Acidic β-lactoglobulin beverage preparation

Abstract

To provide an acidic, packaged, heat-treated beverage preparation comprising whey protein and having improved organoleptic and / or visual properties.SOLUTION: The present invention pertains to a packaged, heat-treated beverage preparation having a pH in the range of 2.0-4.7, the beverage preparation comprising a total amount of protein of 2 to 45% w / w relative to the weight of the beverage, wherein at least 85% w / w of the protein is β lactoglobulin (BLG), and optionally, sweetener, sugar polymers and / or flavour. The beverage preparation is at least pasteurized and sterile.SELECTED DRAWING: None

Description

[Technical Field]

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

[0002] Dietary supplements containing whey protein are commonly used for muscle synthesis, weight management, and maintenance of muscle and body weight. These supplements are targeted at a variety of consumers, including athletes, children, the elderly, and patients who are malnourished or at risk of malnutrition and / or have increased protein requirements.

[0003] Whey protein can be isolated from whey or whey serum. Whey typically contains a mixture of β-lactoglobulin (BLG), α-lactalbumin (ALA), serum albumin, and immunoglobulins, with BLG being the most dominant. Therefore, whey protein concentrate (WPC) contains a mixture of these proteins. Whey protein isolate (WPI) contains less fat and lactose than WPC.

[0004] Beverages containing whey protein are well known. For example, there are acidic, heat-treated beverages that contain whey protein.

[0005] Etzel 2004 (Etzel, MR, 2004, Manufacture and use of dairy protein fraction. American Society for Nutritional Science, pp. 996-1002) describes a beverage containing 2.5 wt% WPI at pH 2-7. They found that the heat-treated beverage was only obtainable with the addition of a flocculant inhibitor. [Overview of the Initiative]

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

[0007] Some of the challenges in incorporating whey protein into acidic heat-treated beverages include the formation of unstable precipitates that settle in the beverage, high viscosity or gel formation, and unpleasant tastes due to a high degree of astringency and / or dry texture.

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

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

[0010] The inventors hereby discover that such packaged, heat-treated beverages can be served within a wide acidic pH range up to pH 4.7, while still retaining low viscosity and optionally low levels of astringency and dry texture. The present invention provides both clear and opaque but stable beverages.

[0011] Therefore, one aspect of the present invention is a packaged heat-treated beverage preparation having a pH in the range of 2.0 to 4.7, wherein the beverage is - Proteins that make up 2-45% w / w of the weight of the beverage, of which at least 85% w / w is BLG, -Optionally, sweeteners and / or flavors This relates to preparations, including those mentioned above.

[0012] Another aspect of the present invention is a method for producing a packaged heat-treated beverage preparation having a pH in the range of 2.0 to 4.7, comprising the following steps: a) -2 to 45% by weight of protein, of which at least 85% is BLG, -Optionally, sweeteners and / or flavors The process of preparing a liquid solution containing, b) The process of packaging the liquid solution and Includes, The liquid solution of step a) and / or the packaged liquid solution of step b) are subjected to a heat treatment including at least pasteurization. Regarding the method.

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

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

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

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

[0017] [Figure 1] This figure shows images of BLG and WPI beverages with a pH of 3.7 and a protein content of 6% w / w, which were heat-treated at 120°C for 20 seconds and 75°C for 15 seconds. [Figure 2] This figure shows images of WPI-B pH 3.0~3.7 at 120°C and BLG pH 3.7 at 120°C / 20 seconds. [Figure 3] This figure shows images of WPI-B pH3.0~3.7 at 75℃ and BLG pH3.7 at 75℃ / 15 seconds. [Figure 4] This figure shows images of WPI-B at pH 3.7 and BLG at pH 3.9, at 75°C / 15 seconds. [Figure 5] This figure shows the turbidity of a BLG beverage preparation treated with 6% UHT (120°C / 20 seconds). [Figure 6] This figure shows the turbidity of a 6% pasteurized (75°C / 15 seconds) BLG beverage composition. [Figure 7] This figure shows the viscosity of a BLG beverage preparation after 6% UHT treatment (120°C / 20 seconds). [Figure 8] This figure shows the yellowness (b*) of a beverage composition treated with 6% UHT (120°C / 20 seconds). [Figure 9] This figure shows the yellowness (b*) of a 6% pasteurized (75°C / 15 seconds) beverage composition. [Figure 10] This figure shows images of a 15% BLG beverage with a pH of 3.7 (left) and a 6% WPI-a with a pH of 3.7 (right) after being heated at 75℃ for 15 seconds. [Figure 11] This figure shows the sensory evaluation of high-protein BLG beverage compositions, as well as images of 6 w / w% and 15 w / w% BLG samples at pH 3.7. [Figure 12] The figure shows high-protein beverage preparations prepared by heating 30%, 27.5%, 25%, and 20% BLG at 75°C for 5 minutes (from left to right). The viscosity remained low even after heating. [Figure 13] This figure shows images of different WPI and BLG samples. [Figure 14] This figure shows the sensory evaluation (on a scale of 0 to 15) of beverages. The results are for WPI pH3.0 at 120℃ / 20 seconds and BLG pH3.7 at 75℃ / 15 seconds. [Figure 15] This figure shows the effect of pH and temperature on sourness. [Figure 16] This figure shows sensory data regarding the astringency of BLG beverages at pH 3.0 (120℃ / 20 seconds) and pH 3.7 (75℃ / 15 seconds). [Figure 17] This figure shows sensory data regarding the dry texture of pH 3.7 beverages at 120°C / 20 seconds and 75°C / 15 seconds. [Figure 18] This figure shows sensory data regarding whey aroma when BLG is maintained in its natural conformation. [Figure 19] The image shows a 6% BLG beverage with a pH of 3.7, heat-treated at 95°C for 5 minutes, and fortified with minerals. [Figure 20] The image shows a 6% BLG beverage with a pH of 3.7, heat-treated at 75°C for 5 minutes, and to which minerals have been added. [Figure 21] This shows the stability of a pH 4.3 milky BLG beverage with and without sucrose added, after heat treatment at 93°C for 4 minutes. [Figure 22] This figure shows an image of an opaque 6% protein BLG beverage prepared by heating at 75°C for 5 minutes at a pH of 4.2-4.5. [Figure 23] This figure shows images of BLG and SPI beverages that have been heat-treated at pH 3.7 at 75°C for 5 minutes. [Figure 24] This figure shows images of BLG and SPI beverages with a pH of 3.7. [Modes for carrying out the invention]

[0018] definition In the context of this invention, the terms "β-lactoglobulin" or "BLG" refer to β-lactoglobulin from, for example, naturally occurring unfolding and / or glycosylated mammalian species, and include naturally occurring genetic variants. The terms further include aggregated BLG, precipitated BLG, and crystalline BLG. When referring to the amount of BLG, it refers to the total amount of BLG, including aggregated BLG. The total amount of BLG is determined according to Example 1.31. The term "aggregated BLG" refers to BLG that is at least partially unfolded and further aggregated with other denatured BLG molecules and / or other denatured whey proteins, typically by hydrophobic interactions and / or covalent bonds.

[0019] BLG is the most dominant protein in bovine whey and milk, present in several genetic variants, with the main ones in milk labeled A and B. BLG is a lipocalin protein that can bind to many hydrophobic molecules, suggesting a role in their transport. BLG has also been shown to bind to iron via siderofoam, potentially playing a role in the fight against pathogens. Homologs of BLG are lacking in human breast milk.

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

[0021] In the context of the present invention, the terms “non-aggregated β-lactoglobulin” or “non-aggregated BLG” also refer to β-lactoglobulin from, for example, naturally occurring unfolding and / or glycosylated mammalian species, and include naturally occurring genetic variants. However, the terms do not include aggregated BLG, precipitated BLG, or crystallized BLG. The amount or concentration of non-aggregated BLG is determined according to Example 1.6.

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

[0023] In the context of this invention, the term "crystal" refers to a solid material in which its constituent elements (atoms, molecules, or ions, etc.) are arranged in a highly ordered microscopic structure, forming a crystal lattice that extends in all directions.

[0024] In the context of this invention, the term “BLG crystal” refers to a non-aggregated, preferably naturally occurring, protein crystal primarily containing BLG, which is arranged in a highly ordered microscopic structure and forms a crystal lattice extending in all directions. BLG crystals can be, for example, monolithic or polycrystalline, and may be, for example, intact crystals, crystal fragments, or a combination thereof. Crystal fragments are formed, for example, when intact crystals are subjected to mechanical shear during processing. Crystal fragments also have the highly ordered microscopic structure of the crystal, but may lack the uniform surface and / or uniform corners or edges of the intact crystal. For example, see Figure 18 of PCT application number PCT / EP2017 / 084553 for many examples of intact BLG crystals, and Figure 13 of PCT application number PCT / EP2017 / 084553 for examples of BLG crystal fragments. In both cases, BLG crystals or fragments can be visually identified using an optical microscope as clearly defined compact and coherent structures. BLG crystals or fragments are usually at least partially transparent. Protein crystals are also known to be birefringent, and this optical property can be used to identify unknown particles with crystalline structures. Amorphous BLG aggregates, on the other hand, usually appear as poorly defined, opaque, and as open or porous clumps of irregular size.

[0025] In the context of this invention, the term "crystallize" refers to the formation of protein crystals. Crystallization can occur spontaneously, for example, or can be initiated by the addition of a crystallizing species.

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

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

[0028] α-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 main structural differences with β-lactoglobulin is that ALA does not have a free thiol group that can function as a starting point for covalent aggregation reactions.

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

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

[0031] In the context of the present invention, the terms “casein macropeptide” or “CMP” refer to hydrophilic peptides derived from the hydrolysis of “κ-CN” or “kappa-casein” from, for example, natural and / or glycosylated mammalian species, residues 106-169, and include naturally occurring genetic variants by aspartate proteinase, such as chymosin.

[0032] 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, and preferably at least 80% w / w, relative to total solids.

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

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

[0035] The term "whey" refers to the liquid phase remaining after the 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 several types of whey, such as "sweet whey," which is a whey product produced by rennet-based precipitation of casein, and "acid whey" or "sour whey," which is a whey product produced by acid-based precipitation of casein. Acid-based precipitation of casein can be achieved, for example, by adding food acids or by bacterial culture.

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

[0037] The terms "whey protein" or "serum protein" refer to proteins present in whey.

