of protein.

[0224] Preferably, the low mineral composition has at least one of the following, preferably two or more, and more preferably all of them:

[0225] - Ash content of up to 0.7% w / w relative to total solids,

[0226] - Total content of calcium and magnesium up to 0.2% w / w relative to total solids,

[0227] - Total content of sodium and potassium up to 0.08% w / w relative to total solids,

[0228] - Total phosphorus content of up to 80 mg per 100 g of protein.

[0229] More preferably, the low mineral composition has at least one of the following, preferably two or more, and more preferably all of them:

[0230] - Ash content of up to 0.5% w / w relative to total solids,

[0231] - Total content of calcium and magnesium up to 0.15% w / w relative to total solids,

[0232] - Total content of sodium and potassium up to 0.06% w / w relative to total solids,

[0233] - Total phosphorus content of up to 50 mg per 100 g of protein.

[0234] It is particularly desirable that the low mineral composition has the following:

[0235] - Ash content of up to 0.5% w / w relative to total solids,

[0236] - Total content of calcium and magnesium up to 0.15% w / w relative to total solids,

[0237] - Total content of sodium and potassium up to 0.06% w / w relative to total solids,

[0238] - Total phosphorus content of up to 50 mg per 100 g of protein.

[0239] The inventors have discovered that the present invention can produce a packaged heat-treated beverage formulation having a very low content of phosphorus and other minerals, such as potassium, which is advantageous for patients suffering from kidney disease or otherwise having impaired kidney function.

[0240] The packaged heat-treated beverage formulation is preferably a low phosphorus beverage formulation.

[0241] The packaged heat-treated beverage formulation is preferably a low-potassium beverage formulation.

[0242] The packaged heat-treated beverage formulation is preferably a low phosphorus and low potassium beverage formulation.

[0243] In the context of the present invention, the term “low phosphorus” relates to a composition having a total phosphorus content of up to 100 mg of phosphorus per 100 g of protein, e.g., a liquid, powder, or other food product. Preferably, the low phosphorus composition has a total phosphorus content of up to 80 mg of phosphorus per 100 g of protein. More preferably, the low phosphorus composition may have a total phosphorus content of up to 50 mg of phosphorus per 100 g of protein. Even more preferably, the low phosphorus composition may have a total phosphorus content of up to 20 mg of phosphorus per 100 g of protein. Even more preferably, the low phosphorus composition may have a total phosphorus content of up to 5 mg of phosphorus per 100 g of protein. The low phosphorus composition according to the present invention may be used as a food ingredient for the production of food products for a group of patients with impaired renal function.

[0244] Accordingly, in some particularly preferred embodiments of the present invention, the packaged heat-treated beverage formulation contains up to 80 mg of phosphorus per 100 g of protein. Preferably, the packaged heat-treated beverage formulation contains up to 30 mg of phosphorus per 100 g of protein. More preferably, the packaged heat-treated beverage formulation contains up to 20 mg of phosphorus per 100 g of protein. Even more preferably, the packaged heat-treated beverage formulation contains up to 10 mg of phosphorus per 100 g of protein. Most preferably, the packaged heat-treated beverage formulation contains up to 5 mg of phosphorus per 100 g of protein.

[0245] The phosphorus content is related to the total amount of elemental phosphorus in the composition and is determined according to Example 1.19.

[0246] 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 potassium per 100 g of protein. Preferably, the low potassium composition has a total potassium content of up to 600 mg potassium per 100 g of protein. More preferably, the low potassium composition may have a total potassium content of up to 500 mg potassium per 100 g of protein. More preferably, the low potassium composition may have a total potassium content of up to 400 mg potassium per 100 g of protein. Even more preferably, the low potassium composition may have a total potassium content of up to 300 mg potassium per 100 g of protein. Even more preferably, the low potassium composition may have a total potassium content of up to 200 mg potassium per 100 g of protein. Even more preferably, the low potassium composition may have a total potassium content of up to 100 mg potassium per 100 g of protein. More preferably, the low potassium composition may have a total potassium content of up to 50 mg of potassium per 100 g of protein, and more preferably, the low potassium composition may have a total potassium content of up to 10 mg of potassium per 100 g of protein.

[0247] The low-potassium composition according to the present invention can be used as a food ingredient for the production of food products for a group of patients with impaired kidney function.

[0248] Accordingly, in some particularly preferred embodiments of the present invention, the packaged heat-treated beverage formulation contains up to 600 mg of potassium per 100 g of protein. More preferably, the packaged heat-treated beverage formulation contains up to 500 mg of potassium per 100 g of protein. More preferably, the packaged heat-treated beverage formulation contains up to 400 mg of potassium per 100 g of protein. More preferably, the packaged heat-treated beverage formulation contains up to 300 mg of potassium per 100 g of protein. Even more preferably, the packaged heat-treated beverage formulation contains up to 200 mg of potassium per 100 g of protein. Even more preferably, the packaged heat-treated beverage formulation contains up to 100 mg of potassium per 100 g of protein. More preferably, the packaged heat-treated beverage formulation contains up to 50 mg of potassium per 100 g of protein, and more preferably, the packaged heat-treated beverage formulation contains up to 10 mg of potassium per 100 g of protein.

[0249] The potassium content is related to the total amount of elemental potassium in the composition and is determined according to Example 1.19.

[0250] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation comprises up to 100 mg phosphorus / 100 g protein and up to 700 mg potassium / 100 g protein, preferably up to 80 mg phosphorus / 100 g protein and up to 600 mg potassium / 100 g protein, more preferably up to 60 mg phosphorus / 100 g protein and up to 500 mg potassium / 100 g protein, more preferably up to 50 mg phosphorus / 100 g protein and up to 400 mg potassium / 100 g protein, or more preferably up to 20 mg phosphorus / 100 g protein and up to 200 mg potassium / 100 g protein, or even more preferably up to 10 mg phosphorus / 100 g protein and up to 50 mg potassium / 100 g protein. In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation contains up to 100 mg phosphorus / 100 g protein and up to 340 mg potassium / 100 g protein.

[0251] A heat-treated beverage formulation containing small amounts of phosphorus and potassium can advantageously be supplemented with carbohydrates and lipids, and the heat-treated beverage formulation preferably further comprises a total amount of carbohydrates in the range between 30% and 60% of the total energy content of the beverage, preferably between 35% and 50%, and a total amount of lipids in the range between 20% and 60% of the total energy content, preferably between 30% and 50%.

[0252] In one embodiment of the present invention, the packaged heat-treated beverage formulation comprises a plurality of vitamins. In an exemplary embodiment, the packaged heat-treated beverage formulation comprises at least 10 vitamins. In an exemplary embodiment, the packaged heat-treated beverage formulation comprises vitamin A, vitamin B1, vitamin B2, vitamin B3, and vitamin B5 ,It includes a plurality of vitamins selected from the group consisting of 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.

[0253] In one embodiment of the present invention, the packaged heat-treated beverage comprises a plurality of vitamins and a plurality of minerals.

[0254] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation contains one or more food acids selected from the group consisting of citric acid, malic acid, tartaric acid, acetic acid, benzoic acid, butyric acid, lactic acid, fumaric acid, succinic acid, ascorbic acid, adipic acid, phosphoric acid, and mixtures thereof.

[0255] In one embodiment of the present invention, the packaged heat-treated beverage formulation further comprises a flavor selected from the group consisting of salts, flavorings, flavor enhancers and / or spices. In a preferred embodiment of the present invention, the flavor comprises chocolate, cocoa, lemon, orange, lime, strawberry, banana, forest fruit flavors, or combinations thereof. The selection of the flavor may vary depending on the beverage to be produced.

[0256] Transparency is a parameter that consumers use to evaluate products. One way to determine the transparency of a liquid food product is by measuring the turbidity of the product, as described in Example 1.7.

[0257] In some embodiments of the packaged heat-treated beverage formulation, it is advantageous for the beverage formulation to be transparent. This may be advantageous, for example, when the beverage is used as a sports drink or as "protein water," in which case it is advantageous for the beverage to resemble water in appearance.

[0258] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a turbidity of up to 200 NTU, and such beverage is transparent.

[0259] Surprisingly, it has been revealed by the inventors that a transparent heat-treated beverage formulation having a turbidity of up to 200 NTU can be obtained by the heat-treated beverage formulation according to the present invention.

[0260] This was revealed when the applied heat treatment was sterilization and pasteurization.

[0261] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation has a turbidity of up to 150 NTU, or preferably up to 100 NTU, or preferably up to 80 NTU, or preferably up to 60 NTU, or more preferably up to 40 NTU, or preferably up to 30 NTU, preferably up to 20 NTU, more preferably up to 10 NTU, more preferably up to 5 NTU, and even more preferably up to 2 NTU.

[0262] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a turbidity of more than 200 NTU, and such beverage is opaque.

[0263] In some embodiments of packaged heat-treated beverage formulations, it is advantageous for the beverage formulation to be opaque. This is advantageous, for example, if the beverage is to resemble milk and have a milky white appearance. The appearance of nutritionally complete supplements is typically opaque.

[0264] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation has a turbidity of greater than 250 NTU. Preferably, the packaged heat-treated beverage formulation has a turbidity of greater than 300 NTU, more preferably a turbidity of greater than 500 NTU, more preferably a turbidity of greater than 1000 NTU, preferably a turbidity of greater than 1500 NTU, and even more preferably a turbidity of greater than 2000 NTU.

[0265] In heat-treated beverage formulations, the amount of insoluble matter is a measure of the beverage's instability and the extent to which precipitated substances settle over time. Beverages containing a large amount of insoluble matter are typically considered unstable.

[0266] In the context of the present invention, a whey protein beverage formulation is considered "stable" if up to 15% of the total protein in a heated sample precipitates upon centrifugation at 3000 g for 5 minutes. Refer to the analysis method in Example 1.10.

[0267] Surprisingly, when BLG is used as a protein source in an amount of at least 85 w / w%, compared to when WPI with low BLG content is used as a protein source, the protein fraction was found to contain up to 15% insoluble matter after centrifugation at 3000 g for 5 minutes, which proves that the beverage formulation is stable.

[0268] Accordingly, in some preferred embodiments of the present invention, the protein fraction of the heat-treated beverage formulation contains up to 15% insoluble material.

[0269] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation contains up to 15% insoluble material.

[0270] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation preferably contains up to 12% insoluble material, more preferably up to 10% insoluble material, even more preferably up to 8% insoluble material, and most preferably up to 6% insoluble material.

[0271] Even lower levels of insoluble material are often desirable, and in some preferred embodiments, the packaged heat-treated beverage formulation contains up to 4% insoluble material, preferably up to 2% insoluble material, more preferably up to 1% insoluble material, and most preferably contains no detectable insoluble material at all.

[0272] Consumers prefer heat-treated beverages to be liquid, easy to drink, and liquid rather than gel.

[0273] One way to determine the viscosity of a beverage formulation is by measuring the viscosity of the beverage as described in Example 1.8.

[0274] In some embodiments of packaged heat-treated beverage formulations, it is advantageous for the beverage formulation to have very low viscosity. This is advantageous when the beverage is used as a sports drink, or in some embodiments of nutritionally complete or nutritionally incomplete supplements.

[0275] Surprisingly, it has been revealed by the inventors that heat-treated beverage formulations, such as pasteurized and even sterilized beverages with an acidic pH, have a viscosity of up to 200 centipoise (cP) measured at 22°C at a shear rate of 100 / s.

[0276] Accordingly, in some preferred embodiments of the present invention, the packaged heat-treated beverage formulation has a viscosity of up to 200 cP.

[0277] Preferably, the viscosity of the packaged heat-treated beverage formulation is up to 150 cP, preferably up to 100 cP, more preferably up to 80 cP, even more preferably up to 50 cP, and most preferably up to 40 cP.

[0278] Even lower viscosity is often desirable, and thus, in some preferred embodiments of the present invention, the viscosity of the packaged heat-treated beverage formulation is up to 20 cP, preferably up to 10 cP, more preferably up to 5 cP, even more preferably up to 3 cP, even more preferably up to 2 cP, and most preferably up to 1 cP. .

[0279] It has been previously found that, in order to produce acidic, clear, heat-treated beverages containing WPI, it is essential to add an anti-aggregation agent to the beverage when the beverage has a pH greater than 3.0, see, for example, Etzel 2004 (Etzel, MR, 2004, Manufacture and use of dairy protein fraction. American Society for Nutritional Science, pp. 996-1002).

[0280] Surprisingly, the inventors discovered that a clear heat-treated beverage containing at least 85% w / w BLG can be produced at a pH higher than 3.0 without the addition of an anti-coagulant.

[0281] Accordingly, in some preferred embodiments of the present invention, the packaged heat-treated beverage formulation does not contain any anti-aggregation agent, or alternatively contains only a trace amount of anti-aggregation agent.

[0282] In the context of the present invention, the term “anti-coagulant” relates to food-grade, non-protein surfactants, e.g., lauryl sulfate, polysorbate, and mono- and / or di-glycerides.

[0283] In some embodiments of the present invention, the packaged heat-treated beverage formulation contains up to 0.1% w / w anti-aggregation agent, preferably up to 0.03% w / w anti-aggregation agent, and most preferably does not contain an anti-aggregation agent. The embodiment is particularly preferred in relation to a clear low-fat beverage.

[0284] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation does not contain polyphenols.

[0285] However, in another preferred embodiment of the present invention, the packaged heat-treated beverage formulation contains polyphenols. Polyphenols, for example, epigallocatechin-3-gallate (EGCG), have been shown to limit the aggregation of whey protein upon heat treatment. Although not necessary for manufacturing the high-protein beverage formulation according to the present invention, these may be used as ingredients.

[0286] Accordingly, it may be desirable for the packaged heat-treated beverage formulation to contain a total amount of polyphenols in the range of 0.01 to 1% w / w, more preferably 0.02 to 0.6% w / w, even more preferably 0.03 to 0.4% w / w, and most preferably 0.04 to 0.2% w / w.

[0287] In some preferred embodiments of the present invention, the polyphenol may include at least EGCG and even consist essentially of it.

[0288] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation comprises a total amount of protein of 4.0 to 35% w / w, preferably 4.0 to 30% w / w, relative to the weight of the beverage.

[0289] In another preferred embodiment of the present invention, the packaged heat-treated beverage formulation comprises a total amount of protein of 5.0 to 45% w / w, more preferably 5.0 to 35% w / w, even more preferably 5.0 to 34% w / w, and most preferably 5.0 to 32% w / w relative to the weight of the beverage.

[0290] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation contains a protein content in an amount of at least 3 to 45% w / w, more preferably 11 to 40% w / w, even more preferably 15 to 38% w / w, and most preferably 20 to 36% w / w.

[0291] In some embodiments of the present invention, it is advantageous for the packaged heat-treated beverage formulation to have a protein content of 2.0 to 10.0% w / w relative to the weight of the beverage.

[0292] Accordingly, in some embodiments of the present invention, the packaged heat-treated beverage formulation preferably comprises a total amount of protein of 2.0 to 10% w / w relative to the weight of the beverage, preferably a total amount of protein of 3.0 to 10% w / w relative to the weight of the beverage, preferably a total amount of protein of 5.0 to 9.0% w / w relative to the weight of the beverage, and preferably a total amount of protein of 6.0 to 8.0% w / w relative to the weight of the beverage.

[0293] In some embodiments of the present invention, it is advantageous for the protein content of the beverage to be high, such as 10.0 to 45.0% w / w relative to the weight of the beverage.

[0294] Accordingly, in some embodiments of the present invention, the packaged heat-treated beverage formulation preferably comprises a total amount of protein of 10.0 to 45.0% w / w relative to the weight of the beverage, preferably a total amount of protein of 10.0 to 20% w / w relative to the weight of the beverage, preferably a total amount of protein of 12 to 30% w / w relative to the weight of the beverage, preferably a total amount of protein of 15 to 25% w / w relative to the weight of the beverage, and preferably a total amount of protein of 18 to 20% w / w relative to the weight of the beverage.

[0295] In another preferred embodiment of the present invention, it is advantageous that the protein content of the packaged heat-treated beverage formulation is 5.0 to 45.0% w / w, preferably 6.0 to 35% w / w, more preferably 7.0 to 34% w / w, even more preferably 8.0 to 32% w / w, and most preferably 10 to 30% w / w relative to the weight of the packaged heat-treated beverage formulation.

[0296] The present invention can surprisingly provide a packaged heat-treated beverage formulation having a protein content of more than 15% w / w, even more than 20% w / w. Accordingly, in some preferred embodiments of the present invention, the packaged heat-treated beverage formulation preferably comprises a total amount of protein of 15 to 45.0% w / w relative to the weight of the beverage formulation, preferably a total amount of protein of 20 to 35% w / w relative to the weight of the beverage formulation, more preferably a total amount of protein of 21 to 34% w / w relative to the weight of the beverage formulation, and even more preferably a total amount of protein of 25 to 32% w / w relative to the weight of the beverage formulation.

[0297] In another preferred embodiment of the present invention, the packaged heat-treated beverage formulation preferably comprises a total amount of protein of 21 to 35% w / w relative to the weight of the beverage formulation, preferably a total amount of protein of 25 to 35% w / w relative to the weight of the beverage formulation, more preferably a total amount of protein of 28 to 35% w / w relative to the weight of the beverage formulation, and even more preferably a total amount of protein of 30 to 35% w / w relative to the weight of the beverage formulation.

[0298] In a further preferred embodiment of the present invention, the packaged heat-treated beverage formulation preferably comprises a total amount of protein of 21 to 33% w / w relative to the weight of the beverage formulation, preferably a total amount of protein of 25 to 33% w / w relative to the weight of the beverage formulation, and more preferably a total amount of protein of 28 to 33% w / w relative to the weight of the beverage formulation.

[0299] The protein in the liquid solution is preferably prepared from mammalian milk, preferably ruminant milk, for example, milk from dairy cows, sheep, goats, buffalo, camels, llamas, mares, and / or deer. Protein derived from cow's milk is particularly preferred. Accordingly, the protein in the liquid solution is preferably cow's milk protein.

[0300] The protein in the liquid solution is preferably whey protein and / or milk serum protein, more preferably bovine whey protein and / or milk serum protein.

[0301] The packaged heat-treated beverage formulation of the present invention is particularly useful as a sports drink, in which case it preferably contains only a limited amount of lipids and / or optionally also a limited amount of carbohydrates.

[0302] In some preferred embodiments of the present invention, the formulation is particularly useful as a sports drink and comprises, for example, a total amount of protein in the range of 2 to 45% w / w with respect to the weight of the drink, preferably 2 to 20% w / w with respect to the weight of the drink, or preferably 2 to 10% w / w with respect to the weight of the drink, most preferably 2 to 6% w / w with respect to the weight of the drink.

[0303] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation is particularly useful as a nutritional supplement for nutritionally incomplete foods and comprises, for example, a total amount of protein in the range of 2 to 45% w / w relative to the weight of the beverage, preferably 2 to 20% w / w relative to the weight of the beverage, or preferably 3 to 10% w / w relative to the weight of the beverage.

[0304] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation is particularly useful as a nutritionally complete nutritional supplement and comprises, for example, a total amount of protein in the range of 4 to 45% w / w or preferably 5 to 20% w / w with respect to the weight of the beverage.

[0305] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation is particularly advantageous for patients with kidney disease or otherwise impaired kidney function.

[0306] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation comprises a total amount of protein in the range of, for example, 2 to 45% w / w relative to the weight of the beverage, preferably 2 to 20% w / w relative to the weight of the beverage, or preferably 3 to 12% w / w relative to the weight of the beverage, or preferably 3 to 10% w / w relative to the weight of the beverage.

[0307] It is particularly desirable for packaged heat-treated beverage formulations to include BLG isolates, for example, in combination with other protein sources, preferably as a primary protein source and, if possible, even as the sole protein source.

[0308] The packaged heat-treated beverage formulation of the present invention may contain macronutrients other than protein. In some embodiments of the present invention, the packaged heat-treated beverage formulation further contains carbohydrates. The total carbohydrate content in the heat-treated beverage formulation of the present invention varies depending on the intended use of the heat-treated beverage formulation.

[0309] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation further comprises at least one source of carbohydrates. In one exemplary embodiment, at least one source of carbohydrates is selected from the group consisting of sucrose, maltodextrin, corn syrup solids, saccharose, maltose, sucromalt, maltitol powder, glycerin, glucose polymer, corn syrup, modified starch, resistant starch, rice-derived carbohydrate, isomaltulose, white sugar, glucose, fructose, lactose, high-fructose corn syrup, honey, sugar alcohol, fructooligosaccharide, soybean fiber, corn fiber, guar gum, konjac powder, polydextrose, fibrosol, and combinations thereof.

[0310] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation comprises a sugar polymer, namely an oligosaccharide and / or polysaccharide.

[0311] In some preferred embodiments, the packaged heat-treated beverage formulation further comprises carbohydrates in a range between 0% and 95% of the total energy content of the formulation, preferably between 10% and 85% of the total energy content of the formulation, preferably between 20% and 75% of the total energy content of the formulation, or preferably between 30% and 60% of the total energy content of the formulation.

[0312] A lower carbohydrate content is often desirable, and thus, in some preferred embodiments of the present invention, the carbohydrate content of a packaged heat-treated beverage formulation is preferably in the range between 0% and 30% of the total energy content of the formulation, more preferably in the range between 0% and 20% of the total energy content of the formulation, and even more preferably in the range between 0% and 10% of the total energy content of the formulation.

[0313] In some preferred embodiments of the present invention, the carbohydrate content of the packaged heat-treated beverage formulation is up to 5% of the total energy content of the formulation, more preferably up to 1% of the total energy content of the formulation, and even more preferably up to 0.1% of the total energy content of the formulation.

[0314] In some preferred embodiments of the present invention, the formulation is particularly useful as a sports drink and contains a total amount of carbohydrates of up to 75%, preferably up to 40%, preferably up to 10%, or preferably up to 5% of the total energy content (E) of the drink.

[0315] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation is particularly useful as a nutritionally incomplete nutritional supplement and contains a total amount of carbohydrates in the range between 75% and 95% of the total energy content (E) of the beverage, preferably 80 to 90 E%.

[0316] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation is particularly useful as a nutritionally complete nutritional supplement and contains a total amount of carbohydrates in the range between 30% and 60% of the total energy content of the beverage, preferably in the range between 35% and 50%.

[0317] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation is particularly advantageous for patients with kidney disease or otherwise impaired kidney function.

[0318] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation comprises a total amount of carbohydrates in the range between 30% and 60% of the total energy content of the beverage, preferably in the range between 35% and 50%.

[0319] In one embodiment of the present invention, the packaged heat-treated beverage formulation further comprises at least one additional ingredient selected from the group consisting of vitamins, flavoring agents, minerals, sweeteners, antioxidants, food acids, lipids, carbohydrates, prebiotics, probiotics, and non-whey proteins.

[0320] In one embodiment of the present invention, the beverage formulation 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 salt, neotame, saccharin, stevia extract, steviol glycoside, e.g., rebaudioside A, or combinations thereof. In some embodiments of the present invention, it is particularly preferred that the sweetener comprises one or more high-intensity sweeteners (HIS) or even consists of them.

[0321] HIS is found in both natural and artificial sweeteners, and typically has a sweetness strength at least 10 times that of sucrose.

[0322] If used, the total amount of HIS is typically in the range of 0.01 to 2% w / w. For example, the total amount of HIS may be in the range of 0.05 to 1.5% w / w. Alternatively, the total amount of HIS may be in the range of 0.1 to 1.0% w / w.

[0323] The choice of sweetener may vary depending on the beverage to be produced; for example, high-intensity sugar sweeteners (e.g., aspartame, acesulfame-K, or sucralose) may be used in beverages where no energy contribution from the sweetener is desired, whereas for beverages with a natural profile, natural sweeteners (e.g., steviol glycoside, sorbitol, or sucrose) may be used.

[0324] Additionally, it may be desirable for the sweetener to include or even consist of one or more polyol sweeteners. Non-limiting examples of useful polyol sweeteners are maltitol, mannitol, lactitol, sorbitol, inositol, xylitol, threitol, galactitol, or combinations thereof. If used, the total amount of polyol sweetener is typically in the range of 1 to 20% w / w. For example, the total amount of polyol sweetener may be in the range of 2 to 15% w / w. Alternatively, the total amount of polyol sweetener may be in the range of 4 to 10% w / w.

[0325] The packaged heat-treated beverage formulation of the present invention may contain macronutrients other than protein. In some embodiments of the present invention, the packaged heat-treated beverage formulation further contains lipids. The total lipid content in the heat-treated beverage formulation of the present invention varies depending on the intended use of the heat-treated beverage formulation.

[0326] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation has a lipid content in the range of 0% to 60% of the total energy content of the formulation, or preferably between 0% to 50% of the total energy content of the formulation, or preferably between 0% to 45% of the total energy content of the formulation, or preferably between 0% to 30% of the total energy content of the formulation, or preferably between 0% to 20% of the total energy content of the formulation, or preferably between 0% to 10% of the total energy content of the formulation, or preferably between 0% to 5% of the total energy content of the formulation.

