BLG-based instant beverage powder

Abstract

The present invention relates to an instant beverage powder product and a method for manufacturing an instant beverage powder product, a liquid food produced from instant beverage powder and a method for manufacturing the liquid food, uses of the liquid food, and a kit comprising an instant beverage powder product.

Description

Technology Field

[0001] The present invention relates to an instant beverage powder product and a method for manufacturing an instant beverage powder product, a liquid food produced from instant beverage powder and a method for manufacturing the liquid food, uses of the liquid food, and a kit comprising an instant beverage powder product. Background Technology

[0002] Nutritional supplements containing whey (milk serum) protein are generally used for muscle synthesis, weight control, and maintaining muscle and body weight. These supplements target different types of consumers, such as sportsmen / women, athletes, children, the elderly, and patients who are malnourished or at risk of malnutrition, as well as those with increased protein requirements. Therefore, consumer perception of nutritional supplements is very important, as consumers must feel a sense of satisfaction when consuming the product.

[0003] Whey protein can be isolated from whey or whey. Whey typically contains a mixture of beta-lactoglobulin (BLG), alpha-lactalbumin (ALA), serum albumin, and immunoglobulin, of which BLG is the most dominant. Therefore, whey protein concentrate (WPC) contains a mixture of these proteins. Whey protein isolate (WPI) contains less fat and lactose than WPC. The isolation of beta-lactoglobulin (BLG) from whey or whey is the subject of numerous publications and typically involves several separation steps and often chromatographic techniques to obtain a purified beta-lactoglobulin product.

[0004] International patent application WO2002 / 056707 (Nestle) describes a balanced powder blend composition having at least one fat or oil source, at least one carbohydrate source, and at least one protein source. This composition is advantageously added to food to supplement the nutritional value of the food, but does not substantially alter the taste of the food.

[0005] WO 2018 / 115520 A1 discloses a method for preparing an isolated beta-lactoglobulin composition for food and / or a composition containing crystallized beta-lactoglobulin based on the crystallization of BLG in a salting-in mode. The crystallized BLG can then be separated from the residual mother liquor.

[0006] WO 2011 / 112695 A1 discloses a nutritional composition and a method for preparing and using the nutritional composition. The nutritional composition comprises whey protein micelles and leucine and provides a sufficient amount of leucine to improve human protein synthesis while also maintaining a low-viscosity fluid matrix and acceptable organoleptic properties.

[0007] WO2011 / 051436 A1 discloses at least partially transparent compositions intended for human or animal consumption and relates to packaging of such compositions. One embodiment of the present invention relates to at least partially transparent containers containing at least partially transparent aqueous non-alcoholic compositions. The containers comprise at least one polarizer that visualizes liquid crystals present in the compositions.

[0008] WO2004 / 049819 A2 discloses a method for improving the functional properties of globular proteins, the method comprising: providing a solution of one or more globular proteins in which the protein(s) are at least partially aggregated in fibrils; and performing one or more of the following steps in a random order: increasing the pH; increasing the salt concentration; concentrating the solution; and changing the solvent quality of the solution. Preferably, the solution of one or more globular proteins is provided by heating at a low pH or by adding a denaturing agent. Also disclosed are the protein additive thus obtained, its use for food and non-food applications, and food and non-foods containing the protein additive.

[0009] WO 2010 / 037736 A1 discloses the isolation of whey proteins and the preparation of whey products and whey isolates. In particular, the present invention relates to the isolation of β-lactoglobulin products and the isolation of α-rich whey protein isolates from whey obtained from animals. The α-rich whey protein isolates provided by the present invention are high in α-lactalbumin and immunoglobulin G in addition to being low in β-lactoglobulin.

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

[0011] The inventors have provided an instant beverage powder product having a high BLG content. The product is shelf-stable and simultaneously an appetite-stimulating food, that is, the appearance and taste of the product attract the attention of customers.

[0012] Accordingly, an embodiment of the present invention comprises at least 1% w / w BLG, preferably at least 5% w / w BLG, wherein:

[0013] i. The degree of crystallinity of the BLG is at least 20%, preferably at least 40%, and / or,

[0014] ii. At least 85% w / w of the total protein is composed of BLG, and

[0015] The invention relates to an instant beverage powder further comprising at least one additional ingredient selected from the group consisting of vitamins, flavoring agents, coloring agents, minerals, sweeteners, antioxidants, edible acids, lipids, carbohydrates, prebiotics, probiotics, antifoaming agents, and non-whey proteins.

[0016] Another aspect of the present invention relates to a method for producing instant beverage powder comprising BLG and at least one optional ingredient, the method comprising the step of obtaining instant beverage powder by blending a dried BLG isolate with at least one additional ingredient selected from the group consisting of vitamins, flavoring agents, coloring agents, minerals, sweeteners, antioxidants, edible acids, lipids, carbohydrates, prebiotics, probiotics, antifoaming agents, and non-whey proteins.

[0017] Another aspect of the present invention relates to a liquid food comprising a liquid and a powder according to the present invention.

[0018] A further aspect of the present invention relates to a method for manufacturing a liquid food according to the present invention, wherein the method

[0019] i. A step of adding instant beverage powder according to the present invention,

[0020] ii. optionally adding at least one additional component, and

[0021] iii. Includes the step of mixing the obtained powder and liquid to form a uniform mixture.

[0022] A further aspect of the present invention relates to an instant beverage powder according to the present invention for use as a nutritional supplement.

[0023] Further aspects of the present invention relate to a kit comprising a powder according to the present invention:

[0024] i. A tool for measuring the above powder, and

[0025] ii. A container having a lid for opening and closing the container;

[0026] Here, the container is intended for mixing the powder with a liquid to form a food, and the container is suitable for drinking the food directly from the container. Brief explanation of the drawing

[0027] Figure 1 is a micrograph of a BLG crystal recovered from feed 3 of Example 3 of PCT application PCT / EP2017 / 084553. Figure 2 shows micrographs of both whole and fragmented BLG crystals obtained from feed 2 of Example 3 of PCT application PCT / EP2017 / 084553. Figure 3 is a photograph of a test tube containing six subsamples of low phosphorus beverages prepared in Example 5. Specific details for implementing the invention

[0028] definition

[0029] In the context of the present invention, the term “beta-lactoglobulin” or “BLG” relates to beta-lactoglobulin from mammalian species in, for example, natural, unfolded, and / or glycosylated forms, and includes naturally occurring genetic variants. The term also includes aggregated BLG, precipitated BLG, and crystalline BLG. When referring to the amount of BLG, the total amount of BLG including aggregated BLG is referenced. The total amount of BLG is determined according to Example 1.31. The term “aggregated BLG” relates to BLG that is at least partially unfolded and also aggregated with other modified BLG molecules and / or other modified whey proteins by hydrophobic interactions and / or covalent bonding.

[0030] BLG is the most dominant protein in bovine whey and exists as several genetic variants, with the major proteins in dairy cow's milk designated as A and B. BLG is a lipocalin protein capable of binding to many hydrophobic molecules, suggesting a role in their transport. BLG has also been shown to bind to iron via siderophores and may play a role in fighting pathogens. Homologues of BLG are not found in human breast milk.

[0031] Bovine BLG is a relatively small protein consisting of approximately 162 amino acid residues with a molecular weight of about 18.3 to 18.4 kDa. Under physiological conditions, it is primarily a dimer but dissociates into monomers at pH below about 3, which are preserved in its natural state as determined using nuclear magnetic resonance spectroscopy. Conversely, BLG also occurs as tetramers, octamers, and other multiple aggregated forms under various natural conditions.

[0032] In the context of the present invention, the term “non-aggregated beta-lactoglobulin” or “non-aggregated BLG” also relates to beta-lactoglobulin from mammalian species in a naturally unfolded and / or glycosylated form, for example, and includes naturally occurring genetic variants. However, the term does not include aggregated BLG, precipitated BLG, or crystallized BLG. The amount or concentration of non-aggregated BLG is determined according to Example 1.6.

[0033] The percentage of non-aggregated BLG to total BLG is (m 총 BLG - m 비응집체 BLG ) / m 총 BLG It is determined by calculating *100%. m 총 BLG is the concentration or amount of BLG determined according to Example 1.31, and m 비-응집된 BLG is the concentration or amount of non-aggregated BLG determined according to Example 1.6.

[0034] In the context of the present invention, the term “crystal” relates to a solid material in which components (e.g., atoms, molecules, or ions) are arranged in a highly ordered microscopic structure to form a crystal lattice that extends in all directions.

[0035] In the context of the present invention, the term “BLG crystal” relates to protein crystals that form a crystal lattice extending in all directions, containing natural BLG arranged in a non-aggregated and preferably highly ordered fine structure. The BLG crystals may be, for example, monolithic or polycrystalline, and may be, for example, intact crystals, crystal fragments, or a combination thereof. Crystal fragments are formed, for example, when intact crystals are subjected to mechanical shearing during processing. Crystal fragments also possess a highly ordered fine crystal structure but may lack the uniform surface and / or uniform edges or corners of the intact crystals. For example, refer to FIG. 1 for examples of multiple intact BLG crystals and FIG. 2 for examples of BLG crystal fragments. In both cases, the BLG crystals or crystal fragments can be visually identified as having a well-defined, dense, and coherent structure using an optical microscope. The BLG crystals or crystal fragments are often at least partially transparent. Protein crystals are also known to be birefringent, and this optical property can be used to identify unknown particles with a crystal structure. On the other hand, amorphous BLG aggregates often appear as insufficiently defined, opaque, and irregularly sized open or porous masses.

[0036] In the context of the present invention, the term "crystallize" relates to the formation of protein crystals. Crystallization may occur spontaneously, for example, or be initiated by the addition of a crystallization seed.

[0037] In the context of the present invention, the term “edible composition” relates to a composition that is safe for human consumption and use as a food ingredient and does not contain problematic amounts of toxic components, such as toluene or other unwanted organic solvents.

[0038] In the context of the present invention, the term “ALA” or “alpha-lactalbumin” relates to alpha-lactalbumin from mammalian species in, for example, natural and / or glycosylated forms, and includes naturally occurring genetic variants. The term also includes aggregated ALA and precipitated BLG. When referring to the amount of ALA, the total amount of ALA including aggregated ALA is referred to, for example. The total amount of ALA is determined according to Example 1.31. The term “aggregated ALA” relates to ALA that is typically at least partially unfolded and aggregated with other modified ALA molecules and / or other modified whey proteins by hydrophobic interactions and / or covalent bonding.