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

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

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

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

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

[0043] Procedure for testing whether a liquid under a specific set of conditions is supersaturated with respect to BLG: a) Transfer 50 ml of the liquid sample to be tested into a centrifuge tube (VWR catalog number 525-0402) with a height of 115 mm, an inner diameter of 25 mm, and a capacity of 50 mL. Throughout steps a) to h), care should be taken to maintain the sample and subsequent fractions under the original physical and chemical conditions of the liquid. b) Immediately centrifuge the sample at 3000g for 3.0 minutes, with a maximum acceleration of 30 seconds and a maximum deceleration of 30 seconds. c) Immediately after centrifugation, transfer as much of the supernatant as possible (without disturbing the pellets if any have formed) to a second centrifuge tube (the same type as in step a). d) Take a small sample of 0.05 mL of the supernatant (small sample A). e) Add 10 mg of BLG crystals with a maximum particle size of 200 microns (at least 98% pure, non-aggregated BLG relative to the total solids) to the second centrifuge tube and stir the mixture. f) Leave the second centrifuge tube at its original temperature for 60 minutes. g) Immediately after step f), the second centrifuge tube is centrifuged at 500g for 10 minutes, and then another small sample of 0.05 mL (small sample B) is taken from the supernatant. h) If present, collect the centrifugation pellet from step g), resuspend it in milliQ water, and immediately examine the suspension under a microscope for the presence of visible crystals. i) Determine the concentration of non-aggregated BLG in small samples A and B using the method outlined in Example 1.6 - express the result as %BLG w / w relative to the total weight of the small samples. The concentration of non-aggregated BLG in small sample A is C BLG,A This is called C, and the concentration of non-aggregated BLG in small sample B is C BLG,B It is called that. j) The liquid from which the sample was taken in step a) is c BLG,B ga c BLG,A If the value was lower than that and crystals were observed in step i), then it was supersaturated (under certain conditions).

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

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

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

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

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

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

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

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

[0052] WPI or SPI preferably contains the following: 90-100% w / w protein relative to total solids. BLG at 15-70% w / w relative to total protein, ALA at 8-50% w / w relative to total protein, and CMP at 0-40% w / w relative to total protein.

[0053] Alternatively, however, WPI or SPI may also preferably contain: 90-100% w / w protein relative to total solids. 30-95% w / w BLG relative to total protein, ALA at 4-35% w / w relative to total protein, and CMP at 0-25% w / w relative to total protein.

[0054] More preferably, WPI or SPI may contain the following: 90-100% w / w protein relative to total solids. 30-90% w / w BLG relative to total protein, ALA at 4-35% w / w relative to total protein, and CMP at 0-25% w / w relative to total protein.

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

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

[0057] The terms “essentially derived from” and “essentially derived from” mean that the claim or feature in question encompasses the specified material or process and any material or process that does not substantially affect the basic and novel properties of the claimed invention.

[0058] In the context of this invention, the phrase "Y and / or X" means "Y" or "X", or "Y and X". Following the same line of logic, "n1, n2, ..., n i-1 and / or n i The phrase "" is either "n1" or "n2" or... or "n i-1 " or "n1" or constituent elements: n1, n2, ...n i-1 and n i This means any combination of the following.

[0059] In the context of the present invention, the terms "dried" or "shrunk" mean that the composition or product contains at most 10% w / w of water, preferably at most 6% w / w, and more preferably even less water.

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

[0061] In the context of this invention, the pH of the powder refers to the pH of 10 g of powder mixed with 90 g of desalted water, and is measured according to Example 1.16.

[0062] In the context of the present invention, a 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 that particular composition, product, or material, unless another criterion (e.g., total solids or total protein) is specifically mentioned.

[0063] In the context of the present invention, the method step "concentrate" and the verb "concentrate" encompass both total solids-based protein concentration and gross weight-based protein concentration with respect to protein concentration. This means, for example, that concentration does not necessarily require an increase in the absolute w / w concentration of protein in the composition, as long as the protein content increases relative to the total solids.

[0064] In the context of this invention, the term "weight ratio" between component X and component Y is used in calculations m X / m Y (In the formula, m X m is the amount (weight) of component X, Y This refers to the value obtained by the amount (weight) of component Y.

[0065] In the context of this invention, the term "at least pasteurization" refers to a heat treatment that has a microbial killing effect of 10 seconds or more at 70°C. The criterion for determining the sterilization effect is Escherichia coli (E. coli) O157:H7.

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

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

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

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

[0070] When liquids such as beverage preparations are sterilized and aseptically packaged in sterile containers, they typically have a shelf life of at least six months at room temperature. Sterilization kills spores and microorganisms that can cause spoilage of the liquid.

[0071] In the context of this invention, the term "energy content" means the total amount of energy contained in the food. Energy content can be measured in kilojoules (kJ) or kilocalories (kcal), and is called calories per unit volume of food, for example, kcal per 100 grams of food. An example is a beverage having an energy content of 350 kcal / 100 grams.

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

[0073] In the context of this invention, the term "nutritionally complete dietary supplement" is understood to mean a food containing proteins, lipids, and carbohydrates, and further containing vitamins, minerals, and trace elements, and having a nutritional profile that matches a complete and healthy diet.

[0074] In the context of this invention, the term "nutritionally incomplete supplement" means a food containing one or more major nutrients and optionally further containing vitamins, minerals, and trace elements. A nutritionally incomplete beverage may contain protein as its sole nutrient, or it may contain, for example, protein and carbohydrates.

[0075] The terms "Foods for Special Medical Purposes (FSMP)" or "Medical Foods" refer to foods intended for oral or tube feeding, used for specific medical disorders, diseases, or conditions with specific nutritional requirements, and used under medical supervision. Medical foods may be nutritionally complete or nutritionally incomplete supplements / beverages.

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

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

[0078] The terms "instant beverage powder" or "instant beverage powder product" refer to powders that can be converted into liquid beverages by adding a liquid such as water.

[0079] In the context of the present invention, the terms “beverage preparation” and “preparation” as used in substance refer to any aqueous liquid that can be ingested as a beverage, for example, by pouring, sipping, or tube feeding.

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

[0081] In the context of this invention, the term "astringency" relates to texture. Astringency is perceived as a contraction of the cheek muscles, leading to increased saliva production. Therefore, astringency is not a taste in itself, but rather a physical texture and time-dependent sensation in the mouth.

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

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

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

[0085] In the context of the present invention, the terms “lipid,” “fat,” and “oil” as used herein are interchangeable 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 they are suitable for human consumption.

[0086] In the context of this invention, the term "transparent" encompasses beverage preparations that have a visually clear appearance, allowing light to pass through and through which a clear image is visible. Transparent beverages have a turbidity of up to 200 NTU.

[0087] In the context of this invention, the term "opaque" includes beverage preparations having a visually indistinct appearance, which have a turbidity greater than 200 NTU.

[0088] One aspect of the present invention is a packaged heat-treated beverage preparation having a pH in the range of 2.0 to 4.7, wherein the beverage is - Proteins that make up 2-45% w / w of the weight of the beverage, of which at least 85% w / w is BLG, -Optionally, sweeteners, sugar polymers and / or flavors This relates to preparations, including those mentioned above.

[0089] Packaged heat-treated beverage preparations containing at least 85% w / w protein are highly beneficial for several reasons. A high BLG content in acidic beverages allows for a higher pH range and lower heating temperatures, while still maintaining clarity and colorlessness, even when high protein concentrations are applied. Surprisingly, BLG beverages were found to have lower astringency, dry texture, acidity, whey aroma, and citric acid flavor compared to WPI beverages containing small amounts of BLG.

[0090] Another advantage of the present invention and the expanded pH range is that it is possible to produce a milky beverage that is still white, not yellowish, and still stable, while having high turbidity and low viscosity.

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

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

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

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

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

[0096] The degree of protein nativeness depends on several factors, including protein concentration, pH, and the temperature and duration of heat treatment.

[0097] The intrinsic tryptophan fluorescence emission ratio R = I330 / I350 is a measure of the nativity of a protein. If R is at least 1.11, the native conformation is dominant; if 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.

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

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

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

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

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

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

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

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

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

[0107] The beverage preparation may optionally contain 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 preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of less than 1.11, thus indicating that the proteins are at least partially unfolded and aggregation is dominant.

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

[0109] Protein denaturation can also be explained by analytical methods other than tryptophan fluorescence. This method is described in Example 1.3.

[0110] In some preferred embodiments of the present invention, the protein fraction of the packaged heat-treated beverage preparation has a 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%.

[0111] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation 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%.

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

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

[0114] Surprisingly, regarding appearance, it was found that using a whey protein beverage in which at least 85% w / w of protein is BLG allowed for an increase in pH during heat treatment, resulting in improvements in visual perception (color and turbidity) and viscosity compared to heat-treated WPI beverages.

[0115] Surprisingly, significant differences in sensory parameters were found between the BLG beverage of the present invention and the beverage produced with WPI. Surprisingly, and advantageously, the BLG beverage was found to have lower levels of astringency, dry texture, sourness, whey aroma, and citric acid flavor compared to the WPI beverage. It was further found that raising the pH of an acidic beverage reduces the amount of sweetener needed to balance the acidity of the beverage, and therefore, the concentration of sweetener required for such a beverage is lower.

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

[0117] These pH ranges are particularly suitable when beverage preparations are pasteurized.

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

[0119] These pH ranges, combined with high-temperature treatments such as sterilization, are particularly suitable for producing clear beverages with low viscosity and improved taste.

[0120] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 4.1 to 4.7, which is particularly suitable for producing a stable beverage having a milky 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.

[0121] The visual appearance of beverage preparations is important to consumers, both for clear and opaque beverages. In particular, for clear, water-like beverages or white, milky beverages, the inventors have found it advantageous to be able to control the color of the beverage, or rather, to control the lack of color in the beverage.

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

[0123] The CIELAB color scale described in Example 1.9 is used to determine the color of the beverage. For example, positive delta-b * The value indicates a color that is yellower than desalinated water, and a negative delta b. * The value indicates a beverage that is bluer than desalted water. Therefore, to obtain a beverage that is neither yellow nor blue, color delta b * Consumers generally prefer values ​​that are close to zero.

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

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

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

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

[0128] These beverages have a higher delta b * It has less yellow color compared to beverages containing WPI that have a higher value and a more yellow color.

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

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

[0131] The protein fraction of a packaged heat-treated beverage preparation exhibits a delta a value in the range of -0.2 to 0.2 on the CIELAB color scale, particularly when the preparation has a turbidity of up to 200 NTU, more preferably up to 40 NTU. * It is typically preferable that the packaged heat-treated beverage preparation has a color value delta a in the range of -0.15 to 0.15, preferably -0.10 to 0.10, on the CIELAB color scale. * It has.

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

[0133] In some embodiments of the present invention, the packaged heat-treated beverage preparation contains multiple minerals. In one exemplary embodiment, the packaged heat-treated beverage preparation contains at least four minerals. In one embodiment, the four minerals are sodium, potassium, magnesium, and calcium.

[0134] The inventors have surprisingly found that, using BLG isolates as defined herein and in Example 2, it is possible to produce heat-treated beverage preparations with high mineral concentrations without compromising viscosity. This makes it possible to produce packaged heat-treated beverage preparations with high mineral content, enabling the production of beverages that are nutritionally complete or nutritionally incomplete dietary supplements.

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

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

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

[0138] In another exemplary embodiment, the packaged heat-treated beverage preparation contains several minerals selected from the group consisting of calcium, iodine, zinc, copper, chromium, iron, phosphorus, magnesium, selenium, manganese, molybdenum, sodium, potassium, and combinations thereof.