[0327] The amount of lipids is determined according to ISO 1211:2010 (determination of fat content - Rose-Gottlieb weight method).

[0328] In some preferred embodiments of the present invention, the lipid content of the packaged heat-treated beverage formulation is up to 3% of the total energy content of the formulation, more preferably up to 1% of the total energy content of the formulation, and even more preferably up to 0.1% of the total energy content of the formulation.

[0329] In some preferred embodiments of the present invention, the formulation is particularly useful as a sports drink and contains, for example, a total amount of lipids of up to 10 E%, preferably up to 1 E%.

[0330] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation is particularly useful as a nutritionally incomplete nutritional supplement, and, for example, contains a total amount of lipids of up to 10%, preferably up to 1 E%, of the total energy content of the beverage.

[0331] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation is particularly useful as a nutritionally complete nutritional supplement and, for example, contains a total amount of lipids in the range of 20 to 50% of the total energy content, preferably between 30 E% and 40 E%.

[0332] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation is particularly advantageous for patients with kidney disease or otherwise impaired kidney function.

[0333] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation comprises, for example, a total amount of lipids in the range of 20 to 60% of the total energy content, preferably between 30 E% and 50 E%.

[0334] The beverage formulation typically contains a total amount of water in the range of 50 to 98% w / w, preferably 45 to 97% w / w, more preferably 40 to 95% w / w, even more preferably 35 to 90% w / w, and most preferably 30 to 85% w / w.

[0335] In some preferred embodiments of the present invention, the beverage formulation contains a total amount of water in the range of 55 to 90% w / w, preferably in the range of 57 to 85% w / w, more preferably in the range of 60 to 80% w / w, even more preferably in the range of 62 to 75% w / w, and most preferably in the range of 65 to 70% w / w.

[0336] In some preferred embodiments of the present invention, the beverage formulation contains a total amount of water in the range of 90 to 98% w / w, preferably in the range of 92 to 97.5% w / w, more preferably in the range of 94 to 97% w / w, even more preferably in the range of 95 to 97% w / w, and most preferably in the range of 96 to 97% w / w. Such embodiments are useful for beverages, for example, clear water.

[0337] In some preferred embodiments of the present invention, the beverage formulation is non-alcoholic, which means that it contains 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 no detectable ethanol.

[0338] The beverage formulation typically contains a total amount of solids in the range of 1 to 45% w / w, preferably 5 to 40% w / w, more preferably 10 to 35% w / w, even more preferably 12 to 30% w / w, and most preferably 16 to 25% w / w.

[0339] In some preferred embodiments of the present invention, the beverage formulation contains a total amount of solids in the range of 10 to 45% w / w, preferably 15 to 43% w / w, more preferably 20 to 40% w / w, even more preferably 25 to 38% w / w, and most preferably 30 to 35% w / w.

[0340] In some preferred embodiments of the present invention, the liquid solution contains a total amount of solids in the range of 1 to 10% w / w, preferably 1.5 to 8% w / w, more preferably 2 to 6% w / w, even more preferably 2 to 5% w / w, and most preferably 2 to 4% w / w. Such embodiments are useful for beverages, for example, clear water.

[0341] Some of the non-solid beverage formulations are preferably water.

[0342] In some preferred embodiments of the present invention, the sum of alpha-lactalbumin (ALA) and caseinomacropeptide (CMP) constitutes at least 40% w / w, 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 protein in the beverage formulation.

[0343] In another preferred embodiment of the present invention, each major non-BLG whey protein is present as a weight percentage relative to total protein, which is up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, most preferably up to 6% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey.

[0344] Even lower concentrations of major non-BLG whey protein may be desirable. Accordingly, in a further preferred embodiment of the present invention, each major non-BLG whey protein is present as a weight percentage relative to total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey.

[0345] In some preferred embodiments of the present invention, ALA constitutes up to 80% w / w, preferably up to 60% w / w, more preferably up to 40% w / w, and most preferably up to 30% w / w of the non-BLG protein in the beverage formulation.

[0346] A lower content of ALA may be desirable, and thus in some preferred embodiments of the present invention, ALA constitutes up to 20% w / w, preferably up to 15% w / w, more preferably up to 10% w / w, and most preferably up to 5% w / w of the non-BLG protein in the beverage formulation.

[0347] The inventors have observed indications that a reduction in lactoferrin and / or lactoperoxidase is particularly advantageous for obtaining a neutral-colored whey protein product.

[0348] Accordingly, in some preferred embodiments of the present invention, lactoferrin is present as a weight percentage relative to total protein, which is up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, most preferably up to 6% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey. Even lower concentrations of lactoferrin may be preferred. Accordingly, in a further preferred embodiment of the present invention, lactoferrin is present as a weight percentage relative to total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey.

[0349] Similarly, in some preferred embodiments of the present invention, lactoperoxidase is present as a weight percentage relative to total protein, which is up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, most preferably up to 6% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey. Even lower concentrations of lactoperoxidase may be preferred. Accordingly, in a further preferred embodiment of the present invention, lactoperoxidase is present as a weight percentage relative to total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey.

[0350] Lactoferrin and lactoperoxidase are quantified according to Example 1.29.

[0351] In one embodiment of the present invention, the packaged heat-treated beverage formulation is a sports drink.

[0352] In one embodiment of the present invention, the packaged heat-treated beverage formulation is a nutritionally complete nutritional supplement.

[0353] In one embodiment of the present invention, the packaged heat-treated beverage formulation is a nutritionally incomplete nutritional supplement.

[0354] In one embodiment of the present invention, the packaged heat-treated beverage formulation is a low phosphorus and low potassium beverage suitable for patients with kidney disease or otherwise impaired kidney function.

[0355] The packaged heat-treated beverage formulation of the present invention is particularly useful as a sports drink, in which case it preferably contains only a limited amount of lipids and / or optionally also a limited amount of carbohydrates.

[0356] In some preferred embodiments of the present invention, the formulation is particularly useful as a sports drink and comprises, for example, the following:

[0357] - A total amount of protein in the range of 2 to 45% w / w relative to the weight of the beverage, preferably 2 to 20% w / w relative to the weight of the beverage, or preferably 2 to 10% w / w relative to the weight of the beverage, most preferably 2 to 6% w / w relative to the weight of the beverage,

[0358] - A total amount of carbohydrates up to 75%, preferably up to 40 E%, preferably up to 10 E%, or preferably up to 5 E% of the total energy content (E) of the beverage, and

[0359] - Total amount of lipids up to 10 E%, preferably up to 1 E%.

[0360] In some preferred embodiments of the present invention, packaged heat-treated beverage formulations are particularly useful as nutritional supplements for nutritionally incomplete foods and include, for example, the following:

[0361] - Total amount of protein in the range of 2 to 45% w / w relative to the weight of the beverage, preferably 2 to 20% w / w relative to the weight of the beverage, or preferably 3 to 10% w / w relative to the weight of the beverage,

[0362] - A total amount of carbohydrates in the range of 70% to 95% of the total energy content (E) of the beverage, preferably 80 to 90% E%, and

[0363] - A total amount of lipids up to 10%, preferably up to 1% of the total energy content of the beverage.

[0364] In some preferred embodiments of the present invention, packaged heat-treated beverage formulations are particularly useful as nutritionally complete nutritional supplements and include, for example, the following:

[0365] - Total amount of protein in the range of 4 to 45% w / w relative to the weight of the beverage, preferably in the range of 5 to 20% w / w relative to the weight of the beverage,

[0366] - A total amount of carbohydrates in the range of 30% to 60% of the total energy content of the beverage, preferably in the range of 35% to 50%, and

[0367] - Total amount of lipids in the range of 20 to 50% of total energy content, preferably in the range between 30 E% and 40 E%.

[0368] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation is particularly advantageous for patients with kidney disease or otherwise impaired renal function. The beverage formulation has very low content of phosphorus and other minerals, such as potassium.

[0369] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation comprises, for example, the following:

[0370] - Total amount of protein in the range of 2 to 45% w / w relative to the weight of the beverage, preferably 2 to 20% w / w relative to the weight of the beverage, or preferably 3 to 12% w / w relative to the weight of the beverage, preferably 3 to 10% w / w relative to the weight of the beverage,

[0371] - A total amount of carbohydrates in the range of 30% to 60% of the total energy content of the beverage, preferably in the range of 35% to 50%, and

[0372] - Total amount of lipids in the range of 20 to 60% of total energy content, preferably in the range between 30 E% and 50 E%.

[0373] Heat-treated beverage formulations are preferably present in suitable containers, e.g., bottles, cartons, bricks, pouches, and / or bags, as described herein.

[0374] The inventors surprisingly discovered that heat-treated beverage formulations exposed to at least some protein denaturation tend to develop color during storage under certain conditions. The inventors subsequently discovered that the color development is caused at least partially by exposure to light (electromagnetic radiation) and that this phenomenon increases as the concentration of BLG increases. The inventors also discovered that the problem can be reduced or even avoided by selecting a container that blocks at least some of the ambient light.

[0375] Accordingly, in some preferred embodiments of the present invention, the container wall has a light transmittance of up to 10%, preferably up to 1%, more preferably up to 0.1%, even more preferably up to 0.01%, and most preferably up to 0.001% at any wavelength in the range of 250 to 500 nm.

[0376] In another preferred embodiment of the present invention, the container wall has an average light transmittance of up to 10%, preferably up to 1%, more preferably up to 0.1%, even more preferably up to 0.01%, and most preferably up to 0.001% in the range of 250 to 500 nm.

[0377] The light transmittance of a container wall is measured by providing a flat piece of the container wall and measuring the light transmittance through the container wall at any relevant wavelength. The measurement is performed using a standard spectrophotometer, and the piece of the container wall is inserted into the optical path (e.g., using a cuvette or similar array) so that the plane of the piece of the container wall is aligned perpendicular to the optical path. The transmittance at wavelength i is T i = I i,이후 / I i,이전 It is calculated as *100%, where, I i,이전 is the light intensity at wavelength i before reaching the container wall, and I i,이후 is the intensity of wavelength i after the light beam of the optical path passes through a piece of the container wall.

[0378] Average light transmittance is the total transmittance measurement T taken within a given wavelength range. i It is calculated by calculating the sum of and dividing the sum by the number of transmittance measurements within a given wavelength range.

[0379] In some preferred embodiments of the present invention, the container wall has a light transmittance of up to 10%, preferably up to 1%, more preferably up to 0.1%, even more preferably up to 0.01%, and most preferably up to 0.001% at any wavelength in the range of 250 to 800 nm.

[0380] In another preferred embodiment of the present invention, the container wall has an average light transmittance of up to 10%, preferably up to 1%, more preferably up to 0.1%, even more preferably up to 0.01%, and most preferably up to 0.001% in the range of 250 to 800 nm.

[0381] Such low light transmittance containers may be manufactured, for example, using pigmented, absorbent-containing, or coated polymers or colored or coated glass, or alternatively, may include a metal layer in the container wall in the form of, for example, aluminum foil. Such light-nontransmitting or low light transmittance containers are known in the food and pharmaceutical industries.

[0382] Non-limiting examples of suitable polymer materials are, for example, polyethylene terephthalate (PET) or PET-like polymers.

[0383] The inventors have also discovered that if a beverage formulation primarily contains BLG in its natural form, color development can be reduced or even avoided. For example, it has been found that reducing the degree of protein denaturation using a less denaturation process reduces color problems.

[0384] Accordingly, in some preferred embodiments of the present invention, at least a portion of the container wall, preferably the entire container, is transparent. In some preferred embodiments of the present invention, at least a portion of the container wall, preferably the entire container wall, has an average light transmittance 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% in the range of 400 to 700 nm.

[0385] One aspect of the present invention relates to a method for producing a packaged heat-treated beverage formulation having a pH in the range of 2 to 4.7, said method comprising the following steps.

[0386] a) Step of providing a liquid solution containing the following:

[0387] - Total amount of protein of 2 to 45 weight% - where, at least 85% of the protein is BLG -

[0388] - Optionally, sweeteners, sugar polymers and / or flavorings

[0389] b) Step of packaging the liquid solution,

[0390] Here, the liquid solution of step a) and / or the packaged liquid solution of step b) are heat-treated, including at least pasteurization.

[0391] The liquid solution of step a) preferably has the same chemical composition as described in the content of the heat-treated beverage formulation, except that the liquid solution lacks final heat treatment. Accordingly, the embodiments and preferences mentioned in the context of the heat-treated beverage formulation apply equally to the liquid solution.

[0392] In some preferred embodiments of the liquid solution 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.

[0393] Since a higher relative amount of BLG is feasible and therefore desirable, in some preferred embodiments of the present 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 about 100% w / w.

[0394] For example, the liquid solution preferably contains 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, and even more preferably at least 99.0%, and most preferably contains BLG in an amount of at least 99.5% w / w relative to total protein, e.g., approximately 100.0% w / w relative to total protein.

[0395] The packaging in step b) may be any suitable packaging technology, and any suitable container may be used to package the liquid solution.

[0396] However, in a preferred embodiment of the present invention, the packaging in step b) is aseptic packaging, that is, the liquid solution is packaged under aseptic conditions. For example, aseptic packaging may be performed using an aseptic filling system and preferably comprises filling the liquid solution into one or more aseptic container(s).

[0397] If the liquid solution is already sterile or contains very few microorganisms before filling, aseptic filling and sealing are particularly desirable.

[0398] Examples of useful containers include, for example, bottles, cardboard boxes, bricks, and / or bags.

[0399] In some preferred embodiments of the present invention, the packaged liquid solution of step b) is heat-treated, including at least pasteurization. Such embodiments are typically referred to as in-vessel heat treatment or retort treatment and involve heating the entire container and its contents to achieve pasteurization or even sterilization. When using in-vessel heat treatment, it is particularly desirable to maintain the temperature in the range of 70 to 82°C, more preferably in the range of 70 to 80°C, and most preferably in the range of 70 to 78°C. In this way, the level of protein unfolding is maintained to a minimum.

[0400] In another preferred embodiment of the present invention, the liquid solution of step a) is heat-treated, including at least pasteurization, and then packaged in step b).

[0401] In a particularly preferred embodiment, the heat treatment includes the step of heating the beverage formulation to a temperature in the range of 70 to 82°C.

[0402] In some preferred embodiments of the present invention, the heat-treatment temperature is in the range of 70 to 80°C, preferably in the range of 70 to 79°C, more preferably in the range of 71 to 78°C, even more preferably in the range of 72 to 77°C, most preferably in the range of 73 to 76°C, e.g., approximately 75°C.

[0403] Preferably, when performed in a temperature range of 70 to 82°C, the heat-treatment duration is 1 second to 30 minutes. The highest exposure time is best suited for the lowest temperature in the temperature range, and vice versa. The lower the pH of the liquid solution, the higher the temperature it can withstand without unfolding.

[0404] In a particularly preferred embodiment of the present invention, the heat treatment provides 70 to 80°C for 1 second to 30 minutes, more preferably 71 to 77°C for 1 minute to 25 minutes, and even more preferably 72 to 76°C for 2 minutes to 20 minutes.

[0405] In some preferred embodiments of the present invention, the heat treatment includes the step of heating to a temperature of 85°C to 95°C for 1 to 3 minutes.

[0406] In some embodiments, a higher temperature may also be desirable, particularly if unfolding and optionally agglomeration of the BLG is required. For example, the heat-treatment temperature 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.

[0407] In some preferred embodiments of the present invention, sterilization includes a temperature in the range of 120 to 150°C for 4 to 30 seconds.

[0408] The heat treatment may include, for example, a temperature in the range of 90 to 130°C and a duration in the range of 5 seconds to 10 minutes. The heat treatment may include, for example, a step of heating at a temperature in the range of 90 to 95°C for a duration of 1 to 10 minutes, for example, at approximately 120°C for approximately 20 seconds. Alternatively, the heat treatment may include a step of heating at a temperature in the range of 115 to 125°C for a duration of 5 to 30 seconds, for example, at approximately 120°C for 20 seconds.

[0409] Alternatively, the heat treatment may be a UHT-type treatment, for example, typically comprising a temperature in the range of 135 to 144°C and a duration in the range of 2 to 10 seconds.

[0410] Alternatively, but also preferably, the heat treatment may include a temperature in the range of 145 to 180°C and a duration in the range of 0.01 to 2 seconds, more preferably a temperature in the range of 150 to 180°C and a duration in the range of 0.01 to 0.3 seconds.

[0411] Implementation of heat treatment may involve the use of equipment such as plate or tubular heat exchangers, scraped surface heat exchangers, or retort systems. Alternatively, and particularly preferably, for heat treatment above 95°C, direct steam-based heating may be used, for example, by direct steam injection, direct steam infusion, or spray cooking. Additionally, such direct steam-based heating is preferably used in conjunction with flash cooling. Suitable examples of implementation of spray cooking are found in WO2009113858A1, which is incorporated herein by all means. Suitable examples of direct steam injection and direct steam infusion are found in WO2009113858A1 and WO 2010 / 085957 A3, which are incorporated herein by all means. General modes of high-temperature processing are found, for example, in the following incorporated herein by reference for all purposes: "Thermal technologies in food processing" ISBN 185573558 X.

[0412] In some preferred embodiments of the present invention, pasteurization is combined with another physical microbial reduction.

[0413] Useful examples of physical microbial reduction include one or more of bacterial filtration, UV radiation, hyperbaric therapy, pulsed electric field therapy, and ultrasound.

[0414] In some particularly preferred embodiments of the present invention, the heat treatment is selected to provide a degree of protein denaturation of up to 50%, preferably up to 20%, more preferably up to 10%, and most preferably up to 5%.

[0415] In addition, the heat treatment is preferably selected to provide an intrinsic tryptophan fluorescence ratio (I330 / I350) of at least 1.11, preferably at least 1.13, more preferably at least 1.15, and even more preferably at least 1.17.

[0416] In some preferred embodiments of the present invention, heat treatment is performed to sterilize, thereby producing a sterilized liquid beverage formulation. Such sterilization can preferably be achieved by combining bacterial filtration and heat treatment, for example, pasteurization. Sterilization may include, for example, heat treatment followed by bacterial filtration, or more preferably, heat treatment followed by bacterial filtration.

[0417] In the context of the present invention, the term “bacterial filtration” relates to filtration performed with a pore size sufficient to contain microorganisms such as bacteria and spores, but not with a pore size that contains natural BLG. Bacterial filtration is often also referred to as sterile filtration and involves microfiltration of the corresponding liquid. Bacterial filtration is typically performed with a membrane having a pore size of up to 1 micron, preferably up to 0.8 microns, more preferably up to 0.6 microns, even more preferably up to 0.4 microns, and most preferably up to 0.2 microns.

[0418] Bacterial filtration may include a membrane having a pore size of, for example, 0.02 to 1 micron, preferably 0.03 to 0.8 microns, more preferably 0.04 to 0.6 microns, even more preferably 0.05 to 0.4 microns, and most preferably 0.1 to 0.2 microns.

[0419] In some preferred embodiments of the present invention, the liquid solution is bacterially filtered and then heat-treated using a temperature of up to 80°C, preferably up to 78°C. The duration of this heat-treatment is preferably selected to be sufficiently long to produce a sterile beverage formulation.

[0420] In another preferred embodiment of the present invention, the liquid solution is bacterially filtered and then heat-treated using a temperature of 81 to 160°C, more preferably 100 to 155°C. The combination of the temperature and duration of this heat-treatment is preferably selected to provide a sterile beverage formulation.

[0421] Depending on the heat-treatment temperature used, it is beneficial to cool the beverage formulation. According to a preferred embodiment of the method of the present invention, after heat-treatment, the heat-treated beverage formulation is cooled at any stage preferably to 0 to 50°C, preferably to 0 to 25°C or preferably to 0 to 20°C or preferably to 0 to 15°C, preferably to 0 to 10°C or preferably to 4 to 8°C or preferably to 2 to 5°C or preferably to 1 to 5°C.

[0422] If the beverage formulation is pasteurized, it is cooled to preferably 0 to 15°C, preferably 1 to 10°C, more preferably 1 to 6°C after heat treatment.

[0423] According to one embodiment of the present invention, pH can generally be adjusted using any acid or base. Those skilled in the art will recognize that there are suitable means for pH adjustment. 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.

[0424] Useful examples of useful bases are hydroxide salts, e.g., sodium hydroxide or potassium hydroxide, carbonate salts or hydrocarbonate salts, carboxylate salts, e.g., citrate salts or lactic acid salts, and combinations thereof. Preferably, a base, e.g. KOH or NaOH, is used to adjust the pH.

[0425] In some preferred embodiments of the present invention, the liquid solution has a pH in the range of 3.0 to 4.3. This pH range is particularly desirable for the production of a clear beverage with low viscosity and improved taste.

[0426] Regarding appearance, surprisingly, it has been found that using a whey protein drink in which at least 85% w / w of the protein is BLG increases the pH during heat treatment, thereby improving both visual perception (color and turbidity) and viscosity compared to a heat-treated WPI drink. Thus, the present invention increases the pH range that allows for the production of a low-viscosity, preferably clear acidic drink containing whey protein.

[0427] Surprisingly, it was found that there are significant differences in sensory parameters between the BLG beverage of the present invention and the beverage produced with WPI. Surprisingly and favorably, it was found that the BLG beverage had lower levels of astringency, dry texture, sourness, whey flavor, and citric acid flavor compared to the WPI beverage. Furthermore, it was found that increasing the pH of the acidic beverage requires less sweetener to balance the acidity of the beverage, and therefore, such beverages require a lower concentration of sweetener.

[0428] In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 3.0 to 4.1, or preferably 3.1 to 4.0 or preferably 3.2 to 3.9, or preferably 3.7 to 3.9, more preferably 3.4 to 3.9, and even more preferably 3.5 to 3.9.

[0429] Accordingly, it is desirable for the liquid solution to have a pH in the range of 3.0 to 4.1, or preferably 3.1 to 4.0 or preferably 3.2 to 3.9, or preferably 3.7 to 3.9, more preferably 3.4 to 3.9, and even more preferably 3.5 to 3.9.

[0430] The inventors have discovered that it is particularly difficult to produce acidic protein drinks at a pH greater than 3.6, especially if the drink must be transparent. However, the present invention makes this possible.

[0431] Accordingly, the pH of the liquid solution may preferably be in the range of 3.7 to 4.3, more preferably in the range of 3.9 to 4.3, and even more preferably in the range of 4.1 to 4.3.

[0432] Alternatively, but also preferably, the pH of the packaged heat-treated beverage formulation may be in the range of 3.7 to 4.1, more preferably in the range of 3.9 to 4.1.

[0433] This pH range is particularly relevant when pasteurizing beverage formulations.

[0434] In some preferred embodiments of the present invention, the liquid solution preferably has a pH in the range of 3.0 to 3.9, or preferably 3.2 to 3.7, or preferably 3.4 to 3.6, or preferably 3.5 to 3.7, or preferably 3.4 to 3.6.

[0435] This pH range combined with high-temperature processing, such as sterilization, is particularly relevant for the production of clear beverages with low viscosity and improved taste.

[0436] In some preferred embodiments of the present invention, the liquid solution has a pH in the range of 4.1 to 4.7, and this pH range is particularly relevant for the production of a stable beverage having a milky white appearance and high turbidity while still having low viscosity. In some embodiments of the present invention, the pH range is 4.2 to 4.6. In some other embodiments of the present invention, the pH range is 4.2 to 4.5.

[0437] In some preferred embodiments of the present invention, the liquid solution contains a total amount of protein of 4.0 to 35% w / w, preferably 4.0 to 30% w / w, more preferably 5.0 to 30% w / w, relative to the weight of the beverage.

[0438] In another preferred embodiment of the present invention, the liquid solution comprises a total amount of protein of 5.0 to 45% w / w, more preferably 5.0 to 35% w / w, even more preferably 5.0 to 34% w / w, and most preferably 5.0 to 32% w / w relative to the liquid solution.

[0439] In some preferred embodiments of the present invention, the liquid solution contains a protein content in an amount of at least 3 to 45% w / w, more preferably 11 to 40% w / w, even more preferably 15 to 38% w / w, and most preferably 20 to 36% w / w.

[0440] In some embodiments of the present invention, it is advantageous for the liquid solution to have a protein content of 2.0 to 10.0% w / w relative to the weight of the solution.

[0441] Accordingly, in some embodiments of the present invention, the liquid solution preferably comprises a total amount of protein of 2.0 to 10% w / w relative to the weight of the liquid solution, preferably a total amount of protein of 3.0 to 10% w / w relative to the weight of the liquid solution, preferably a total amount of protein of 5.0 to 9.0% w / w relative to the weight of the liquid solution, and preferably a total amount of protein of 6.0 to 8.0% w / w relative to the weight of the liquid solution.

[0442] In some cases, it is advantageous for the protein content of the liquid solution to be high, such as 10.0 to 45.0% w / w relative to the weight of the liquid solution.