[0039] Alpha-lactalbumin (ALA) is a protein present in the milk of almost all mammalian species. ALA forms the regulatory subunit of the lactose synthase (LS) heterodimer, and β-1,4-galactosyltransferase (beta4Gal-T1) forms the catalytic component. Together, these proteins enable LS to produce lactose by transferring the galactose moiety to glucose. One of the major structural differences from beta-lactoglobulin is that ALA does not possess any free thiol groups that can act as initiators for covalent aggregation reactions.

[0040] In the context of the present invention, the term “non-aggregated ALA” also relates to ALA from mammalian species in a naturally unfolded and / or glycosylated form, for example, and includes naturally occurring genetic variants. However, the term does not include aggregated ALA or precipitated ALA. The amount or concentration of non-aggregated BLG is determined according to Example 1.6.

[0041] The percentage of non-aggregated ALA to total ALA is (m 총 ALA - m 비응집체 ALA ) / m 총 ALA It is determined by calculating *100%. m 총 ALAis the concentration or amount of ALA determined according to Example 1.31, and m 비-응집된 ALA is the concentration or amount of non-aggregated ALA determined according to Example 1.6.

[0042] In the context of the present invention, the term “caseinomacropeptide” or “CMP” comprises a naturally occurring genetic variant of an aspartate proteinase, for example, chymosin, derived from the hydrolysis of “κ-CN” or “kappa-casein” from mammalian species in a natural and / or glycosylated form, relating to residues 106 to 169.

[0043] 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.

[0044] In the context of the present invention, the term "BLG isolate powder" relates to BLG isolate in powder form and preferably to free-flowing powder.

[0045] In the context of the present invention, the term "BLG isolate liquid" relates to BLG isolate in liquid form and preferably to an aqueous liquid.

[0046] The term "whey" refers to the liquid phase remaining after casein in milk has been precipitated and removed. Casein precipitation can be achieved, for example, by acidifying the milk and / or using rennet enzymes. There are various types of whey, such as "sweet whey," 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 edible acids or bacterial culture.

[0047] The term "whey" refers to the liquid remaining when casein and milk fat globules have been removed from milk, for example, by microfiltration or large-pore ultrafiltration. Whey may also be referred to as "ideal whey."

[0048] The terms "whey protein" or "serum protein" relate to proteins present in whey.

[0049] In the context of the present invention, the term “whey protein” relates to a protein found in whey or whey. A whey protein may be a subset of protein species found in whey or whey, may even be a single whey protein species, or may be a complete set of protein species found in whey or / and whey.

[0050] In the context of the present invention, the major non-BLG proteins of the standard whey protein concentrate from sweet whey are ALA, CMP, bovine serum albumin, immunoglobulin, osteopontin, lactoferrin, and lactoperoxidase. In the context of the present invention, the weight percentage of the major non-BLG whey proteins of the standard whey protein concentrate from sweet whey is as follows:

[0051] 18% w / w amount of ALA relative to total protein,

[0052] 18% w / w amount of CMP relative to total protein,

[0053] 4% w / w amount of BSA relative to total protein,

[0054] 5% w / w amount of casein species relative to total protein,

[0055] 6% w / w of total protein, immunoglobulin

[0056] 0.5% w / w amount of osteopontin relative to total protein,

[0057] 0.1% w / w amount of lactoferrin relative to total protein, and

[0058] 0.1% w / w amount of lactoperoxidase relative to total protein.

[0059] The term casein relates to casein proteins found in milk and includes natural micellar casein found in raw milk, individual casein species, and casein salts.

[0060] In the context of the present invention, a liquid that is “supersaturated” or “supersaturated with respect to BLG” comprises a concentration of dissolved non-aggregated BLG higher than the saturation point of non-aggregated BLG in the liquid under given physical and chemical conditions. The term “supersaturation” is well known in the field of crystallization (see, for example, Gerard Coquerela, “Crystallization of molecular systems from solution: phase diagrams, supersaturation and other basic concepts”, Chemical Society Reviews, p. 2286–2300, Issue 7, 2014), and supersaturation can be determined by a number of different measurement techniques (for example, by spectroscopy or particle size analysis). In the context of the present invention, supersaturation with respect to BLG is determined by the following procedure.

[0061] Procedure for testing whether a liquid becomes supersaturated with respect to BLG under a specific set of conditions:

[0062] a) Transfer a 50 ml sample of the liquid to be tested to a centrifuge tube (VWR Catalog No. 525-0402) having a height of 115 mm, an inner diameter of 25 mm, and a capacity of 50 mL. During steps a) through h), care must be taken to maintain the sample and its subsequent fractions under the original physical and chemical conditions of the liquid.

[0063] b) Immediately centrifuge the sample at 3000 g for 3.0 minutes with a maximum acceleration of 30 seconds and a maximum deceleration of 30 seconds.

[0064] c) Immediately after centrifugation, transfer as much of the supernatant as possible to a second centrifugation tube (of the same type as in step a) (if a pellet has formed, do not disturb the pellet).

[0065] d) Take a 0.05 mL subsample (subsample A) of the supernatant.

[0066] e) Add 10 mg of BLG crystals having a particle size of up to 200 microns (at least 98% pure, non-aggregated BLG relative to total solids) to the second centrifuge tube and shake the mixture.

[0067] f) Leave the second centrifuge tube at its original temperature for 60 minutes.

[0068] g) Immediately after step f), centrifuge the second centrifuge tube at 500 g for 10 minutes, then take another 0.05 mL subsample (subsample B) of the supernatant.

[0069] h) If the centrifugation pellet from step g) is present, recover it, resuspend it in milliQ water, and immediately inspect the suspension for the presence of microscopic crystals.

[0070] i) Determine the concentration of non-aggregated BLG in subsamples A and B using the method summarized in Example 1.6—the results are expressed as % BLG w / w relative to the total weight of the subsamples. The concentration of non-aggregated BLG in subsample A is C BLG, A It is referred to as, and the concentration of non-aggregated BLG in subsample B is C BLG, B It is referred to as.

[0071] j) The liquid from which the sample of step a) was taken is c BLG, B a c BLG, A If a crystal is observed at a lower level and at step i), it is supersaturated (under specific conditions).

[0072] In the context of the present invention, the terms “liquid” and “solution” include both compositions without particulate matter and compositions containing a combination of liquid and solid and / or semi-solid particles, such as protein crystals or other protein particles. Accordingly, the “liquid” or “solution” may be a suspension or even a slurry. However, the “liquid” and “solution” are preferably pumpable.

[0073] In the context of the present invention, the terms "whey protein concentrate (WPC)" and "whey protein concentrate (SPC)" relate to dry or aqueous compositions containing a total amount of protein of 20 to 89% w / w with respect to total solids.

[0074] WPC or SPC preferably contains the following:

[0075] 20 to 89% w / w protein relative to total solids,

[0076] 15 to 70% w / w BLG relative to total protein,

[0077] 8 to 50% w / w ALA relative to total protein, and

[0078] 0 to 40% w / w CMP for protein.

[0079] Alternatively, but also preferably, WPC or SPC may contain the following:

[0080] 20 to 89% w / w protein relative to total solids,

[0081] 15 to 90% w / w BLG of total protein,

[0082] 4 to 50% w / w ALA relative to total protein, and

[0083] 0 to 40% w / w CMP for protein.

[0084] Preferably, WPC or SPC contains the following:

[0085] 20 to 89% w / w protein relative to total solids,

[0086] 15 to 80% w / w BLG relative to total protein,

[0087] 4 to 50% w / w ALA relative to total protein, and

[0088] 0 to 40% w / w CMP for protein.

[0089] More preferably, WPC or SPC contains the following:

[0090] 70 to 89% w / w protein relative to total solids,

[0091] 30 to 90% w / w BLG relative to total protein,

[0092] 4 to 35% w / w ALA relative to total protein, and

[0093] 0 to 25% w / w CMP for protein.

[0094] SPC typically contains no CMP or only trace amounts of CMP.

[0095] The terms "whey protein isolate (WPI)" and "whey protein isolate (SPI)" relate to dry or aqueous compositions containing a total amount of protein of 90 to 100% w / w with respect to total solids.

[0096] WPI or SPI preferably contains the following:

[0097] 90 to 100% w / w protein relative to total solids,

[0098] 15 to 70% w / w BLG relative to total protein,

[0099] 8 to 50% w / w ALA relative to total protein, and

[0100] 0 to 40% w / w CMP relative to total protein.

[0101] Alternatively, but also preferably, WPI or SPI may contain the following:

[0102] 90 to 100% w / w protein relative to total solids,

[0103] 30 to 95% w / w BLG of total protein,

[0104] 4 to 35% w / w ALA relative to total protein, and

[0105] 0 to 25% w / w CMP relative to total protein.

[0106] More preferably, WPI or SPI may contain the following:

[0107] 90 to 100% w / w protein relative to total solids,

[0108] 30 to 90% w / w BLG relative to total protein,

[0109] 4 to 35% w / w ALA relative to total protein, and

[0110] 0 to 25% w / w CMP relative to total protein.

[0111] SPI typically contains no CMP or only trace amounts of CMP.

[0112] In the context of the present invention, the term “additional protein” means a protein that is not BLG. The additional protein present in the whey protein solution comprises one or more of the non-BLG proteins typically found in whey or whey. Non-limiting examples of such proteins are alpha-lactalbumin, bovine serum albumin, immunoglobulin, caseinomacropeptide (CMP), osteopontin, lactoferrin, and milk fat globule membrane protein.

[0113] The terms "essentially made" and "essentially made" mean that the claim or feature includes materials or steps specified and that do not substantially affect the basic and novel characteristic(s) of the claimed invention.

[0114] In the context of the present invention, the phrase "Y and / or X" means "Y" or "X" or "Y and X". Following the same logic, the phrase "n1, n2, ..., n i-1 , and / or n i " is " n1" or " n2" or ... or "n i-1 " or "n i " or means any combination of components: n1, n2,...n i-1 , and n i .

[0115] In the context of the present invention, the term “dry” or “dried” means that the composition or product contains up to 10% w / w, preferably up to 6% w / w, and more preferably less than this amount of water.

[0116] In the context of the present invention, the term "physical microbial reduction" relates to physical interactions with the composition that reduce the total amount of viable microorganisms in the composition. The term does not include the addition of chemicals that kill microorganisms. The term also does not include heat exposure to which the sprayed droplets are exposed during spray-drying, but includes possible preheating prior to spray-drying.

[0117] In the context of the present invention, the pH of the powder is 10 g mixed in 90 g of deionized water. It refers to the pH of the powder and is measured according to Example 1.16.