[0139] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation contains up to 150 mM KCl and up to 150 mM CaCl2, or the packaged heat-treated beverage preparation contains up to 130 mM KCl and up to 130 mM CaCl2, or the packaged heat-treated beverage preparation contains up to 110 mM KCl and up to 110 mM CaCl2, or the packaged heat-treated beverage preparation contains up to 100 mM KCl and up to 100 mM CaCl2, or preferably the packaged heat-treated beverage preparation contains up to 80 mM KCl and up to 80 mM CaCl2, or preferably the packaged heat-treated beverage preparation contains up to 50 mM KCl and up to 50 mM CaCl2, or preferably the packaged heat-treated beverage preparation contains up to 40 mM KCl and up to 40 mM CaCl2.

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

[0141] In the context of the present invention, the term "low mineral content" refers to compositions having at least one, preferably two, and more preferably all of the following: For example, liquids, beverages, powders, or other foods: - Ash content of up to 1.2% w / w relative to total solids, -Total calcium and magnesium content of up to 0.3% w / w relative to total solids, -Total sodium and potassium content of up to 0.10% w / w relative to total solids, -Total phosphorus content of up to 100mg per 100g of protein.

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

[0143] More preferably, the low-mineral composition comprises at least one, preferably two or more, and more preferably all of the following: - Ash content of up to 0.5% w / w relative to total solids, -Total calcium and magnesium content of up to 0.15% w / w relative to total solids, -Total sodium and potassium content of up to 0.06% w / w relative to total solids, -Total phosphorus content of up to 50mg per 100g of protein.

[0144] It is particularly preferable that the low-mineral composition has the following: - Ash content of up to 0.5% w / w relative to total solids, -Total calcium and magnesium content of up to 0.15% w / w relative to total solids, -Total sodium and potassium content of up to 0.06% w / w relative to total solids, -Total phosphorus content of up to 50mg per 100g of protein.

[0145] The inventors have found that the present invention makes it possible to prepare packaged, heat-treated beverage preparations that have very low levels of phosphorus and other minerals such as potassium, which are beneficial for patients with kidney disease or impaired kidney function.

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

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

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

[0149] In the context of the present invention, the term "low phosphorus" refers to a composition having a total phosphorus content of up to 100 mg of phosphorus per 100 g of protein, for example, a liquid, powder, or other food. 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 compositions according to the present invention may be used as food ingredients for producing foods for a group of patients with impaired renal function.

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

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

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

[0153] The low-potassium composition according to the present invention can be used as a food ingredient for producing foods for a group of patients with impaired renal function.

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

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

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

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

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

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

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

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

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

[0163] In some embodiments of packaged heat-treated beverage preparations, it is beneficial that the beverage preparation is transparent. This may be advantageous, for example, when the beverage is used as a sports drink or "protein water," in which case it is beneficial that the appearance of the beverage resembles water.

[0164] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a turbidity of up to 200 NTU, and such a beverage is clear.

[0165] Surprisingly, the inventors have discovered that the heat-treated beverage preparation according to the present invention can yield a clear heat-treated beverage preparation having a turbidity of up to 200 NTU.

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

[0167] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation has a turbidity of up to 150 NTU, preferably up to 100 NTU, preferably up to 80 NTU, preferably up to 60 NTU, more preferably up to 40 NTU, 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.

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

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

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

[0171] The amount of insoluble substances in a heat-treated beverage preparation is a measure of the beverage's instability and the extent to which sedimentation occurs over time. Beverages with a large amount of insoluble substances are typically considered unstable.

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

[0173] Surprisingly, compared to using WPI with a low BLG content as the protein source, using BLG at an amount of at least 85 w / w% resulted in the protein fraction containing up to 15% insoluble material after centrifugation at 3000g for 5 minutes, demonstrating the stability of the beverage preparation.

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

[0175] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation contains up to 15% insoluble material.

[0176] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation 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.

[0177] Furthermore, even lower levels of insoluble matter are generally preferred, and in some preferred embodiments, the packaged heat-treated beverage preparation contains up to 4% insoluble matter, preferably up to 2% insoluble matter, more preferably up to 1% insoluble matter, and most preferably no detectable insoluble matter at all.

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

[0179] One method for determining the viscosity of a beverage preparation is to measure the viscosity of the beverage, as described in Example 1.8.

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

[0181] Surprisingly, the inventors have discovered that beverage preparations with an acidic pH, subjected to heat treatment such as pasteurization, and further subjected to sterilization, can have a maximum viscosity of 200 centipoise (cP) when measured at 22 degrees Celsius with a shear rate of 100 cubic centimeters per second.

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

[0183] Preferably, the viscosity of the packaged heat-treated beverage preparation 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.

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

[0185] It has been previously found that adding a flocculation inhibitor to a beverage is essential for producing acidic, clear, heat-treated beverages containing WPI with a pH above 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).

[0186] Surprisingly, the inventors have found that a transparent heat-treated beverage containing at least 85% w / w of BLG can be produced even at a pH higher than 3.0 without adding a flocculation inhibitor.

[0187] Thus, in some preferred embodiments of the present invention, the packaged heat-treated beverage preparation does not contain a flocculation inhibitor or alternatively contains only a trace amount of a flocculation inhibitor.

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

[0189] In some embodiments of the present invention, the packaged heat-treated beverage preparation contains a flocculation inhibitor at a maximum of 0.1% w / w, preferably at a maximum of 0.03% w / w, and most preferably does not contain a flocculation inhibitor. These embodiments are particularly preferred for transparent low-fat beverages.

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

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

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

[0193] 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 based on the weight of the beverage.

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

[0195] The packaged heat-treated beverage preparation of the present invention is particularly useful as a sports beverage, in which case it preferably also contains an optionally very limited amount of lipids and / or an optionally limited amount of carbohydrates.

[0196] In some preferred embodiments of the present invention, the preparation is particularly useful as a sports beverage, for example containing a total amount of protein in the range of 2 to 45% w / w, preferably 2 to 20% w / w, or preferably 2 to 10% w / w, most preferably 2 to 6% w / w based on the weight of the beverage.

[0197] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation is particularly useful as a nutritionally incomplete dietary supplement, for example containing a total amount of protein in the range of 2 to 45% w / w, preferably 2 to 20% w / w, or preferably 3 to 10% w / w based on the weight of the beverage.

[0198] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation is particularly useful as a nutritionally complete dietary supplement, for example containing a total amount of protein in the range of 4 to 45% w / w, or preferably 5 to 20% w / w based on the weight of the beverage.

[0199] In some preferred embodiments of the present invention, packaged heat-treated beverage preparations are particularly advantageous for patients with kidney disease or impaired kidney function.

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

[0201] It is particularly preferable that the packaged heat-treated beverage preparation contains BLG isolate in combination with other protein sources, preferably as the primary protein source, and if possible as the sole protein source.

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

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

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

[0205] Even lower carbohydrate content is generally preferred, and therefore, in some preferred embodiments of the present invention, it is preferably in the range of 0 to 30% of the total energy content of the preparation, more preferably in the range of 0 to 20% of the total energy content of the preparation, and even more preferably in the range of 0 to 10% of the total energy content of the preparation.

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

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

[0208] In some preferred embodiments of the present invention, a packaged heat-treated beverage preparation is particularly useful as a nutritionally complete dietary supplement and contains a total amount of carbohydrates in the range of 30-60%, preferably in the range of 35-50%, of the total energy content of the beverage.

[0209] In some preferred embodiments of the present invention, packaged heat-treated beverage preparations are particularly advantageous for patients with kidney disease or impaired kidney function.

[0210] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation contains 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 50E%.

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

[0212] In one embodiment of the present invention, the liquid solution further contains 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 glycosides such as rebaudioside A, or combinations thereof. In some embodiments of the present invention, it is particularly preferred that the sweetener contains or consists of one or more high-intensity sweeteners (HIS).

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

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

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

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

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

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

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

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

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

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

[0223] In some preferred embodiments of the present invention, packaged heat-treated beverage preparations are particularly advantageous for patients with kidney disease or impaired kidney function.

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

[0225] In some preferred embodiments of the present invention, the sum of α-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 powdered non-BLG protein.

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

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

[0228] The inventors have observed indications that the reduction of lactoferrin and / or lactoperoxidase is particularly advantageous for obtaining color-neutral whey protein products.

[0229] Therefore, in some preferred embodiments of the present invention, lactoferrin is present at a weight percentage of total protein, up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, and most preferably up to 6% of total protein in a standard whey protein concentrate from sweet whey. Even lower concentrations of lactoferrin may be desired. Therefore, in a more preferred embodiment of the present invention, lactoferrin is present at a weight percentage of total protein, up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of total protein in a standard whey protein concentrate from sweet whey.

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

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

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

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

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

[0235] In one embodiment of the present invention, a packaged, heat-treated beverage preparation is a low-phosphorus and low-potassium beverage suitable for patients with kidney disease or impaired kidney function.

[0236] The packaged heat-treated beverage preparation of the present invention is particularly useful as a sports drink, in which case it preferably also contains optionally a very limited amount of lipids and / or optionally a limited amount of carbohydrates.

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

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

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

[0240] In some preferred embodiments of the present invention, the packaged heat-treated beverage preparation is particularly advantageous for patients with kidney disease or impaired kidney function. The beverage preparation has very low levels of other minerals such as phosphorus and potassium.

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

[0242] One aspect of the present invention is a method for producing a packaged heat-treated beverage preparation having a pH in the range of 2 to 4.7, comprising the following steps: a) -2 to 45% by weight of protein, of which at least 85% is BLG, -Optionally, sweeteners, sugar polymers and / or flavors The process of preparing a liquid solution containing, b) The process of packaging the liquid solution Includes, The liquid solution of step a) and / or the packaged liquid solution of step b) are subjected to a heat treatment including at least pasteurization. Regarding the method.

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

[0244] Since an even higher relative amount of BLG is achievable and 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 is BLG.

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

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

[0247] Aseptic filling and sealing are particularly preferred when the liquid solution is already sterile or contains very few microorganisms before filling.

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

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

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

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

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

[0253] Preferably, the duration of the heat treatment is 1 second to 30 minutes when performed in the temperature range of 70 to 82 degrees Celsius. The maximum exposure time is optimal at the lowest temperature in the temperature range, and vice versa. The lower the pH of the liquid solution, the more it can withstand higher temperatures without unfolding.

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

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

[0256] In some embodiments, higher temperatures may be preferred, particularly when unfolding and optionally aggregation of the BLG are also 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.

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

[0258] 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, heating to a temperature in the range of 90 to 95°C for 1 to 10 minutes, or, for example, heating to approximately 120°C for approximately 20 seconds. Alternatively, the heat treatment may include heating to a temperature in the range of 115 to 125°C for 5 to 30 seconds, or, for example, heating to approximately 120°C for approximately 20 seconds.

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

[0260] Alternatively, however, the heat treatment may also 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.

[0261] The heat treatment may be carried out using equipment such as plate or multi-tube heat exchangers, scraped surface heat exchangers, or retort systems. Alternatively, direct steam heating may be used, for example, using direct steam injection, direct steam injection, or spray cooking, which is particularly preferred for heat treatment above 95°C. Furthermore, such direct steam heating is preferably used in combination with flash cooling. A suitable example of the implementation of spray cooking can be found in International Publication No. 2009113858, which is incorporated herein by reference for all purposes. A suitable example of the implementation of direct steam injection and direct steam injection can be found in International Publication No. 2009113858 and International Publication No. 2010 / 085957, which are incorporated herein by reference for all purposes. General aspects of high-temperature treatment can be found, for example, in "Thermal technologies in food processing," ISBN 185573558 X, which is incorporated herein by reference for all purposes.