[0443] Accordingly, in some embodiments of the present invention, the liquid solution preferably comprises a total amount of protein of 10.0 to 45.0% w / w relative to the weight of the liquid solution, preferably a total amount of protein of 10.0 to 20% w / w relative to the weight of the liquid solution, preferably a total amount of protein of 12 to 30% w / w relative to the weight of the liquid solution, preferably a total amount of protein of 15 to 25% w / w relative to the weight of the liquid solution, and preferably a total amount of protein of 18 to 20% w / w relative to the weight of the liquid solution.

[0444] In another preferred embodiment of the present invention, it is advantageous that the protein content of the liquid solution is 5.0 to 45.0% w / w, preferably 6.0 to 35% w / w, more preferably 7.0 to 34% w / w, even more preferably 8.0 to 32% w / w, and most preferably 10 to 30% w / w relative to the weight of the liquid solution.

[0445] The present invention can surprisingly provide a packaged heat-treated beverage formulation having a protein content exceeding 15% w / w, or even 20% w / w. Accordingly, in some preferred embodiments of the present invention, the liquid solution preferably comprises a total amount of protein of 15 to 45.0% w / w relative to the weight of the liquid solution, preferably a total amount of protein of 20 to 35% w / w relative to the weight of the liquid solution, more preferably a total amount of protein of 21 to 34% w / w relative to the weight of the liquid solution, and even more preferably a total amount of protein of 25 to 32% w / w relative to the weight of the liquid solution.

[0446] In another preferred embodiment of the present invention, the liquid solution preferably comprises a total amount of protein of 21 to 35% w / w relative to the weight of the liquid solution, preferably a total amount of protein of 25 to 35% w / w relative to the weight of the liquid solution, more preferably a total amount of protein of 28 to 35% w / w relative to the weight of the liquid solution, and even more preferably a total amount of protein of 30 to 35% w / w relative to the weight of the liquid solution.

[0447] In a further preferred embodiment of the present invention, the liquid solution preferably comprises a total amount of protein of 21 to 33% w / w relative to the weight of the liquid solution, preferably a total amount of protein of 25 to 33% w / w relative to the weight of the liquid solution, and more preferably a total amount of protein of 28 to 33% w / w relative to the weight of the liquid solution.

[0448] The protein in the liquid solution is preferably prepared from mammalian milk, preferably ruminant milk, for example, milk from dairy cows, sheep, goats, buffalo, camels, llamas, mares, and / or deer. Protein derived from cow's milk is particularly preferred. Accordingly, the protein in the liquid solution is preferably cow's milk protein.

[0449] The protein in the liquid solution is preferably whey protein and / or milk serum protein, more preferably bovine whey protein and / or milk serum protein.

[0450] In some preferred embodiments of the present invention, the sum of alpha-lactalbumin (ALA) and caseinomacropeptide (CMP) constitutes at least 40% w / w, 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 protein in the liquid solution.

[0451] In another preferred embodiment of the present invention, each major non-BLG whey protein is present as a weight percentage relative to total protein, which is up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, most preferably up to 6% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey.

[0452] Even lower concentrations of major non-BLG whey protein may be desirable. Accordingly, in a further preferred embodiment of the present invention, each major non-BLG whey protein is present as a weight percentage relative to total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey.

[0453] In some preferred embodiments of the present invention, ALA constitutes up to 80% w / w, preferably up to 60% w / w, more preferably up to 40% w / w, and most preferably up to 30% w / w of the non-BLG protein in the beverage formulation.

[0454] A lower content of ALA may be desirable, and thus in some preferred embodiments of the present invention, ALA constitutes up to 20% w / w, preferably up to 15% w / w, more preferably up to 10% w / w, and most preferably up to 5% w / w of the non-BLG protein in the beverage formulation.

[0455] In some preferred embodiments of the present invention, the liquid solution has a pH in the range of 3.0 to 3.9 and a total amount of protein of 10 to 34% w / w, more preferably 12 to 30% w / w, and even more preferably 15 to 25% w / w relative to the weight of the liquid solution.

[0456] In another preferred embodiment of the present invention, the liquid solution has a pH in the range of 3.7 to 3.9 and a total amount of protein of 10 to 34% w / w, more preferably 12 to 30% w / w, and even more preferably 15 to 25% w / w relative to the weight of the liquid solution.

[0457] In a further preferred embodiment of the present invention, the liquid solution has the following:

[0458] - pH in the range of 3.0 to 3.9, preferably 3.7 to 3.9,

[0459] - A total amount of protein of 10 to 34% w / w, more preferably 12 to 30% w / w, and even more preferably 15 to 25% w / w relative to the weight of the liquid solution, and

[0460] - An intrinsic tryptophan fluorescence emission ratio of at least 1.13, more preferably at least 1.15, even more preferably at least 1.17, and most preferably at least 1.19 (I330 nm / I350 nm).

[0461] In a further preferred embodiment of the present invention, the liquid solution has the following:

[0462] - pH in the range of 3.0 to 3.9, preferably 3.7 to 3.9,

[0463] - A total amount of protein of 10 to 34% w / w, more preferably 12 to 30% w / w, and even more preferably 15 to 25% w / w relative to the weight of the liquid solution, and

[0464] - Protein denaturation of up to 10%, preferably up to 5%, more preferably up to 1%.

[0465] As mentioned above, surprisingly, the inventors discovered that a clear heat-treated beverage containing at least 85% w / w BLG can be produced at a pH higher than 3.0 without the addition of an anti-coagulant.

[0466] Accordingly, in some preferred embodiments of the present invention, the liquid solution does not contain any anti-coagulant, or alternatively contains only a trace amount of anti-coagulant.

[0467] In some embodiments of the present invention, the liquid solution contains up to 0.1% w / w anti-aggregation agent, preferably up to 0.03% w / w anti-aggregation agent, and most preferably does not contain an anti-aggregation agent. The embodiment is particularly preferred in relation to a clear and low-fat beverage.

[0468] In some preferred embodiments of the present invention, the liquid solution does not contain polyphenols.

[0469] However, in another preferred embodiment of the present invention, the liquid solution contains polyphenols. Accordingly, it may be desirable for the liquid solution to contain a total amount of polyphenols in the range of 0.01 to 1% w / w, more preferably 0.02 to 0.6% w / w, even more preferably 0.03 to 0.4% w / w, and most preferably 0.04 to 0.2% w / w.

[0470] In some preferred embodiments of the present invention, the polyphenol may include at least EGCG and even consist essentially of it.

[0471] It is particularly desirable for the liquid solution to contain BLG isolates, for example, in combination with other protein sources, preferably as a major protein source and, if possible, even as the only protein source.

[0472] The BLG isolate is preferably a BLG isolate powder, or the liquid BLG isolate contains water and solid BLG isolate powder in an amount ranging from 1 to 50% w / w.

[0473] Preferably, the beta-lactoglobulin (BLG) isolate powder prepared by spray-drying 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 comprises the following:

[0474] - At least 30% w / w of total protein,

[0475] - At least 85% w / w of total protein in BLG, and

[0476] - Up to 10% w / w of water.

[0477] The BLG isolated powder preferably has one or more of the following:

[0478] - At least 0.2 g / cm³ 3 The bulk density of,

[0479] - At least 1.11 intrinsic tryptophan fluorescence emission ratio (I330 / I350),

[0480] - Up to 10% protein denaturation,

[0481] - Thermal stability of up to 200 NTU at pH 3.9, and

[0482] - Up to 1,000 colony-forming units / g.

[0483] BLG isolated powder is preferably an edible composition.

[0484] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of 2 to 4.9. Such powder is particularly useful for acidic food products and especially acidic beverages.

[0485] In another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5.

[0486] In some preferred embodiments of the present invention, the BLG isolate powder contains 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.

[0487] A higher protein content may be required, and in some preferred embodiments of the present invention, the BLG isolate powder contains 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.

[0488] Total protein is measured according to Example 1.5.

[0489] In some preferred embodiments of the present invention, the BLG isolate powder contains 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, even more preferably at least 98% w / w, and most preferably at least 99.5% w / w relative to total protein.

[0490] In some preferred embodiments of the present invention, the sum of alpha-lactalbumin (ALA) and caseinomacropeptide (CMP) constitutes at least 40% w / w, 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 protein in the powder.

[0491] In another preferred embodiment of the present invention, each major non-BLG whey protein is present as a weight percentage relative to total protein, which is up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, most preferably up to 6% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey.

[0492] Even lower concentrations of major non-BLG whey protein may be desirable. Accordingly, in a further preferred embodiment of the present invention, each major non-BLG whey protein is present as a weight percentage relative to total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey.

[0493] The inventors have observed indications that a reduction in lactoferrin and / or lactoperoxidase is particularly advantageous for obtaining a neutral-colored whey protein product.

[0494] Accordingly, in some preferred embodiments of the present invention, lactoferrin is present as a weight percentage relative to total protein, which is up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, most preferably up to 6% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey. Even lower concentrations of lactoferrin may be preferred. Accordingly, in a further preferred embodiment of the present invention, lactoferrin is present as a weight percentage relative to total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey.

[0495] Similarly, in some preferred embodiments of the present invention, lactoperoxidase is present as a weight percentage relative to total protein, which is up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, most preferably up to 6% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey. Even lower concentrations of lactoperoxidase may be preferred. Accordingly, in a further preferred embodiment of the present invention, lactoperoxidase is present as a weight percentage relative to total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of its weight percentage relative to total protein in a standard whey protein concentrate from sweet whey.

[0496] Lactoferrin and lactoperoxidase are quantified according to Example 1.29.

[0497] In some preferred embodiments of the present invention, the BLG isolate powder has a moisture content 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.

[0498] In some preferred embodiments of the present invention, the BLG isolate powder contains 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%. The BLG isolate powder may contain, for example, carbohydrates, for example, lactose, oligosaccharides and / or hydrolysis products of lactose (i.e., glucose and galactose), sucrose, and / or maltodextrin.

[0499] In some preferred embodiments of the present invention, the BLG isolate powder contains 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.

[0500] The inventors have discovered that it may be advantageous to control the mineral content to achieve some of the desired properties of BLG isolated powder.

[0501] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolate powder is up to 10 mmol / g protein. Preferably, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolate 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.

[0502] In another preferred embodiment of the present invention, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolate powder is at most 1 mmol / g protein. Preferably, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolate powder is at most 0.6 mmol / g protein, more preferably at most 0.4 mmol / g protein, even more preferably at most 0.2 mmol / g protein, and most preferably at most 0.1 mmol / g protein.

[0503] In another preferred embodiment of the present invention, the sum of the amounts of Mg and Ca in the BLG isolated powder is up to 5 mmol / g protein. Preferably, the sum of the amounts of Mg and Ca in the BLG isolated powder is up to 3 mmol / g protein, more preferably up to 1.0 mmol / g protein, and even more preferably up to 0.5 mmol / g protein.

[0504] In another preferred embodiment of the present invention, the sum of the amounts of Mg and Ca in the BLG isolated powder is at most 0.3 mmol / g protein. Preferably, the sum of the amounts of Mg and Ca in 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.

[0505] The inventors have discovered that a low phosphorus / low potassium variant of BLG isolate powder, which is particularly useful for patients with kidney disease, can be used. To make such a product, the BLG isolate powder must have equally low phosphorus and potassium content.

[0506] Accordingly, in some preferred embodiments of the present invention, the BLG isolate powder has a total phosphorus content of up to 100 mg of phosphorus per 100 g of protein. Preferably, the BLG isolate powder has a total phosphorus content of up to 80 mg of phosphorus per 100 g of protein. More preferably, the BLG isolate powder has a total phosphorus content of up to 50 mg of phosphorus per 100 g of protein. Even more preferably, the BLG isolate powder has a total phosphorus content of up to 20 mg of phosphorus per 100 g of protein. The BLG isolate powder has a total phosphorus content of up to 5 mg of phosphorus per 100 g of protein.

[0507] In some preferred embodiments of the present invention, the BLG isolate powder contains up to 600 mg of potassium per 100 g of protein. More preferably, the BLG isolate powder contains up to 500 mg of potassium per 100 g of protein. More preferably, the BLG isolate powder contains up to 400 mg of potassium per 100 g of protein. More preferably, the BLG isolate powder contains up to 300 mg of potassium per 100 g of protein. Even more preferably, the BLG isolate powder contains up to 200 mg of potassium per 100 g of protein. Even more preferably, the BLG isolate powder contains up to 100 mg of potassium per 100 g of protein. Even more preferably, the BLG isolate powder contains up to 50 mg of potassium per 100 g of protein, and even more preferably, the BLG isolate powder contains up to 10 mg of potassium per 100 g of protein.

[0508] The phosphorus content is related to the total amount of elemental phosphorus in the composition and is determined according to Example 1.19. Similarly, the potassium content is related to the total amount of elemental potassium in the composition and is determined according to Example 1.19.

[0509] In some preferred embodiments of the present invention, the BLG isolate powder contains up to 100 mg phosphorus and up to 700 mg potassium per 100 g of protein, preferably up to 80 mg phosphorus and up to 600 mg potassium per 100 g of protein, more preferably up to 60 mg phosphorus and up to 500 mg potassium per 100 g of protein, more preferably up to 50 mg phosphorus and up to 400 mg potassium per 100 g of protein, or more preferably up to 20 mg phosphorus and up to 200 mg potassium per 100 g of protein, or even more preferably up to 10 mg phosphorus and up to 50 mg potassium per 100 g of protein. In some preferred embodiments of the present invention, the BLG isolate powder contains up to 100 mg of phosphorus and up to 340 mg of potassium per 100 g of protein.

[0510] The low phosphorus and / or low potassium composition according to the present invention can be used as a food ingredient for the production of food products for a group of patients with impaired kidney function.

[0511] The inventors have found that for some applications, for example, acidic food products and particularly acidic beverages, it is particularly advantageous to have an acidic BLG isolated powder having a pH of up to 4.9, more preferably up to 4.3. This applies particularly to high-protein, clear acidic beverages.

[0512] In the context of the present invention, a transparent liquid has a turbidity of up to 200 NTU as measured according to Example 1.7.

[0513] Accordingly, in some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of 2 to 4.9. Preferably, the BLG isolate powder has a pH in the range of 2.5 to 4.7, more preferably 2.8 to 4.3, even more preferably 3.2 to 4.0, and most preferably 3.4 to 3.9. Alternatively, but also preferably, the BLG isolate powder may have a pH in the range of 3.6 to 4.3.

[0514] The inventors have discovered that having pH-neutral BLG isolated powder is particularly advantageous for some applications, for example, pH-neutral food products and especially pH-neutral beverages. This applies particularly to high-protein transparent or opaque pH-neutral beverages.

[0515] Accordingly, in some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5. Preferably, the powder has a pH in the range of 6.1 to 8.5, more preferably 6.2 to 8.0, even more preferably 6.3 to 7.7, and most preferably 6.5 to 7.5.

[0516] In another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 5.0 to 6.0. Preferably, the powder has a pH in the range of 5.1 to 5.9, more preferably 5.2 to 5.8, even more preferably 5.3 to 5.7, and most preferably 5.4 to 5.6.

[0517] Advantageously, the BLG isolate powder used in the present invention has a concentration of at least 0.20 g / cm³ 3 , preferably at least 0.30 g / cm³ 3 , more preferably at least 0.40 g / cm³ 3 , more preferably at least 0.45 g / cm³ 3 , more preferably at least 0.50 g / cm³ 3, most preferably at least 0.6 g / cm³ 3 It can have a bulk density.

[0518] Low-density powders, such as freeze-dried BLG isolates, are fluffy and easily enter the air at the production site during use. This is problematic because it increases the risk of cross-contamination of freeze-dried powders with other food products, and dusty environments are known to cause hygiene issues. In extreme cases, dusty environments also increase the risk of dust explosions.

[0519] The high-density variant of the present invention is easy to handle and has a low tendency to enter the surrounding air.

[0520] An additional advantage of the high-density variant of the present invention is that it takes up less space during transport and thus increases the weight of the BLG isolate powder that can be transported in a single volume unit.

[0521] An additional advantage of the high-density variant of the present invention is that other powdered food ingredients, for example, powdered sugar (approximately 0.56 g / cm³) 3 bulk density), granulated sugar (approximately 0.71 g / cm³) 3 bulk density), powdered citric acid (approximately 0.77 g / cm³) 3 When used in a powder mixture with bulk density, the tendency to separate is small.

[0522] The BLG isolated powder of the present invention has a content of 0.2 to 1.0 g / cm³ 3 Range, preferably 0.30 to 0.9 g / cm³ 3 Range, more preferably 0.40 to 0.8 g / cm³ 3 Range, more preferably 0.45 to 0.75 g / cm³ 3 Range, more preferably 0.50 to 0.75 g / cm³ 3 Range, most preferably 0.6 to 0.75 g / cm³ 3 It can have a bulk density within a range.

[0523] The bulk density of the powder is measured according to Example 1.17.

[0524] The inventors have discovered that it is advantageous to maintain the natural form of BLG, and have observed indications that when BLG is used in acidic beverages, increased unfolding of BLG causes an increased level of dry texture.

[0525] The intrinsic tryptophan fluorescence emission ratio (I330 / I350) measures the degree of unfolding of BLG, and the inventors found that less dryness is observed at a high intrinsic tryptophan fluorescence emission ratio associated with less or no unfolding of BLG. The intrinsic tryptophan fluorescence emission ratio (I330 / I350) is measured according to Example 1.1.

[0526] In some preferred embodiments of the present invention, the BLG isolate powder has an intrinsic tryptophan fluorescence emission ratio (I330 / I350) of at least 1.11.

[0527] In some preferred embodiments of the present invention, the BLG isolate powder has an intrinsic tryptophan fluorescence emission 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, most preferably at least 1.19.

[0528] If the BLG isolated powder contains a significant amount of non-protein material, it is desirable to isolate the protein fraction before measuring the intrinsic tryptophan fluorescence emission ratio. Accordingly, in some preferred embodiments of the present invention, the protein fraction of the BLG isolated powder has an intrinsic tryptophan fluorescence emission ratio of at least 1.11.

[0529] In some preferred embodiments of the present invention, the protein fraction of the BLG isolate powder has an intrinsic tryptophan fluorescence emission 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, most preferably at least 1.19.

[0530] The protein fraction can be separated from the BLG isolate powder by, for example, dissolving the BLG isolate powder in deionized water and dialyzing or ultrafiltration-based volume filtration of the solution using a protein-retaining filter. If the BLG isolate powder contains interfering levels of lipids, such lipids can be removed, for example, by microfiltration. Both lipids and small molecules can be removed from the protein fraction by combining the microfiltration and ultrafiltration / volume filtration steps.

[0531] It is often desirable that a significant amount of BLG in the BLG isolate 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 isolate powder is non-aggregated BLG.

[0532] In some preferred embodiments of the present invention, the BLG isolate powder has a protein denaturation degree 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%.

[0533] However, for example, if an opaque beverage is required, it may also be desirable for the BLG isolate powder to have a significant level of protein denaturation. Accordingly, in another preferred embodiment of the present invention, the BLG isolate powder has a degree of protein denaturation of at least 11%, preferably at least 20%, more preferably at least 40%, even more preferably at least 50%, even more preferably at least 75%, and most preferably at least 90%.

[0534] If the BLG isolated powder has a significant level of protein denaturation, it is often desirable to maintain a low level of insoluble protein material, that is, precipitated protein material that precipitates into the beverage during storage. The level of insoluble material is measured according to Example 1.10.

[0535] In some preferred embodiments of the present 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 be more preferable that the BLG isolated powder does not contain any insoluble protein material at all.

[0536] The inventors have discovered that the thermal stability of the BLG isolated powder at pH 3.9 is an excellent indicator of its usefulness for clear high-protein beverages. Thermal stability at pH 3.9 is measured according to Example 1.2.

[0537] It is particularly desirable that the BLG isolated powder has a thermal stability of up to 200 NTU at pH 3.9, 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, and the BLG isolated powder preferably has a thermal stability of up to 10 NTU at pH 3.9, preferably up to 8 NTU, more preferably up to 4 NTU, and even more preferably up to 2 NTU.

[0538] It is desirable to keep the microbial content of BLG isolated powder to a minimum. However, since the process for reducing microorganisms tends to result in protein unfolding and denaturation, it is a difficult task to obtain both a high degree of protein naturalness and a low microbial content. The present invention enables obtaining a very low microbial content while maintaining a high level of BLG naturalness.

[0539] Accordingly, in some preferred embodiments of the present invention, the BLG isolate powder contains up to 15,000 colony-forming units (CFU) / g. Preferably, the BLG isolate powder contains up to 10,000 CFU / g. More preferably, the BLG isolate powder contains up to 5,000 CFU / g. Even more preferably, the BLG isolate powder contains up to 1,000 CFU / g. Even more preferably, the BLG isolate powder contains up to 300 CFU / g. Most preferably, the BLG isolate powder contains up to 100 CFU / g, for example, up to 10 CFU / g. In a particularly preferred embodiment, the powder is sterile. Sterile BLG isolate powder can be produced by combining several physical microbiome reduction processes, such as microfiltration and heat treatment at an acidic pH, during the production of BLG isolate powder.

[0540] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of i) 2 to 4.9, ii) 6.1 to 8.5, or iii) 5.0 to 6.0, and comprises:

[0541] - Total protein in an amount of at least 30% w / w, preferably at least 80% w / w, more preferably at least 90% w / w,

[0542] - At least 85% w / w, preferably at least 90% w / w, of beta-lactoglobulin (BLG) relative to total protein,

[0543] - Up to 6% w / w of water,

[0544] - Up to 2% w / w, preferably up to 0.5% w / w of lipids,

[0545] The above BLG isolated powder has the following:

[0546] - At least 1.11 intrinsic tryptophan fluorescence emission ratio (I330 / I350),

[0547] - Up to 10% protein denaturation, and

[0548] - Thermal stability of up to 200 NTU at pH 3.9.

[0549] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of i) 2 to 4.9 or ii) 6.1 to 8.5 and comprises:

[0550] - Total protein in an amount of at least 30% w / w, preferably at least 80% w / w, more preferably at least 90% w / w,

[0551] - At least 85% w / w, preferably at least 90% w / w, of total protein, more preferably at least 94% w / w of total protein, amount of beta-lactoglobulin (BLG),

[0552] - Up to 6% w / w of water,

[0553] - Up to 2% w / w, preferably up to 0.5% w / w of lipids,

[0554] The above BLG isolated powder has the following:

[0555] - At least 1.11 intrinsic tryptophan fluorescence emission ratio (I330 / I350),

[0556] - Protein denaturation of up to 10%, preferably up to 5%, and

[0557] - Thermal stability of up to 70 NTU at pH 3.9, preferably up to 50 NTU, more preferably up to 40 NTU.

[0558] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of i) 2 to 4.9 or ii) 6.1 to 8.5 and comprises:

[0559] - At least 30% w / w of total protein,

[0560] - At least 85% w / w, preferably at least 90% w / w, of beta-lactoglobulin (BLG) relative to total protein,

[0561] - Up to 6% w / w of water,

[0562] The above BLG isolated powder has the following:

[0563] - At least 0.2 g / cm³ 3 The bulk density of,

[0564] - At least 1.11 intrinsic tryptophan fluorescence emission ratio (I330 / I350),

[0565] - Up to 10% protein denaturation, and

[0566] - Thermal stability of up to 200 NTU at pH 3.9.

[0567] In another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 2 to 4.9 and comprises:

[0568] - at least 80% w / w, preferably at least 90% w / w, more preferably at least 94% w / w of total protein,

[0569] - At least 85% w / w, preferably at least 90% w / w, of total protein, and more preferably at least 94% w / w of total protein, amount of beta-lactoglobulin (BLG).

[0570] - Up to 6% w / w of water,

[0571] - Up to 2% w / w, preferably up to 0.5% w / w of lipids,

[0572] The above BLG isolated powder has the following:

[0573] - At least 0.2 g / cm³ 3 , preferably at least 0.3 g / cm³ 3 , more preferably at least 0.4 g / cm³ 3 The bulk density of,

[0574] - At least 1.11 intrinsic tryptophan fluorescence emission ratio (I330 / I350),

[0575] - Protein denaturation of up to 10%, preferably up to 5%, more preferably up to 2%, and

[0576] - Thermal stability of up to 50 NTU at pH 3.9, preferably up to 30 NTU, more preferably up to 10 NTU.

[0577] In another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5 and comprises:

[0578] - at least 80% w / w, preferably at least 90% w / w, more preferably at least 94% w / w of total protein,

[0579] - At least 85% w / w, preferably at least 90% w / w, of total protein, and more preferably at least 94% w / w of total protein, amount of beta-lactoglobulin (BLG).

[0580] - Up to 6% w / w of water,

[0581] - Up to 2% w / w, preferably up to 0.5% w / w of lipids,

[0582] The above BLG isolated powder has the following:

[0583] - At least 0.2 g / cm³ 3 , preferably at least 0.3 g / cm³ 3 , more preferably at least 0.4 g / cm³ 3 The bulk density of,

[0584] - Protein denaturation of up to 10%, preferably up to 5%, more preferably up to 2%, and

[0585] - Thermal stability of up to 50 NTU at pH 3.9, preferably up to 30 NTU, more preferably up to 10 NTU.