[0118] In the context of the present invention, the weight percentage (% w / w) of a component of a particular composition, product, or material means the weight percentage of that component relative to the weight of the particular composition, product, or material, unless other references (e.g., total solids or total protein) are specifically mentioned.

[0119] In the context of the present invention, the process step "concentration" and the verb "to concentrate" relate to the concentration of protein and include both the concentration of protein based on total solids and the concentration of protein based on total weight. This means that, for example, as long as the protein content increases relative to total solids, the concentration does not necessarily require an increase in the absolute concentration w / w of the protein in the composition.

[0120] In the context of the present invention, the term "weight ratio" between component X and component Y is m X / m Y It refers to the value obtained by calculation, where m X is the amount (weight) of component X, and m Y is the amount (weight) of component Y.

[0121] In the context of the present invention, the term "at least pasteurization" relates to a heat treatment having a microbial killing effect equal to or greater than that of a heat treatment at 70°C for 10 seconds. The criteria for determining the microbial killing effect are E. Coli ( E. coli ) O157:H7.

[0122] In the context of the present invention, the term “whey protein feed” relates to a whey protein source from which liquid BLG isolates are derived. The whey protein feed has a lower content of BLG relative to total protein than the liquid BLG isolates and is typically WPC, WPI, SPC, or SPI.

[0123] In the context of the present invention, the term "BLG-rich composition" relates to a BLG-rich composition produced by isolating BLG from a whey protein feed. The BLG-rich composition typically contains the same whey protein as the whey protein feed, but BLG is present at a significantly higher concentration relative to the total protein than in the whey protein feed. The BLG-rich composition may be prepared from the whey protein feed, for example, by chromatography, protein crystallization, and / or membrane-based protein fractionation. The BLG-rich composition contains an amount of BLG of at least 85% w / w, preferably at least 90% w / w, relative to the total protein. In some cases, the BLG-rich composition may be used directly as a liquid BLG isolate. However, additional processing is often required to convert the BLG-rich composition into a liquid BLG isolate.

[0124] In the context of the present invention, the term "whey protein solution" is used to describe a specific aqueous whey protein composition that is supersaturated relative to BLG in a salting mode and is useful for producing BLG crystals.

[0125] In the context of the present invention, the term “sterile state” means that the sterile composition or product contains no arbitrarily viable microorganisms and therefore there is no microbial growth during storage at room temperature. The sterile composition is in a sterile state.

[0126] When a liquid, such as a beverage preparation, is sterilized and packaged in a sterile container under aseptic conditions, it typically has a shelf life of at least six months at room temperature. Sterilization treatment kills spores and microorganisms that can cause the liquid to spoil.

[0127] In the context of the present invention, the term "energy content" refers to the total energy content contained in a food. Energy content may be measured in kilojoules (kJ) or kilocalories (kcal) and is referred to as calories per serving of food, for example, kcal per 100 grams of food. An example is an instant beverage powder having an energy content of 350 kcal per 100 grams of instant beverage powder.

[0128] The total energy content of a food includes the energy contribution from all macronutrients present in the food, such as energy from protein, lipids, and carbohydrates. The energy distribution from macronutrients within the food can be calculated based on the amount of macronutrients within the food and their contribution to the total energy content of the food. The energy distribution may be referred to as the energy percentage (E%) of the total energy content of the food. For example, in the case of an instant beverage powder containing 20 E% protein, 50 E% carbohydrates, and 30 E% lipids, this means that 20% of the total energy comes from protein, 50% of the total energy comes from carbohydrates, and 30% of the total energy comes from fat (lipids).

[0129] In the context of the present invention, the term "nutritional supplement" relates to a food containing one or more macronutrients, such as protein, lipids, and / or carbohydrates, and optionally containing vitamins and minerals. The nutritional supplement may be complete or incomplete.

[0130] The term "nutritionally complete nutritional supplement" is understood as a food containing protein, lipids, and carbohydrates, and additionally containing vitamins, minerals, and trace elements, wherein the food has a nutritional profile consistent with a complete and healthy diet.

[0131] The term "nutritionally incomplete supplement" refers to a food containing one or more macronutrients and optionally additionally containing vitamins, minerals, and trace elements. A nutritionally incomplete supplement may contain protein as the sole nutrient, or may contain protein, lipids, and carbohydrates.

[0132] The term "Food for Special Medical Purposes (FSMP)" or "medical food" refers to food intended for oral intake or tube administration, used for specific medical disorders, diseases, or conditions that have unique nutritional requirements and are used under medical supervision. Medical foods may be nutritionally complete or nutritionally incomplete supplements.

[0133] The term "nutrient" refers to a substance used by an organism for survival, growth, and reproduction. Nutrients can be macronutrients or micronutrients. Macronutrients are nutrients that provide energy when consumed, such as proteins, lipids, and carbohydrates. Micronutrients are nutrients such as vitamins, minerals, and trace elements.

[0134] The term "instant beverage powder" or "instant beverage powder product" refers to powder that can be converted into a liquid beverage by adding a liquid, such as water.

[0135] In the context of the present invention, the terms "beverage formulation" and "formulation" as substantially used relate to any water-based liquid that can be consumed as a beverage, for example, by pouring, drinking, or supplying via tube.

[0136] In the context of the present invention, the term "protein fraction" relates to proteins of the composition, for example, proteins of powder or beverage formulations.

[0137] In the context of the present invention, the term “astringency” relates to mouthfeeling. Astringency is felt like the contraction of cheek muscles and increases saliva production. Therefore, astringency is not a taste in itself, but a physical mouthfeel and a time-dependent sensation in the mouth.

[0138] In the context of the present invention, the term "dry texture" relates to a sensation in the mouth that feels as if the mouth and teeth are drying out, thereby minimizing saliva production. Therefore, dry texture is not a taste in itself, but a physical texture and a time-dependent sensation in the mouth.

[0139] In the context of the present invention, the term “mineral” as used herein refers to any one of major minerals, trace or minor minerals, other minerals, and combinations thereof, unless otherwise specified. Major minerals include calcium, phosphorus, potassium, sulfur, sodium, chlorine, and magnesium. Trace or minor minerals include iron, cobalt, copper, zinc, molybdenum, iodine, selenium, and manganese, and other minerals include chromium, fluorine, boron, lithium, and strontium.

[0140] In the context of the present invention, the terms “lipid,” “fat,” and “oil” as used herein are used interchangeably to refer to lipid materials derived from or processed from plants or animals, unless otherwise specified. These terms also include synthetic lipid materials insofar as such synthetic materials are suitable for human consumption.

[0141] In the context of the present invention, the term "transparent" includes beverage formulations that have a noticeably clear appearance, allow light to pass through, and display a distinct image. The transparent beverage has a turbidity of up to 200 NTU.

[0142] In the context of the present invention, the term "opaque" includes beverage formulations that are not noticeably clear in appearance and have a turbidity of more than 200 NTU.

[0143] In the context of the present invention, the term “mother liquor” relates to a whey protein solution remaining after BLG has been crystallized and at least partially removed. The mother liquor may still contain some BLG crystals, but generally may contain only small BLG crystals that have been spared from separation.

[0144] The term "instant beverage powder" or "instant beverage powder product" refers to powder that can be converted into a liquid beverage by adding a liquid, such as water.

[0145] details

[0146] Consumers are aware of the general concept of nutritional supplements. Nutritional supplements must stimulate the appetite in taste and appearance; otherwise, consumers will reject them. Additionally, consumers value natural products free of additives. A more important parameter for consumers is the product's shelf life.

[0147] An embodiment of the present invention comprises at least 1% w / w BLG, preferably at least 5% w / w BLG, wherein:

[0148] i. The degree of crystallinity of the BLG is at least 20%, preferably at least 40%, and / or,

[0149] ii. At least 85% w / w of the total protein is composed of BLG, and

[0150] The present invention relates to an instant beverage powder comprising at least one additional ingredient selected from the group consisting of vitamins, flavoring agents, coloring agents, minerals, sweeteners, antioxidants, edible acids, lipids, carbohydrates, prebiotics, probiotics, antifoaming agents, and non-whey proteins.

[0151] The BLG source used in the instant beverage powder may be a BLG isolate or a BLG isolate powder as described in this patent application. In some preferred embodiments of the present invention, the BLG source contributes at least 90% w / w, more preferably at least 95% w / w, even more preferably at least 98% w / w of the total protein of the instant beverage powder, and most preferably contributes to all the protein of the instant beverage powder.

[0152] In some preferred embodiments of the present invention, the BLG source is BLG isolated powder and is the only protein source in instant beverage powder.

[0153] In some preferred embodiments of the present invention, instant beverage powder is prepared by drying and blending BLG isolate powder and other ingredients.

[0154] In another preferred embodiment of the present invention, the instant beverage powder is prepared by using at least one ingredient in a dissolved form and then performing a drying step. The drying step forms, for example, part of a wet-granulation process or a spray-drying step.

[0155] The BLG of the instant beverage powder of the present invention preferably has a low degree of denaturation, such as up to 10%, preferably up to 4%, more preferably up to 1%, even more preferably up to 0.4%, and even more preferably up to 0.1%. Most preferably, the BLG is not denatured at all. For the instant beverage powder, it is advantageous for the BLG to have a low degree of denaturation because it reduces the tendency for foaming to occur when mixed with liquid.

[0156] The instant beverage powder preferably has a protein denaturation of up to 10%, preferably up to 4%, more preferably up to 1%, even more preferably up to 0.4%, and even more preferably up to 0.1%. Most preferably, the protein is not denatured at all.

[0157] In one embodiment of the present invention, the instant beverage powder comprises 1 to 90% w / w BLG. In a preferred embodiment of the present invention, the instant beverage powder comprises 30 to 90% w / w BLG, more preferably BLG in the range of 40 to 90% w / w, or even more preferably BLG in the range of 50 to 90% w / w.

[0158] In another preferred embodiment of the present invention, the instant beverage powder comprises 10 to 97% w / w BLG. In a preferred embodiment of the present invention, the instant beverage powder comprises 30 to 96% w / w BLG, more preferably BLG in the range of 40 to 95% w / w, or even more preferably BLG in the range of 50 to 94% w / w.

[0159] In one embodiment of the present invention, the instant beverage powder comprises 1 to 50% w / w BLG. In a preferred embodiment of the present invention, the instant beverage powder comprises 2 to 45% w / w BLG, more preferably BLG in the range of 3 to 40% w / w, or even more preferably BLG in the range of 3 to 35% w / w.

[0160] In a preferred embodiment of the present invention, the instant beverage powder contains BLG of at least 85% w / w of the total amount of protein.