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

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

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

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

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

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

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

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

[0270] Useful examples of useful bases include hydroxide salts, such as sodium hydroxide or potassium hydroxide; carbonates or bicarbonates; carboxylates, such as citrates or lactates; and combinations thereof. Preferably, a base such as KOH or NaOH is used to adjust the pH.

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

[0272] Surprisingly, regarding appearance, it was found that using a whey protein beverage in which at least 85% w / w of protein is BLG allowed for an increase in pH during heat treatment, resulting in improvements in visual perception (color and turbidity) and viscosity compared to heat-treated WPI beverages.

[0273] Surprisingly, significant differences in sensory parameters were found between the BLG beverage of the present invention and the beverage produced with WPI. Surprisingly, and advantageously, the BLG beverage was found to have lower levels of astringency, dry texture, sourness, whey aroma, and citric acid flavor compared to the WPI beverage. It was further found that raising the pH of an acidic beverage reduces the amount of sweetener needed to balance the acidity of the beverage, and therefore, the concentration of sweetener required for such a beverage is lower.

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

[0275] These pH ranges are particularly suitable when beverage preparations are pasteurized.

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

[0277] These pH ranges, combined with high-temperature treatments such as sterilization, are particularly suitable for producing clear beverages with low viscosity and improved taste.

[0278] In some preferred embodiments of the present invention, the liquid solution has a pH in the range of 4.1 to 4.7, which is particularly suitable for producing a stable beverage having a milky 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.

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

[0280] 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.

[0281] Therefore, in some embodiments of the present invention, the liquid solution preferably contains 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.

[0282] In some embodiments of the present invention, it is advantageous that the protein content of the liquid solution is high, such as 10.0 to 45.0% w / w relative to the weight of the liquid solution.

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

[0284] It is particularly preferable that the liquid solution contains a BLG isolate in combination with other protein sources, preferably as the primary protein source, and if possible as the sole protein source.

[0285] The BLG isolate is preferably BLG isolate powder, or the liquid BLG isolate contains water and a solid amount of BLG isolate powder ranging from 1 to 50% w / w.

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

[0287] The BLG isolate powder preferably has one or more of the following: -At least 0.2 g / cm³ 3 bulk density, - At least an intrinsic tryptophan fluorescence emission ratio of 1.11 (I330 / I350), -Maximum protein denaturation degree of 10%, -Thermal stability at pH 3.9 with a maximum of 200 NTU, and - Up to 1000 colony-forming units / g.

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

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

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

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

[0292] In some cases, an even higher protein content may be required, and in some preferred embodiments of the present invention, the BLG isolate powder contains 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 of total protein.

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

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

[0295] In some preferred embodiments of the present invention, the sum of α-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 powdered non-BLG protein.

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

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

[0298] The inventors have observed indications that the reduction of lactoferrin and / or lactoperoxidase is particularly advantageous for obtaining color-neutral whey protein products.

[0299] Therefore, in some preferred embodiments of the present invention, lactoferrin is present at a weight percentage of total protein, up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, and most preferably up to 6% of total protein in a standard whey protein concentrate from sweet whey. Even lower concentrations of lactoferrin may be desired. Therefore, in a more preferred embodiment of the present invention, lactoferrin is present at a weight percentage of total protein, up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of total protein in a standard whey protein concentrate from sweet whey.

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

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

[0302] In some preferred embodiments of the present invention, the BLG isolate powder has a water 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.

[0303] In some preferred embodiments of the present invention, the BLG isolate powder contains 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% of carbohydrates. The BLG isolate powder may contain carbohydrates such as lactose, oligosaccharides and / or hydrolyzed lactose products (i.e., glucose and galactose), sucrose and / or maltodextrin.

[0304] 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.

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

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

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

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

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

[0310] The inventors have found that a low-phosphorus / low-potassium variant of BLG isolate powder is particularly useful for patients with kidney disease. To produce such a product, the BLG isolate powder must have equally low levels of phosphorus and potassium.

[0311] Therefore, in some preferred embodiments of the present invention, the BLG isolate powder has a total phosphorus content of up to 100 mg 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.

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

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

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

[0315] The low-phosphorus and / or low-potassium composition according to the present invention can be used as a food ingredient for producing foods for a group of patients with impaired renal function.

[0316] The inventors have found that having an acidic BLG isolate powder having a pH of up to 4.9, and more preferably up to 4.3, is particularly advantageous for several applications, such as acidic foods, especially acidic beverages. This is particularly applicable to high-protein, clear acidic beverages.

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

[0318] Therefore, 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, however also preferably, the BLG isolate powder may have a pH in the range of 3.6 to 4.3.

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

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

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

[0322] Advantageously, the BLG isolate powder used in this invention contains 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 may have a bulk density.

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

[0324] The high-density variant of the present invention is easier to handle and less likely to flow into the surrounding air.

[0325] An additional advantage of the high-density variants of the present invention is that they take up less space during transport, thereby increasing the weight of BLG isolate powder that can be transported in one volume.

[0326] Furthermore, an advantage of the high-density variant of the present invention is that these are, for example, powdered sugar (bulk density approximately 0.56 g / cm³). 3 ), granulated sugar (bulk density approximately 0.71 g / cm³) 3 ), powdered citric acid (bulk density approximately 0.77 g / cm³) 3 It is less likely to separate when used in powder mixtures with other powdered food ingredients such as ).

[0327] The BLG isolate powder of the present invention is 0.2 to 1.0 g / cm³. 3 The range is preferably 0.30 to 0.9 g / cm³. 3 The range is, more preferably 0.40 to 0.8 g / cm³. 3 The range, more preferably 0.45 to 0.75 g / cm³ 3 The range, more preferably 0.50 to 0.75 g / cm³ 3 The range is most preferably 0.6 to 0.75 g / cm³. 3 It can have a bulk density within this range.

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

[0329] The inventors found it advantageous to maintain the natural conformation of BLG and observed that when BLG is used in acidic beverages, increased unfolding of BLG results in an increased level of dry texture.

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

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

[0332] In some preferred embodiments of the present invention, the BLG isolate powder has an intrinsic tryptophan fluorescence 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, and most preferably at least 1.19.

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

[0334] In some preferred embodiments of the present invention, the protein fraction of the BLG isolate powder has an intrinsic tryptophan fluorescence 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, and most preferably at least 1.19.

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

[0336] It is generally preferable that a substantial portion of the 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.

[0337] 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%.

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

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

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

[0341] The inventors found that the thermal stability of BLG isolate powder at pH 3.9 is an excellent indicator of its usefulness in clear, high-protein beverages. Thermal stability at pH 3.9 is measured according to Example 1.2.

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

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

[0344] Therefore, 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 particularly preferred embodiments, the powder is sterile. Sterile BLG isolate powder can be prepared, for example, by combining several physical microbial reduction methods during the production of the BLG isolate powder, such as microfiltration at an acidic pH and heat treatment.

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

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

[0347] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of i) 2 to 4.9 or ii) 6.1 to 8.5. - At least 30% w / w of total protein, -Beta-lactoglobulin (BLG) in an amount of at least 85% w / w, preferably at least 90% w / w, relative to total protein. - Up to 6% w / w of water Includes, The BLG isolate powder is -At least 0.2 g / cm³ 3 bulk density, - At least an intrinsic tryptophan fluorescence emission ratio of 1.11 (I330 / I350), - A maximum of 10% protein denaturation, and - Thermal stability at pH 3.9 with a maximum of 200 NTU It has.

[0348] In another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 2 to 4.9. -Total protein in an amount of at least 80% w / w, preferably at least 90% w / w, and more preferably at least 94% w / w, -β-lactoglobulin (BLG) in an amount of at least 85% w / w, preferably at least 90% w / w, and more preferably at least 94% w / w of total protein, - Up to 6% w / w of water, - A maximum of 2% w / w of lipids, preferably a maximum of 0.5% w / w. Includes, The BLG isolate powder is -At least 0.2 g / cm³ 3 Preferably at least 0.3 g / cm³ 3 , more preferably at least 0.4 g / cm³ 3 bulk density, - At least an intrinsic tryptophan fluorescence emission ratio of 1.11 (I330 / I350), - A protein denaturation degree of up to 10%, preferably up to 5%, more preferably up to 2%, and - Thermal stability at pH 3.9 up to 50 NTU, preferably up to 30 NTU, and more preferably up to 10 NTU. It has.

[0349] In yet another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5. -Total protein in an amount of at least 80% w / w, preferably at least 90% w / w, and more preferably at least 94% w / w, -β-lactoglobulin (BLG) in an amount of at least 85% w / w, preferably at least 90% w / w, and more preferably at least 94% w / w of total protein, - Up to 6% w / w of water, - A maximum of 2% w / w of lipids, preferably a maximum of 0.5% w / w. Includes, The BLG isolate powder is -At least 0.2 g / cm³ 3 Preferably at least 0.3 g / cm³ 3 , more preferably at least 0.4 g / cm³ 3 bulk density, - A protein denaturation degree of up to 10%, preferably up to 5%, more preferably up to 2%, and - Thermal stability at pH 3.9 up to 50 NTU, preferably up to 30 NTU, and more preferably up to 10 NTU. It has.

[0350] In a more preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5. -Total protein in an amount of at least 80% w / w, preferably at least 90% w / w, and more preferably at least 94% w / w, -β-lactoglobulin (BLG) in an amount of at least 85% w / w, preferably at least 90% w / w, and more preferably at least 94% w / w of total protein, - Up to 6% w / w of water, - A maximum of 2% w / w of lipids, preferably a maximum of 0.5% w / w. Includes, The BLG isolate powder is -At least 0.2 g / cm³ 3 Preferably at least 0.3 g / cm³ 3 , more preferably at least 0.4 g / cm³ 3 bulk density, - A protein denaturation degree of up to 10%, preferably up to 5%, more preferably up to 2%, and - Thermal stability at pH 3.9 up to 50 NTU, preferably up to 30 NTU, and more preferably up to 10 NTU. It has.

[0351] In a more preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 5.0 to 6.0. -Total protein in an amount of at least 80% w / w, preferably at least 90% w / w, and more preferably at least 94% w / w, -β-lactoglobulin (BLG) in an amount of at least 85% w / w, preferably at least 90% w / w, and more preferably at least 94% w / w of total protein, - Up to 6% w / w of water, - A maximum of 2% w / w of lipids, preferably a maximum of 0.5% w / w. Includes, The BLG isolate powder is -At least 0.2 g / cm³ 3 Preferably at least 0.3 g / cm³ 3 , more preferably at least 0.4 g / cm³ 3 bulk density, - A maximum protein denaturation degree of 10%, preferably a maximum of 5%, more preferably a maximum of 2%. -Thermal stability at pH 3.9 of up to 50 NTU, preferably up to 30 NTU, and more preferably up to 10 NTU, and - Preferably a BLG crystallinity of less than 10% It has.