[0586] In a further preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5 and comprises the following:

[0587] - at least 80% w / w, preferably at least 90% w / w, more preferably at least 94% w / w of total protein,

[0588] - At least 85% w / w, preferably at least 90% w / w, of total protein, and more preferably at least 94% w / w of total protein, amount of beta-lactoglobulin (BLG).

[0589] - Up to 6% w / w of water,

[0590] - Up to 2% w / w, preferably up to 0.5% w / w of lipids,

[0591] The above BLG isolated powder has the following:

[0592] - At least 0.2 g / cm³ 3 , preferably at least 0.3 g / cm³ 3 , more preferably at least 0.4 g / cm³ 3 The bulk density of,

[0593] - Protein denaturation of up to 10%, preferably up to 5%, more preferably up to 2%, and

[0594] - Thermal stability of up to 50 NTU at pH 3.9, preferably up to 30 NTU, more preferably up to 10 NTU.

[0595] In a further preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 5.0 to 6.0 and comprises:

[0596] - at least 80% w / w, preferably at least 90% w / w, more preferably at least 94% w / w of total protein,

[0597] - At least 85% w / w, preferably at least 90% w / w, of total protein, and more preferably at least 94% w / w of total protein, amount of beta-lactoglobulin (BLG).

[0598] - Up to 6% w / w of water,

[0599] - Up to 2% w / w, preferably up to 0.5% w / w of lipids,

[0600] The above BLG isolated powder has the following:

[0601] - At least 0.2 g / cm³ 3 , preferably at least 0.3 g / cm³ 3 , more preferably at least 0.4 g / cm³ 3 The bulk density of,

[0602] - Protein denaturation of up to 10%, preferably up to 5%, more preferably up to 2%,

[0603] - Thermal stability of up to 50 NTU at pH 3.9, preferably up to 30 NTU, more preferably up to 10 NTU, and

[0604] - Preferably, BLG crystallinity of less than 10%.

[0605] BLG isolate powder containing BLG in an amount of at least 85% w / w relative to total protein is typically provided by a method comprising the following steps:

[0606] a) A step of providing a liquid BLG isolate having the following:

[0607] i) pH in the range of 2 to 4.9,

[0608] ii) pH in the range of 6.1 to 8.5, or

[0609] iii) pH in the range of 5.0 to 6.0

[0610] The above liquid BLG isolate contains BLG in an amount of at least 85% w / w relative to total protein, step

[0611] b) Optionally, a step of physically reducing microorganisms in the liquid BLG isolate,

[0612] c) A step of drying the liquid BLG isolated material, preferably spray-drying.

[0613] BLG isolates are preferably prepared from mammalian milk, preferably ruminant milk, for example, milk from dairy cows, sheep, goats, buffalo, camels, llamas, mares, and / or deer. Proteins derived from cow's milk are particularly preferred. Thus, BLG is preferably bovine BLG.

[0614] Liquid BLG isolates can be provided in a number of different ways.

[0615] Typically, the provision of a liquid BLG isolate comprises or even consists of the step of isolating BLG from a whey protein feed to provide a BLG-rich composition by one or more of the following methods:

[0616] - Crystallization or precipitation of BLG by salt treatment,

[0617] - Crystallization or precipitation of BLG by salting out,

[0618] - Ion exchange chromatography, and

[0619] - Separation of whey protein by ultrafiltration.

[0620] A particularly preferred method for providing a BLG-rich composition is by crystallization of BLG, preferably by salting or alternatively by salting out.

[0621] The whey protein source is preferably WPC, WPI, SPC, SPI, or a combination thereof.

[0622] The term "whey protein feedstock" relates to a BLG-rich composition and subsequently a composition from which a liquid BLG isolate is derived.

[0623] In some embodiments of the present invention, the preparation of the BLG-rich composition comprises or even consists of high salt BLG crystallization in the pH range of 3.6 to 4.0 in accordance with US No. 2,790,790 A1.

[0624] In another embodiment of the present invention, the preparation of the BLG-rich composition comprises or is even made by 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).

[0625] However, in a particularly preferred embodiment of the present invention, the BLG-rich composition is prepared by crystallization at pH 5 to 6 under salt-soluble conditions as described in PCT application No. PCT / EP2017 / 084553, incorporated herein by reference for all purposes.

[0626] In some preferred embodiments of the present invention, the BLG-rich composition is an edible BLG composition according to PCT / EP2017 / 084553 containing at least 90% BLG relative to total protein and preferably containing BLG crystals.

[0627] If the BLG-rich composition isolated from a whey protein feed does not yet possess the necessary characteristics to be used as a liquid BLG isolate, it may be processed in one or more steps selected from the following group as part of providing a liquid BLG isolate:

[0628] - Desalination,

[0629] - Mineral addition,

[0630] - Dilution,

[0631] - Concentration,

[0632] - Physical microbial reduction, and

[0633] - pH adjustment.

[0634] Non-limiting examples of desalination include, for example, dialysis, gel filtration, UF / dialysis filtration, NF / dialysis filtration, and ion exchange chromatography.

[0635] Non-limiting examples of mineral addition include the addition of salts permitted in water-soluble foods, e.g., salts of Na, K, Ca, and / or Mg. Such salts may be, e.g., phosphate salts, chloride salts, or salts of food acids, e.g., citrate salts or lactate salts. Minerals may be added in solid, suspended, or dissolved form.

[0636] Non-limiting examples of dilution include, for example, the addition of liquid diluents, such as water, demineralized water, or aqueous solutions of minerals, acids, or bases.

[0637] Non-limiting examples of concentration include, for example, evaporation, reverse osmosis, nanofiltration, ultrafiltration, and combinations thereof.

[0638] If the concentration needs to increase the protein concentration relative to the total solids, it is desirable to use a concentration step such as ultrafiltration or alternatively dialysis. If it is not necessary to increase the protein concentration relative to the total solids, methods such as evaporation, nanofiltration, and / or reverse osmosis may be useful.

[0639] Non-limiting examples of physical microbial reduction include, for example, heat treatment, bacterial filtration, UV radiation, high-pressure treatment, pulsed electric field treatment, and ultrasound. These methods are well known to those skilled in the art.

[0640] Non-limiting examples of pH adjustment include, for example, the addition of a base and / or acid, preferably a base and / or acid acceptable to food. It is particularly desirable to use an acid and / or base 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.

[0641] In some preferred embodiments of the present invention, the liquid solution has a color value delta b* in the range of -0.10 to +0.51 on the CIELAB color scale, particularly if the formulation has a turbidity of up to 200 NTU, more preferably up to 40 NTU.

[0642] In another preferred embodiment of the present invention, the liquid solution has a color value delta b* in the range of 0.0 to 0.40 on the CIELAB color scale, preferably in the range of +0.10 to +0.25.

[0643] The liquid solution of the present invention may contain macronutrients other than protein.

[0644] In some embodiments of the present invention, the liquid solution further comprises carbohydrates. The total carbohydrate content in the liquid solution of the present invention varies depending on the intended use of the final heat-treated beverage formulation.

[0645] In some preferred embodiments of the present invention, the liquid solution further comprises at least one carbohydrate source. In one exemplary embodiment, at least one carbohydrate source is selected from the group consisting of sucrose, maltodextrin, corn syrup solids, sucromalt, glucose polymer, corn syrup, modified starch, resistant starch, rice-derived carbohydrate, isomaltulose, white sugar, glucose, fructose, lactose, galactose, maltose, dextrose, high-fructose corn syrup, honey, sugar alcohol, fructooligosaccharide, soybean fiber, corn fiber, guar gum, konjac powder, polydextrose, fibersol, and combinations thereof.

[0646] In some preferred embodiments of the present invention, the liquid solution comprises a sugar polymer, namely an oligosaccharide and / or a polysaccharide.

[0647] In some preferred embodiments, the liquid solution further comprises carbohydrates in a range between 0% and 95% of the total energy content of the liquid solution, preferably between 10% and 85% of the total energy content of the liquid solution, preferably between 20% and 75% of the total energy content of the liquid solution, or preferably between 30% and 60% of the total energy content of the liquid solution.

[0648] A lower carbohydrate content is often desirable, and thus, in some preferred embodiments of the present invention, the carbohydrate content of the liquid solution is preferably in the range between 0% and 30% of the total energy content of the formulation, more preferably in the range between 0% and 20% of the total energy content of the formulation, and even more preferably in the range between 0% and 10% of the total energy content of the formulation.

[0649] In some preferred embodiments of the present invention, the carbohydrate content of the liquid solution is up to 5% of the total energy content of the liquid solution, more preferably up to 1% of the total energy content of the liquid solution, and even more preferably up to 0.1% of the total energy content of the liquid solution.

[0650] In one embodiment of the present invention, the liquid solution further comprises at least one additional component selected from the group consisting of vitamins, flavoring agents, minerals, sweeteners, antioxidants, food acids, lipids, carbohydrates, prebiotics, probiotics, and non-whey proteins.

[0651] 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 salt, neotame, saccharin, stevia extract, steviol glycoside, e.g., rebaudioside A, or combinations thereof. In some embodiments of the present invention, it is particularly preferred that the sweetener comprises one or more high-intensity sweeteners (HIS) or even consists of them.

[0652] HIS is found in both natural and artificial sweeteners, and typically has a sweetness strength at least 10 times that of sucrose.

[0653] If used, the total amount of HIS is typically in the range of 0.01 to 2% w / w. For example, the total amount of HIS may be in the range of 0.05 to 1.5% w / w. Alternatively, the total amount of HIS may be in the range of 0.1 to 1.0% w / w.

[0654] The choice of sweetener may vary depending on the beverage to be produced; for example, high-intensity sweeteners (e.g., aspartame, acesulfame-K, or sucralose) may be used in beverages where no energy contribution from the sweetener is desired, whereas for beverages with a natural profile, natural sweeteners (e.g., steviol glycoside, sorbitol, or sucrose) may be used.

[0655] Alternatively, or additionally, carbohydrate sweeteners may be used.

[0656] Additionally, it may be desirable for the sweetener to include or even consist of one or more polyol sweeteners. Non-limiting examples of useful polyol sweeteners are maltitol, mannitol, lactitol, sorbitol, inositol, xylitol, threitol, galactitol, or combinations thereof. If used, the total amount of polyol sweetener is typically in the range of 1 to 20% w / w. For example, the total amount of polyol sweetener may be in the range of 2 to 15% w / w. Alternatively, the total amount of polyol sweetener may be in the range of 4 to 10% w / w.

[0657] The liquid solution of the present invention may contain macronutrients other than protein. In some embodiments of the present invention, the liquid solution further contains lipids. The total lipid content in the final heat-treated beverage formulation of the present invention varies depending on the intended use of the heat-treated beverage formulation.

[0658] In some preferred embodiments of the present invention, the liquid solution has a lipid content in the range between 0% and 60% of the total energy content of the liquid solution, or preferably between 0% and 50% of the total energy content of the liquid solution, or preferably between 0% and 45% of the total energy content of the liquid solution, or preferably between 0% and 30% of the total energy content of the liquid solution, or preferably between 0% and 20% of the total energy content of the liquid solution, or preferably between 0% and 10% of the total energy content of the liquid solution, or preferably between 0% and 5% of the total energy content of the liquid solution.

[0659] The amount of lipids is determined according to ISO 1211:2010 (Determination of fat content - Rose-Gottlieb Gravimetric Method).

[0660] In some preferred embodiments of the present invention, the lipid content of the liquid solution is up to 3% of the total energy content of the liquid solution, more preferably up to 1% of the total energy content of the liquid solution, and even more preferably up to 0.1% of the total energy content of the liquid solution.

[0661] The liquid solution typically contains a total amount of water in the range of 50 to 99% w / w, preferably 45 to 97% w / w, more preferably 40 to 95% w / w, even more preferably 35 to 90% w / w, and most preferably 30 to 85% w / w.

[0662] In some preferred embodiments of the present invention, the liquid solution contains a total amount of water in the range of 55 to 90% w / w, preferably in the range of 57 to 85% w / w, more preferably in the range of 60 to 80% w / w, even more preferably in the range of 62 to 75% w / w, and most preferably in the range of 65 to 70% w / w.

[0663] In some preferred embodiments of the present invention, the liquid solution contains a total amount of water in the range of 90 to 99% w / w, preferably in the range of 92 to 98.5% w / w, more preferably in the range of 94 to 98% w / w, even more preferably in the range of 95 to 98% w / w, and most preferably in the range of 96 to 98% w / w. Such embodiments are useful for beverages, for example, such as clear water.

[0664] In some preferred embodiments of the present invention, the liquid solution is non-alcoholic, which means that it contains 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 no detectable ethanol.

[0665] The liquid solution typically contains a total amount of solid in the range of 1 to 45% w / w, preferably 5 to 40% w / w, more preferably 10 to 35% w / w, even more preferably 12 to 30% w / w, and most preferably 16 to 25% w / w.

[0666] In some preferred embodiments of the present invention, the liquid solution contains a total amount of solid in the range of 10 to 45% w / w, preferably 15 to 43% w / w, more preferably 20 to 40% w / w, even more preferably 25 to 38% w / w, and most preferably 30 to 35% w / w.

[0667] In some preferred embodiments of the present invention, the liquid solution contains a total amount of solids in the range of 1 to 10% w / w, preferably 1.5 to 8% w / w, more preferably 2 to 6% w / w, even more preferably 2 to 5% w / w, and most preferably 2 to 4% w / w. Such embodiments are useful for beverages, for example, clear water.

[0668] The non-solid liquid part of the solution is preferably water.

[0669] The inventors have discovered that it may be advantageous to control the mineral content to achieve some of the desired properties of a packaged heat-treated beverage formulation. In some embodiments of the present invention, the packaged heat-treated beverage formulation 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.

[0670] The inventors have surprisingly discovered that when the BLG isolate is used as defined herein, a heat-treated beverage formulation having a high mineral concentration without viscosity impairment can be produced in Example 2. This provides the possibility that a packaged heat-treated beverage formulation can be produced with a high mineral content, and that a beverage that is a nutritionally complete or nutritionally incomplete supplement can be produced.

[0671] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg and Ca in the liquid solution is in the range of 0 to 750 mM, preferably in the range of 100 to 600 mM or preferably in the range of 200 to 500 mM.

[0672] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg and Ca in the liquid solution is up to 750 mM.

[0673] In another preferred embodiment of the present invention, the sum of the amounts of Na, K, Mg and Ca in the liquid solution is up to 600 mM, preferably up to 500 mM, or preferably up to 400 mM, or preferably up to 300 mM, or preferably up to 200 mM, preferably up to 170 mM, most preferably up to 150 mM, or preferably up to 130 mM, or preferably up to 100 mM, or preferably up to 80 mM, or preferably up to 60 mM, or preferably up to 40 mM, or preferably up to 30 mM, or preferably up to 20 mM, or preferably up to 10 mM, or preferably up to 5 mM, or preferably up to 1 mM.

[0674] In another exemplary embodiment, the liquid solution comprises a plurality of minerals selected from the group consisting of calcium, iodine, zinc, copper, chromium, iron, phosphorus, magnesium, selenium, manganese, molybdenum, sodium, potassium, and combinations thereof.

[0675] In some preferred embodiments of the present invention, the liquid solution comprises up to 150 mM KCl and up to 150 mM CaCl2, or the liquid solution comprises up to 130 mM KCl and up to 130 mM CaCl2, or the liquid solution comprises up to 110 mM KCl and up to 110 mM CaCl2, or the liquid solution comprises up to 100 mM KCl and up to 100 mM CaCl2, preferably the liquid solution comprises up to 80 mM KCl and up to 80 mM CaCl2, preferably the liquid solution comprises up to 50 mM KCl and up to 50 mM CaCl2, or preferably the liquid solution comprises up to 40 mM KCl and up to 40 mM CaCl2.

[0676] In another preferred embodiment of the present invention, the liquid solution is a low-mineral solution.

[0677] In the context of the present invention, the term “low mineral” relates to a composition, e.g., a liquid, beverage, powder, or other food product having at least one, preferably two, and more preferably all of the following:

[0678] - Up to 1.2% w / w ash content relative to total solids,

[0679] - Total content of calcium and magnesium up to 0.3% w / w relative to total solids,

[0680] - Total content of sodium and potassium up to 0.10% w / w relative to total solids,

[0681] - Total phosphorus content of up to 100 mg of phosphorus per 100 g of protein.

[0682] Preferably, the low mineral composition has at least one of the following, preferably two or more, and more preferably all of them:

[0683] - Ash content of up to 0.7% w / w relative to total solids,

[0684] - Total content of calcium and magnesium up to 0.2% w / w relative to total solids,

[0685] - Total content of sodium and potassium up to 0.08% w / w relative to total solids,

[0686] - Total phosphorus content of up to 80 mg per 100 g of protein.

[0687] More preferably, the low mineral composition has at least one of the following, preferably two or more, and more preferably all of them:

[0688] - Ash content of up to 0.5% w / w relative to total solids,

[0689] - Total content of calcium and magnesium up to 0.15% w / w relative to total solids,

[0690] - Total content of sodium and potassium up to 0.06% w / w relative to total solids,

[0691] - Total phosphorus content of up to 50 mg per 100 g of protein.

[0692] It is particularly desirable that the low mineral composition has the following:

[0693] - Ash content of up to 0.5% w / w relative to total solids,

[0694] - Total content of calcium and magnesium up to 0.15% w / w relative to total solids,

[0695] - Total content of sodium and potassium up to 0.06% w / w relative to total solids,

[0696] - Total phosphorus content of up to 50 mg per 100 g of protein.

[0697] The inventors have discovered that the present invention can produce a packaged heat-treated beverage formulation having a very low content of phosphorus and other minerals, such as potassium, which is advantageous for patients suffering from kidney disease or otherwise having impaired kidney function.

[0698] The liquid solution is preferably a low-phosphorus solution.

[0699] The liquid solution is preferably a low potassium solution.

[0700] The liquid solution is preferably a low phosphorus and low potassium solution.

[0701] In the context of the present invention, the term “low phosphorus” relates to a composition having a total phosphorus content of up to 100 mg of phosphorus per 100 g of protein, e.g., a liquid, powder, or other food product. Preferably, the low phosphorus composition has a total phosphorus content of up to 80 mg of phosphorus per 100 g of protein. More preferably, the low phosphorus composition may have a total phosphorus content of up to 50 mg of phosphorus per 100 g of protein. Even more preferably, the low phosphorus composition may have a total phosphorus content of up to 20 mg of phosphorus per 100 g of protein. Even more preferably, the low phosphorus composition may have a total phosphorus content of up to 5 mg of phosphorus per 100 g of protein. The low phosphorus composition according to the present invention may be used as a food ingredient for the production of food products for a group of patients with impaired renal function.

[0702] Accordingly, in some particularly preferred embodiments of the present invention, the liquid solution contains up to 80 mg of phosphorus per 100 g of protein. Preferably, the liquid solution contains up to 30 mg of phosphorus per 100 g of protein. More preferably, the liquid solution contains up to 20 mg of phosphorus per 100 g of protein. Even more preferably, the liquid solution contains up to 10 mg of phosphorus per 100 g of protein. Most preferably, the liquid solution contains up to 5 mg of phosphorus per 100 g of protein.

[0703] The phosphorus content is related to the total amount of elemental phosphorus in the composition and is determined according to Example 1.19.

[0704] 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 potassium per 100 g of protein. Preferably, the low potassium composition has a total potassium content of up to 600 mg potassium per 100 g of protein. More preferably, the low potassium composition may have a total potassium content of up to 500 mg potassium per 100 g of protein. More preferably, the low potassium composition may have a total potassium content of up to 400 mg potassium per 100 g of protein. Even more preferably, the low potassium composition may have a total potassium content of up to 300 mg potassium per 100 g of protein. Even more preferably, the low potassium composition may have a total potassium content of up to 200 mg potassium per 100 g of protein. Even more preferably, the low potassium composition may have a total potassium content of up to 100 mg potassium per 100 g of protein. More preferably, the low potassium composition may have a total potassium content of up to 50 mg of potassium per 100 g of protein, and more preferably, the low potassium composition may have a total potassium content of up to 10 mg of potassium per 100 g of protein.

[0705] low potassium according to the present invention The composition can be used as a food ingredient for the production of food products for a group of patients with impaired kidney function.

[0706] Accordingly, in some particularly preferred embodiments of the present invention, the liquid solution contains up to 600 mg of potassium per 100 g of protein. More preferably, the liquid solution contains up to 500 mg of potassium per 100 g of protein. More preferably, the liquid solution contains up to 400 mg of potassium per 100 g of protein. More preferably, the liquid solution contains up to 300 mg of potassium per 100 g of protein. Even more preferably, the liquid solution contains up to 200 mg of potassium per 100 g of protein. Even more preferably, the liquid solution contains up to 100 mg of potassium per 100 g of protein. Even more preferably, the liquid solution contains up to 50 mg of potassium per 100 g of protein, and even more preferably, the liquid solution contains up to 10 mg of potassium per 100 g of protein.

[0707] The potassium content is related to the total amount of elemental potassium in the composition and is determined according to Example 1.19.

[0708] In some preferred embodiments of the present invention, the liquid solution comprises up to 100 mg phosphorus / 100 g protein and up to 700 mg potassium / 100 g protein, preferably up to 80 mg phosphorus / 100 g protein and up to 600 mg potassium / 100 g protein, more preferably up to 60 mg phosphorus / 100 g protein and up to 500 mg potassium / 100 g protein, more preferably up to 50 mg phosphorus / 100 g protein and up to 400 mg potassium / 100 g protein, or more preferably up to 20 mg phosphorus / 100 g protein and up to 200 mg potassium / 100 g protein, or even more preferably up to 10 mg phosphorus / 100 g protein and up to 50 mg potassium / 100 g protein. In some preferred embodiments of the present invention, the packaged heat-treated beverage formulation comprises up to 100 mg phosphorus / 100 g protein and up to 340 mg potassium / 100 g protein.

[0709] A liquid solution containing small amounts of phosphorus and potassium can advantageously be supplemented with carbohydrates and lipids, and the heat-treated beverage formulation preferably further comprises a total amount of carbohydrates in the range between 30% and 60% of the total energy content of the liquid solution, preferably between 35% and 50%, and a total amount of lipids in the range between 20% and 60% of the total energy content, preferably between 30% and 50%.

[0710] In one embodiment of the present invention, the liquid solution comprises a plurality of vitamins. In an exemplary embodiment, the liquid solution comprises at least 10 vitamins. In an exemplary embodiment, the liquid solution comprises vitamin A, vitamin B1, vitamin B2, vitamin B3, and vitamin B5 , It includes a plurality of vitamins selected from the group consisting of 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.

[0711] In one embodiment of the present invention, the liquid solution comprises a plurality of vitamins and a plurality of minerals.

[0712] In some preferred embodiments of the present invention, the liquid solution contains one or more food acids selected from the group consisting of citric acid, malic acid, tartaric acid, acetic acid, benzoic acid, butyric acid, lactic acid, fumaric acid, succinic acid, ascorbic acid, adipic acid, phosphoric acid, and mixtures thereof.

[0713] In one embodiment of the present invention, the liquid solution further comprises a flavoring selected from the group consisting of salts, flavorings, flavor enhancers, and / or spices. In a preferred embodiment of the present invention, the flavoring comprises chocolate, cocoa, lemon, orange, lime, strawberry, banana, forest fruit flavorings, or combinations thereof. The selection of the flavoring may vary depending on the beverage to be produced.

[0714] One embodiment of the present invention relates to the use of a protein solution comprising a total amount of protein of 2 to 45% w / w, preferably 3 to 35% w / w, relative to the weight of the solution, wherein at least 85 w / w %, preferably at least 90% w / w, of the protein is BLG to control the turbidity of a heat-treated acidic beverage formulation having a pH in the range of 2.0 to 4.7.

[0715] Another aspect of the present invention relates to the use of a protein solution comprising a total amount of protein of 2 to 45% w / w relative to the weight of the solution, wherein at least 85 w / w%, preferably at least 90 w / w%, of the protein is BLG to control the astringent taste of a heat-treated acidic beverage formulation having a pH in the range of 2.0 to 4.7.

[0716] Another aspect of the present invention relates to a packaged heat-treated beverage formulation as defined herein, for use in a method of treating a disease associated with poor protein absorption.

[0717] Another aspect of the present invention relates to the use of packaged heat-treated beverage formulations as defined herein as dietary supplements.

[0718] In a preferred embodiment of the present invention, a packaged heat-treated beverage formulation as defined herein is used as a dietary supplement and is consumed before, during, or after exercise.

[0719] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0720] - A total amount of protein of 2 to 45% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0721] - Optionally, sweeteners, sugar polymers and / or flavorings,

[0722] Here:

[0723] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.11, and

[0724] - The lipid content is up to 5% of the total energy content of the formulation.

[0725] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0726] - A total amount of protein of 2 to 10% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0727] - Optionally, sweeteners and / or flavorings,

[0728] Here:

[0729] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.11, and

[0730] - The lipid content is up to 5% of the total energy content of the formulation.

[0731] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0732] - A total amount of protein of 10 to 45% w / w, preferably 10 to 35% w / w, relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0733] - Optionally, sweeteners and / or flavorings,

[0734] Here:

[0735] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.11, and

[0736] - The lipid content is up to 5% of the total energy content of the formulation.