[0161] In one embodiment of the present invention, the instant beverage powder comprises at least 85% w / w BLG with respect to total protein, e.g., at least 86% w / w BLG with respect to total protein, at least 87% w / w BLG with respect to total protein, at least 88% w / w BLG with respect to total protein, and at least 89% w / w BLG with respect to total protein.

[0162] In one embodiment of the present invention, the instant beverage powder comprises at least 91% w / w BLG with respect to total protein, e.g., at least 92% w / w BLG with respect to total protein, at least 93% w / w BLG with respect to total protein, at least 94% w / w BLG with respect to total protein, at least 95% w / w BLG with respect to total protein, at least 96% w / w BLG with respect to total protein, at least 97% w / w BLG with respect to total protein, at least 98% w / w BLG with respect to total protein, or at least 99% w / w BLG with respect to total protein.

[0163] In some preferred embodiments of the instant beverage powder 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.

[0164] Since a higher relative amount of BLG is feasible and desirable, in some preferred embodiments of the present invention, at least 94% w / w of the protein in the instant beverage powder 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 of the protein is BLG.

[0165] For example, instant beverage powder preferably contains at least 97.5% w / w, preferably at least 98.0% w / w, more preferably at least 98.5% w / w, and even more preferably at least 99.0% of BLG with respect to total protein, and most preferably contains at least 99.5% w / w with respect to total protein, for example, about 100.0% w / w with respect to total protein.

[0166] The protein in the instant beverage powder is preferably produced from mammalian milk, preferably ruminant milk, for example, the milk of dairy cows, sheep, goats, buffalo, camels, llamas, horses, and / or deer. Protein derived from cow's milk is particularly preferred. Therefore, the protein in the instant beverage powder is preferably bovine milk protein.

[0167] The protein in the instant beverage powder is preferably whey protein or whey protein, and more preferably bovine whey protein or whey protein.

[0168] The intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) is a measure of the degree of unfolding of BLG, and the inventors found that the intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) was measured according to Example 1.1 at a high BLG tryptophan fluorescence emission ratio that is associated with low unfolding of BLG or unrelated to unfolding.

[0169] In some preferred embodiments of the present invention, the instant beverage powder has an intrinsic tryptophan fluorescence emission ratio of at least 1.11 (I330 nm / I350 nm).

[0170] In some preferred embodiments of the present invention, the instant beverage powder has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.12, preferably at least 1.13, more preferably at least 1.15, even more preferably at least 1.17, and most preferably at least 1.19.

[0171] If the instant beverage powder contains a significant amount of non-protein material, it is desirable to isolate the protein fraction before measuring the intrinsic tryptophan fluorescence emission ratio. Accordingly, in some preferred embodiments of the present invention, the protein fraction of the instant beverage powder has an intrinsic tryptophan fluorescence emission ratio of at least 1.11.

[0172] In some preferred embodiments of the present invention, the protein fraction of the instant beverage powder has an intrinsic tryptophan fluorescence emission ratio (I330 nm / I350 nm) of at least 1.12, preferably at least 1.13, more preferably at least 1.15, even more preferably at least 1.17, and most preferably at least 1.19.

[0173] The protein fraction can be separated from the instant beverage powder, for example, by dissolving the instant beverage powder in deionized water and dialysis or ultrafiltration-based filtration of the solution using a filter that retains the protein.

[0174] In some preferred embodiments of the present invention, the degree of crystallization of BLG in the instant beverage powder is at least 20%, preferably at least 40%, more preferably at least 60%, even more preferably at least 80%, and most preferably at least 90%. Having a degree of crystallization of at least 20% of BLG means that a significant amount of BLG exists in the instant beverage powder in the form of dried BLG crystals.

[0175] The inventors have discovered that a crystallinity of at least 20% of BLG is advantageous because it means that the protein exists in a form with a higher density than traditional WPI. This provides the instant beverage powder with a higher overall bulk density, results in less dust, and makes it easier for the end user to handle. The inventors have also observed a reduced tendency for particle separation in dry-blended instant beverage powders containing, for example, carbohydrate powder and / or edible acid powder in addition to protein powder.

[0176] The present invention makes it possible to provide a low-carbohydrate instant beverage powder having both sweetness and high protein content.

[0177] Accordingly, in some preferred embodiments of the present invention, the instant beverage powder comprises the following:

[0178] - Energy content in the range of 320 to 380 kcal per 100 grams of powder, preferably in the range of 350 to 370 kcal per 100 grams,

[0179] - Energy contribution from protein in the range of 90 to 100 E%, preferably in the range of 95 to 100 E%,

[0180] - BLG in an amount of at least 85% w / w, preferably at least 90% w / w, and more preferably at least 94% w / w, relative to total protein,

[0181] - Contribution of energy from carbohydrates in the range of 0 to 10 E%, preferably in the range of 0 to 5 E%,

[0182] - Total amount of high-intensity sweetener in the range of 0.01 to 4% w / w, preferably in the range of 0.05 to 3%,

[0183] - Having a bulk density of at least 0.45 g / mL, preferably at least 0.50 g / mL, more preferably at least 0.6 g / mL.

[0184] The protein of the instant beverage powder preferably has a pH in the range of i) 2 to 4.9, ii) 6.1 to 8.5, or iii) 5.0 to 6.0, and is provided by a BLG isolate powder comprising the following:

[0185] - At least 90% w / w, preferably at least 95% w / w amount of total protein,

[0186] - An amount of BLG of at least 85% w / w of total protein, preferably at least 90% w / w of total protein, more preferably at least 94% w / w of total protein;

[0187] The above BLG isolated powder has the following:

[0188] - A bulk density of at least 0.45 g / mL, preferably at least 0.50 g / mL, more preferably at least 0.6 g / mL, and

[0189] - One or more of the following:

[0190] - an intrinsic tryptophan fluorescence emission ratio (I330 / I350) of at least 1.11, preferably at least 1.13, more preferably at least 1.15,

[0191] - Protein denaturation of up to 10%, preferably at least 5%,

[0192] - Thermal stability of up to 200 NTU at pH 3.9, and

[0193] - Up to 1,000 colony-forming units / g.

[0194] In one embodiment of the present invention, the instant beverage powder further comprises at least one additional ingredient selected from the group consisting of vitamins, flavoring agents, coloring agents, minerals, sweeteners, antioxidants, edible acids, lipids, carbohydrates, prebiotics, probiotics, and non-whey proteins.

[0195] Additional ingredients ensure that the instant beverage powder contains desired nutrients, namely nutrients specifically formulated for weight gain, for patients who are malnourished or at risk of malnutrition or suffering from kidney disease, or can be used as a nutritional supplement by, for example, sportsmen or athletes.

[0196] In a preferred embodiment of the present invention, the instant beverage powder is vitamin A, vitamin B1 (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), and vitamin B5 (pantothenic acid). ,It may include a vitamin selected from the group consisting of vitamin B6 (pyridoxine), vitamin B7 (biotin), vitamin B9 (folic acid), and vitamin B12 (cobalamin), vitamin C, vitamin D, vitamin E, vitamin K, choline, inositol, salts thereof, derivatives thereof, and combinations thereof.

[0197] In one embodiment, the instant beverage powder may include a flavoring agent selected from the group consisting of salt, flavorings, flavor enhancers 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.

[0198] In one embodiment, the instant beverage powder may contain a mineral selected from the group consisting of boron, calcium, chromium, copper, iodine, iron, magnesium, manganese, molybdenum, nickel, phosphorus, potassium, sodium, selenium, silicon, tin, vanadium, zinc, or a combination thereof.

[0199] Instant beverage powder may additionally contain salts and minerals typically present in products derived from whey or milk. The mineral content of the instant beverage powder is typically indicated by the ash content of the food ingredient or product. In a preferred embodiment, the instant beverage powder may contain an antioxidant selected from the group consisting of beta-carotene, vitamin C, vitamin E, selenium, or combinations thereof.

[0200] In one embodiment of the present invention, the instant beverage powder may comprise one or more sweeteners, such as a carbohydrate sweetener, a polyol, and / or a high-intensity sweetener. The instant beverage powder may comprise, for example, a total amount of a carbohydrate sweetener in the range of 0.001 to 20% w / w with respect to the total weight of the instant beverage powder. Alternatively, the instant beverage powder may comprise a total amount of a carbohydrate sweetener in the range of 0.1 to 15% w / w with respect to the total weight of the food.

[0201] In one embodiment of the present invention, the instant beverage powder comprises at least one high-intensity sweetener. In one embodiment, the at least one high-intensity sweetener is selected from the group consisting of aspartame, cyclamate, sucralose, acesulfame salt, neotame, saccharin, stevia extract, steviol glycoside, e.g., rebaudioside A, or combinations thereof. In some embodiments of the present invention, it is particularly preferred that the sweetener comprises one or more high-intensity sweeteners (HIS) or even consists of them.

[0202] High-intensity sweeteners are found in both natural and artificial sweeteners, and typically, the sweetener intensity is at least 10 times that of sucrose.

[0203] If used, the total amount of high-intensity sweetener is typically in the range of 0.01 to 4% w / w. For example, the total amount of high-intensity sweetener may be in the range of 0.05 to 3% w / w. Alternatively, the total amount of high-intensity sweetener may be in the range of 0.1 to 2.0% w / w.

[0204] The choice of sweetener may vary depending on the consumers of the beverage and product to be produced, for example, it may be adjusted according to the specific diagnosis of the patient. High-intensity sugar sweeteners (e.g., aspartame, acesulfame-K, or sucralose) may be used in beverages where the contribution of energy from the sweetener is not desired, whereas for beverages with a natural profile, natural sweeteners (e.g., steviol glycoside, sorbitol, or sucrose) may be used.

[0205] Additionally, it may be desirable for the sweetener to include or even consist of one or more polyol sweeteners. Non-limiting examples of useful polyol sweeteners are maltitol, mannitol, lactitol, sorbitol, inositol, xylitol, threitol, galactitol, or combinations thereof. If used, the total amount of polyol sweetener is typically in the range of 1 to 40% w / w. For example, the total amount of polyol sweetener may be in the range of 2 to 30% w / w. Alternatively, the total amount of polyol sweetener may be in the range of 4 to 20% w / w.

[0206] In one embodiment of the present invention, the instant beverage powder may comprise one or more of the following:

[0207] i. Sweeteners, e.g., sugar sweeteners and / or sugar-free sweeteners,

[0208] ii. Flavoring agents,

[0209] iii. At least one edible acid, e.g., citric acid or other suitable edible acid,

[0210] iv. The sum of the amounts of Na, K, Mg, and Ca in instant beverages is up to 10 mmol / g protein, and

[0211] Here, a 10% w / w solution of the powder in deionized water has a pH in the range of 2 to 8.