[0352] BLG isolate powder containing at least 85% w / w of BLG relative to total protein is typically obtained through the following process: a) i) pH in the range of 2 to 4.9 ii) pH in the range of 6.1 to 8.5, iii) pH in the range of 5.0 to 6.0 A liquid BLG isolate having the following steps: b) Optionally, a step of subjecting the liquid BLG isolate to physical microbial reduction, c) A step of drying the liquid BLG isolate, preferably by spray drying. It is provided by a method that includes the following.

[0353] BLG isolates are preferably prepared from mammalian milk, preferably from the milk of ruminant animals such as cows, sheep, goats, buffalo, camels, llamas, horses, and / or deer. Bovine milk-derived proteins are particularly preferred. Therefore, BLG is preferably bovine BLG.

[0354] Liquid BLG isolates can be provided in several different ways.

[0355] Typically, the provision of a liquid BLG isolate includes, or comprises, isolating BLG from a whey protein feedstock to obtain a BLG concentrated composition by one or more of the following methods: - Crystallization or precipitation of BLG by salt dissolution, - Crystallization or precipitation of BLG by salting out, - Ion exchange chromatography, and - Fractionation of whey protein by ultrafiltration.

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

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

[0358] The term "whey protein supply material" refers to compositions derived from BLG concentrates and subsequent liquid BLG isolates.

[0359] In some embodiments of the present invention, the preparation of the BLG concentrated composition includes or comprises high-salt BLG crystallization in the pH range of 3.6 to 4.0 according to U.S. Patent No. 2,790,790.

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

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

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

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

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

[0365] Non-limiting examples of mineral addition include the addition of soluble food-acceptable salts, such as salts of Na, K, Ca, and / or Mg. Such salts may be, for example, phosphates, chlorides, or salts of food acids, such as citrates or lactates. Minerals may be added in solid, suspension, or soluble form.

[0366] 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.

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

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

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

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

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

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

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

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

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

[0376] Even lower carbohydrate content is generally preferred, and therefore, in some preferred embodiments of the present invention, it is preferably in the range of 0 to 30% of the total energy content of the preparation, more preferably in the range of 0 to 20% of the total energy content of the preparation, and even more preferably in the range of 0 to 10% of the total energy content of the preparation.

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

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

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

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

[0381] The choice of sweetener may depend on the beverage being manufactured. High-intensity sugar sweeteners (e.g., aspartame, acesulfame K, or sucralose) may be used in beverages where an energy contribution from the sweetener is undesirable, while natural sweeteners (e.g., steviol glycosides, sorbitol, or sucrose) may be used in beverages with a natural profile.

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

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

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

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

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

[0387] In some embodiments of the present invention, a packaged heat-treated beverage preparation contains multiple minerals. In one exemplary embodiment, the liquid solution contains at least four minerals. In one embodiment, the four minerals are sodium, potassium, magnesium, and calcium.

[0388] The inventors have surprisingly found that, using BLG isolates as defined herein and in Example 2, it is possible to produce heat-treated beverage preparations with high mineral concentrations without compromising viscosity. This makes it possible to produce packaged heat-treated beverage preparations with high mineral content, enabling the production of beverages that are nutritionally complete or nutritionally incomplete dietary supplements.

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

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

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

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

[0393] In some preferred embodiments of the present invention, the liquid solution contains up to 150 mM KCl and up to 150 mM CaCl2, or the liquid solution contains up to 130 mM KCl and up to 130 mM CaCl2, or the liquid solution contains up to 110 mM KCl and up to 110 mM CaCl2, or the liquid solution contains up to 100 mM KCl and up to 100 mM CaCl2, or preferably the liquid solution contains up to 80 mM KCl and up to 80 mM CaCl2, or preferably the liquid solution contains up to 50 mM KCl and up to 50 mM CaCl2, or preferably the liquid solution contains up to 40 mM KCl and up to 40 mM CaCl2.

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

[0395] In the context of the present invention, the term "low mineral content" refers to compositions having at least one, preferably two, and more preferably all of the following: For example, liquids, beverages, powders, or other foods: - Ash content of up to 1.2% w / w relative to total solids, -Total calcium and magnesium content of up to 0.3% w / w relative to total solids, -Total sodium and potassium content of up to 0.10% w / w relative to total solids, -Total phosphorus content of up to 100mg per 100g of protein.

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

[0397] More preferably, the low-mineral composition comprises at least one, preferably two or more, and more preferably all of the following: - Ash content of up to 0.5% w / w relative to total solids, -Total calcium and magnesium content of up to 0.15% w / w relative to total solids, -Total sodium and potassium content of up to 0.06% w / w relative to total solids, -Total phosphorus content of up to 50mg per 100g of protein.

[0398] It is particularly preferable that the low-mineral composition has the following: - Ash content of up to 0.5% w / w relative to total solids, -Total calcium and magnesium content of up to 0.15% w / w relative to total solids, -Total sodium and potassium content of up to 0.06% w / w relative to total solids, -Total phosphorus content of up to 50mg per 100g of protein.

[0399] The inventors have found that the present invention makes it possible to prepare packaged, heat-treated beverage preparations that have very low levels of phosphorus and other minerals such as potassium, which are beneficial for patients with kidney disease or impaired kidney function.

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

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

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

[0403] In the context of the present invention, the term "low phosphorus" refers to a composition having a total phosphorus content of up to 100 mg of phosphorus per 100 g of protein, for example, a liquid, powder, or other food. 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 compositions according to the present invention may be used as food ingredients for producing foods for a group of patients with impaired renal function.

[0404] Therefore, 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.

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

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

[0407] The low-potassium composition according to the present invention can be used as a food ingredient for producing foods for a group of patients with impaired renal function.

[0408] Therefore, 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.

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

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

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

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

[0413] In one embodiment of the present invention, the liquid solution contains a plurality of vitamins and a plurality of minerals.

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

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

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

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

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

[0419] Another aspect of the present invention relates to the use of packaged heat-treated beverage preparations provided herein as dietary supplements.

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

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

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

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

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

[0425] In a preferred embodiment of the present invention, the packaged heat-treated beverage preparation has a pH in the range of 2.0 to 4.2, preferably 3.0 to 3.9, or preferably 3.2 to 3.7, and the beverage is Proteins that make up 2-10% w / w of the total weight of the beverage, of which at least 85% w / w, preferably at least 90% w / w, are BLG. -Optionally, sweeteners and / or flavors. Includes, The packaged heat-treated beverage preparation has a turbidity of up to 200 NTU, preferably up to 40 NTU.

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

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

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

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

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

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

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

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

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

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

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

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

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

[0439] In some embodiments of the present invention, the heat-treated beverage has a shelf life of at least 6 months at 25°C. - Edible BLG compositions as defined in PCT / EP2017 / 084553, providing a total amount of at least 1% (w / w), preferably at least 5% (w / w), of BLG. - Sweeteners, such as sugar sweeteners and / or non-sugar sweeteners, - At least one food acid, e.g., citric acid or other suitable food acid, -Optionally, flavorings and - Up to 80mg phosphorus / 100g protein Includes, It has a pH in the range of 2.5 to 4.0.

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

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

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

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

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

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

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

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

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

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

[0450] After allowing the pH-adjusted mixture to stand for 30 minutes, transfer 25 mL of the mixture to a 30 mL thin-walled glass test tube. Heat this to 75.0°C for 300 seconds by immersing it in a water bath at 75.0°C. Immediately after heating, transfer the glass test tube to an ice bath to cool to 1–5°C, and measure the turbidity of the heat-treated sample according to Example 1.7.

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

[0452] 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: - A first aqueous solution containing 5.0% (w / w) total protein and having a pH of 7.0 or 3.0, and A second aqueous solution containing -5.0% (w / w) total protein and having a pH of 4.6. pH adjustment is performed using 3% (w / w) NaOH (aqueous solution) or 5% (w / w) HCl (aqueous solution).

[0453] Total protein content of the first aqueous solution (P pH7.0又は3.0 ) is determined according to Example 1.5.

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

[0455] Example 1.4 Determination of protein denaturation (by pH 4.6 acidic precipitation) using reverse-phase UPLC analysis BLG samples (such as unheated reference and heated BLG beverage compositions) were diluted to 2% with MQ water. 5 mL of the protein solution, 10 mL of Milli-Q water, 4 mL of 10% acetic acid, and 6 mL of 1.0 M NaOAc were mixed and stirred for 20 minutes to precipitate and aggregate the denatured protein near pH 4.6. The solution was filtered through a 0.22 μm filter to remove aggregates and non-native proteins.

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

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

[0458] Gradient: 0 - 6.00 minutes 24 - 45% B; 6.00 - 6.50 minutes 45 - 90% B; 6.50 - 7.00 minutes 90% B; 7.00 - 7.50 minutes 90 - 24% B and 7.50 - 10.00 minutes 24% B. The concentration of native bLG in the sample was determined using the area of the BLG peak relative to the protein standard (Sigma L0130) (5-level calibration curve).

[0459] Samples were further diluted and reinjected if outside the linear range.

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

[0461] Example 1.6: Determination of non-aggregated BLG, ALA and CMP The contents of non-aggregated α-lactalbumin (ALA), β-lactoglobulin (BLG) and caseinomacropeptide (CMP) were analyzed by HPLC at 0.4 mL / min, respectively. 25 μL of the filtered sample was equilibrated with the eluent (465 g of Milli-Q water, 417.3 g of acetonitrile and 1 mL of trifluoroacetic acid) and injected into two TSKgel 3000PWxl (7.8 mm × 30 cm, Tosohass, Japan) columns connected in series with a binding precolumn PWxl (6 mm × 4 cm, Tosohass, Japan) using a UV detector at 210 nm.

[0462] Native α-lactalbumin (C α ), β-lactoglobulin (C β ) and caseinomacropeptide (C CMPThe 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.

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

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

[0465] Turbidity is measured in turbidimetric turbidity units (NTU).

[0466] 20 mL of beverage / sample was added to an NTU glass and placed in a Turbiquant® 3000IR turbidimeter. The NTU value was measured after stabilization and the process was repeated twice.

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

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

[0469] Unless otherwise specified, viscosity is 100 seconds. -1 The shear rate is provided in centipoise (cP). A higher measured cP value indicates higher viscosity.

[0470] Alternatively, viscosity can be estimated using Viscoman by Gilson, approximately 300 seconds. -1 It is reported as the shear rate.

[0471] Example 1.9: Color determination The color was measured using a chroma meter (Konica Minolta, CR-400). 15g of the sample was added to a small petri dish (55x14.2mm, VWR catalog number 391-0895) to avoid bubble formation. The protein content of the sample was standardized to 6.0 w / w% protein or less.

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

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

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

[0475] Standardize the sample to 6.0 w / w% protein or less.

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

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

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

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

[0480] Calculate protein loss:

number

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

[0482] Calculate the amount of insoluble protein:

number

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

[0484] Participants' mouths were rinsed between each sample using crackers, white tea, melon, and water.

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

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

[0487] Attributes (see the table above) were evaluated on a 15cm scale, with 0 = low intensity and 15 = high intensity.

[0488] Statistical analysis was performed using Panelcheck software with a three-way ANOVA test for multiple replicates. Samples were fixed, and panels were randomly configured.