[0737] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0738] - A total amount of protein of 2 to 45% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0739] - Optionally, sweeteners and / or flavorings,

[0740] - Packaged heat-treated beverage formulations have a turbidity of up to 200 NTU, preferably up to 40 NTU.

[0741] In another preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises:

[0742] - A total amount of protein of 2 to 45% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0743] - Optionally, sweeteners and / or flavorings,

[0744] - Packaged heat-treated beverage formulations have a turbidity of up to 200 NTU, preferably up to 40 NTU.

[0745] In a further preferred embodiment of the present invention, a packaged heat-treated beverage formulation has a pH in the range of 3.5 to 4.7, preferably 3.7 to 4.3, more preferably 3.7 to 4.1, and said beverage formulation comprises:

[0746] - A total amount of protein of 2 to 45% w / w, preferably 5.0 to 35% w / w, more preferably 6.0 to 32% w / w relative to the weight of the beverage,

[0747] - At least 88% w / w, preferably at least 90% w / w, more preferably at least 92% w / w of protein is BLG.

[0748] - Total amount of lipids up to 5% w / w, preferably up to 1% w / w, more preferably up to 0.2% w / w,

[0749] - Optionally, sweeteners and / or flavorings,

[0750] The above beverage formulation has the following:

[0751] - Viscosity of up to 100 cP, preferably up to 20 cP, more preferably up to 10 cP,

[0752] - an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.13, preferably at least 1.15, more preferably at least 1.16 and

[0753] - Optionally, turbidity of up to 50 NTU, preferably up to 20 NTU, more preferably up to 10 NTU.

[0754] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0755] - A total amount of protein of 2 to 10% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0756] - Optionally, sweeteners and / or flavorings,

[0757] Packaged heat-treated beverage formulations have a turbidity of up to 200 NTU, preferably up to 40 NTU.

[0758] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0759] - A total amount of protein of 10 to 45% w / w, preferably 10 to 20% w / w, relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0760] - Optionally, sweeteners and / or flavorings,

[0761] Packaged heat-treated beverage formulations have a turbidity of up to 200 NTU, preferably up to 40 NTU.

[0762] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0763] - A total amount of protein of 2 to 45% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0764] - Optionally, sweeteners and / or flavorings,

[0765] Here:

[0766] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.11, and

[0767] - The protein fraction of the beverage formulation has a color value delta b* in the range of -0.10 to +0.51 on the CIELAB color scale, where,

[0768] Delta b* = b measured at room temperature 6.0 w / w % 단백질로 표준화된 샘플 * - b 탈염수 *am.

[0769] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0770] - A total amount of protein of 2 to 10% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0771] - Optionally, sweeteners and / or flavorings,

[0772] Here:

[0773] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.11, and

[0774] - The protein fraction of the beverage formulation has a color value delta b* in the range of -0.10 to +0.51 on the CIELAB color scale, where,

[0775] Delta b* = b measured at room temperature 6.0 w / w % 단백질로 표준화된 샘플 * - b 탈염수 *am.

[0776] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0777] - A total amount of protein of 10 to 45% w / w, preferably 10 to 20% w / w, relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0778] - Optionally, sweeteners and / or flavorings,

[0779] Here:

[0780] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.11, and

[0781] - The protein fraction of the beverage formulation has a color value delta b* in the range of -0.10 to +0.51 on the CIELAB color scale, where,

[0782] Delta b* = b measured at room temperature 6.0 w / w % 단백질로 표준화된 샘플 * - b 탈염수 *am.

[0783] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0784] - A total amount of protein of 2 to 45% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0785] - Optionally, sweeteners and / or flavorings,

[0786] Here:

[0787] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.11, and

[0788] - The sum of the amounts of Na, K, Mg and Ca is up to 750 mM, preferably up to 400 mM, preferably up to 200 mM.

[0789] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0790] - A total amount of protein of 2 to 10% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0791] - Optionally, sweeteners and / or flavorings,

[0792] Here:

[0793] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.11, and

[0794] - The sum of the amounts of Na, K, Mg and Ca is up to 750 mM, preferably up to 400 mM, preferably up to 200 mM.

[0795] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0796] - A total amount of protein of 10 to 45% w / w, preferably 10 to 20% w / w, relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0797] - Optionally, sweeteners and / or flavorings,

[0798] Here:

[0799] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.11, and

[0800] - The sum of the amounts of Na, K, Mg and Ca is up to 750 mM, preferably up to 400 mM, preferably up to 200 mM.

[0801] In a preferred embodiment of the present invention, a packaged, heat-treated, opaque beverage formulation has a pH in the range of 3.0 to 4.7, preferably 3.9 to 4.6, more preferably 4.0 to 4.5, and said beverage comprises:

[0802] - A total amount of protein of 2 to 45% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0803] - Optionally, sweeteners and / or flavorings,

[0804] Here:

[0805] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.11, and / or,

[0806] - Here, the protein fraction has a protein denaturation of up to 5% and / or,

[0807] - The lipid content exceeds 5% of the total energy content of the preparation.

[0808] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 3.0 to 4.7, preferably 3.9 to 4.6, more preferably 4.0 to 4.5, and the beverage comprises the following:

[0809] - A total amount of protein of 2 to 45% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0810] - Optionally, sweeteners and / or flavorings,

[0811] Here:

[0812] - Turbidity is greater than 200 NTU, preferably greater than 1000 NTU and / or,

[0813] - The viscosity is up to 200 cP.

[0814] In a preferred embodiment of the present invention, a packaged, heat-treated, opaque beverage formulation has a pH in the range of 3.0 to 4.7, preferably 3.9 to 4.6, more preferably 4.0 to 4.5, and said beverage comprises:

[0815] - A total amount of protein of 2 to 10% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0816] - Optionally, sweeteners and / or flavorings,

[0817] Here:

[0818] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio of at least 1.11 (I330 nm / I350 nm) and / or

[0819] - Here, the protein fraction has a protein denaturation degree of more than 5% and / or,

[0820] - The lipid content is up to 5% of the total energy content of the formulation.

[0821] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 3.0 to 4.7, preferably 3.9 to 4.6, more preferably 4.0 to 4.5, and the beverage comprises the following:

[0822] - A total amount of protein of 2 to 10% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0823] - Optionally, sweeteners and / or flavorings,

[0824] Here:

[0825] - Turbidity is greater than 200 NTU, preferably greater than 1000 NTU and / or

[0826] - The viscosity is up to 200 cP.

[0827] In a preferred embodiment of the present invention, a packaged, heat-treated, opaque beverage formulation has a pH in the range of 3.0 to 4.7, preferably 3.9 to 4.6, more preferably 4.0 to 4.5, and said beverage comprises:

[0828] - A total amount of protein of 10 to 45% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0829] - Optionally, sweeteners and / or flavorings,

[0830] Here:

[0831] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.11 and / or

[0832] - Here, the protein fraction has a protein denaturation of up to 5% and / or,

[0833] - The lipid content exceeds 5% of the total energy content of the preparation.

[0834] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 3.0 to 4.7, preferably 3.9 to 4.6, more preferably 4.0 to 4.5, and the beverage comprises the following:

[0835] - A total amount of protein of 10 to 45% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0836] - Optionally, sweeteners and / or flavorings,

[0837] Here:

[0838] - Turbidity is greater than 200 NTU, preferably greater than 1000 NTU and / or,

[0839] - The viscosity is up to 200 cP.

[0840] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0841] - A total amount of protein of 5 to 34% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0842] - Optionally, sweeteners and / or flavorings,

[0843] Here:

[0844] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.13, and

[0845] - The lipid content exceeds 5% of the total energy content of the preparation.

[0846] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 2.0 to 4.7, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage comprises the following:

[0847] - A total amount of protein of 5 to 10% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0848] - Optionally, sweeteners and / or flavorings,

[0849] Here:

[0850] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.13, and

[0851] - The lipid content is up to 5% of the total energy content of the formulation.

[0852] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 3.7 to 4.3, preferably 3.9 to 4.3 or preferably 4.1 to 4.3, and the beverage comprises:

[0853] - A total amount of protein of 5 to 10% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0854] - Optionally, sweeteners and / or flavorings,

[0855] Here:

[0856] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.13, and

[0857] - The lipid content is up to 5% of the total energy content of the formulation.

[0858] In a preferred embodiment of the present invention, the packaged heat-treated beverage formulation has a pH in the range of 3.7 to 4.3, preferably 3.9 to 4.3 or preferably 4.1 to 4.3, and the beverage comprises:

[0859] - A total amount of protein of 10 to 35% w / w relative to the weight of the beverage - where at least 85% w / w, preferably at least 90% w / w, of the protein is BLG - and

[0860] - Optionally, sweeteners and / or flavorings,

[0861] Here:

[0862] - The protein fraction of the beverage formulation has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.13, and

[0863] - The lipid content is up to 5% of the total energy content of the formulation.

[0864] In some embodiments of the present invention, the heat-treated beverage has a shelf life of at least 6 months at 25°C and comprises the following, having a pH in the range of 2.5 to 4.0:

[0865] - Edible BLG as defined in PCT / EP2017 / 084553 for providing a total amount of at least 1% (w / w), preferably at least 5% (w / w) of BLG,

[0866] - Sweeteners, e.g., sugar sweeteners and / or non-sugar sweeteners,

[0867] - At least one food acid, for example, citric acid or other suitable food acid,

[0868] - Optionally, flavoring agents, and

[0869] - Up to 80 mg phosphorus / 100 g protein.

[0870] In a preferred embodiment of the present invention, the invention relates to the use of a protein solution comprising a total amount of protein of 3 to 30% w / w relative to the weight of the solution, wherein at least 85 w / w %, preferably at least 90 w / w % of protein is BLG to control the turbidity of a heat-treated acidic beverage formulation having a pH in the range of 3.0 to 4.5.

[0871] In a preferred embodiment of the present invention, the invention relates to the use of a protein solution comprising a total amount of protein of 3 to 30% w / w relative to the weight of the solution, wherein at least 85 w / w%, preferably at least 90 w / w%, of the protein is BLG to control the astringent taste of a heat-treated acidic beverage formulation having a pH in the range of 2.0 to 4.0.

[0872] A preferred embodiment of the present invention relates to a heat-treated beverage formulation obtainable by one or more methods described herein.

[0873] It should be noted that the embodiments and features described in the context of one aspect of the present invention apply to other aspects of the present invention as well.

[0874] All patent and non-patent literature cited in this application is incorporated herein by reference in its entirety.

[0875] The present invention will now be described in more detail in the following non-limiting embodiments.

[0876] Examples

[0877] Example 1: Analysis Method

[0878] Example 1.1: Determination of protein naturalness by intrinsic tryptophan fluorescence

[0879] Tryptophan (Trp) fluorescence spectroscopy is a well-described tool for monitoring protein folding and unfolding. Trp residues embedded within natural proteins typically exhibit the highest fluorescence emission around 330 nm compared to when they are located at higher solvent exposure sites, such as in unfolded proteins. In unfolded proteins, the wavelength for Trp fluorescence emission typically shifts to higher wavelengths and is often measured around 350 nm. Here, the inventors utilize this shift to monitor thermally induced unfolding by calculating the ratio between fluorescence emission at 330 nm and 350 nm to investigate the effect of heating temperature.

[0880] The analysis includes the following steps:

[0881] The beverage composition was diluted to 0.6 mg / ml in MQ water.

[0882] ㆍ Transfer 300 μl of the sample to a white 96-well plate to prevent air bubbles, or transfer 3 mL to a 10 mm quartz cuvette.

[0883] The tryptophan fluorescence emission intensity between 310 nm and 400 nm was recorded from the top by excitation at 295 using a 5 nm slit.

[0884] ㆍ Samples were measured at 22°C using a Cary Eclipse fluorescence spectrophotometer equipped with a plate reader accessory (G9810A) or a single cuvette holder.

[0885] The luminescence intensity ratio was calculated by dividing the fluorescence luminescence intensity measured at 330 nm by the luminescence intensity at 350 nm (R = I330 / I350) and used as a measure of protein nativity.

[0886] o At least R in 1.11 describes the dominant natural BLG form, and

[0887] o R less than 1.11 reports at least partial unfolding and aggregation.

[0888] Example 1.2: Thermal stability at pH 3.9

[0889] Thermal stability at pH 3.9:

[0890] Thermal stability at pH 3.9 is a measure of the ability of a protein composition to remain clear during long-term pasteurization at pH 3.9.

[0891] Thermal stability at pH 3.9 is determined by the steps of: mixing a sample of powder or liquid to be tested with water (or alternatively, if it is a dilute liquid, concentrating it by low-temperature evaporation) to form an aqueous solution containing 6.0% w / w protein with a pH of 3.9, and adjusting the pH to 3.9 with the minimum required amount of 0.1 M NaOH or 0.1 M HCl.

[0892] After leaving the pH-adjusted mixture for 30 minutes, transfer 25 mL of the mixture to a 30 mL thin-walled glass test tube. Immerse the tube in a water bath at 75.0°C and heat it to 75.0°C for 300 seconds. Immediately after heating, transfer the glass test tube to an ice bath to cool it to 1 to 5°C, and measure the turbidity of the heat-treated sample according to Example 1.7.

[0893] Example 1.3: Determination of the degree of protein denaturation of a whey protein composition

[0894] It is known that denatured whey protein has lower solubility at pH 4.6 than at pH values ​​below or above pH 4.6, and therefore, 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 where the protein in solution is stable.

[0895] More specifically, in the case of whey protein, the whey protein composition to be analyzed (e.g., powder or aqueous solution) is converted as follows:

[0896] - A first aqueous solution containing 5.0% (w / w) total protein and having a pH of 7.0 or 3.0, and

[0897] - A second aqueous solution containing 5.0% (w / w) total protein and having a pH of 4.6.

[0898] pH adjustment is performed using 3% (w / w) NaOH (aqueous) or 5% (w / w) HCl (aqueous).

[0899] Total protein content of the first aqueous solution (P pH 7.0 또는 3.0 ) is determined according to Example 1.5.

[0900] The second aqueous solution is stored at room temperature for 2 hours, and then centrifuged at 3000 g for 5 minutes. A sample of the supernatant is recovered and analyzed according to Example 1.5 to obtain the supernatant (S pH4.6 Provides the protein concentration of ).

[0901] The degree of protein denaturation, D, of the whey protein composition is calculated as follows:

[0902] D =((P pH 7.0 또는 3.0 -S pH 4.6 ) / P pH 7.0 또는 3.0 )*100%

[0903] Example 1.4 Determination of protein denaturation using reverse-phase UPLC analysis (using pH 4.6 acid precipitation).

[0904] BLG samples (e.g., non-heated reference and heated BLG beverage compositions) were diluted to 2% in 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 allow precipitation aggregation of the denatured protein at a pH of around 4.6. The solution was filtered through a 0.22 μm filter to remove aggregates and non-natural proteins.

[0905] All samples were diluted to the same degree by adding polished water.

[0906] For each sample, an equal volume was loaded into a UPLC system equipped with a UPLC column (protein BEH C4; 300 Å; 1.7 μm; 150 x 2.1 mm) and detected at 214 nm.

[0907] The sample was executed using the following conditions:

[0908] Buffer A: Milli-Q water, 0.1% w / w TFA

[0909] Buffer B: HPLC-grade acetonitrile, 0.1% w / w TFA

[0910] Flow: 0.4 ml / min

[0911] Gradient: 0 to 6.00 min 24 to 45% B; 6.00 to 6.50 min 45 to 90% B; 6.50 to 7.00 min 90% B; 7.00 to 7.50 min 90 to 24% B and 7.50 to 10.00 min 24% B.

[0912] The concentration of natural BLG in the sample (5-level calibration curve) was determined using the area of ​​the BLG peak relative to the protein standard (Sigma L0130).

[0913] The sample was further diluted, and reinjected when it fell outside the linear range.

[0914] Example 1.5: Determination of Total Protein

[0915] The total protein content (net protein) of the sample is determined by the following:

[0916] 1) ISO 8968-1 / 2|IDF 020-1 / 2- Determination of Total Nitrogen in Milk Samples - Determination of Nitrogen Content - Part 1 / 2: Determination of Nitrogen Content Using the Kjeldahl Method

[0917] 2) ISO 8968-4|IDF 020-4- Determination of Non-Protein Nitrogen of Milk Samples - Determination of Nitrogen Content - Part 4: Determination of Non-Protein Nitrogen Content.

[0918] 3) Total amount of protein (m 총 질소 - m 비-단백질-질소 Calculated as )*6.38.

[0919] Example 1.6: Determination of non-aggregated BLG, ALA, and CMP

[0920] The content of non-aggregated alpha-lactalbumin (ALA), beta-lactoglobulin (BLG), and caseinomacropeptide (CMP), respectively, was analyzed by HPLC at 0.4 mL / min. 25 μL of filtered samples were injected into two TSKgel3000PWxl (7.8 mm 30 cm, Tosohass, Japan) columns connected in series with an attached pre-column PWxl (6 mm x 4 cm, Tosohass, Japan) equilibrated in an eluent (consisting of 465 g Milli-Q water, 417.3 g acetonitrile, and 1 mL trifluoroacetic acid), and a UV detector was used at 210 nm.

[0921] Natural alpha-lactalbumin (C 알파 ), beta-lactoglobulin (C 베타 ), and caseinomacropeptide (C CMP The quantitative determination of the content of ) was performed by comparing the peak area obtained for the corresponding standard protein with the peak area of ​​the sample.

[0922] The total amount of additional protein (non-BLG protein) was determined by subtracting the amount of BLG from the amount of total protein (determined according to Example 1.5).

[0923] Example 1.7: Determination of Turbidity

[0924] Turbidity is the cloudiness or haziness of a fluid caused by a large number of particles that are generally invisible to the naked eye, similar to smoke in the air.

[0925] Turbidity is measured in NTU (nephelometric turbidity units).

[0926] 20 mL of beverage / sample was added to an NTU glass and placed in a Turbiquant® 3000 IR turbidimeter. The NTU value was measured after stabilization and repeated twice.

[0927] Example 1.8: Determination of Viscosity

[0928] The viscosity of the beverage formulation was measured using a rheometer (Anton Paar, Physica MCR301).

[0929] 3.8 mL of the sample was added to cup DG26.7. The sample was equilibrated to 22°C, and then 50 seconds -1 Pre-shear for 30 seconds, followed by 30 seconds of equilibrium time and 1 s -1 and 200s -1 and 1s -1 A shear rate sweep was performed between them.

[0930] Viscosity is 100s unless otherwise specified. -1 It is expressed in units of centipoise (cP) at the shear rate. The higher the measured cP value, the higher the viscosity.

[0931] Alternatively, viscosity was estimated using Viscoman (manufacturer: Gilson) and was approximately 300s -1 It was reported at a shear rate of .

[0932] Example 1.9: Determination of color

[0933] Color was measured using a Chroma Meter (Konica Minolta, CR-400). A 15 g sample was placed in a small Petri dish (55 x 14.2 mm, VWR Cat# 391-0895) to prevent bubble formation. The protein content of the sample was standardized to a protein of 6.0 w / w % or less.

[0934] The colorimeter was calibrated with a white calibration plate (No. 19033177). The illuminant was set to D65 and the observer was set to 2 degrees. The hue (CIELAB color space, a*-, b*-, L*- values) was measured as the average of three individual readings at different locations on the Petri dish with the lid covering the suspension.

[0935] Demineralized water standards have the following values:

[0936] L* 39.97±0.3

[0937] a* 0.00±0.06

[0938] b* -0.22±0.09

[0939] The measurement was converted into a delta / difference value based on the demineralized water measurement.

[0940] Delta L* = L 6.0 w / w % 단백질로 표준화된 샘플 * - L 탈염수 *, measured at room temperature

[0941] Delta a* = a 6.0 w / w % 단백질로 표준화된 샘플 * - a 탈염수 *, measured at room temperature

[0942] Delta b* = b 6.0 w / w % 단백질로 표준화된 샘플 * - b 탈염수 *, measured at room temperature

[0943] The sample is standardized to less than 6.0 w / w % protein.

[0944] The L*a*b* color space (also referred to as the CIELAB space) is one of the uniform color spaces defined by the International Commission on Illumination (CIE) in 1976 and was used to quantitatively report brightness and hue (ISO 11664-4:2008(E) / CIE S 014- 4 / E:2007).

[0945] In this space, L* represents brightness (values ​​from 0 to 100), the darkest black at L* = 0, and the brightest white at L* = 100.

[0946] Color channels a* and b* represent true neutral gray values ​​at a* = 0 and b* = 0. The a* axis represents the green-red component, where green is in the negative direction and red is in the positive direction. The b* axis represents the blue-yellow component, where blue is in the negative direction and yellow is in the positive direction.

[0947] Example 1.10 Beverage Stability Test / Insoluble Protein Material

[0948] When less than 15% of the total protein precipitates from a heated sample during centrifugation at 3000 g for 5 minutes, the whey protein drink composition is considered stable.

[0949] Approximately 20 g of the sample was added to a centrifuge tube and centrifuged at 3000 g for 5 minutes.

[0950] Protein recovery was quantified using Kjeldahl analysis of the protein before centrifugation and the supernatant after centrifugation. Refer to Example 1.5.

[0951] Protein loss is calculated as follows:

[0952]

[0953] Additionally, this parameter is sometimes referred to as the level of insoluble protein material and can be used to analyze both liquid and powder samples. If the sample is a powder, 10 g of powder is suspended in 90 g of deionized water and hydrated at 22°C for 1 hour under gentle stirring. Approximately 20 g of the sample (e.g., liquid sample or suspended powder sample) was added to a centrifuge tube and centrifuged at 3000 g for 5 minutes. Before centrifugation (P 총 ) Protein and after centrifugation (P 3000xg Protein recovery according to Example 1.5 was quantified using Kjeldahl analysis of the supernatant.

[0954] The amount of insoluble protein material is calculated as follows:

[0955]

[0956] Example 1.11: Sensory Evaluation

[0957] The heat-treated beverage formulations underwent descriptive sensory evaluation. The beverage formulations were heat-treated using a plate heat exchanger.

[0958] Mix 1 volume of sample with 1 volume of water, compare with non-heated whey protein isolate, and also use lactic acid and citric acid to form an attribute list prior to the final tasting session:

[0959]

[0960] Participants' mouths were cleaned between each sample using crackers, white tea, melon, and water.

[0961] A 15 mL test sample was placed in a small cup at ambient temperature (20 to 25℃).

[0962] Test samples were provided three times to 10 individuals in each of three different blocks in a random order.

[0963] The properties (see table above) were evaluated on a 15 cm scale with 0 = low strength and 15 = high strength.

[0964] Statistical analysis was performed in 'Panelcheck' software using a 3-way ANOVA test with multiple replications. The samples were fixed and the panel was randomized.

[0965] Significant differences between samples were evaluated using Bonferroni correction, which implies the minimum significant difference value (pair comparison of groups related to the character).

[0966] Example 1.12: Determination of transparency by imaging

[0967] A photograph of the beverage formulation was taken by placing a sample in a turbidity NTU measuring vial that came into contact with a piece of paper containing the text 'lorem ipsum'. The vial was photographed using a smartphone, and the inventors evaluated whether the text could be clearly observed through the vial.

[0968] Example 1.13: Determination of Ash Content

[0969] The ash content of food products is determined according to NMKL 173:2005 "Ashes in food, weight measurement".

[0970] Example 1.14: Determination of Conductivity

[0971] The "conductivity" (sometimes referred to as "specific conductivity") of an aqueous solution is a measure of the solution's ability to conduct electricity. Conductivity can be determined, for example, by measuring the AC resistance of the solution between two electrodes, and the results are typically provided in millisiemens per centimeter (mS / cm). Conductivity can be measured, for example, according to the EPA (the US Environmental Protection Agency) Method No. 120.1.

[0972] Conductivity values ​​mentioned herein have been normalized to 25°C unless otherwise specified.

[0973] Conductivity is measured using a conductivity meter (WTW Cond 3210 with Tetracon 325 electrodes).

[0974] The system is calibrated as described in the manual before use. The electrode is thoroughly rinsed in the same type of medium where the measurement is to be performed to avoid local dilution. The electrode is lowered into the medium so that the area where the measurement takes place is completely submerged. Then, the electrode is shaken to remove any trapped air. Then, the electrode is held in place until a stable value is obtained and can be recorded from the display.

[0975] Example 1.15: Determination of total solids in the solution

[0976] The total solids of the solution are [NMKL 110 2 nd It can be determined according to [Edition, 2005 (Total solids (Water) - Gravimetric determination in milk and milk products)]. NMKL is an abbreviation for "Nordic Food Methodology Committee (Nordisk Metodikkomite for Naeringsmidler)".

[0977] The water content of the solution can be calculated as 100% - the relative amount of total solids (% w / w).

[0978] Example 1.16: Determination of pH

[0979] All pH values ​​are measured using a pH glass electrode and normalized to 25°C.

[0980] The pH glass electrode (with temperature compensation) is carefully rinsed and compensated before use.

[0981] If the sample is in liquid form, the pH is measured directly in the liquid solution at 25°C.