[0212] The pH of the instant beverage powder can be measured by dissolving 10 g of instant beverage powder in 90 ml of deionized water at room temperature, as described in Example 1.16.

[0213] The inventors have discovered that it is advantageous to use low phosphorus / low potassium BLG isolated powder in instant beverage powders, for example, instant beverage powders that are particularly useful for patients with kidney disease.

[0214] By adding sweeteners, flavorings, and / or edible acids, the taste of the product can be designed so that the instant beverage powder attracts the attention of consumers. In one embodiment of the present invention, the consumer may be a patient for whom the flavor, sweetener, and acid profile of the instant beverage powder are adjusted to the patient's needs and diagnosis.

[0215] In a preferred embodiment of the present invention, the instant beverage powder may include a high-intensity sweetener and a flavoring agent. In a more preferred embodiment of the present invention, the instant beverage powder comprises 0.001 to 0.05% w / w of sucralose and 0.01 to 0.2% w / w of a flavor selected from chocolate, cocoa, lemon, orange, lime, strawberry, banana, forest fruit flavors, or combinations thereof.

[0216] In one embodiment of the present invention, the instant beverage powder comprises an antifoaming agent. The antifoaming agent may be selected from antifoaming agents suitable for food. The antifoaming agent may be selected from oil-based antifoaming agents, water-based antifoaming agents, silicone-based antifoaming agents, EP / PO-based antifoaming agents, or a combination thereof.

[0217] The instant beverage powder has a moisture content of up to 6% w / w. In one embodiment of the present invention, the instant beverage powder contains up to 5% w / w water, preferably up to 4% w / w water, more preferably up to 3% w / w water, and even more preferably up to 2% w / w water.

[0218] Lowering the moisture content of the powder can increase the storage stability of the instant beverage powder.

[0219] The inventors have discovered that it may be advantageous to control the mineral content to achieve some of the desired properties of instant beverage powder.

[0220] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg, and Ca in the instant beverage powder is up to 10 mmol / g protein. Preferably, the sum of the amounts of Na, K, Mg, and Ca in the instant beverage powder is up to 6 mmol / g protein, more preferably up to 4 mmol / g protein, and even more preferably up to 2 mmol / g protein.

[0221] In another preferred embodiment of the present invention, the sum of the amounts of Na, K, Mg, and Ca in the instant beverage powder is up to 1 mmol / g protein. Preferably, the sum of the amounts of Na, K, Mg, and Ca in the instant beverage powder is up to 0.6 mmol / g protein, more preferably up to 0.4 mmol / g protein, even more preferably up to 0.2 mmol / g protein, and most preferably up to 0.1 mmol / g protein.

[0222] In one embodiment of the present invention, the instant beverage powder comprises dry BLG crystals obtainable by one or more methods described in PCT / EP2017 / 084553, for example. The instant beverage powder containing BLG crystals may have a bulk density of at least 0.30 g / mL, preferably at least 0.4 g / mL.

[0223] The inventors have observed that instant beverage powder in which at least a portion of the BLG is in crystalline form has a higher density than a similar instant BLG composition without BLG crystals. Accordingly, in some preferred embodiments of the present invention, the instant beverage powder has a bulk density of at least 0.30 g / mL, preferably at least 0.40 g / mL. Preferably, the instant beverage powder has a bulk density of at least 0.45 g / mL. More preferably, the instant beverage powder has a bulk density of at least 0.50 g / mL. Even more preferably, the instant beverage powder has a bulk density of at least 0.6 g / mL. The instant beverage powder may have, for example, a bulk density of at least 0.7 g / mL.

[0224] The instant beverage powder of the present invention preferably has a bulk density in the range of 0.3 to 1.0 g / mL, preferably in the range of 0.40 to 0.9 g / mL, more preferably in the range of 0.45 to 0.8 g / mL, even more preferably in the range of 0.45 to 0.75 g / mL, even more preferably in the range of 0.50 to 0.75 g / mL, and most preferably in the range of 0.6 to 0.75 g / mL.

[0225] The bulk density of the powder can be measured according to Example 1.17.

[0226] The total protein content and energy content of the instant beverage powder of the present invention vary depending on the intended use of the instant beverage powder. The energy content of the instant beverage powder is in the range of 200 to 500 kcal per 100 grams of powder.

[0227] In the case of instant beverage powder, the contribution of energy from protein may be at least 7 E%, preferably at least 25 E%, more preferably at least 30 E%, and even more preferably at least 40 E%.

[0228] In a preferred embodiment of the present invention, the contribution of energy from the protein is in the range of 10 to 30 E%, preferably in the range of 10 to 15 E%, or more preferably in the range of 11 E%. Alternatively, the contribution of energy from the protein is in the range of 15 to 25 E%, preferably in the range of 18 to 22 E%.

[0229] In a preferred embodiment of the present invention, the contribution of energy from protein is in the range of 7 to 25 E%, preferably in the range of 10 to 25 E%, more preferably in the range of 15 to 20 E%, or even more preferably the instant beverage powder contains 15 E% from protein or 20 E% from protein, or the contribution of energy from protein is in the range of 8 to 15 E%.

[0230] In another preferred embodiment of the present invention, the contribution of energy from the protein is at least 50 E%, preferably at least 60 E% or at least 70 E% or more preferably at least 80 E%. In a preferred embodiment of the present invention, the contribution of energy from the protein is in the range of 80 to 100 E%, preferably in the range of 90 to 100 E% or more preferably in the range of 95 to 100 E%.

[0231] In a preferred embodiment of the present invention, the contribution of energy from protein is in the range of 30 to 80 E%, preferably in the range of 60 to 80 E%. In another embodiment of the present invention, the contribution of energy from protein is in the range of 30 to 40 E%, or more preferably, the instant beverage powder contains 33 E% from protein.

[0232] Instant beverage powder can be used as a nutritional supplement for weight gain, for example, to treat patients who are malnourished or at risk of malnutrition, for patients with kidney disease, or, for example, by sportsmen or athletes before, during, or after exercise.

[0233] The instant beverage powder of the present invention may contain macronutrients other than protein. The instant beverage powder may contain carbohydrates and / or lipids in addition to protein. The total lipid content in the instant beverage powder of the present invention varies depending on the intended use of the instant beverage powder. In one embodiment of the present invention, the contribution of energy from lipids is in the range of 0 to 60 E%.

[0234] In a preferred embodiment of the present invention, the contribution of energy from lipids is in the range of 0 to 5 E% from lipids, preferably in the range of 0 to 3 E% or more preferably in the range of 0 to 2 E%.

[0235] Even less lipid may be desirable, and thus, in a preferred embodiment of the present invention, the contribution of energy from lipid is in the range of 0 to 1 E% from lipid, preferably in the range of 0 to 0.1 E% or more preferably in the range of 0 to 0.01 E%.

[0236] In a preferred embodiment of the present invention, the contribution of energy from lipids is in the range of 30 to 60 E%, preferably in the range of 30 to 50 E%, or more preferably, the instant beverage powder comprises 35 E% from lipids, 45 E% from lipids, or 50 E% from lipids. Alternatively, the contribution of energy from lipids is in the range of 25 to 45 E%.

[0237] In a preferred embodiment of the present invention, the contribution of energy from lipids is in the range of 15 to 20 E%, preferably in the range of 16 to 18 E%, or more preferably, the instant beverage powder contains 16 E% from lipids.

[0238] Instant beverage powder can be used as a nutritional supplement for weight gain, for example, to treat patients who are malnourished or at risk of malnutrition, for patients with kidney disease, or, for example, by sportsmen or athletes before, during, or after exercise.

[0239] In one embodiment of the present invention, the instant beverage powder may include carbohydrates in addition to protein. The energy contribution of carbohydrates to the total energy of the instant beverage powder may be in the range of 0 to 90 E%.

[0240] Carbohydrates may be selected from sugars, oligosaccharides, or polysaccharides. Examples of sugars are mono-, disaccharides, and polyols. Examples of oligosaccharides are malto-oligosaccharides, such as maltodextrin, or other oligosaccharides, such as raffinose, stachyose, or fructo-oligosaccharides. Examples of polysaccharides are starches, such as amylose, amylopectin, or modified starch and non-starch polysaccharides, such as dietary fiber, cellulose, pectin, and hydrocolloids. In a preferred embodiment of the present invention, carbohydrates are selected from maltodextrin, saccharose, or glucose syrup.

[0241] The total carbohydrate content in the instant beverage powder of the present invention varies depending on the intended use of the instant beverage powder. In the case of instant beverage powder used for sportsmen or athletes, carbohydrates in the form of sugars may be added to increase the sportsman's immediate energy, or carbohydrates in the form of slow carbohydrates or dietary fiber may be added to extend satiety.

[0242] In a preferred embodiment of the present invention, the contribution of energy from carbohydrates is in the range of 70 to 90 E%, preferably in the range of 75 to 85 E%, or more preferably, the instant beverage powder contains 89 E% from carbohydrates.

[0243] In a preferred embodiment of the present invention, the contribution of energy from carbohydrates is in the range of 30 to 50 E%, preferably in the range of 35 to 45 E%, or more preferably, the instant beverage powder comprises 35 E% from carbohydrates, 45 E% from carbohydrates, or 50 E% from carbohydrates. Alternatively, the contribution of energy from carbohydrates is in the range of 40 to 60 E%, e.g., in the range of 45 to 55 E%.

[0244] In another preferred embodiment of the present invention, the contribution of energy from carbohydrates is in the range of 0 to 20 E%. In a preferred embodiment of the present invention, the contribution of energy from carbohydrates is in the range of 0 to 10 E%, preferably in the range of 0 to 5 E%.

[0245] In a more preferred embodiment of the present invention, the contribution of energy from carbohydrates is in the range of 0 to 4 E%, more preferably 0 to 1 E%, and even more preferably 0 to 0.2 E%.

[0246] In another preferred embodiment of the present invention, the contribution of energy from carbohydrates is in the range of 3 to 20 E%. In a preferred embodiment of the present invention, the contribution of energy from carbohydrates is in the range of 4 to 15 E%. In another embodiment of the present invention, the contribution of energy from carbohydrates is in the range of 45 to 55 E%.

[0247] Instant beverage powder can be used as a nutritional supplement for weight gain, for example, to treat patients who are malnourished or at risk of malnutrition, for patients with kidney disease, or, for example, by sportsmen or athletes before, during, or after exercise.