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

[0490] Example 1.12: Determination of transparency by imaging The beverage preparation was photographed by placing the sample in a turbidity NTU measurement vial that was in contact with a piece of paper inscribed with the text "lorem ipsen". The inventors used a smartphone to photograph the vial and evaluated whether the text was clearly visible through the vial.

[0491] Example 1.13: Measurement of Ash Content The ash content of food is determined according to NMKL 173:2005, "Method of Ash Weight in Food."

[0492] Example 1.14: Determination of conductivity The "electrical conductivity" (sometimes called "specific conductivity") of an aqueous solution is a measure of the solution's ability to conduct electricity. Conductivity can be determined, for example, by measuring the AC resistance of the solution between two electrodes, and the result is typically expressed in units of millisiemens / cm (mS / cm). Conductivity can be measured, for example, according to EPA (U.S. Environmental Protection Agency) Method 120.1.

[0493] The conductivity values ​​mentioned herein are normalized to 25°C unless otherwise specified.

[0494] Conductivity is measured using a conductivity meter (WTW Cond 3210 equipped with a tetracon 325 electrode).

[0495] The system should be calibrated as described in the manual before use. The electrodes should be thoroughly rinsed in the same type of medium in which the measurement will be performed to avoid localized dilution. Lower the electrodes into the medium so that the area to be measured is completely submerged. Then, agitate the electrodes to remove any trapped air. Finally, keep the electrodes still until stable values ​​can be acquired and recorded from the display.

[0496] Example 1.15: Measurement of total solids content of a solution The total solids content of a solution can be determined according to NMKL 110, 2nd edition, 2005 (Total Solids (Water) - Weight Determination of Milk and Dairy Products). NMKL is an abbreviation for the "Nordic Commission on European Standard Analytical Methods and Food Analysis."

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

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

[0499] The pH glass electrode (with temperature compensation) should be carefully rinsed and calibrated before use.

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

[0501] If the sample is in powder form, dissolve 10 grams of the powder in 90 ml of desalted water at room temperature while stirring vigorously. Then, measure the pH of the solution at 25°C.

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

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

[0504] This method allows us to define the following three types of densities: • Packing density: This is the mass of the powder divided by the volume of the powder after it has been transferred to the specified graduated cylinder. • Loosening density is the mass obtained by dividing the volume of powder after 100 taps under the conditions specified in this standard by its volume. • Bulk density: This is the mass obtained by dividing the volume of the powder by the volume of the powder after 625 taps under the conditions specified in this standard.

[0505] These methods use a special graduated cylinder, 250 ml, graduated from 0 to 250 ml, weighing 190 ± 15 g (J. Engelsmann AG67059 Ludwigshafen / Rh) and a Stampf volumetric instrument (e.g., J. Engelsmann AG).

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

[0507] Pre-processing: Store the sample to be measured at room temperature.

[0508] Next, the sample is thoroughly mixed by repeatedly rotating and swirling the container (to avoid grinding the particles). The container should not be filled more than two-thirds full.

[0509] procedure: Measure 100.0 ± 0.1 g of powder and transfer it to a graduated cylinder. Read the volume V0 in ml.

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

[0511] Secure the graduated cylinder to the Stampf volumetric instrument and tap it 100 times. Smooth the surface with a spatula and measure the volume V. 100 Read it in ml.

[0512] Change the number of tabs to 625 (including 100 taps). After tapping, flatten the surface and create volume V. 625 Read it in ml.

[0513] Density calculation: The looseness density and bulk density, expressed in g / ml, are calculated according to the following formula: Bulk density = M / V (In the formula, M represents the sample weighed in grams, and V represents the volume after 625 taps in ml).

[0514] Example 1.18: Determining the water content of powder The water content of food is measured according to ISO 5537:2004 (Milking powder - Measurement of water content (reference method)). NMKL is an abbreviation for the "Nordic Commission on European Standard Analytical Methods and Food Analysis".

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

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

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

[0518] Pre-processing: Measure out a certain amount of powder and transfer it to a microwave decomposition tube. Add 35 mL of 1 M HNO. Decompose the sample with microwaves according to the microwave instructions. Place the decomposed tube in a ventilation chamber, remove the lid, and allow the volatile fumes to evaporate.

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

[0520] A blind sample is prepared by diluting a mixture of 110 mL of 1 M HNO3 and 0.5 mL of yttrium solution in 2% HNO3 to a final volume of 100 mL using Milli-Q water.

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

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

[0523] Example 1.21: Determination of the degree of crystallinity of BLG in liquid The degree of crystallinity of BLG in a liquid with a pH in the range of 5-6 is determined using the following method. a) Transfer 10 mL of the liquid sample to a Maxi-Spin filter equipped with a CA membrane with a pore size of 0.45 microns. b) Immediately rotate the filter at 1500g for 5 minutes, keeping the centrifuge at 2°C. c) Add 2 mL of cold Milli-Q water (2°C) to the holding liquid side of the spin filter. Immediately, with the centrifuge still cooled to 2°C, spin the filter at 1500 g for 5 minutes. Collect the permeate (permeate A), measure its volume, and determine the BLG concentration via HPLC using the method outlined in Example 1.31. d) Add 4 mL of 2M NaCl to the retaining liquid side of the filter, stir quickly, and let the mixture stand at 25°C for 15 minutes. e) Immediately spin the filter with 1500g for 5 minutes and collect the permeate (permeate B). f) Determine the total BLG weights of permeate A and permeate B using the method outlined in Example 1.31, and convert these results to total BLG weights rather than weight percentages. The weight of BLG in permeate A is m 透過液A It is called and the weight of BLG in permeate B is m 透過液B It is called that. g) Determine the degree of crystallinity of the liquid related to BLG as follows: Crystallinity=m 透過液B / (m 透過液A +m 透過液B ) * 100%

[0524] Example 1.22: Determination of the degree of crystallinity of BLG in dry powder This method is used to determine the degree of crystallinity of BLG in the dried powder. a) Mix 5.0 grams of the powder sample with 20.0 grams of cold Milli-Q water (2°C) and let stand at 2°C for 5 minutes. b) Transfer the liquid sample to a Maxi-Spin filter equipped with a 0.45 micron CA membrane. c) Immediately spin the filter at 1500g for 5 minutes, keeping the centrifuge at 2°C. d) Add 2 mL of cold Milli-Q water (2°C) to the holding liquid side of the spin filter, immediately spin the filter at 1500 g for 5 minutes, collect the permeate (permeate A), measure the volume, determine the BLG concentration via HPLC using the method outlined in Example 1.31, and convert the result to the total weight of BLG instead of weight %. The weight of BLG in permeate A is m 透過液A It is called that. f) Next, the degree of crystallinity of BLG in the powder is calculated using the following formula:

number

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

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

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

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

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

[0530] The crystalline material is separated, and X-ray crystal structure analysis is performed to confirm the existence of a crystalline structure. Preferably, the crystal lattice (space group and unit cell dimensions) is also confirmed to correspond to the lattice of a BLG crystal.

[0531] The chemical composition of the separated crystalline material was analyzed to confirm that the solid was mainly composed of BLG.

[0532] Example 1.25: Determining the total amount of lactose The total amount of lactose is determined according to ISO 5765-2:2002 (IDF 79-2:2002) "Powdered milk, dry ice mixtures and processed cheese - Determination of lactose content - Part 2: Enzymatic method using the galactose portion of lactose".

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

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

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

[0536] Approximately 500 μl of the sample was transferred to the prism surface of the instrument, and the measurement was started. The measured value was read and recorded.

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

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

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

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

[0541] The separation method is size exclusion chromatography (SEC), which uses 6M guanidine HCl buffer as both the sample solvent and the HPLC mobile phase. Mercaptoethanol is used as a reducing agent to reduce the disulfide (SS) of the protein or protein aggregate to create an unfolding monomer structure.

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

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

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

[0545] Equipment / Materials: 1. HPLC pump 515 (Waters) with manual seal cleaning 2. HPLC Pump Controller Module II (Waters) 3. Autosampler 717 (Waters) 4. Dual absorbance detector 2487 (Waters) 5. Computer software capable of generating quantitative reports (Empower 3, Waters) 6. Analysis columns: Two TSK-GEL G3000SWXL (7.8 x 300 mm, P / N: 08541) Guard column: TSK-Guard Column SWxL (6.0 x 40mm, P / N: 08543) 7. Ultrasonic bath (Branson 5200) 25mm syringe filter with 8.0.2μm cellulose acetate membrane (514-0060, VWR)

[0546] procedure: Mobile phase: A. Stock buffer solution 1. Weigh 6.6g of Na2HPO45, 3.5g of NaH2PO4, and 2.9g of EDTA into a 1000mL beaker. Dissolve in 800 mL of water. 2. Measure the pH and adjust it to 7.5 ± 0.1 if necessary using HCl (to lower the pH) or NaOH (to raise the pH). 3. Transfer to a 1000 mL volumetric flask and dilute with water to the specified volume.

[0547] B.6M guanidine HCl mobile phase 1. Weigh 1146g of guanidine HCl into a 2000mL beaker and add 200mL of stock buffer (A). 2. Dilute this solution with water to approximately 1600 mL while mixing with a magnetic stirring rod (50°C). 3. Adjust the pH to 7.5 ± 0.1 with NaOH. 4. Transfer to a 2000 mL volumetric flask and dilute with water to the specified volume. 5.0.Filter the solvent using a solvent filtration apparatus equipped with a 22 μm membrane filter.

[0548] Calibration Standard The calibration standard for each protein to be quantified is adjusted as follows: 1. 10 mL of approximately 25 mg of protein reference standard. Measure the substance accurately (to the nearest 0.01 mg) into a volumetric flask and dissolve it in 10 mL of water. This will be the protein stock standard solution (S1) for the protein. 2. Pipette 200 μl of S1 into a 20 ml volumetric flask and dilute with the mobile phase to the specified volume. This is the low-usage standard solution WS1. 3. Pipette 500 μL of S1 into a 10 mL volumetric flask and dilute with the mobile phase to the specified volume. This is the standard solution WS2. 4. Pipette 500 μL of S1 into a 5 mL volumetric flask and dilute with the mobile phase to the specified volume. This is the standard solution WS3. 5. Pipette 750 μL of S1 into a 5 mL volumetric flask and dilute with the mobile phase to the specified volume. This is the standard solution WS4. 6. Pipette 1.0 mL of S1 into a 5 mL volumetric flask and dilute with the mobile phase to the specified volume. This is the highly used standard solution WS5. 7. Using a graduated disposable pipette, transfer 1.5 mL of WS1-5 into separate vials. Add 10 μL of 2-mercaptoethanol to each vial and cap. Vortex the solution for 10 seconds. Leave the standard setup at ambient temperature for about 1 hour. 8.0.Filter the standard using a 22 μm cellulose acetate syringe filter.