[0982] If the sample is a powder, dissolve 10 grams of powder in 90 ml of deionized water at room temperature while stirring vigorously. Then, the pH of the solution is measured at 25°C.

[0983] Example 1.17: Determination of loose density and bulk density

[0984] The density of a dry powder is defined as the relationship between the weight and volume of the powder analyzed using a special Stampf volumetric meter (i.e., a measuring cylinder) under specified conditions. Density is typically expressed in g / ml or kg / L.

[0985] In this method, a sample of the dried powder is placed in a measuring cylinder. After a specified number of taps, the volume of the product is read and the density is calculated.

[0986] Three types of density can be defined by this method:

[0987] ■ Poured density, calculated by dividing the mass by the volume of the powder after transferring to the specified measuring cylinder

[0988] ■ Loose density, calculated by dividing the mass by the volume of the powder after 100 taps according to the conditions specified in this standard.

[0989] ■ Bulk density, calculated by dividing the mass by the volume of the powder after 625 taps according to the conditions specified in this standard.

[0990] The above method uses a special measuring cylinder, 250 ml, graduated from 0 to 250 ml, with a weight of 190 ± 15 g (J. Engelsmann AG 67059 Ludwigshafen / Rh) and a Stampf volumetric measuring instrument, e.g., J. Engelsmann A. G.

[0991] The loose density and bulk density of the dried product are determined by the following procedure.

[0992] Preprocessing:

[0993] The sample to be measured is stored at room temperature.

[0994] Next, rotate and turn the container repeatedly to thoroughly mix the sample (to prevent particle crushing). The container is not filled more than 2 / 3 full.

[0995] procedure:

[0996] Weigh 100.0 ± 0.1 grams of powder and transfer it to a measuring cylinder. Read the volume V0 in ml units.

[0997] If 100 g of powder does not fit into the cylinder, the amount must be reduced to 50 or 25 g.

[0998] Secure the measuring cylinder to the Stampf volumetric meter and tap it 100 times. Flatten the surface with a spatula and volume V 100 It is read in ml units.

[0999] Change the number of taps to 625 (including 100 taps). After tapping, flatten the surface, and volume V 625 It is read in ml units.

[1000] Density calculation:

[1001] Calculate the loose density and bulk density in g / ml according to the following formula:

[1002] Bulk density = M / V

[1003] Here, M represents the weighed sample in grams, and V represents the volume after 625 taps in ml.

[1004] Example 1.18: Determination of moisture content of powder

[1005] The moisture content of food products is determined according to ISO 5537:2004 (milk powder - determination of moisture content (reference method)). NMKL is an abbreviation for the "Nordic Food Methodology Committee (Nordisk Metodikkomite for Naeringsmidler)".

[1006] Example 1.19: Determination of amounts of calcium, magnesium, sodium, potassium, and phosphorus (ICP-MS method)

[1007] The total amount of calcium, magnesium, sodium, potassium, and phosphorus is determined using a procedure to first decompose the sample using microwave digestion, and then the total amount of mineral(s) is determined using an ICP device.

[1008] device:

[1009] The microwave is from the manufacturer (Anton Paar) and the ICP is the Optima 2000DV from the manufacturer (PerkinElmer Inc.).

[1010] ingredient:

[1011] 1 M HNO3

[1012] Yttrium in 2% HNO3

[1013] Standards suitable for calcium, magnesium, sodium, potassium, and phosphorus in 5% HNO3

[1014] Preprocessing:

[1015] Weigh 0.2 grams of the powder sample or 1 g of the liquid sample and transfer the powder to a microwave digestion tube. Add 5 mL of 1 M HNO3. Dilute the sample in the microwave according to the microwave instructions. Place the digested tube in a fume cupboard, remove the lid, and allow the volatile fumes to evaporate.

[1016] Measurement Procedure:

[1017] Transfer the pretreated sample to DigiTUBE using a known amount of Milli-Q water. Add 2% HNO3 yttrium solution to the digestion tube (approx. 0.25 mL per 50 mL diluted sample) and dilute to a known volume using Milli-Q water. Analyze the sample in ICP using the procedure described by the manufacturer.

[1018] A blind sample is prepared by using Milli-Q water to dilute a mixture of 10 mL of 1 M HNO3 and 0.5 mL of yttrium solution in 2% HNO3 to a final volume of 100 mL.

[1019] Prepare at least three standard samples with concentrations in parentheses, with the expected sample concentrations.

[1020] The detection limit for liquid samples is 0.005 g / 100 g sample for Ca, Na, K and Phosphorus, and 0.0005 g / 100 g sample for Mg. The detection limit for powder samples is 0.025 g / 100 g sample for Ca, Na, K and Pho, and 0.0005 g / 100 g sample for Mg.

[1021] In cases where the detection limit of Pho is below, the value of the detection limit is used in the examples to demonstrate the maximum amount of Pho existing in the worst-case scenario.

[1022] Example 1.20: Determination of Furosine Value:

[1023] The furosine value is determined as described in the literature ["Maillard Reaction Evaluation by Furosine Determination During Infant Cereal Processing", Guerra-Hernandez et al, Journal of Cereal Science 29(1999) 171-176], and the total amount of protein is determined according to Example 1.5. The furosine value is reported in mg of furosine per 100 g of protein.

[1024] Example 1.21: Determination of the degree of crystallinity of BLG in liquid

[1025] The following method is used to determine the degree of crystallization of BLG in a liquid having a pH in the range of 5 to 6.

[1026] a) Transfer a 10 mL sample of the liquid to a Maxi-Spin filter with a 0.45 micron pore size CA membrane.

[1027] b) Immediately spin the filter at 1500 g for 5 minutes. Maintain the centrifuge at 2°C.

[1028] c) Add 2 mL of cold Milli-Q water (2°C) to the retentate side of the spin filter, immediately centrifuge the filter at 1500 g for 5 minutes while keeping the centrifuge cooled to 2°C, collect the permeate (permeate A), measure the volume, and determine the BLG concentration by HPLC using the method summarized in Example 1.31.

[1029] d) Add 4 mL of 2 M NaCl to the retaining surface of the filter, shake rapidly, and let the mixture sit at 25°C for 15 minutes.

[1030] e) Immediately rotate the filter at 1500 g for 5 minutes and collect the permeate (permeate B).

[1031] f) Determine the total weight of BLG in permeate A and permeate B using the method summarized in Example 1.31, and convert the result into the total weight of BLG instead of weight percentage. The weight of BLG in permeate A is m 투과액 A It is referred to as, The weight of BLG in permeate B is m 투과액 B It is referred to as.

[1032] g) For BLG, the degree of crystallinity of the liquid is determined as follows:

[1033] Degree of crystallization = m 투과액 B / (m 투과액 A + m 투과액 B )*100%

[1034] Example 1.22: Determination of the degree of crystallinity of BLG in dry powder

[1035] This method is used to determine the degree of crystallinity of BLG in dry powder.

[1036] a) Mix 5.0 grams of powder sample with 20.0 grams of cold Milli-Q water (2°C) and let it stand for 5 minutes at 2°C.

[1037] b) Transfer a sample of the liquid to a Maxi-Spin filter with a 0.45 micron CA membrane.

[1038] c) Immediately spin the filter at 1500 g for 5 minutes. Maintain the centrifuge at 2°C.

[1039] d) Add 2 mL of cold Milli-Q water (2°C) to the retention side of the spin filter, immediately spin the filter at 1500 g for 5 minutes, collect the permeate (Permeate A), measure its volume, determine the BLG concentration via HPLC using the method summarized in Example 1.31, and convert the result to a result relative to the total weight of BLG instead of a weight percentage. The weight of BLG in Permeate A is m 투과액 A It is referred to as.

[1040] f) Next, the degree of crystallinity of BLG in the powder is calculated using the following formula:

[1041]

[1042] Here, m BLG 총 is the total amount of BLG in the powder sample of step a).

[1043] If the total amount of BLG in the powder sample is unknown, it can be determined by suspending another 5 g powder sample (from the same powder source) in 20.0 g of Milli-Q water, adding aqueous NaOH to adjust the pH to 7.0, leaving the mixture at 25°C for 1 hour under stirring, and finally determining the total amount of BLG in the powder sample using Example 1.31.

[1044] Example 1.23: Determination of UF permeate conductivity

[1045] Transfer 15 mL of the sample to an Amicon Ultra-15 centrifuge filter device with a 3 kDa cutoff (3000 NMWL) and centrifuge at 4000 g for 20 to 30 minutes, or until a sufficient volume of UF permeate for conductivity measurement accumulates at the bottom of the filter device. Conductivity is measured immediately after centrifugation. Sample handling and centrifugation are performed at the temperature of the sample source.

[1046] Example 1.24: Detection of dried BLG crystals in powder

[1047] The presence of dried BLG crystals in the powder can be confirmed in the following way:

[1048] A sample of the powder to be analyzed was resuspended, gently mixed in deionized water at a temperature of 4°C in a weight ratio of 2 parts water to 1 part powder, and rehydrated at 4°C for 1 hour.

[1049] The rehydrated sample is examined under a microscope to confirm the presence of crystals, and preferably, birefringence is detected using plane polarization.

[1050] To prove the existence of a crystal structure, it is desirable to isolate the crystalline material and process it by X-ray crystallography, and also to verify whether the crystal lattice (space group and unit cell dimensions) matches the crystal lattice of the BLG crystal.

[1051] Analyze the chemical composition of the separated crystalline material to verify whether its solid is mainly composed of BLG.

[1052] Example 1.25: Determination of the total amount of lactose

[1053] The total amount of lactose is determined according to: ISO 5765-2:2002 (IDF 79-2: 2002) "Dried milk, dried ice-mixes and processed cheese - Determination of lactose content - Part 2: Enzymatic method utilizing the galactose moiety of the lactose".

[1054] Example 1.26: Determination of Total Amount of Carbohydrates:

[1055] The amount of carbohydrates is determined using the Sigma Aldrich Total Carbohydrate Test Kit (Cat MAK104-1KT), in which carbohydrates are hydrolyzed and converted into furfural and hydroxyfurfural, which are converted into chromagen monitored by a spectrophotometer at 490 nm.

[1056] Example 1.27: Determination of Total Amount of Lipids

[1057] The amount of lipids is determined according to ISO 1211:2010 (determination of fat content - Rose-Gottlieb weight method).

[1058] Example 1.28: Determination of Brix

[1059] Brix measurements were performed using a PAL-α digital handheld refractometer (Atago) calibrated for polished water (water filtered by reverse osmosis to obtain a conductivity of up to 0.05 mS / cm).

[1060] Approximately 500 μl of sample was transferred to the prism surface of the instrument and measurement was started. The measured value was read and recorded.

[1061] The Brix of a whey protein solution is proportional to the content of total solids (TS), and TS (% w / w) is approximately Brix * 0.85.

[1062] Example 1.29 Determination of Lactoferrin and Lactoperoxidase

[1063] The concentration of lactoferrin is determined by ELISA immunoassay as summarized by the literature [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)].

[1064] The concentration of lactoperoxidase is determined using a commercially available bovine lactoperoxidase kit.

[1065] Example 1.30: Determination of the number of colony-forming units

[1066] The determination of the number of colony-forming units per gram of sample is performed according to: ISO 4833-1:2013(E): Microbiology of food and animal feed - Horizontal method for microbial counting - Colony counting technique at 30°C.

[1067] Example 1.31: Determination of the total amount of BLG, ALA, and CMP

[1068] 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 of the composition. Unlike the method of Example 1.6, this method also measures proteins present in aggregated form and thus provides a measurement of the total amount of protein species in the composition.

[1069] The separation mode is size exclusion chromatography (SEC), and the method uses 6 M guanidine HCl buffer as both the sample solvent and the HPLC mobile phase. Mercaptoethanol is used as a reducing agent to reduce disulfides (SS) in proteins or protein aggregates to produce unfolded monomer structures.

[1070] Sample preparation is easily achieved by dissolving 10 mg of protein equivalent in the mobile phase.

[1071] Two TSK-GEL G3000SWXL (7.7 mm x 30.0 cm) columns (GPC columns) and a guard column are arranged in series to achieve proper separation of major proteins in the raw material.

[1072] The eluted analyte is detected and quantified by UV detection (280 nm).

[1073] Equipment / Materials:

[1074] 1. HPLC Pump 515 (Waters) with Manual Seal Cleaning Function

[1075] 2. HPLC Pump Controller Module II (Waters)

[1076] 3. Autosampler 717 (Waters)

[1077] 4. Dual absorbance detector 2487 (Waters)

[1078] 5. Computer software capable of generating quantitative reports (Empower 3, Waters)

[1079] 6. Analysis columns: 2 TSK-GEL G3000SWXL (7.8 x 300 mm, P / N: 08541).

[1080] Guard Column: TSK-Guard Column SWxL(6.0 x 40 mm, P / N: 08543).

[1081] 7. Ultrasonic Bath (Branson 5200)

[1082] 8. 25 mm syringe filter with 0.2 μm cellulose acetate membrane (514-0060, VWR)

[1083] procedure:

[1084] Lee Dong-sang:

[1085] A. Stock buffer solution.

[1086] 1. Weigh 56.6 g of Na2HPO4, 3.5 g of NaH2PO4, and 2.9 g of EDTA into a 1000 mL beaker. Dissolve them in 800 mL of water.

[1087] 2. Measure the pH and, if necessary, adjust it to 7.5 ± 0.1 using HCl (to decrease pH) or NaOH (to increase pH).

[1088] 3. Transfer to a 1000 mL volumetric flask and dilute the volume with water.

[1089] B. 6 M guanidine HCl mobile phase.

[1090] 1. Weigh 1146 g of guanidine HCl into a 2000 mL beaker and add 200 mL of stock buffer solution (A).

[1091] 2. Dilute this solution with water to about 1600 mL while mixing using a magnetic stirring bar (50°C).

[1092] 3. Adjust the pH to 7.5 ± 0.1 using NaOH.

[1093] 4. Transfer to a 2000 mL volumetric flask and dilute the volume with water.

[1094] 5. It is filtered using a solvent filtration device equipped with a 0.22 μm membrane filter.

[1095] Correction standard.

[1096] The calibration standards for each protein to be quantified are prepared in the following manner:

[1097] 1. Accurately weigh about 25 mg of protein reference standard (to the right of 0.01 mg) into a 10 mL volumetric flask and dissolve it in 10 mL of water.

[1098] This is a standard protein stock solution (S1) of protein.

[1099] 2. Pipette 200 μl of S1 into a 20 mL volumetric flask and dilute the volume with the mobile phase.

[1100] This is the low working standard solution WS1.

[1101] 3. Pipette 500 μL of S1 into a 10 mL volumetric flask and dilute the volume with the mobile phase.

[1102] This is standard solution WS2.

[1103] 4. Pipette 500 μL of S1 into a 5 mL volumetric flask and dilute the volume with the mobile phase. .

[1104] This is standard solution WS3.

[1105] 5. Pipette 750 μL of S1 into a 5 mL volumetric flask and dilute the volume with the mobile phase.

[1106] This is standard solution WS4.

[1107] 6. Pipette 1.0 mL of S1 into a 5 mL volumetric flask and dilute the volume with the mobile phase.

[1108] This is a high working standard solution WS5.

[1109] 7. Using a graduated disposable pipette, transfer 1.5 mL of WS1 to 5 into a separate vial.

[1110] Add 10 μL of 2-mercaptoethanol to each vial and cap. Vortex the solution for 10 seconds.

[1111] Maintain the standard at ambient temperature for about 1 hour.

[1112] 8. Filter the standard using a 0.22 μm cellulose acetate syringe filter.

[1113] The purity of the protein is measured using Kjeldahl (N x 6.38) and HPLC as the area % of the standard solution WS5.

[1114] Protein (mg) = "Standard protein weight" (mg) x P1 x P2

[1115] P1 = P%(Kil'dal)

[1116] P2 = Protein Area % (HPLC)

[1117] Sample manufacturing

[1118] 1. Weigh 25 mg of the protein equivalent of the original sample into a 25 mL volumetric flask.

[1119] 2. Add approximately 20 mL of mobile phase and dissolve the sample for about 30 minutes.

[1120] 3. Add the mobile phase to the volume, and add 167 μL of 2-mercaptoethanol to 25 mL of the sample solution.

[1121] 4. After sonicating for about 30 minutes, keep the sample at ambient temperature for about 1½ hours.

[1122] 5. Mix the solution and filter it using a 0.22 μl cellulose acetate syringe filter.

[1123] HPLC System / Column

[1124] Column equilibration

[1125] 1. Connect the GPC guard column and two GPC analysis columns in series.

[1126] New columns are typically shipped with phosphate-salt buffer.

[1127] 2. Gradually flow water through the new column at a rate of 0.1 mL / min to 0.5 mL / min over 30 to 60 minutes.

[1128] Continue flushing for about 1 hour.

[1129] 3. Gradually decrease the flow rate from 0.5 mL / min to 0.1 mL / min and replace the mobile phase in the reservoir.

[1130] 4. To prevent pressure shock and maintain at 0.5 mL / min, gradually increase the pump flow rate from 0.1 mL / min to 0.5 mL / min within 30 to 60 minutes.

[1131] 5. Inject 10 samples to saturate the column and wait until the peak elutes.

[1132] This will help with column conditioning.

[1133] This step is performed without having to wait for each injection to be completed before the next injection.

[1134] 6. Equilibrate with the mobile phase for at least 1 hour.

[1135] Calculation of results

[1136] The quantitative determination of the content of proteins to be quantified, e.g., alpha-lactalbumin, beta-lactoglobulin, and caseinomacropeptide, is performed by comparing the peak area obtained for the corresponding standard protein with the peak area of ​​the sample. The results are reported as g of the specific protein per 100 g of the original sample, or as a weight percentage of the specific protein relative to the weight of the original sample.

[1137] Example 2: Production of spray-dried, acidic BLG isolate powder

[1138] whey protein supply

[1139] Lactose-depleted UF retained from sweet whey from a standard cheese production process was filtered through a 1.2 micron filter and fat-reduced through a Synder FR membrane before being used as a feed for the BLG crystallization process. The chemical composition of the feed can be seen in Table A. The inventors note that all weight percentages of specific proteins, such as BLG and ALA, mentioned in this example relate to the weight percentage of non-aggregated protein relative to total protein.

[1140] conditioning

[1141] Sweet whey feed was filtered with a feed concentration of 21% Total Solids (TS) ± 5 using polished water (water filtered by reverse osmosis to obtain a conductivity of up to 0.05 mS / cm) as the normal volume filtration medium, 46 mil spacers, and a feed pressure of 1.5 to 3.0 bar using a Koch HFK-328 type membrane (70 m 2 It was conditioned at an ultrafiltration setting at 20°C using a membrane. Subsequently, HCl was added to adjust the pH to approximately 5.5. Volumetric filtration was continued over a period of 20 minutes until the conductivity drop of the retained material was less than 0.1 mS / cm. Subsequently, the permeate flow rate was 1.43 L / h / m² 2 The retention was concentrated until it was less than [amount]. The first sample of the concentrated retention was taken and centrifuged at 3000 g for 5 minutes. The supernatant of the first sample was used to determine the BLG yield.

[1142] crystallization

[1143] The concentrated retention was transferred to a 300 L crystallization tank and seeded with pure BLG crystalline material made from rehydrated and spray-dried BLG crystals. Afterwards, the seeded whey protein solution was cooled from 20°C to approximately 6°C over approximately 10 hours to allow BLG crystals to form and grow.

[1144] After cooling, a sample of the crystal-containing whey protein solution (second sample) was taken, and the BLG crystals were separated by centrifugation at 3000 g for 5 minutes. The supernatant and crystal pellet from the second sample were analyzed by HPLC as described below. The crystallization yield was calculated as summarized below and was determined to be 57%.

[1145] [Table A]

[1146] Chemical composition of the supply

[1147]

[1148] Determination of BLG Yield Using HPLC:

[1149] The supernatants of the first and second samples were diluted to the same degree by adding polished water, and the diluted supernatants were filtered through a 0.22 μm filter. For each filtered and diluted supernatant, an equal volume was loaded into an HPLC system equipped with a Phenomenex Jupiter® 5 μm C4 300 Å, LC column 250 x 4.6 mm, Ea., and detected at 214 nm.

[1150] The sample was executed using the following conditions:

[1151] Buffer A: MilliQ water, 0.1% w / w TFA

[1152] Buffer B: HPLC-grade acetonitrile, 0.085% w / w TFA

[1153] Flow: 1 mL / min

[1154] Column temperature: 40℃

[1155] Gradient: 0 to 30 min 82 to 55% A and 18 to 45% B; 30 to 32 min 55 to 10% A and 45 to 90% B; 32.5 to 37.5 min 10% A and 90% B; 38 to 48 min 10 to 82% A and 90 to 18% B.

[1156] Data Processing:

[1157] Since both supernatants were treated in the same manner, the relative yield can be calculated by directly comparing the areas of the BLG peaks. Since the crystals contain only BLG and all samples were treated in the same manner, the concentration of alpha-lactalbumin (ALA) and, consequently, the area of ​​ALA must 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.

[1158]

[1159] The relative yield is calculated by the following formula:

[1160]

[1161] Acid dissolution of BLC crystals

[1162] The remaining material from the crystallization tank was separated using a decanter at a feed flow rate of 75 L / h, 64 spacers, and 350 g, 2750 RPM, and 150 RPM Diff. before separating the feed mixed with polished water in a 1:2 ratio. Subsequently, the BLG crystal / solid phase from the decanter was mixed with polished water to form a thinner slurry before adding phosphoric acid or HCl to rapidly dissolve the crystals and lower the pH to approximately 3.0.

[1163] After dissolving the BLG crystals, the pure BLG protein liquid was concentrated to 15 Brix at the same UF setting used to prepare the feed for crystallization, and the pH was adjusted to a final pH of approximately 3.8. Subsequently, the liquid BLG isolate was heated to 75°C for 5 minutes and then cooled to 10°C. Heat treatment was found to reduce the microbial load from 137,000 CFU / g before heat treatment to <1,000 CFU / g after heat treatment. Heat treatment did not cause any protein denaturation, and the intrinsic tryptophan fluorescence ratio (I330 nm / I350 nm) was determined to be 1.20, which indicated the natural form of the BLG molecule.

[1164] BLG was dried in a pilot plant spray dryer with an inlet temperature of 180°C and an outlet temperature of 75°C. The resulting powder sampled at the outlet had a moisture content of approximately 4% w / w, and the chemical composition of the powder is shown in Table B. A sample of the dried powder was dissolved, the degree of protein denaturation was determined to be 1.5%, and the intrinsic tryptophan fluorescence emission ratio (I330 / I350) was measured as 1.20.

[1165] [Table B]

[1166] Composition of BLG isolated powder (BDL = below detection limit)

[1167]

[1168] The bulk density (625 tab) of the spray-dried powder is 0.2 to 0.3 g / cm³ 3 It was estimated as.

[1169] Example 3: Preparation of a standard whey protein drink

[1170] Dried BLG isolate protein powder containing ≥85% BLG based on protein is dispersed in up to about 95% of deionized water required to reach the desired final protein concentration. Acidic BLG isolate powder is produced as summarized in Example 2, while pH 5.5 BLG isolate powder is produced as summarized in Example 2 of PCT / EP2017 / 084553.

[1171] As described in PCT / EP2017 / 084553, the dissolution of BLG substances may be aided by the addition of an acid (selected from one or more food-grade acids such as phosphoric acid, hydrochloric acid, citric acid, malic acid, or salts in their dissolved or powdered forms). If the pH decreases during dissolution due to the addition of acid, the pH should preferably not exceed the desired target pH (i.e., avoid unnecessary titration with acid and / or base).

[1172] Optionally, other ingredients including minerals, sweeteners, flavorings, stabilizers, emulsifiers, or sources of fats and carbohydrates may be added. The process used includes the following steps:

[1173] o If fat is included, the first step is to heat the oil to 70°C in a water bath.

[1174] o Step of mixing with an emulsifier (typically up to 0.2 w / w % in the final recipe); for example, the emulsifier Grindsted Citrem LR10, a citric acid ester, is recommended for phosphate-free applications. Allow cooling to 60°C (to avoid / reduce potential denaturation / aggregation when mixed with protein).

[1175] o Step of slowly mixing with preheated water (60℃).

[1176] o A step of adding all powder ingredients in a pre-mixed form (to avoid 'fisheyes'); this includes the pre-mixing of carbohydrates and proteins.

[1177] o Optionally, a step of adding minerals (NaCl, KCl, CaCl2, and MgCl2) to achieve Na, K, Ca, and Mg concentrations that comply with Foods for Special Medical Purposes (FSMP) requirements (the target is the middle of the acceptable range). The minerals are dissolved in demineralized water and added to reach the desired concentrations of Na, K, Ca, and Mg. Other food-grade minerals that comply with FMSP may be additionally used and added in dissolved or powder form.

[1178] o If necessary, a step of finally adjusting the pH using up to 10% phosphoric acid (or other food-grade acid) or up to 10% NaOH.

[1179] o Step of adding the remaining water to reach the desired protein concentration and

[1180] o A step in which the composition is optionally homogenized ('upstream homogenization').