[0248] The instant beverage powder of the present invention contains protein, and in addition to protein, may contain lipids and / or carbohydrates depending on the intended use of the instant beverage powder.

[0249] In a preferred embodiment of the present invention, the instant powder further comprises vitamins, minerals, and trace elements. The instant beverage powder may be used as a nutritional supplement for weight gain, for example, to treat patients who are malnourished or at risk thereof, for patients suffering from kidney disease, or for example, by sportsmen or athletes.

[0250] The energy content of the instant beverage powder may be in the range of 200 to 400 kcal per 100 grams of powder. In a preferred embodiment of the present invention, the energy content of the instant beverage powder is in the range of 300 to 400 kcal per 100 grams of powder, more preferably the range of 320 to 380 kcal per 100 grams of powder, or most preferably the range of 350 to 370 kcal per 100 grams of powder.

[0251] In a preferred embodiment of the present invention, the instant beverage supplement has an energy distribution such as 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids. In a more preferred embodiment of the present invention, the instant beverage supplement has an energy distribution of 10 to 15 E% protein, 75 to 85 E% carbohydrates, and 0 to 1 E% lipids. In a preferred embodiment of the present invention, the instant beverage supplement has an energy distribution of 11 E% protein, 89 E% carbohydrates, and 0 E% lipids.

[0252] In one embodiment of the present invention, the instant beverage powder comprises protein, carbohydrates, and lipids, and optionally comprises vitamins, minerals, and trace elements. The instant beverage powder may be designed so that the recommended daily intake of the beverage powder provides the recommended daily intake of vitamins, minerals, and trace elements. However, this is not a mandatory requirement.

[0253] Such instant beverage powders can be useful as nutritional supplements when consumers are interested in nutritional supplements that have a ratio of macronutrients that reflects a healthy diet in terms of energy distribution, macronutrients, and micronutrients, for example, when nutritional supplements are provided under the supervision of a healthcare professional.

[0254] The energy content of the instant beverage powder is in the range of 400 to 500 kcal per 100 grams of powder. In a preferred embodiment of the present invention, the energy content of the instant beverage powder is in the range of 410 to 490 kcal per 100 grams of powder, more preferably the range of 420 to 480 kcal per 100 grams of powder, or most preferably the range of 440 to 460 kcal per 100 grams of powder.

[0255] In a preferred embodiment of the present invention, the instant beverage supplement has an energy distribution such as 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids. In a more preferred embodiment of the present invention, the instant beverage supplement has an energy distribution of 10 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, or more preferably 15 to 20 E% protein, 35 to 45 E% carbohydrates, and 35 to 50 E% lipids. In an even more preferred embodiment of the present invention, the instant beverage supplement has an energy distribution of 15 E% protein, 35 E% carbohydrates, and 50 E% lipids, an energy distribution of 20 E% protein, 45 E% carbohydrates, and 35 E% lipids, or an energy distribution of 8 to 15 E% protein, 40 to 47 E% carbohydrates, and 45 E% lipids.

[0256] In a preferred embodiment of the present invention, the instant powder further comprises vitamins, minerals, and trace elements.

[0257] Instant beverage powder can be used as a nutritional supplement for weight gain, for example, to treat patients who are malnourished or at risk of malnutrition, for patients with kidney disease, or, for example, by sportsmen or athletes before, during, or after exercise.

[0258] In one embodiment of the present invention, the instant beverage powder comprises protein, carbohydrates, and lipids. Such instant beverage powder may be useful as a nutritional supplement when protein intake is a consumer's top priority, for example, when the consumer wishes to supplement a regular meal. The energy content of the instant beverage powder is in the range of 200 to 500 kcal per 100 grams of powder. In a preferred embodiment of the present invention, the energy content of the instant beverage powder is in the range of 200 to 350 kcal per 100 grams of powder, or more preferably, the energy content of the instant beverage powder is in the range of 200 to 300 kcal per 100 grams of powder.

[0259] In a preferred embodiment of the present invention, the instant beverage supplement has an energy distribution such as 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids. In a more preferred embodiment of the present invention, the instant beverage supplement has an energy distribution of 90 to 98 E% protein, 0 to 10 E% carbohydrates, and 0 to 3 E% lipids. In an even more preferred embodiment of the present invention, the instant beverage supplement has an energy distribution of 95 to 98 E% protein, 0 to 5 E% carbohydrates, and 0 to 2 E% lipids.

[0260] In one embodiment of the present invention, the instant powder further comprises vitamins, minerals, and trace elements. In a preferred embodiment of the present invention, the instant powder further comprises vitamins, minerals, and trace elements. The instant beverage powder may be used as a nutritional supplement for weight gain, for example, to treat patients who are malnourished or at risk thereof, for patients suffering from kidney disease, or for example, by sportsmen or athletes.

[0261] In one embodiment of the present invention, the instant beverage powder comprises protein, carbohydrates, and lipids. The energy content of the instant beverage powder is in the range of 200 to 420 kcal per 100 grams of powder. In a preferred embodiment of the present invention, the energy content of the instant beverage powder is in the range of 300 to 420 kcal per 100 grams of powder, more preferably the energy content of the instant beverage powder is in the range of 320 to 380 kcal per 100 grams of powder, and even more preferably the energy content of the instant beverage powder is in the range of 350 to 370 kcal per 100 grams of powder.

[0262] In a preferred embodiment of the present invention, the instant beverage supplement has an energy distribution such as 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids. In a more preferred embodiment of the present invention, the instant beverage supplement has an energy distribution of 60 to 80 E% protein, 4 to 15 E% carbohydrates, and 16 to 18 E% lipids. In an even more preferred embodiment of the present invention, the instant beverage supplement has an energy distribution of 30 to 40 E% protein, 45 to 55 E% carbohydrates, and 12 to 18 E% lipids, for example, 33 E% protein, 46 E% carbohydrates, and 15 E% lipids.

[0263] Alternatively, the energy content of the instant beverage powder may be in the range of 150 to 250 kcal per 100 grams of powder with an energy distribution such as 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, or preferably 15 to 25 E% protein, 45 to 55 E% carbohydrates, and 30 to 40 E% lipids.

[0264] In one embodiment of the present invention, the instant powder further comprises vitamins, minerals, and trace elements.

[0265] In a preferred embodiment of the present invention, the instant powder further comprises vitamins, minerals, and trace elements. The instant beverage powder may be used as a nutritional supplement for weight gain, for example, to treat patients who are malnourished or at risk thereof, for patients suffering from kidney disease, or for example, by sportsmen or athletes.

[0266] In a preferred embodiment of the present invention, the instant beverage powder may further comprise a flavoring agent, a coloring agent, a sweetener, an antioxidant, an edible acid, a lipid, a carbohydrate, a prebiotic, a probiotic, or a non-whey protein.

[0267] For convenience, the instant beverage powder may be sold as a kit comprising the instant powder of the present invention, a tool for measuring said powder, and a container having a lid for opening and closing the container, wherein said container is for mixing said powder with a liquid to form a food, and said container is suitable for drinking the food directly from the container. Examples of useful containers are, for example, bottles, cartons, bricks, pouches and / or bags.

[0268] A consumer purchasing the kit will obtain all the items necessary to easily prepare the liquid food according to the present invention. The measuring tool ensures that the consumer measures an accurate amount of instant powder relative to the amount of water in the container.

[0269] In one embodiment of the present invention, the tool for measuring instant powder is a spoon and the container is a beverage bottle. In one embodiment of the present invention, the container has an internal marking indicating how much liquid is filled into the container. In one embodiment of the present invention, the lid has an opening suitable for drinking the liquid food directly from the container and for closing while mixing the liquid food.

[0270] The pH of the instant beverage powder is important because the taste of the product made from the instant beverage powder varies depending on the pH of the product. The pH of the powder can be determined by measuring the pH of the instant beverage powder in a 10% w / w solution in deionized water at 25°C, as described in Example 1.16. In one embodiment of the present invention, the pH of the instant beverage powder in a 10% w / w solution in deionized water is in the range of 2 to 8 at 25°C.

[0271] In one embodiment of the present invention, the pH is in the range of 2.0 to 4.9, e.g., 2.5 to 4.7, more preferably 2.8 to 4.3, even more preferably 3.2 to 4.0, and most preferably 3.4 to 3.9. Alternatively, but also preferably, the instant beverage powder may have a pH in the range of 3.6 to 4.3.

[0272] Alternatively, the pH of the instant beverage powder in a 10% w / w solution in deionized water is in the range of 5.0 to 6.0 at 25°C, and 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.

[0273] Alternatively, the pH of the instant beverage powder is 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.

[0274] However, one aspect of the present invention relates to the use of instant beverages as defined herein as food ingredients.

[0275] One aspect of the present invention relates to a packaged instant beverage powder product comprising a container containing the instant beverage powder product described herein.

[0276] In one embodiment of the present invention, the instant beverage powder product is optionally sealed in a container packed with an inert gas.

[0277] A wide range of different containers may be used to store instant beverage powder products. For example, the containers may be selected from the group consisting of bottles, cans, bags, pouches, and sachets.

[0278] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, and the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein.

[0279] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2 to 8.

[0280] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2.0 to 4.9.

[0281] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 5.0 to 6.0.

[0282] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 6.1 to 8.5.

[0283] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2 to 8, and the instant beverage powder further contains vitamins, minerals and trace elements.

[0284] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2.0 to 4.9, and the instant beverage powder further contains vitamins, minerals and trace elements.

[0285] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 5.0 to 6.0, and the instant beverage powder further contains vitamins, minerals and trace elements.

[0286] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 6.1 to 8.5, and the instant beverage powder further contains vitamins, minerals and trace elements.

[0287] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2 to 8, and the instant beverage powder further contains vitamins, minerals and trace elements, wherein the sum of the amounts of Na, K, Mg and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0288] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2.0 to 4.9, and the instant beverage powder further contains vitamins, minerals and trace elements, wherein the sum of the amounts of Na, K, Mg and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0289] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 5.0 to 6.0, and the instant beverage powder further contains vitamins, minerals and trace elements, wherein the sum of the amounts of Na, K, Mg and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0290] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 500 kcal per 100 grams of powder, the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 6.1 to 8.5, and the instant beverage powder further contains vitamins, minerals and trace elements, wherein the sum of the amounts of Na, K, Mg and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0291] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein.

[0292] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2 to 8.

[0293] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2.0 to 4.9.

[0294] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 5.0 to 6.0.

[0295] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 6.1 to 8.5.