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

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

[0551] HPLC system / column Column equilibration 1. Connect the GPC Guard column and the two GPC analysis columns in series. A new column is typically placed in a phosphate buffer. 2. Gradually introduce water into a new column at a rate of 0.1–0.5 mL / min over 30–60 minutes. Continue playing for about an hour. 3. Gradually reduce the flow rate from 0.5 mL / min to 0.1 mL / min, replacing the mobile phase in the reservoir. 4. To avoid pressure shock, gradually increase the pump flow rate to 0.1-0.5 mL / min over 30-60 minutes, and then maintain it at 0.5 mL / min. 5. Inject 10 samples into the column to saturate it, and wait for the peaks to elute. This helps with column conditioning. This process can be performed without having to wait for each injection to complete before the next injection. 6. Equilibrium with the mobile phase for at least 1 hour.

[0552] Calculation of the result The quantitative determination of the content of proteins to be quantified, such as α-lactalbumin, β-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.

[0553] Example 2: Preparation of spray-dried acidic BLG isolate powder Whey protein supply ingredients Lactose-depleted UF (ultra-fine) retained liquid derived from sweet whey from a standard cheese-making process was filtered through a 1.2-micron filter and fat was reduced via a Synder FR membrane before use as a feedstock for the BLG crystallization method. The chemical composition of the feedstock is shown in Table A. Note that all weight percentages of specific proteins such as BLG and ALA mentioned in this example refer to the weight percentage of non-aggregated proteins relative to total protein.

[0554] adjustment Sweet whey feed material is supplied to a 46-mill spacer at a feed material pressure of 1.5-3.0 bar using a Koch HFK-328 type membrane (70m). 2Using a membrane, polishing water (water filtered by reverse osmosis to obtain a maximum conductivity of 0.05 mS / cm) was used as the diafiltration medium, and the feed material concentration was adjusted to a total solids (TS) of 21% ± 5 at an ultrafiltration setting of 20°C. Then, HCl was added to adjust the pH to approximately 5.5. Diafiltration was continued until the decrease in conductivity of the retained solution was less than 0.1 mS / cm over 20 minutes. The permeate flow was then measured at 1.43 L / hour / m 2 The holding solution was concentrated until it was less than [amount missing]. A first sample of the concentrated holding solution was taken and subjected to centrifugation at 3000g for 5 minutes. The supernatant of the first sample was used to determine the BLG yield.

[0555] crystallization The concentrated holding solution was transferred to a 300L crystallization tank, and pure BLG crystal material made from BLG crystals that had been rehydrated and spray-dried was seeded. Subsequently, the seeded whey protein solution was cooled from 20°C to approximately 6°C for about 10 hours to form and grow BLG crystals.

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

[0557] [Table A]

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

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

[0560] Data processing: Since both supernatants were processed using the same method, the relative yield can be calculated by directly comparing the areas of the BLG peaks. Because the crystals contain only BLG and all samples were processed using the same method, the concentration of α-lactalbumin (ALA), and therefore the ALA area, should be the same in all samples. Therefore, the ALA area before and after crystallization is used as a correction factor (cf) when calculating the relative yield.

number

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

number

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

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

[0564] 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 water content of approximately 4% w / w, and the chemical composition of the powder is shown in the table. The dried powder sample was dissolved, the degree of protein denaturation was determined to be 1.5%, and the intrinsic tryptophan fluorescence emission ratio (I330 / I350) was measured to be 1.20.

[0565] [Table B]

[0566] The bulk density (625 taps) of the spray-dried powder is 0.2-0.3 g / cm³. 3 It was estimated that...

[0567] Example 3: Preparation of a typical whey protein beverage A dried BLG isolate protein powder containing more than 85% BLG is dispersed in approximately 75% desalted water, which is necessary to reach the desired final protein concentration.

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

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

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

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

[0572] Add the remaining water to reach the desired protein concentration, and optionally homogenize the composition.

[0573] For comparison, whey protein isolate was used to replace more than 85% of the BLG product in the preparation of the reference sample, while maintaining the remaining steps.

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

[0575] Example 4: Heat treatment of whey protein composition The beverage was heat-treated using a plate heat exchanger (manufacturer: OMVE HTST / UHT pilot plant HT320-20) equipped with a 10 μm bonded microfiber filter element, code 12-57-60k (Headline filter), by heating at 120°C for 20 seconds (high temperature, short duration (HTST), resulting in denaturation of BLG) or by holding at 75°C for 15 seconds to 5 minutes (BLG remains in its natural state). Other heat treatment conditions may be applied.

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

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

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

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

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

[0581] Using the same procedure, a WPI beverage was prepared from WPI powder.

[0582] Table 1 below shows the composition of the BLG powder used in the preparation of the beverage, and for comparison, the composition of WPI is also listed.

[0583] [Table 1]

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

[0585] The results are shown in Table 2 and Figure 1 below. [Table 2]

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

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

[0588] The viscosity remained low.

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

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

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

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

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

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

[0595] Surprisingly, the inventors found that when the BLG beverage preparation was sterilized using UHT (Figure 2), it remained clear even at pH 3.7, and when pasteurized (Figures 3 and 4), where the WPI was opaque under those conditions, the pH could exceed 3.7 (pH 3.9-4.1). These findings were further supported by turbidity measurements shown in Figure 5 (UHT) and Figure 6 (pasteurized), where the WPI remained below 40 NTU even at pH 3.7 and 3.9, respectively, where the WPI significantly exceeded 40 NTU.

[0596] In BLG beverage preparations, the viscosity remains low after UHT treatment. This low viscosity demonstrates that the beverage sample is easily drinkable. Viscosity increases dramatically, especially at high pH values, when WPI is used (Figure 7).

[0597] The authors further investigated the yellowness (b) of heat-treated WPI beverages containing small amounts of BLG (both UHT and pasteurized). * We found that the value significantly exceeded BLG up to at least pH 3.7. See Figures 8 (UHT) and 9 (pasteurization).

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

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

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

[0601] The solution was thermally treated at 75–120°C for a duration of 15 seconds to 5 minutes, as described in Table 3, and immediately cooled on ice.

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

[0603] [Table 3]

[0604] result: The results are shown in Table 3 and Figures 10 to 12 above.

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

[0606] Figure 11 shows the sensory evaluation of the high-protein BLG beverage composition, as well as images of the 6 w / w% and 15 w / w% BLG samples at pH 3.7; both samples are clear.

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

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

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

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

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

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

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

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

[0615] Example 8: Whey protein beverage preparation with improved taste BLG and WPI samples were prepared. The composition of the samples is shown below.

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

[0617] The samples were analyzed by a sensory panel of 10 people (see Example 1.11). The WPI samples were particularly yellow at high pH values ​​and had higher b * The sample had a value and exhibited high turbidity. The analytical data is presented in Table 3.

[0618] [Table 4]

[0619] The visual appearance of the samples in Table 4 is shown in Figure 13.

[0620] The sensory evaluation data is shown in Figures 14 to 18.

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

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

[0623] Figure 15 shows that raising the pH to 3.7 before heat treatment reduces the acidity of the BLG beverage at both 120°C and 75°C while maintaining the clarity and low color of the product. This was not possible with WPI, as a clear and clarified beverage cannot be produced at pH 3.7, as seen in Table 2 and Figure 1.

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

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

[0626] Figure 18 demonstrates that maintaining BLG in its natural state by using heating at 75°C / 15 seconds at pH 3.7, which is not possible to produce a clear, clarified, colorless WPI beverage, reduces the whey aroma.

[0627] A clear beverage could not be produced using WPI (Whey Protein Isolate) heat-treated at 75°C for 15 seconds at a pH of 3.7. See also Figure 3.

[0628] Example 9: Low-color sweetened BLG beverage preparation A 6% w / w BLG beverage was prepared. Please refer to the following BLG powder composition. The beverage was prepared as described in Example 3. [Table 4-2]

[0629] The prepared BLG beverage contained 6% protein and had pH values ​​of 3.7 and 4.3.

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

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

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

[0633] [Table 5]

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

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

[0636] [Table 6] [Table 7]

[0637] result: Table 7 shows that both BLG beverages containing additives and BLG beverages without additives remain low viscosity, clear, and essentially colorless.

[0638] Example 10: Exemplary method for a clarified BLG beverage preparation containing added minerals The BLG powder used in this example had a pH of 5.5 and contained approximately 96% w / w of protein as BLG (and 0.4% w / w of protein as ALA).

[0639] Acidic BLG isolate powder was prepared according to Example 2, and the beverage preparation was prepared according to Example 5.

[0640] High-temperature heat treatment of beverage preparations: A 6% BLG beverage preparation with a pH of 3.7 was prepared. KCl and CaCl2 were added in liquid form from a 1M stock solution. These were heat-treated at less than 95°C for 5 minutes.

[0641] result: The results are summarized in Table 8 and Figure 19 below.

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

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

[0644] Gelation was observed with the addition of 30 mM KCl (turbid gel).

[0645] Gelation was observed upon the addition of 220 mM CaCl (clarified gel).

[0646] The mineral additions in Table 8 significantly exceed the difference between the BLG and WPI products, clearly suggesting that protein composition is more important than the difference in minerals compared to WPI. The sample remained clear (see Figure 19) and had low viscosity within the limits shown in Table 8 below.

[0647] [Table 8]

[0648] Low-temperature heat treatment of beverage preparations A 6% BLG beverage preparation with a pH of 3.7 was prepared. KCl and CaCl2 were added in liquid form from a 1M stock solution. These were heat-treated at a pasteurization temperature of 75°C for 5 minutes.

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

[0650] Figure 20 shows an image of a 6% BLG beverage with a pH of 3.7, which has been heat-treated at 75°C for 5 minutes and to which minerals have been added. A: Added minerals 0mM, B: Added KCl 100 mM, C: Added CaCl2 100mM, D: Added KCl 100mM and added CaCl2 100mM

[0651] Even when 100 mM KCl or 100 mM CaCl2 was added to the beverage composition before heating, the beverage preparation remained clear. See Figure 20. Furthermore, even when both 100 mM KCl and 100 mM CaCl2 were added, the viscosity remained surprisingly low.

[0652] [Table 9]

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

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

[0655] The results are shown in Tables 10 and 11 and Figure 21 below.

[0656] [Table 10]

[0657] [Table 11]

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

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

[0660] The results presented in Tables 10 and 11 and Figure 21 demonstrate that high-end pH levels, such as pH 4.3, enable the production of dairy beverages, which is preferable in some embodiments of the present invention, for example, when consumers prefer whey protein beverages with a milky appearance. It was also found that low viscosity was achieved at pH 4.3 in both sucrose-containing and sucrose-free preparations.

[0661] The color remained neutral. This is particularly preferred by consumers who prefer dairy beverages not to have a yellowish tint. * When the value is high, a yellowish color can be observed.

[0662] Even after centrifugation at 3000×g for 5 minutes, the protein loss was less than 15%, indicating that the beverage was stable, as evidenced by its high turbidity.

[0663] Due to gelation and high viscosity, it was not possible to produce a milky 6 w / w% protein WPI beverage based on WPI-A or WPI-B at pH 4.3, and this applied to both WPI samples with and without sucrose added.

[0664] Example 12: Milk whey protein beverage, prolonged low-temperature heat treatment Exemplary method for producing a dairy beverage containing BLG at different pH levels: Dissolve BLG powder in tap water and adjust the pH to 4.2-4.5 using 10% phosphoric acid according to Example 3. When the preparation was thermally treated at 75°C for 5 minutes, it had a protein content of 6% w / w. The BLG beverage is produced based on BLG powder having a pH of 3.9, containing approximately 92% w / w of protein as BLG and approximately 0.42% w / w of protein as ALA.