[1181] o Step of heat-treating the composition.

[1182] o Random homogenization step ('downstream homogenization').

[1183] For comparison, whey protein isolates replace ≥ 85% BLG products when making reference samples while preserving the remaining steps.

[1184] The sample was stored at 20°C in a dark environment.

[1185] Example 4: Heat treatment of whey protein composition

[1186] Heat treatment of the beverage was performed by heating at 120°C for 20 seconds (High Temperature, Short Time (HTST), resulting in BLG denaturation) or at 75°C for a holding time of 15 seconds to 5 minutes (BLG remains in its natural state) using a plate heat exchanger (manufacturer: OMVE HTST / UHT Pilot Plant HT320-20) equipped with a 10 μm coupled microfiber filter element, code 12-57-60k (headline filter). Other heat treatment conditions may also be applied.

[1187] The heat-treated beverage composition was tapped into a 100 mL sterile bottle at 75 to 85°C, then immediately sealed and placed on ice.

[1188] In other experiments, heat treatment was performed by transferring the whey protein source into thin-walled glass vials containing 15 to 30 mL of sample. The vials were immersed in a pre-equilibrated water bath at a target temperature in the range of 75°C to 95°C for 1 to 5 minutes, and then cooled on ice.

[1189] Example 5: Production of heat-treated beverage formulations

[1190] In this example, LG beverages and WPI beverages containing 6% protein and having a pH of 3.7 were prepared.

[1191] The BLG beverage was prepared by dissolving pH 5.5 BLG isolated powder (as described in Example 7 of PCT / EP2017 / 084553) in deionized water at 10°C. 10% H3PO4 was slowly added to the solution. The final pH was adjusted to pH 3.7.

[1192] The solution was heat-treated at 120°C for 20 seconds using a plate heat exchanger, or at 75°C for a holding time of 15 seconds to 5 minutes as described in Example 4. The beverage was tapped to provide a heat-sterilized whey protein beverage composition.

[1193] WPI beverages use the same procedure but are manufactured from WPI powder.

[1194] Table 1 below presents the composition of BLG powder used for the manufacture of beverage formulations, and the composition of WPI is also listed for comparison.

[1195] [Table 1]

[1196] Composition of BLG powder (pH 5.5 powder) and WPI powder

[1197]

[1198] A beverage formulation containing BLG and WPI, with a pH of 3.7 and a protein content of 6% w / w, was heat-treated at 120°C for 20 seconds and at 75°C for 15 seconds, wherein 95.9 w / w % of the protein was BLG. In the WPI beverage (WPI-B), 57 w / w % of the protein was BLG. The turbidity (Example 1.7), viscosity (Example 1.8), and color (Example 1.9) of the different samples were analyzed.

[1199] The results are presented in Table 2 and Figure 1 below.

[1200] [Table 2]

[1201]

[1202] conclusion:

[1203] The turbidity of the BLG sample remained low at 75°C, whereas the WPI sample had high turbidity. The WPI sample was also opaque. Refer to Fig. 1.

[1204] The sterilized BLG sample had a turbidity of 7.0 NTU compared to WPI, which had a turbidity of 263 NTU.

[1205] Viscosity was also kept low.

[1206] Therefore, a clear beverage with a protein content of approximately 96 w / w% and a BLG content at pH 3.7 can be produced, whereas this is not possible with an opaque WPI sample under the same conditions.

[1207] Example 6a: Demonstrated that the accessible pH range for clear whey protein drinks can be extended.

[1208] A BLG sample was prepared in which approximately 92 w / w % of the 6 w / w % protein was BLG, and for comparison, two different WPI samples were prepared containing approximately 60 w / w % (WPI-A) and 57 w / w % (WPI-B) of BLG, respectively.

[1209] The 6 w / w % whey protein compositions were prepared as described in Example 3, with the final pH adjusted using 10% phosphoric acid to obtain selected pH values ​​between 3.0 and 3.9, respectively (BLG isolate powder was prepared according to Example 2). In one embodiment of the experiment, samples adjusted to pH levels between 3.0 and 3.9 were UHT treated at 120°C for 20 seconds, tapped, sealed, and cooled. In another embodiment of the experiment, pH 3.0 and 3.9 samples were pasteurized at 75°C for 15 seconds as described in Example 4.

[1210] The turbidity (Example 1.7), viscosity (Example 1.8), color (Example 1.9), and visual appearance (Example 1.12) of different samples were analyzed.

[1211] The results are presented in Figures 2 to 10.

[1212] result:

[1213] Figure 2 shows images of WPI-B at pH 3.0 to 3.7 and BLG beverages at pH 3.7, heat-treated at 120°C for 20 seconds. Figure 3 shows images of WPI-B at pH 3.0 to 3.7, heat-treated at 75°C, and BLG at pH 3.7, heat-treated at 75°C for 15 seconds. Figure 4 shows images of WPI-B at pH 3.7 and BLG beverages at pH 3.9, heated at 75°C for 15 seconds.

[1214] Surprisingly, the inventors discovered that the BLG beverage formulation remains visually clear at pH 3.7 when sterilized in UHT (Fig. 2), and can even exceed pH 3.7 (pH 3.9 to 4.1) when the WPI is pasteurized under opaque conditions (Figs. 3 and 4). These findings are further supported by turbidity measurements, as shown in Figs. 5 (UHT) and 6 (pasteurization), where the WPI remains below 40 NTU even at pH 3.7 and 3.9, respectively, where the WPI significantly exceeds 40 NTU.

[1215] The viscosity of the BLG beverage formulation remains low upon UHT treatment. The low viscosity demonstrates that the beverage sample was easy to drink. In particular, when using WPI at high pH values, the viscosity increases dramatically (Fig. 7).

[1216] The inventors also found that the yellowness (b*-value) of a heat-treated WPI beverage containing a small amount of BLG (both UHT and pasteurization) significantly exceeds that of BLG up to at least pH 3.7. Refer to Figs. 8 (UHT) and 9 (pasteurization).

[1217] conclusion:

[1218] Using a whey protein drink in which at least 85% w / w of protein is BLG enables at least two significant opportunities to provide consumers with a whey protein drink having desired characteristics:

[1219] 1. Increase pH during heat treatment to improve visual perception (color, turbidity) and viscosity compared to WPI.

[1220] 2. Pasteurization further expands the accessible pH range while maintaining the benefits of 1).

[1221] Example 6b: Demonstrated that the accessible pH range for clear whey protein drinks can be extended.

[1222] In this example, BLG beverages containing 6 wt% and 12 wt% protein and having pH values ​​of 2.7, 3.0, 3.3, and 3.7 were prepared as described in Example 3. The BLG powder used for the preparation of the beverage is presented in Table 3, and the powder contains 98.2 w / w% protein as BLG. The amounts of Na, K, Ca, and Mg are below detection levels.

[1223] [Table 3]

[1224] Composition of BLG isolated powder (BDL = below detection limit)

[1225]

[1226] The final pH of the beverage was adjusted to pH 2.7, 3.0, 3.3, and 3.7 using 1 M phosphoric acid. The beverage was heat-treated at 75°C or 95°C for 5 minutes using a water bath as summarized in Example 4.

[1227] Naturalness was analyzed by the turbidity of the protein (Example 1.7) and the unique tryptophan fluorescence emission ratio R=I330 / I350 of different samples (Example 1.1).

[1228] The results are presented in Table 4.

[1229] [Table 4]

[1230] Analysis data of high-protein beverages (6 and 12 w / w %) prepared from BLG at pH 2.7 to 3.7 while heating at 75°C and 95°C for 5 minutes.

[1231]

[1232] result:

[1233] The results clearly demonstrate that clear BLG beverages containing 6 w / w % or 12 w / w % protein with a turbidity of less than 16 NTU can be produced. The beverages were heat-treated at 75°C for 5 minutes or at 95°C for 5 minutes. The beverages showed no aggregation or sedimentation upon visual inspection. In contrast, the WPI sample of Example 6a (see Fig. 4) shows turbidity after heat treatment at 75°C for 15 seconds at pH 3.7, with the WPI sample (6 w / w % protein) showing turbidity.

[1234] In addition, it was also found that, surprisingly, both beverages containing 6 w / w% or 12 w / w% protein in a pH range of 2.7 to 3.7, heat-treated at 75°C for 5 minutes, had excellent natural forms, as evidenced by a surprisingly high Trp fluorescence ratio of 1.17 to 1.18.

[1235] Example 6c: Color stability of a beverage after storage at 20°C for 6 months.

[1236] In this example, BLG beverages containing 6 wt% protein and having pH values ​​of 3.0 and 3.7 were prepared as described in Example 3. The BLG powder used for the preparation of the beverage is listed in Table 3.

[1237] The final pH of the beverage was adjusted to pH 3.0 and 3.7 using 10% phosphoric acid.

[1238] The beverage was heat-treated at 75°C for 15 seconds using a plate heat exchanger, or UHT treated at 120°C for 20 seconds as summarized in Example 4.

[1239] After heat treatment, the beverage was stored in a dark place at 20°C for 6 months.

[1240] The colors of different samples (Example 1.9) were analyzed after 0 days and 6 weeks, and after 3 and 6 months.

[1241] The results are presented in Figures 25 and 26.

[1242] result:

[1243] The results confirm that color development in BLG beverages during storage was found to be slower in pasteurized samples (75°C / 15 seconds) compared to UHT-treated BLG beverages (120°C / 20 seconds). Surprisingly, it was revealed that after 6 months of storage, the color (yellowness) in both pasteurized and UHT-treated BLG beverages was even lower than that of freshly prepared WPI beverages (WPI-A and WPI-B). Figures 8 (UHT) and 9 (pasteurized) showed b*-values ​​of 0.33 to 0.43 at pH 3.0 and b*-values ​​of 0.7 to 1.15 at pH 3.7 in freshly prepared WPI beverages. This is particularly useful for beverages containing at least 6 w / w % BLG as a protein source for whey protein beverages, which are useful for producing colorless beverages.

[1244] Example 7: Preparation of Heat-Sterilized High-Protein Drink Using BLG

[1245] A BLG sample was prepared in which approximately 92 w / w% of the protein was BLG (0.42 w / w% was ALA), and for comparison, a WPI sample was prepared using WPI-A in which approximately 60 w / w% of the protein was BLG (8 w / w% was ALA), and the WPI powder had a pH of 3.3.

[1246] A BLG isolated powder product (from Example 2, the pH of the powder was 3.9) was dispersed in tap water to produce a beverage with protein concentrations ranging from 6.0, 10, 15, 20, 25, and 32 w / w %, adjusted to pH 3.7 using 10% phosphoric acid.

[1247] A BLG isolate powder containing 98.2 w / w % of protein as BLG (see Table 3 above) was dispersed in deionized water to produce a beverage with a protein concentration of 12 w / w %. The pH was adjusted to 3.7 using 1 M HCl.

[1248] The solution was heat-treated at 75 to 120°C for a duration of 15 seconds to 5 minutes according to Example 4 and as described in Table 5, and immediately cooled on ice.

[1249] The viscosity of different samples (Example 1.8), the naturalness of the protein determined by the intrinsic tryptophan fluorescence emission ratio R=I330 / I350 (Example 1.1), visual appearance (Example 1.12), and turbidity (Example 1.7) were analyzed.

[1250] [Table 5]

[1251] Analysis data of high-protein beverages prepared from BLG at pH 3.7 while heating at 75℃, 90℃, and 120℃

[1252]

[1253] result:

[1254] The results are presented in Table 5 and Figures 10 to 12 above.

[1255] Figure 10 shows images of a clear and translucent 15 w / w % BLG beverage at pH 3.7 heated at 75°C / 15 seconds (left) and an opaque 6% WPI-A at pH 3.7 heated at 75°C / 15 seconds (right).

[1256] Figure 11 shows the sensory evaluation of the high-protein BLG beverage composition and images of 6 w / w % and 15 w / w % BLG samples at pH 3.7, and both samples are clear.

[1257] Figure 12 shows high-protein beverage formulations prepared by heating BLG beverages with protein contents of 32 w / w %, 27.5 w / w %, 25 w / w %, and 20 w / w % (from left to right) at 75°C for 5 minutes, and all samples were low viscosity and liquid.

[1258] The inventors surprisingly discovered that all solutions remain at a low viscosity even when heated at 75°C for up to 5 minutes, which suggests that there is little to no denaturation.

[1259] The viscosity observed in the high protein was typical of non-aggregated natural protein (flow behavior described by Inthavong, Kharlamova, Nicolai, Chassenieux, & Nicolai, 2016), exhibiting about 10 cP at 200 g / l.

[1260] It was confirmed by tryptophan fluorescence spectroscopy that BLG maintains its natural form, as demonstrated by having an intrinsic tryptophan luminescence ratio (I330 / I350) of at least 1.11 when heated gently (75°C), whereas more severe heating caused denaturation, as indicated by an intrinsic tryptophan luminescence ratio (I330 / I350) of less than 1.11.

[1261] RP-HPLC analysis confirmed the denaturation of the 6% BLG beverage heated at 75°C for 5 minutes and the 41% denaturation when heated at 95°C for 5 minutes.

[1262] It was found that the viscosity was low even after heating.

[1263] It was found that BLG beverage formulations can be heated above the denaturation temperature. However, heating at 95°C / 5 min causes gelation of BLG beverages containing more than 16 w / w % of protein, whereas 12 w / w % at 95°C / 5 min, 10 w / w % at 90°C / 5 min, and 6 w / w % at 120°C / 15 sec remain liquid. As evidenced by the decrease in the intrinsic tryptophan luminescence ratio (I330 / I350), at least partial denaturation / aggregation occurs under these heating conditions.

[1264] However, as evidenced by the intrinsic tryptophan luminescence ratio (I330 / I350) of 1.17, it was found that in a BLG beverage formulation containing 12 w / w % protein, when heat-treated at 75°C / 5 min, BLG remained in its natural form and the beverage was also clear and in liquid form.

[1265] To great surprise to the inventors, the sensory panel (see Example 1.11 and Fig. 11 for analysis) did not detect a significant difference in the dry texture of the 6 and 15% BLG beverage formulations heated at 75°C.

[1266] Example 8: Whey protein drink formulation with improved taste

[1267] BLG samples and WPI samples were prepared. The composition of the samples is shown below. The BLG isolate powder used is produced according to Example 2.

[1268]

[1269] The samples were analyzed by a sensory panel of 10 people (see Example 1.11). The WPI samples were yellower and had higher b*-values, and they were more turbid than the BLG beverages, especially at higher pH values. The analysis data are presented in Table 6.

[1270] [Table 6]

[1271] Analysis data of whey protein drinks prepared from BLG at pH 3.0 and pH 3.7 while heating at 75℃ and 120℃.

[1272]

[1273] Turbidity (NTU), 100s -1( Viscosity and color values ​​b*, L*, and a* in cP.

[1274] The visual appearance of the sample in Table 6 is shown in Fig. 13.

[1275] Data from sensory evaluation are shown in Figures 14 to 18.

[1276] To calculate Delta b*, the following formula is used:

[1277] Delta b* = b measured at room temperature 6.0 w / w % 단백질로 표준화된 샘플 * - b 탈염수 *

[1278] To calculate delta a*, the following formula is used:

[1279] Delta a* = a measured at room temperature 6.0 w / w % 단백질로 표준화된 샘플 * - a 탈염수 *

[1280] To calculate Delta L*, the following formula is used:

[1281] Delta L* = L measured at room temperature 6.0 w / w % 단백질로 표준화된 샘플 * - L 탈염수 *

[1282] The color values ​​for demineralized water are as follows:

[1283] L*=39.97, a*=0 and b*=-0.22.

[1284] result:

[1285] Significant differences in taste were observed between beverages produced with WPI-A and BLG by utilizing the opportunity to increase pH and decrease heating temperature while maintaining transparency and colorless characteristics. The BLG beverage had lower astringency, dry texture, sourness, whey flavor, and citric acid flavor compared to the WPI beverage shown in Figure 14.

[1286] Figure 15 shows that by increasing the pH to 3.7 before heat treatment, the acidic taste of the BLG beverage decreases at both 120°C and 75°C, while maintaining the transparency and low color of the product. This was not possible with WPI, as can be seen in Table 2 and Figure 1, because a clear and transparent beverage cannot be produced at pH 3.7.

[1287] Figure 16 demonstrates a significant reduction in astringent taste when both temperature and pH are changed from pH 3.0, 120℃ / 20 sec to pH 3.7, 75℃ / 15 sec.

[1288] Figure 17 demonstrates a significant reduction in dry texture by lowering the heating temperature from 120°C / 20 seconds to 75°C / 15 seconds (natural at 75°C versus denatured at 120°C).

[1289] Figure 18 demonstrates that when BLG is kept in its natural state using heating at 75°C / 15 seconds at pH 3.7, the whey flavor is reduced when a clear and clear colorless WPI beverage cannot be produced.

[1290] It was not possible to produce a clear beverage with WPI at pH 3.7 heat-treated at 75℃ / 15 seconds. See also Fig. 3.

[1291] Example 9a: Low-color sweetened BLG beverage formulation

[1292] A 6% w / w BLG beverage was prepared, and the composition of the BLG powder used is referenced below. The beverage was prepared as described in Example 3.

[1293]

[1294] The manufactured BLG beverage contained 6% protein and had a pH of 3.7.

[1295] 8 w / w % sucrose was used as the carbohydrate sucrose. Tests were also performed on the high-intensity sweetener sucralose. Samples were heat-treated in a water bath at 93°C for 4 minutes, and then cooled in an ice bath.

[1296] The visual appearance (Example 1.12), color (Example 1.9), turbidity (Example 1.7), and viscosity (Example 1.8) of different samples were analyzed.

[1297] The results are presented in Table 7 below.

[1298] [Table 7]

[1299] Sucrose added to 6 w / w % protein BLG samples

[1300]

[1301] *Viscoman was used.

[1302] result:

[1303] It was found that sweetened BLG beverages could be produced by using 8 w / w % sucrose as a sweetener and heat-treating them at 93°C for 4 minutes. The addition of 8 w / w % sucrose had only a weak effect on viscosity, turbidity, and transparency (see Table 5), and color was not affected by the addition of sucrose.

[1304] For example, a BLG beverage was prepared having additives typically found in commercial beverages such as sports nutrition foods. This included a 6% w / w protein BLG beverage at pH 3.7 that was heat-treated at 75°C for 5 minutes. Refer to Table 8 below for the composition.

[1305] [Table 8]

[1306] Examples of commercially available products.

[1307]

[1308] [Table 9]

[1309] The result of two recipes

[1310]

[1311] result:

[1312] In Table 9, it can be seen that both the BLG beverage with additives and the BLG beverage without additives maintained low viscosity, were transparent, and were essentially colorless.

[1313] Example 9b: Sweetened low-color clear BLG beverage

[1314] In this example, BLG beverages containing 2 w / w%, 6 w / w%, and 10 wt% protein and having a pH of 3.7 were prepared as described in Example 3. The BLG powder used for the preparation of the beverage contained 98.2 w / w% protein as BLG. Refer to Table 3 (Example 6b). The BLG beverage was also sweetened using sucrose as a carbohydrate source at a final concentration of 5 or 18 w / w% sucrose. The beverage was heat-treated in a water bath at 75°C or 95°C for 5 minutes and subsequently cooled over ice.

[1315] In addition, a beverage containing 10 w / w % protein, 17 w / w % carbohydrates, and a pH of 3.7 was prepared using the BLG powder from Table 3. These samples were heat-treated at 120°C for 20 seconds using a plate heat exchanger according to Example 4.

[1316] The viscosity (Example 1.8), turbidity (Example 1.7), color (Example 1.9), the naturalness of the protein determined by the intrinsic tryptophan fluorescence emission ratio R=I330 / I350 (Example 1.1), and the insoluble protein material (Example 1.10) of different samples were analyzed.

[1317] The results are presented in Table 10 below.

[1318] [Table 10]

[1319] A clear, sweetened, heat-treated protein drink containing BLG at pH 3.7

[1320]

[1321] result:

[1322] The results demonstrate that a stable (<15% insoluble protein material) 2 w / w % beverage containing 18 w / w % carbohydrates in the form of sucrose can be prepared at pH 3.7 by heat-treating at both 75°C and 95°C for 5 minutes.

[1323] The viscosity and turbidity of the 2 w / w % BLG beverage were surprisingly low, and the beverage was colorless, exhibiting delta b* values ​​of 0.29 and 0.31 after heating at 75°C and 95°C, respectively. Heating at 75°C produced a beverage composed primarily of natural whey protein, as evidenced by an I330 / 350 ratio of 1.19. Furthermore, surprisingly, heating at 95°C for 5 minutes also resulted in the production of primarily natural protein, as indicated by a Trp fluorescence ratio of 1.13.

[1324] Table 10 also demonstrates that stable, colorless BLG beverages with low viscosity and turbidity can be produced with 6 w / w % protein (containing 5 or 18 w / w % sucrose) and even 10 w / w % protein (containing 5 w / w % sucrose).

[1325] In addition, it is demonstrated that the BLG of these beverages remains in its natural state when heat-treated at 75°C for 5 minutes, as evidenced by a Trp fluorescence ratio greater than 1.11, and that the beverages are at least partially unfolded / aggregated when heated at 95°C for 5 minutes, as evidenced by a Trp fluorescence ratio greater than 1.11.

[1326] For a 6 w / w % BLG beverage containing 17% sucrose that was heat-treated by sterilizing at 120°C for 20 seconds, both viscosity and turbidity were low, and it was colorless with a delta b* value of 0.47.

[1327] All beverages remained remarkably clear and maintained minimal yellowness even when the concentrations of both protein and carbohydrate sources were increased. Refer to Table 10. This clearly demonstrates that it is possible to manufacture BLG beverages with an energy contribution from carbohydrates ranging from a maximum of 33 E% to at least 90 E%.

[1328] Example 10a: Exemplary process for manufacturing a clear BLG beverage containing added minerals

[1329] The BLG powder used in this example had a pH of 5.5 and contained about 96% w / w of protein as BLG (and 0.4% w / w of protein as ALA).

[1330] Acidic BLG isolated powder was prepared according to Example 2, and a beverage formulation was prepared according to Example 5.

[1331] High-temperature heat treatment of beverage formulations:

[1332] A 6% BLG beverage formulation with a pH of 3.7 was prepared. KCl and CaCl2 were added in liquid form from a 1 M stock solution. These were heat-treated at 95°C for 5 minutes.

[1333] result:

[1334] The results are summarized in Table 11 and Figure 19 below.

[1335] Figure 19 shows an image of a 6% BLG beverage with added minerals, heat-treated at 95°C for 5 minutes at pH 3.7.

[1336] A: 0 mM added minerals

[1337] B: 15 mM added CaCl2

[1338] C: 20 mM added KCl

[1339] D: 10 mM added KCl and 15 mM CaCl2.

[1340] The turbidity of a BLG beverage formulation with added minerals (0 to 20 mM KCl, 0 to 15 mM CaCl2, or 10 mM CaCl2 and 10 mM) was maintained at less than 30 NTU when heated at 95°C for 5 minutes at pH 3.7.

[1341] Gelation was observed in KCl with 30 mM added (cloudy gel).

[1342] Gelation was observed in CaCl2 with 20 mM added (clear gel).

[1343] The results clearly suggest that protein composition is more important than the mineral difference for WPI, as the amount of added minerals in Table 10 significantly exceeds the difference between the BLG and WPI product(s).

[1344] The sample remained clear (see Fig. 19) and had low viscosity within the limits of Table 11 below:

[1345] [Table 11]

[1346] Viscosity and turbidity of BLG beverage after addition of minerals (CaCl2 and KCl), heated at 95°C for 5 minutes at pH 3.7

[1347]

[1348] *Viscoman was used.

[1349] Low-temperature heat treatment of beverage formulations

[1350] A 6% BLG beverage formulation with a pH of 3.7 was prepared. KCl and CaCl2 were added in liquid form from a 1 M stock solution. These were heat-treated at a pasteurization temperature of 75°C for 5 minutes.

[1351] result.

[1352] The inventors surprisingly discovered that exceptionally high mineral concentrations are permissible when using a pasteurization temperature (75°C, 5 min). Refer to Table 12 below.

[1353] Figure 20 shows an image of a 6% BLG beverage with a pH of 3.7 that has been heat-treated at 75°C for 5 minutes and has minerals added.

[1354] A: 0 mM added minerals,

[1355] B: 100 mM added KCl,

[1356] C: 100 mM added CaCl2,

[1357] D: 100 mM added KCl and 100 mM added CaCl2

[1358] Even when 100 mM KCl or 100 mM CaCl2 was added to the beverage composition before heating, the beverage formulation remained clear. Refer to Fig. 20. Additionally, the viscosity was surprisingly low even when both 100 mM KCl and 100 mM CaCl2 were added.

[1359] [Table 12]

[1360] Viscosity and turbidity of BLG beverage after addition of minerals (CaCl2 and KCl), heated at 75°C for 5 minutes at pH 3.7

[1361]

[1362] *Viscoman was used.