[0296] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2 to 8, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0297] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2.0 to 4.9, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0298] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 5.0 to 6.0, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0299] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 6.1 to 8.5, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0300] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2 to 8, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0301] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2.0 to 4.9, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0302] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 5.0 to 6.0, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0303] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 400 to 500 kcal per 100 grams of powder, and the energy distribution is in the range of 7 to 25 E% protein, 30 to 50 E% carbohydrates, and 30 to 55 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 6.1 to 8.5, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0304] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder comprises at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein.

[0305] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2 to 8.

[0306] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2.0 to 4.9.

[0307] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 5.0 to 6.0.

[0308] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 6.1 to 8.5.

[0309] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2 to 8, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0310] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2.0 to 4.9, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0311] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 5.0 to 6.0, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0312] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 6.1 to 8.5, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0313] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2 to 8, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0314] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2.0 to 4.9, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0315] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 5.0 to 6.0, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0316] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 400 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 70 to 90 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 6.1 to 8.5, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0317] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein.

[0318] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2 to 8.

[0319] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 95% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2.0 to 4.9.

[0320] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 5.0 to 6.0.

[0321] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 6.1 to 8.5.

[0322] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2 to 8, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0323] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2.0 to 4.9, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0324] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 5.0 to 6.0, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0325] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 6.1 to 8.5, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0326] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2 to 8, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0327] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2.0 to 4.9, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0328] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 5.0 to 6.0, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0329] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 350 kcal per 100 grams of powder, and the energy distribution is in the range of 80 to 98 E% protein, 0 to 20 E% carbohydrates, and 0 to 5 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 6.1 to 8.5, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0330] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein.

[0331] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2 to 8.

[0332] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2.0 to 4.9.

[0333] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 5.0 to 6.0.

[0334] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 6.1 to 8.5.

[0335] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2 to 8, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0336] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2.0 to 4.9, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0337] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 5.0 to 6.0, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0338] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 6.1 to 8.5, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0339] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2 to 8, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0340] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2.0 to 4.9, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0341] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 5.0 to 6.0, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0342] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 200 to 420 kcal per 100 grams of powder, and the energy distribution is in the range of 30 to 80 E% protein, 3 to 20 E% carbohydrates, and 15 to 20 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 6.1 to 8.5, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0343] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein.

[0344] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2 to 8.

[0345] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2.0 to 4.9.

[0346] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 5.0 to 6.0.

[0347] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 6.1 to 8.5.

[0348] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2 to 8, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0349] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 2.0 to 4.9, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0350] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 5.0 to 6.0, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0351] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, the pH of the powder is in the range of 6.1 to 8.5, and the instant beverage powder further contains vitamins, minerals, and trace elements.

[0352] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2 to 8, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0353] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 2.0 to 4.9, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0354] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 5.0 to 6.0, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0355] In one embodiment of the present invention, the instant beverage powder has an energy content in the range of 150 to 250 kcal per 100 grams of powder, and the energy distribution is in the range of 10 to 30 E% protein, 40 to 60 E% carbohydrates, and 25 to 45 E% lipids, wherein the instant beverage powder contains at least 85% w / w BLG with respect to total protein, preferably at least 90% w / w BLG with respect to total protein, and the pH of the powder is in the range of 6.1 to 8.5, and the instant beverage powder further contains vitamins, minerals, and trace elements, wherein the sum of the amounts of Na, K, Mg, and Ca of the instant beverage powder is at most 10 mmol / g protein.

[0356] In some preferred embodiments of the present invention, the food is a dry food comprising carbohydrates and proteins, e.g., a bar or instant beverage powder, said dry food comprising at least 1% w / w BLG, preferably at least 5% w / w BLG, wherein:

[0357] i) The degree of crystallinity of BLG is at least 20%, preferably at least 40%, and / or,

[0358] ii) At least 90% w / w of the total protein consists of BLG.

[0359] In some particularly preferred embodiments of the present invention, the food is a low-phosphorus food containing up to 40 mg of phosphorus per 100 g of protein.

[0360] Non-limiting examples of foods are, for example, dairy products, candy, beverages, instant drinks, protein bars, enteric nutritional compositions, and bakery products.

[0361] In another preferred embodiment of the present invention, the food is an instant beverage powder comprising or even essentially consisting of the following:

[0362] - An edible BLG composition as defined herein in powder form to provide a total amount of BLG of at least 1% w / w, preferably at least 5% w / w, wherein the edible BLG composition has a degree of crystallization of at least 20% of BLG, and

[0363] - Sweeteners in powder form, e.g., sugar sweeteners and / or sugar-free sweeteners,

[0364] - Arbitrarily, flavoring agent,

[0365] - At least one edible acid in powder form, e.g., citric acid or other suitable edible acid, and

[0366] - Up to 80 mg of protein per 100 g;

[0367] Here, a 10% solution of instant beverage powder in deionized water has a pH in the range of 2.5 to 4.0.

[0368] In some preferred embodiments of the present invention, the edible BLG composition comprises the following:

[0369] - Up to 6% w / w water,

[0370] - At least 80% total protein relative to total solids,

[0371] - At least 95% BLG of total protein;

[0372] The above edible BLG composition is:

[0373] - It is a dry powder,

[0374] - It has a bulk density of at least 0.50 g / mL, preferably at least 0.60 g / mL.

[0375] In another preferred embodiment of the present invention, the edible BLG composition comprises the following:

[0376] - Up to 6% w / w water,

[0377] - At least 80% total protein relative to total solids,

[0378] - At least 95% BLG of total protein;

[0379] The above edible BLG composition is as follows:

[0380] - It is a dry powder,

[0381] - Having a bulk density of at least 0.50 g / mL, preferably at least 0.60 g / mL, and

[0382] - It has a crystallinity of at least 20%, preferably at least 40% of BLG.

[0383] In a further preferred embodiment of the present invention, the edible BLG composition comprises the following:

[0384] - Up to 6% w / w water,

[0385] - At least 80% total protein relative to total solids,

[0386] - At least 95% BLG of total protein,

[0387] - Up to 80 mg of protein per 100 g;

[0388] The above edible BLG composition is as follows:

[0389] - It is a dry powder.

[0390] In a more preferred embodiment of the present invention, the edible BLG composition comprises the following:

[0391] - Up to 6% w / w water,

[0392] - At least 90% total protein relative to total solids,

[0393] - At least 97% BLG of total protein,

[0394] - Up to 50 mg of protein per 100 g;

[0395] The above edible BLG composition is as follows:

[0396] - It is a dry powder.

[0397] In another preferred embodiment of the present invention, the edible BLG composition comprises the following:

[0398] - Up to 6% w / w water,

[0399] - At least 80% total protein based on total solids, preferably at least 90% total protein based on total solids,

[0400] - 30 to 70% BLG relative to total protein,

[0401] - 8 to 25% w / w ALA relative to total protein;

[0402] The above edible BLG composition is as follows:

[0403] - It is a dry powder,

[0404] - It has a crystallinity of at least 20%, preferably at least 40% of BLG.

[0405] In one embodiment of the present invention, a liquid food is prepared from instant beverage powder. By using the instant beverage powder according to the present invention, it is possible to obtain a liquid food in a very short time.

[0406] Accordingly, one aspect of the present invention relates to a method for manufacturing a liquid food according to the present invention, said method

[0407] i. A step of adding instant beverage powder according to the present invention,

[0408] ii. A step of optionally adding at least one additional component, and

[0409] iii. The method includes the step of mixing the obtained powder and liquid to form a uniform mixture.

[0410] In one embodiment, the liquid is selected from the group consisting of water, dairy products, fruit juices, vegetable juices, beverages, and combinations thereof. In one embodiment, the additional ingredient is selected from fruits or vegetables.

[0411] When mixing powder and liquid, mixing can be performed by shaking. After shaking, the instant beverage powder was left for 1 / 2 to 2 minutes to completely dissolve the liquid food. One advantage of the instant beverage powder is that it dissolves easily, forms a uniform solution, and maintains a uniform solution, meaning that practically no separation occurs.

[0412] Generally, a problem associated with instant beverage powders is that foam is generated when making liquid food from the powder. The instant beverage powder of the present invention tends not to generate foam when making liquid food from the powder.

[0413] A liquid food comprising a liquid and a powder according to the present invention can be prepared by mixing the instant beverage powder of the present invention with a liquid. In one embodiment of the present invention, the instant beverage powder may comprise up to 40 grams of the powder per 100 grams of the liquid, for example, up to 30 grams of the powder per 100 grams of the liquid. In a preferred embodiment of the present invention, the liquid food comprises 1 to 30 grams of the powder per 100 grams of the liquid, more preferably 1 to 20 grams of the powder, even more preferably 1 to 10 grams of the powder, or preferably 1 to 5 grams of the powder, or even more preferably 2.5 to 5 grams of the powder.

[0414] In one embodiment of the present invention, the liquid food comprises 5 to 25 grams of the powder per 100 grams of liquid, preferably 5 to 25 grams of the powder per 100 grams of liquid, more preferably 10 to 15 grams of the powder, even more preferably 11 to 14 grams of the powder, or more preferably 11 to 12 grams of the powder.

[0415] In one embodiment of the present invention, the food has an energy content in the range of 30 to 300 kcal per 100 grams of food, preferably in the range of 30 to 100 kcal per 100 grams of food, more preferably in the range of 40 to 90 kcal per 100 grams of food, or even more preferably in the range of 40 to 70 kcal per 100 grams of food.

[0416] Alternatively, the food has an energy content in the range of 100 to 300 kcal per 100 grams of food, preferably in the range of 100 to 250 kcal per 100 grams of food, or more preferably in the range of 125 to 225 kcal per 100 grams of food.

[0417] Liquid foods may include liquids selected from the group consisting of water, dairy products, fruit juices, vegetable juices, beverages, and combinations thereof.

[0418] The appearance of liquid foods, for example, beverages made from instant beverage powder, is very important to consumers. Transparency is a parameter that consumers use to evaluate products. One way to determine the transparency of a liquid food is to measure the turbidity of the product described in Example 1.7.

[0419] In some embodiments of a beverage prepared from instant beverage powder, it is advantageous for the beverage to be transparent. This may be advantageous, for example, when the beverage is used as a sports drink or as "protein water," in which case it is advantageous for the beverage to resemble water in appearance.

[0420] In a preferred embodiment of the present invention, a beverage prepared from instant beverage powder has a turbidity of up to 200 NTU, and such a beverage is transparent and / or translucent.