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

[0666] The results are shown in Table 12 and Figure 22 below.

[0667] 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 4.2–4.5.

[0668] [Table 12]

[0669] result: Beverages with a pH of 4.2 to 4.5 were found to have a milky, opaque appearance and high turbidity, while still exhibiting low viscosity.

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

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

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

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

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

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

[0676] [Table 13]

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

[0678] Furthermore, the SPI beverage is more b than the BLG sample. *The value was high, and therefore it had a yellowish color.

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

[0680] The inventors were surprised to find that a 6% nutritional composition containing 100 mM added KCl and 100 mM added CaCl2 remained liquid (viscosity approximately 1 cP) even after heating at 75°C for 5 minutes, indicating that BLG beverages (85% or more) can tolerate surprisingly high mineral concentrations present during sterilization by pasteurization at 75°C with a maximum holding time of at least 5 minutes (Example 10).

[0681] Since the thermal stability of whey protein is typically compromised by high mineral dosages, the inventors further investigated the opportunity to produce a nutritionally complete acidic BLG beverage and a sterile nutritional beverage containing more than 85% BLG, carbohydrate source, fat source, and minerals in a combination that meets current FSMP (Foods for Special Medical Purposes) requirements.

[0682] Dissolve the protein and mix it with lipids and carbohydrates in the ratios shown in Table 14, based on the energy distribution.

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

[0684] To meet FSMP requirements and produce nutritionally complete dietary supplements, additional vitamins may be added to the beverage.

[0685] [Table 14]

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

[0687] In certain embodiments, (1) 40 mM KCl and 14 mM CaCl2, or (2) 80 mM KCl and 28 mM CaCl2 were added, either together with additional components or (3) without further mineral additions as shown in Table 14.

[0688] The solution was homogenized at 200 bar.

[0689] The solution was thermally treated by immersing it in a 75°C or 95°C water bath for 5 minutes, and then cooled on ice.

[0690] [Table 15]

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

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

[0693] Example 15: Low-phosphoprotein beverage Using the purified BLG product from Example 3 (crystal preparation obtained from supply material 3), four low-phosphorus beverage samples were prepared. All dry components were mixed with desalinated water to obtain 10 kg of each sample, which was then hydrated at 10°C for 1 hour. [Table 15-2]

[0694] The sample is exposed to 90°C for 180 seconds and then aseptically packed into a sterile container.

[0695] Packaged beverages have a shelf life of at least one year at ambient temperature.

[0696] Because all the ingredients used to prepare the five beverages are low in phosphorus, the resulting beverages have a much lower phosphorus content than 80 mg / 100g protein. Therefore, four of the beverages are suitable for use as protein drinks for patients with kidney disease.

[0697] [Claim 1] Packaged heat-treated beverage preparations having a pH in the range of 2 to 4.7, comprising the following: - Of the total amount of protein, at least 85% w / w of which is β-lactoglobulin (BLG) relative to the weight of the beverage, -Optionally, sweeteners, sugar polymers, and / or flavors. [Claim 2] A packaged heat-treated beverage preparation according to claim 1, which is at least pasteurized. [Claim 3] A sterile, packaged, heat-treated beverage preparation according to claim 1. [Claim 4] A packaged heat-treated beverage preparation according to any one of claims 1 to 3, wherein the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of at least 1.11. [Claim 5] A packaged heat-treated beverage preparation according to any one of claims 1 to 4, wherein the protein fraction of the beverage preparation has an intrinsic tryptophan fluorescence emission ratio (I330nm / I350nm) of less than 1.11. [Claim 6] A packaged heat-treated beverage preparation according to any one of claims 1 to 5, wherein the protein fraction has a maximum degree of protein denaturation of 10%. [Claim 7] A packaged heat-treated beverage preparation according to any one of claims 1 to 6, having a maximum degree of protein denaturation of 10%. [Claim 8] A packaged heat-treated beverage preparation according to any one of claims 1 to 7, having a pH in the range of 3.0 to 4.3. [Claim 9] The protein fraction of the beverage preparation corresponds to a delta b color value in the range of -0.10 to +0.51 on the CIELAB color scale. * It has delta b * = Measured at room temperature, b 6.0w / w%タンパク質に標準化された試料 * -b 脱塩水 * A packaged heat-treated beverage preparation according to any one of claims 1 to 8. [Claim 10] Delta b is a color value in the range of -0.10 to +0.51 on the CIELAB color scale. * It has delta b * = Measured at room temperature, b 6.0w / w%タンパク質に標準化された試料 * -b 脱塩水 * A packaged heat-treated beverage preparation according to any one of claims 1 to 9. [Claim 11] A packaged heat-treated beverage preparation according to any one of claims 1 to 10, wherein the sum of the amounts of Na, K, Mg, and Ca is at most 750 mM. [Claim 12] A packaged heat-treated beverage preparation according to any one of claims 1 to 11, having a turbidity of up to 200 NTU. [Claim 13] A packaged heat-treated beverage preparation according to any one of claims 1 to 12, having a turbidity of more than 200 NTU. [Claim 14] A packaged heat-treated beverage preparation according to any one of claims 1 to 13, wherein the protein fraction contains up to 15% insoluble material after centrifugation of 3000 g for 5 minutes. [Claim 15] A packaged heat-treated beverage preparation according to any one of claims 1 to 14, having a viscosity of up to 200 cP centipoise when measured at 22 degrees Celsius with a shear rate of 100 psi / second. [Claim 16] A packaged heat-treated beverage preparation according to any one of claims 1 to 15, which does not contain a flocculation inhibitor. [Claim 17] A packaged heat-treated beverage preparation according to any one of claims 1 to 16, comprising a total amount of protein of 4.0 to 30% w / w relative to the weight of the beverage. [Claim 18] A packaged heat-treated beverage preparation according to any one of claims 1 to 17, further comprising carbohydrates in the range of 0 to 95% of the total energy content of the preparation. [Claim 19] A packaged heat-treated beverage preparation according to any one of claims 1 to 18, further comprising a lipid content of 0 to 60% of the total energy content of the preparation. [Claim 20] A packaged heat-treated beverage preparation according to any one of claims 1 to 19, wherein each major non-BLG whey protein is present in a weight percentage of total protein, up to 20%, preferably up to 15%, more preferably up to 10%, even more preferably up to 6%, and most preferably up to 4%, of its weight percentage of total protein in a standard whey protein concentrate from sweet whey. [Claim 21] A packaged heat-treated beverage preparation according to any one of claims 1 to 20, comprising a BLG isolate. [Claim 22] A method for producing a packaged heat-treated beverage preparation having a pH in the range of 2 to 4.7, comprising the following steps: a) -2 to 45% by weight of protein, of which at least 85% is BLG. -Optionally, sweeteners, sugar polymers and / or flavors, A step of preparing a liquid solution containing the following: b) A step of packaging the liquid solution, The liquid solution in step a) and / or the packaged liquid solution in step b) are subjected to a heat treatment including at least pasteurization. [Claim 23] A protein solution containing a total amount of protein at a ratio of 2-45% w / w relative to the weight of the solution is used to control the turbidity of a heat-treated acidic beverage preparation having a pH in the range of 2.0-4.7, wherein at least 85% w / w% of the protein is BLG. [Claim 24] A protein solution containing a total amount of protein at a ratio of 2-45% w / w relative to the weight of the solution is used to control the astringency of a heat-treated acidic beverage preparation having a pH in the range of 2.0-4.7, wherein at least 85% w / w% of the protein is BLG. [Claim 25] A packaged, heat-treated beverage preparation according to any one of claims 1 to 21, for use in a method for treating a disease related to protein malabsorption. [Claim 26] Use of a packaged heat-treated beverage preparation according to any one of claims 1 to 21 as a nutritional supplement. [Claim 27] Use of the packaged heat-treated beverage preparation according to claim 26, which is consumed before, during, or after exercise.

Claims

1. 1. A packaged thermally treated beverage preparation having a pH in the range of 2 to 4.7, comprising: - a total amount of 10-45% w / w of proteins relative to the weight of the beverage, at least 85% w / w of which is beta-lactoglobulin (BLG); - Optionally, sweeteners, sugar polymers and / or flavors.

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

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

4. 4. The packaged heat-treated beverage preparation according to claim 1, wherein the protein fraction has a degree of protein denaturation of up to 10%.

5. 5. The packaged heat-treated beverage preparation according to any one of claims 1 to 4, having a degree of protein denaturation of up to 10%.

6. A packaged heat-treated beverage preparation according to any one of claims 1 to 5, having a pH in the range of 3.0 to 4.

3.

7. 7. The packaged heat-treated beverage preparation according to any one of claims 1 to 6, wherein the sum of the amounts of Na, K, Mg and Ca is at most 750 mM.

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

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

10. A packaged heat-treated beverage preparation according to any one of claims 1 to 9, wherein the protein fraction contains at most 15% insoluble material after centrifugation at 3000g for 5 minutes.

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

12. A packaged heat-treated beverage preparation according to any one of claims 1 to 11, which does not contain any anti-agglomeration agent.

13. A packaged heat-treated beverage preparation according to any one of claims 1 to 12, comprising a total amount of protein of 10 to 30% w / w by weight of the beverage.

14. A packaged thermally treated beverage preparation according to any one of claims 1 to 13, further comprising carbohydrates in the range of 0-95% of the total energy content of the preparation.

15. A packaged thermally treated beverage preparation according to any one of claims 1 to 14, further having a lipid content of 0 to 60% of the total energy content of the preparation.

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

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

18. A packaged heat-treated beverage preparation described in any one of claims 1 to 17, wherein at least 90 weight percent of the protein is beta-lactoglobulin.

19. A packaged heat-treated beverage preparation described in any one of claims 1 to 18, wherein at least 92 weight percent of the protein is beta-lactoglobulin.

20. 1. A method for producing a packaged thermally treated beverage preparation having a pH in the range of 2 to 4.7, comprising the steps of: a) - 10 to 45% by weight of a total amount of proteins, of which at least 85% are BLGs; optionally sweeteners, sugar polymers and / or flavors, providing a liquid solution comprising: b) packaging the liquid solution; The liquid solution of step a) and / or the packaged liquid solution of step b) are subjected to a heat treatment including at least pasteurization.

21. Use of a protein solution containing a total amount of protein of 2 to 45% w / w based on the weight of the solution, for controlling the turbidity of a heat-treated acidic beverage preparation having a pH in the range of 2.0 to 4.7, wherein at least 85 w / w% of the protein is BLG.

22. Use of a protein solution containing 2-45% w / w total amount of protein based on the weight of the solution to control astringency in a heat-treated acidic beverage preparation having a pH in the range of 2.0-4.7, wherein at least 85% w / w of the protein is BLG.

23. 20. A packaged thermally treated beverage preparation according to any one of claims 1 to 19 for use in a method for treating a disease associated with protein malabsorption.

24. Use of a packaged heat-treated beverage preparation according to any one of claims 1 to 19 as a dietary supplement.

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

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