[1363] Example 10b: BLG beverage with added minerals

[1364] The BLG powder used in this example had a pH of 3.79 and contained about 98.2% w / w of protein as BLG (see Table 3 of Example 6b). The content of the minerals sodium, potassium, calcium, magnesium, and phosphorus in the powder was all below the detection limit.

[1365] High-temperature heat treatment of beverage formulations:

[1366] BLG beverages containing 6 wt% protein and having pH values ​​of 2.7, 3.0, 3.3, and 3.7 were prepared as described in Example 3. The BLG beverages each contained minerals NaCl, KCl, CaCl2, and MgCl2 added from stock solutions dissolved in deionized water at 5, 3, and 1 M concentrations, respectively. The mineral concentrations are listed in Table 13 below.

[1367] [Table 13]

[1368] Minerals added to BLG beverages

[1369]

[1370] *24 kcal / 100 ml: The calculation is based on the following assumptions: 4 kcal / g protein, 9 kcal / g fat, 4 kcal / g carbohydrates

[1371] The beverage was heat-treated at 95°C for 5 minutes using a water bath as summarized in Example 4.

[1372] The turbidity of different samples (Example 1.7), the naturalness of the protein determined by the intrinsic tryptophan fluorescence emission ratio R=I330 / I350 (Example 1.1), and the insoluble protein material (Example 1.10) were analyzed.

[1373] The results are presented in Table 14 below.

[1374] [Table 14]

[1375] Turbidity, naturalness, and insoluble particles of BLG beverages after addition of minerals (NaCl, KCl, CaCl2, and MgCl2), heated at 95°C for 5 minutes at a pH of 2.7 to 3.7

[1376]

[1377] result:

[1378] The results indicate that stable, clear, acidic heat-treated beverages can be produced after heat treatment at 95°C for 5 minutes. The heat-treated BLG beverages contained both 6 w / w% protein and minerals (Na, K, Ca, and Mg), with turbidity of less than 13 NTU and the amount of insoluble particles being less than 3% at pH 2.7 to 3.7. This was also possible with Na, K, Ca, and Mg in the final product, summarized at 27.9 to 28.2 mM (refer to minimum concentrations in Table 13).

[1379] Low-temperature heat treatment of mineral-fortified beverage formulations

[1380] BLG beverages containing 6% by weight and 12% by weight of protein and having pH values ​​of 2.7, 3.0, 3.3, and 3.7 were prepared as described in Example 3.

[1381] The BLG beverages contained minerals NaCl, KCl, CaCl2, and MgCl2 added from salt stock solutions at 5, 3, and 1 M, respectively, to deionized water. The mineral additions are shown in Table 15 below.

[1382] [Table 15]

[1383] Minerals added to BLG beverages.

[1384]

[1385] *24 kcal / 100 ml and 48 kcal / 100 ml: Calculations are based on the following assumptions: 4 kcal / g protein, 9 kcal / g fat, 4 kcal / g carbohydrates

[1386] The beverage was heat-treated at 75°C for 5 minutes using a water bath as summarized in Example 4.

[1387] The turbidity of different samples (Example 1.7) and the naturalness of the protein determined by the intrinsic tryptophan fluorescence emission ratio R=I330 / I350 (Example 1.1) were analyzed.

[1388] The results are presented in Table 16 below.

[1389] [Table 16]

[1390] Turbidity and naturalness of 6 w / w % and 12 w / w % BLG beverages after addition of minerals (NaCl, KCl, CaCl2, and MgCl2, see Table 14), heated at 75°C for 5 minutes at pH 2.7 to 3.7

[1391]

[1392] result

[1393] The results indicate that a clear, acidic heat-treated beverage can be produced after heat treatment at 75°C for 5 minutes. The beverage showed no aggregation or sedimentation upon visual inspection. The heat-treated BLG beverage contained both 6 w / w% protein and minerals (Na, K, Ca, and Mg), both of which had turbidity of less than 10 NTU at pH 2.7 to 3.7. This was also possible with the Na, K, Ca, and Mg of the final product, summarized as 27.9 to 28.9 mM (refer to mineral concentrations in Table 15).

[1394] Surprisingly, it was found that natural (Trp fluorescence ratio 1.17 to 1.20) and clear (less than 10 NTU) BLG beverages could also be produced with Na, K, Ca, and Mg, summarized as 27.9 to 28.9 mM at 6 w / w % protein and 50 to 52.6 mM at 12 w / w % protein by heat treatment at 75°C for 5 minutes. See Table 16.

[1395] Also surprisingly, the 12 w / w % protein BLG beverage at pH 3.0 had a turbidity of 2.8 NTU despite higher mineral concentrations of Na, K, Ca, and Mg, summarized as 50.6 mM (Sample J), ​​which was found to be lower than the turbidity of 8.63 NTU measured in the 6% WPI beverage at pH 3.0 containing low concentrations of minerals (Na, K, Ca, and Mg) in amounts below detection levels. Refer to Table 6 of Example 8. The detection levels of the different minerals are as follows: Na: 0.005 g / 100 g = 2.2 mM, K: 0.005 g / 100 g = 1.3 mM, Ca: 0.005 / 100 g = 1.2 mM, and Mg: 0.0005 / 100 g = 0.2 mM, summarized as 4.9 mM.

[1396] The surprising results offer many new possibilities for the production of clear beverages containing additives that affect protein stability, such as minerals. By using a low heat treatment temperature of 75°C, the protein structure is maintained in its natural state, and increased mineral tolerance is achieved, because minerals only increase / induce protein aggregation when the protein unfolds.

[1397] Example 11a: Milky white whey protein drink, high temperature heat-treated

[1398] An exemplary process for producing an opaque milky white beverage containing BLG and optionally a carbohydrate source. BLG powder is dissolved in tap water, adjusted to pH according to Example 3, and heat-treated at 93°C for 4 minutes. The BLG beverage contained about 92% w / w of protein as BLG and about 0.42% w / w of protein as ALA, and the beverage is produced based on acidic BLG isolate powder with a pH of 3.9 (Example 2).

[1399] A 6% BLG beverage with a pH of 4.3 was prepared. 8% sucrose was added as a carbohydrate source. Turbidity, viscosity, color, and transparency were measured according to the procedures described in Examples 1.7, 1.8, and 1.9, and beverage stability was measured as in Example 1.10.

[1400] The results are presented in Tables 17 and 18 and Figure 21 below.

[1401] [Table 17]

[1402] Stability of a milky white beverage containing BLG, heat-treated at 93℃ / 4 min, 6% protein, and pH 4.3

[1403]

[1404] *Viscoman was used.

[1405] [Table 18]

[1406] Stability of a milky white BLG beverage containing sucrose, heat-treated at 93°C / 4 min, 6% protein, and pH 4.3

[1407]

[1408] A WPI sample containing 6% protein and a pH of 4.3 was prepared. The WPI sample was heat-treated at 94°C for 5 minutes. 0% sucrose or 8% sucrose was added to WPI-A sample, while 0% sucrose or 6% sucrose was added to WPI-B sample.

[1409]

[1410] result:

[1411] Figure 21 shows the stability of milky white BLG beverages with and without sucrose, at pH 4.3, heat-treated at 93°C for 4 minutes. A: 0% sucrose (before centrifugation), B: 8% sucrose (before centrifugation), C: 0% sucrose (after centrifugation), D: 8% sucrose (after centrifugation)

[1412] The results presented in Tables 17 and 18 and Figure 21 demonstrate that a maximum pH of 4.3 enables the production of milky white beverages, which is desirable in some embodiments of the present invention, for example, when consumers prefer whey protein beverages with a milky white appearance. Furthermore, it was found that viscosity is low in both formulations with and without sucrose at a pH of 4.3.

[1413] The color also remained in the middle. This is particularly preferred by consumers who prefer milky white beverages that do not have a yellow tint. If the b* value is high and positive, a yellow color appears.

[1414] In addition, the beverage was found to be stable, as evidenced by the fact that the protein decreased by less than 15% and the turbidity was high even after centrifugation at 3000x g for 5 minutes.

[1415] Milky white 6 w / w % protein WPI beverages based on WPI-A or WPI-B with a pH of 4.3 could not be produced because they gelled and had high viscosity, and this applied to both WPI samples with and without added sucrose.

[1416] Example 11b: Milky white whey protein drink, high temperature heat-treated

[1417] Carbohydrate-free milky white hot-processed beverage:

[1418] The BLG powder used in this example had a pH of 3.79 and contained about 98.2% w / w of protein as BLG (see Table 3 of Example 6b).

[1419] BLG beverages containing 6 wt% protein and having pH values ​​of 4.2 and 4.4 were prepared using 0.75 M NaOH as described in Example 3.

[1420] The beverage was heat-treated at 95°C for 5 minutes using a water bath as summarized in Example 4.

[1421] The turbidity (Example 1.7), viscosity (Example 1.8), the naturalness of the protein determined by the intrinsic tryptophan fluorescence emission ratio R=I330 / I350 (Example 1.1), color (Example 1.9), and insoluble protein material (Example 1.10) of different samples were analyzed.

[1422] The results are presented in Table 19 below.

[1423] [Table 19]

[1424] Milky white 6 wt% BLG beverage heat-treated at 95°C for 5 minutes at pH 4.2 and 4.4

[1425]

[1426] To calculate Delta b*, the following formula is used:

[1427] Delta b* = b measured at room temperature 6.0 w / w % 단백질로 표준화된 샘플 * - b 탈염수 *

[1428] To calculate delta a*, the following formula is used:

[1429] Delta a* = a measured at room temperature 6.0 w / w % 단백질로 표준화된 샘플 * - a 탈염수 *

[1430] To calculate Delta L*, the following formula is used:

[1431] Delta L* = L measured at room temperature 6.0 w / w % 단백질로 표준화된 샘플 * - L 탈염수 *

[1432] The color values ​​for demineralized water are as follows:

[1433] L*=39.97, a*=0 and b*=-0.22.

[1434] result:

[1435] The results presented in Table 19 demonstrate that even a maximum pH of 4.4 enables the production of milky white beverages, which is desirable in some embodiments of the present invention, for example, when consumers prefer whey protein beverages with a milky white appearance.

[1436] Surprisingly, it was found that a stable milky white BLG beverage (turbidity of at least 11,000 NTU) with low viscosity (less than 2 cP) can be successfully produced by heat-treating the beverage at 95°C for 5 minutes, even at a pH value of 4.4.

[1437] As evidenced by the intrinsic tryptophan luminescence ratio (I330 / I350) of 0.99 at pH 4.2 and 1.01 at pH 4.4, heat treatment of the beverage at 95°C for 5 minutes results in at least partial denaturation / aggregation at both pH 4.2 and 4.4.

[1438] In contrast, Example 11a states that a milky white 6 w / w % protein WPI beverage based on WPI-A or WPI-B with a pH of 4.3 cannot be produced because it gels and has high viscosity.

[1439] Milky white, high-temperature processed beverage containing a carbohydrate source:

[1440] The BLG powder used in this example had a pH of 3.79 and contained about 98.2% w / w of protein as BLG (see Table 3 of Example 6b).

[1441] BLG beverages containing 2 wt% and 6 wt% protein were prepared as described in Example 3 by using sucrose as a carbohydrate source, sweetening to a final sucrose concentration of 5 or 18 w / w% sucrose, and adjusting the pH to 4.2.

[1442] The BLG beverage was heat-treated at 95°C for 5 minutes using a plate heat exchanger as summarized in Example 4.

[1443] The turbidity (Example 1.7), viscosity (Example 1.8), the naturalness of the protein determined by the intrinsic tryptophan fluorescence emission ratio R=I330 / I350 (Example 1.1), color (Example 1.9), and insoluble protein material (Example 1.10) of different samples were analyzed.

[1444] The results are presented in Table 20 below. The energy percentage (%E) of protein and carbohydrates is calculated.

[1445] [Table 20]

[1446] Milky white 2 wt% and 6 wt% BLG beverages containing carbohydrates, heat-treated at 95°C for 5 minutes at pH 4.2,

[1447]

[1448] result:

[1449] The results presented in Table 20 demonstrate that heat treatment at 95°C for 5 minutes at a maximum pH of 4.2 enables the production of milky white beverages containing both protein and carbohydrates, where carbohydrates account for 45.5%, 75%, and 90% of the total beverage energy content, respectively. All three beverages had high turbidity exceeding 1276 NTU and low viscosity of 10.3 cP or less. All beverages were stable with less than 1.6% insoluble protein material. At least partial unfolding / aggregation occurred in all three beverages containing both protein and carbohydrates. This is evidenced by an intrinsic tryptophan luminescence ratio (I330 / I350) of 1.03 or less.

[1450] All beverages had a milky white, opaque appearance.

[1451] Example 12a: Milky white whey protein drink, long-term low-temperature heat treatment.

[1452] An exemplary process for producing milky white beverages containing BLG at different pH values. BLG powder is dissolved in tap water and adjusted to a pH of 4.2 to 4.5 using 10% phosphoric acid according to Example 3. The formulation was heat-treated at 75°C for 5 minutes, and the protein content was 6% w / w. The BLG beverage is produced based on BLG powder containing approximately 92% w / w of protein as BLG and 0.42% w / w as ALA, with a pH of 3.9.

[1453] Turbidity, viscosity, color, and visual transparency were measured according to the procedures described in Examples 1.7, 1.8, 1.9, and 1.12.

[1454] The results are presented in Table 21 and Figure 22 below.

[1455] Figure 22 shows an image of an opaque 6% protein BLG beverage prepared by heating at 75°C for 5 minutes at a pH of 4.2 to 4.5.

[1456] [Table 21]

[1457] Properties of opaque BLG beverages at pH 4.2 to 4.5 after heating at 75℃ for 5 minutes

[1458]

[1459] result:

[1460] It was found that at pH 4.2 to 4.5, the beverage had a milky white, opaque appearance and high turbidity, but still had low viscosity.

[1461] Example 12b: Milky white whey protein drink, prolonged low-temperature heat treatment.

[1462] In this example, BLG beverages containing 6 wt% protein and having pH values ​​of 4.2, 4.4, and 4.6 were prepared as described in Example 3. The BLG powder used for the preparation of the beverages contained 98.2 w / w% protein as BLG. Refer to Table 3 of Example 6b.

[1463] The final pH was adjusted to 4.2, 4.4, and 4.6 using 0.75 M NaOH. The beverages were heat-treated at 75°C for 5 minutes using a water bath as summarized in Example 4.

[1464] The turbidity (Example 1.7), viscosity (Example 1.8), amount of insoluble particles (Example 1.10), and color (Example 1.9) of different samples were analyzed.

[1465] The results are presented in Table 21.

[1466] [Table 21]

[1467] Properties of opaque BLG beverages at pH 4.2 to 4.6 after heating at 75℃ for 5 minutes

[1468]

[1469] result:

[1470] Surprisingly, a stable (<7% insoluble particles) milky white beverage with low viscosity can be produced by heat treatment at 75°C for up to 5 minutes at pH 4.2, pH 4.4, and pH 4.6, and it was found that in Example 6a, it exhibited lower viscosity than WPI-A and WPI-B beverages at pH 3.7.

[1471] Example 12c: Milky white whey protein drink containing carbohydrates , long-term low-temperature heat treatment

[1472] The BLG powder used in this example had a pH of 3.79 and contained about 98.2% w / w of protein as BLG (see Table 3 of Example 6b).

[1473] BLG beverages containing 6 wt% and 10 wt% protein were sweetened using sucrose as a carbohydrate source with a final sucrose concentration of 5 or 18 w / w%. The pH was adjusted to 4.2 using 0.75 M NaOH, and the beverage was prepared as described in Example 3.

[1474] The BLG beverage was heat-treated at 75°C for 5 minutes using a water bath as summarized in Example 4.

[1475] The turbidity (Example 1.7), viscosity (Example 1.8), and insoluble protein substances (Example 1.10) of different samples were analyzed.

[1476] The results are presented in Table 22 below. Energy percentages of protein and carbohydrates ( %E) is calculated.

[1477] [Table 22]

[1478] Milky white 6 wt% and 10 wt% BLG beverages containing carbohydrates, heat-treated at 75°C for 5 minutes at pH 4.2.

[1479]

[1480] result:

[1481] The results presented in Table 22 demonstrate that heat treatment at a maximum pH of 4.2 and 75°C for 5 minutes enables the production of a milky white beverage containing both protein and carbohydrates.

[1482] Surprisingly, the milky white beverage was found to be stable with less than 0.7% insoluble particles. The milky white beverage had low viscosity at pH 4.2 and contained carbohydrates of 33.3%, 45.5%, and 75% of the total energy content.

[1483] Example 13: Colorless whey protein drink containing > 85% BLG

[1484] A beverage formulation was prepared in which approximately 92% w / w of the protein was BLG and approximately 0.42% w / w of the protein was ALA (the pH of the BLG-isolated powder was 3.9). Refer to Example 3.

[1485] For comparison, a whey protein sample containing SPI (serum protein isolate) containing approximately 80% w / w BLG and approximately 4% w / w ALA was prepared (the pH of the SPI powder was 6.7).

[1486] The sample had a protein content of 6% w / w. The pH of the beverage was adjusted to pH 3.7.

[1487] The turbidity, viscosity, color, and transparency of the formulation were measured according to the procedures described in Examples 1.7, 1.8, and 1.9, and the beverage stability was measured as in Example 1.10.

[1488] The results are presented in Table 23 and Figures 23 and 24 below.

[1489] [Table 23]

[1490] Properties of BLG and SPI beverages with different heat treatments.

[1491]

[1492] Viscosity was measured in viscomen (Example 1.8).

[1493] result:

[1494] It was found that the viscosity of SPI (approx. 80% BLG, approximately 4% ALA) increased more due to heat treatment compared to the BLG formulation at pH 3.7.

[1495] In addition, the SPI beverage had a higher b* value and was therefore more yellow than the BLG sample.

[1496] Example 14a: Nutritional whey protein drink containing ≥85% BLG, carbohydrate source and fat source

[1497] Example 14a describes an exemplary process for preparing a heat-sterilized beverage formulation in which at least 85% w / w of the protein is BLG.

[1498] The inventors surprisingly discovered that a 6% nutritional composition containing 100 mM added KCl and 100 mM added CaCl2 remains liquid even after heating at 75°C for 5 minutes (viscosity at about 1 cP), and that a BLG beverage (≥85%) accommodates a surprisingly large mineral concentration present during heat treatment, which is pasteurized at 75°C and maintained for at least 5 minutes (Example 10a).

[1499] Because the thermal stability of whey protein often suffers from high mineral dosages, the inventors have therefore further investigated the opportunity to produce a nutritionally complete acidic BLG beverage to produce a sterilized nutritional beverage containing ≥85% BLG, a carbohydrate source, a fat and a mineral source in a combination that meets current FSMP (Food for Special Medical Purposes) requirements.

[1500] Dissolve the protein and mix the lipids and carbohydrates in an example ratio based on the energy distribution as described in Table 24.

[1501] Food-grade acids and minerals were selected to meet the requirements for Food for Special Purposes Products (FSMP).

[1502] Vitamins may be additionally supplied to meet FSMP requirements and produce a nutritionally complete supplement.

[1503] [Table 24]

[1504] Composition of an exemplary nutritional composition comprising sources of protein, carbohydrates, and fats.

[1505]

[1506] A 6 w / w % BLG nutritional beverage containing 13.5 w / w % sucrose and 4.7 w / w % rapeseed oil was mixed at 70°C. The composition of protein, fat, and carbohydrates is selected to meet medical nutrition recommendations.

[1507] In a characteristic mode, (1) 40 mM KCl and 14 mM CaCl2 or (2) 80 mM KCl and 28 mM CaCl2 were added with additional ingredients as shown in Table 24 or (3) without the addition of additional minerals.

[1508] The solution was homogenized at 200 bar.

[1509] The solution was heat-treated by immersing it in a water bath at 75°C or 95°C for 5 minutes and cooled on ice.

[1510] [Table 25]

[1511] Nutritional composition containing BLG, carbohydrates, fats, and added mineral sources

[1512]

[1513] result:

[1514] It was found that opaque beverages can be produced by heating BLG with fat and carbohydrate sources at 75°C and 95°C.

[1515] At 75°C, it maintains its natural state (the Trp fluorescence ratio was 1.18 despite being composed of fat), but at 95°C, it causes denaturation (Trp fluorescence). Viscosity is maintained at a low level. Since it was possible to maintain its natural form, it enables the administration of minerals important for medical nutrition (FSMP requirements). Furthermore, the ability of the nutritional composition to remain in a liquid state in the presence of selected minerals clearly suggests feasibility for use in medical nutrition.

[1516] Example 14b: Nutritional whey protein drink containing carbohydrates, fats, and added mineral sources

[1517] Example 14b describes a process for producing a milky white beverage containing BLG, a carbohydrate source, a fat source, and added minerals that comply with FSMP requirements.

[1518] Since the thermal stability of whey protein often suffers from high mineral content, this example investigated the opportunity to produce a nutritionally complete acidic BLG beverage that meets current FSMP (Food for Special Medical Purposes) requirements by sterilizing a nutritional beverage containing 6 to 16.7% w / w of BLG source (98.2% purity), 2.7 to 7.3% w / w of fat source (rapeseed oil), and 7.5 to 18% w / w of sucrose (as described in Table 26 below).

[1519] Mineral composition using food-grade acids and salts to reach pH and pH 3.5, and Na, K, Ca of the final beverage, and The levels of Mg...

Claims

Claim 1 A packaged heat-treated beverage preparation having a pH in the range of 2 to 4.7, wherein the beverage comprises a protein in the range of 2 to 45 w / w relative to the total weight of the beverage, and optionally, a sweetener, a sugar polymer, or a flavoring, wherein at least 85 w / w of the protein is beta-lactoglobulin (BLG), and the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio of at least 1.11, or the packaged heat-treated beverage preparation has a protein denaturation of up to 10%, wherein the emission ratio is determined by dividing the fluorescence emission intensity at 330 nm by the fluorescence emission intensity at 350 nm, and wherein the preparation is at least pasteurized. Claim 2 In paragraph 1, a packaged heat-treated beverage preparation in a sterile state. Claim 3 delete Claim 4 A packaged heat-treated beverage formulation having a pH in the range of 3.0 to 4.3, according to claim 1 or 2. Claim 5 In claim 1 or 2, the protein fraction of the beverage formulation has a color value delta b* in the range of -0.10 to +0.51 on the CIELAB color scale, wherein delta b* = b, measured at room temperature. 6.0 w / w% 단백질로 표준화된 샘플 * - b 탈염수 Phosphorus, packaged heat-treated beverage preparation. Claim 6 In claim 1 or 2, the beverage formulation has a color value delta b* in the range of -0.10 to +0.51 on the CIELAB color scale, wherein delta b* = b, measured at room temperature. 6.0 w / w% 단백질로 표준화된 샘플 * - b 탈염수 Phosphorus, packaged heat-treated beverage preparation. Claim 7 A packaged heat-treated beverage formulation according to claim 1 or 2, wherein the sum of the amounts of Na, K, Mg and Ca is at most 750 mM. Claim 8 A packaged heat-treated beverage formulation having a turbidity of up to 200 NTU, in accordance with claim 1 or 2. Claim 9 A packaged heat-treated beverage formulation according to claim 1 or 2, wherein the viscosity measured at 22°C at a shear rate of 100 / s is up to 200 cP centipoise. Claim 10 A packaged heat-treated beverage formulation according to claim 1 or 2, comprising 4.0 to 35% w / w of protein relative to the total weight of the beverage. Claim 11 In claim 10, a packaged heat-treated beverage formulation comprising 4.0 to 30% w / w of protein relative to the total weight of the beverage. Claim 12 A packaged heat-treated beverage formulation according to claim 1 or 2, wherein at least 90% w / w of the protein is beta-lactoglobulin (BLG). Claim 13 A packaged heat-treated beverage formulation according to claim 12, wherein at least 92% w / w of the protein is beta-lactoglobulin (BLG). Claim 14 A method for manufacturing a packaged heat-treated beverage formulation having a pH in the range of 2 to 4.7, comprising: a) providing a liquid solution having a pH in the range of 2 to 4.7, comprising 2 to 45 weight% of protein relative to the total weight of the beverage, - optionally, a sweetener, a sugar polymer, or a flavoring, wherein at least 85% of the protein is BLG; b) packaging the liquid solution, wherein the protein fraction of the liquid solution has an intrinsic tryptophan fluorescence emission ratio of 1.11 or more, or the liquid solution has a protein denaturation degree of up to 10%, and the emission ratio is determined by dividing the fluorescence emission intensity at 330 nm by the fluorescence emission intensity at 350 nm; and wherein the liquid solution of step a) or the packaged liquid solution of step b) undergoes heat treatment including at least pasteurization. Claim 15 A packaged heat-treated beverage preparation for use in administration to a patient suffering from protein malabsorption, according to claim 1 or 2. Claim 16 A packaged heat-treated beverage preparation for use as a dietary supplement, according to claim 1 or 2. Claim 17 In paragraph 16, a packaged heat-treated beverage preparation consumed before, during, or after exercise. Claim 18 delete Claim 19 delete Claim 20 delete Claim 21 delete Claim 22 delete Claim 23 delete Claim 24 delete Claim 25 delete Claim 26 delete Claim 27 delete

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