[0421] In some preferred embodiments of the present invention, a beverage prepared from instant beverage powder has a turbidity of up to 150 NTU, or preferably up to 100 NTU, or preferably up to 80 NTU, or preferably up to 60 NTU, or more preferably up to 40 NTU, or preferably up to 30 NTU, preferably up to 20 NTU, more preferably up to 10 NTU, more preferably up to 5 NTU, and even more preferably up to 2 NTU.

[0422] In a preferred embodiment of the present invention, a beverage prepared from instant beverage powder has a turbidity of more than 200 NTU, and such a beverage is opaque.

[0423] In some embodiments of beverages prepared from instant beverage powder, it is advantageous for the beverage to be opaque. This is advantageous, for example, if the beverage is to have a milky white appearance similar to milk. The appearance of nutritionally complete supplements is typically opaque.

[0424] In some preferred embodiments of the present invention, a beverage prepared from instant beverage powder has a turbidity of greater than 250 NTU. Preferably, the beverage has a turbidity of greater than 300 NTU, more preferably greater than 500 NTU, more preferably greater than 1000 NTU, preferably greater than 1500 NTU, and even more preferably greater than 2000 NTU.

[0425] The color of a product is very important to consumers. Instant beverage powder products containing whey protein have a slightly yellowish tint. However, when using instant beverage powder containing BLG with a crystallinity of at least 20% or instant beverage powder in which at least 90% of the protein consists of BLG, the color of the product is substantially less yellow, and the product appears whiter. Therefore, there is no need to add color to the instant beverage powder to mask the yellow color.

[0426] One method of measuring the color of a product is to use the CIELAB color space, which represents color as three numbers; L* is for brightness, and a* and b* are for the green-red and blue-yellow components. Example 1.9 describes a method for measuring L*, a*, and b* values ​​for a liquid food.

[0427] In one embodiment of the present invention, the protein fraction of the liquid food has a color value delta b* in the range of -0.10 to +0.51 on the CIELAB color scale, wherein delta b* = b measured at room temperature. 6.0 w / w % 단백질로 표준화된 샘플 * - b 탈염수 *am.

[0428] Another aspect of the present invention relates to a method for producing instant beverage powder comprising BLG and at least one optional ingredient, the method comprising the step of obtaining instant beverage powder by blending a dried BLG isolate with at least one additional ingredient selected from the group consisting of vitamins, flavoring agents, coloring agents, minerals, sweeteners, antioxidants, edible acids, lipids, carbohydrates, prebiotics, probiotics, antifoaming agents, and non-whey proteins.

[0429] In one embodiment of the present invention, the BLG of the BLG source is coated with an organic acid. For example, if the BLG source is a powder, this means that the powder is coated with an organic acid. When manufacturing instant beverage powders containing protein, lecithin is typically used to improve the solubility of the protein. However, lecithin is a source of phosphorus. Therefore, it is desirable to find another method to improve the solubility of instant beverage powders containing protein.

[0430] The inventors have discovered that the solubility of instant beverage powder is improved by coating BLG crystals or powder particles of a BLG source with one or more organic acids. The organic acid or salt of an organic acid may be selected from the group consisting of pyruvate, aconitate, citrate, iso-citrate, ketoglutarate, succinyl-CoA, succinate, fumarate, maleate, oxaloacetate, tartrate, acetate, tannic acid, benzoic acid, maleic acid, and lactate. In a preferred embodiment of the present invention, BLG crystals are coated with an organic acid or salt of an organic acid selected from the group consisting of pyruvate, citrate, iso-citrate, ketoglutarate, succinate, fumarate, maleate, oxaloacetate, tartrate, acetate, maleic acid, and lactate and salts thereof.

[0431] In a preferred embodiment of the present invention, the BLG crystals of the BLG source are coated with a citrate selected from the group consisting of, for example, trisodium citrate, potassium citrate, and calcium citrate.

[0432] In a preferred embodiment of the present invention, BLG from a BLG source is coated with an organic acid using spray-drying or a fluid bed. It is particularly preferred that dried BLG crystals are coated with an organic acid or a salt thereof, for example, using spray-drying or a fluid bed.

[0433] A BLG source may be obtained from a whey protein feed in which BLG has been isolated as crystals. One method for preparing BLG isolates is described in International Patent Application No. PCT / EP2017 / 084553, which is incorporated herein by reference. BLG isolates may be prepared as described on page 6, lines 23 to 32 of filed PCT / EP2017 / 084553, wherein an edible composition comprising BLG in a crystallized and / or isolated form corresponds to the BLG isolates of the present invention. In a preferred embodiment, BLG isolates are prepared by the method described on page 39, lines 15 to 34 of filed PCT / EP2017 / 084553. In another preferred embodiment, the BLG isolate is prepared by the method described on page 41, lines 1 to 24 of filed PCT / EP2017 / 084553.

[0434] In some preferred embodiments of the present invention, the instant beverage powder comprises or even consists of a BLG isolate powder containing dried BLG crystals, said BLG isolate powder is coated with an organic acid and / or a salt of an organic acid. The weight ratio between the weight of the BLG isolate and the total weight of the sum of the organic acid and the deprotonated organic acid is preferably 5 to 100, more preferably 8 to 60, even more preferably 10 to 40, and most preferably 12 to 30.

[0435] One aspect of the present invention relates to a method for preparing BLG isolate powder coated with an organic acid and / or a salt of an organic acid, said method comprising the following steps:

[0436] - A step of providing a BLG isolate powder to be coated, preferably comprising or even consisting of dried BLG crystals obtained by the BLG crystallization process described herein, and

[0437] - A step of applying an organic acid and / or a salt of an organic acid to the BLG isolate powder to be coated, preferably in an amount sufficient to coat the BLG isolate powder but without dissolving it,

[0438] - A step of evaporating residual moisture from the arbitrarily coated BLG isolated powder.

[0439] The BLG isolate powder to be coated preferably has both high protein content and high BLG purity. The BLG isolate powder to be coated preferably has a degree of crystallization of at least 20%, preferably at least 40%, more preferably at least 60%, and even more preferably at least 80% of BLG.

[0440] The organic acid and / or salt of the organic acid to be coated in an amount sufficient to provide a weight ratio between the weight of the BLG isolated powder and the total weight of the sum of the organic acid and the deprotonated organic acid is preferably 5 to 100, more preferably 8 to 60, even more preferably 10 to 40, and most preferably 12 to 30.

[0441] The organic acid and / or salt of the organic acid is preferably applied to the BLG isolate powder in a fluidized bed system by spraying the organic acid and / or salt of the organic acid, preferably in a dissolved form, onto the fluidized bed to coat the BLG isolate powder. During operation, the temperature is preferably in the range of 5 to 70°C, more preferably in the range of 50 to 65°C, for example, preferably about 60°C.

[0442] After the application of an organic acid and / or a salt of an organic acid, the coated BLG isolate can be treated to evaporate additional moisture until the moisture content is preferably up to 6% w / w, more preferably up to 5% w / w.

[0443] The organic acid is preferably an edible organic acid, that is, a so-called edible acid.

[0444] In some preferred embodiments of the present invention, the BLG source used to produce instant beverage powder has a solid content of at least 20% w / w. Preferably, the BLG source has a solid content of at least 30% w / w, more preferably, the BLG source has a solid content of at least 40% w / w, and even more preferably, the BLG source has a solid content of at least 50% w / w, for example, at least 60% w / w.

[0445] In another preferred embodiment of the present invention, the BLG source used to produce instant beverage powder has a solid content in the range of 20 to 80% w / w. Preferably, the BLG source has a solid content in the range of 30 to 70% w / w. More preferably, the BLG source has a solid content in the range of 40 to 65% w / w. Even more preferably, the BLG source has a solid content in the range of 50 to 65% w / w, for example, approximately 60% w / w.

[0446] The BLG source is preferably a BLG isolate powder, or the liquid BLG isolate contains water and a solid content of BLG isolate powder in an amount ranging from 1 to 50% w / w. It is particularly preferable that the BLG source is a BLG isolate powder.

[0447] Preferably, the beta-lactoglobulin (BLG) isolate powder prepared by spray-drying has a pH in the range of i) 2 to 4.9, ii) 6.1 to 8.5, or iii) 5.0 to 6.0, and comprises the following:

[0448] - At least 30% w / w of total protein,

[0449] - At least 85% w / w of BLG relative to total protein, and

[0450] - Up to 10% w / w of water.

[0451] The BLG isolated powder preferably has one or more of the following:

[0452] - At least 0.2 g / cm³ 3 The bulk density of,

[0453] - At least 1.11 intrinsic tryptophan fluorescence emission ratio (I330 / I350),

[0454] - Up to 10% protein denaturation,

[0455] - Thermal stability of up to 200 NTU at pH 3.9, and

[0456] - Up to 1,000 colony-forming units / g.

[0457] The BLG isolate powder is preferably an edible composition. In some preferred embodiments of the present invention, the BLG isolate powder is an edible BLG composition as defined herein.

[0458] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of 2 to 4.9. Such powder is particularly useful for acidic foods and especially acidic beverages.

[0459] In another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5.

[0460] In some preferred embodiments of the present invention, the BLG isolate powder contains an amount of total protein 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.

[0461] A higher protein content may be required, and in some preferred embodiments of the present invention, the BLG isolate powder contains a total protein in an amount of at least 85% w / w, preferably at least 90% w / w, at least 92% w / w, more preferably at least 94% w / w, and even more preferably at least 95% w / w.

[0462] Total protein is measured according to Example 1.5.

[0463] In some preferred embodiments of the present invention, the BLG isolate powder contains an amount of BLG of at least 92% w / w, preferably at least 95% w / w, more preferably at least 97% w / w, even more preferably at least 98% w / w, and most preferably at least 99.5% w / w of total protein.

[0464] In some preferred embodiments of the present invention, the sum of alpha-lactalbumin (ALA) and caseinomacropeptide (CMP) constitutes at least 40% w / w, preferably at least 60% w / w, more preferably at least 70% w / w, and most preferably at least 90% w / w of the non-BLG protein in the powder.

[0465] In another preferred embodiment of the present invention, each major non-BLG whey protein is present as a weight percentage of total protein, which is up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, and most preferably up to 6% of its weight percentage of total protein in a standard whey protein concentrate from sweet whey.

[0466] Even lower concentrations of major non-BLG whey proteins may be desirable. Accordingly, in a further preferred embodiment of the present invention, each major non-BLG whey protein is present as a weight percentage of total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of its weight percentage of total protein in a standard whey protein concentrate from sweet whey.

[0467] The inventors have seen indications that a reduction in lactoferrin and / or lactoperoxidase is particularly advantageous for obtaining color-neutral whey protein products.

[0468] Accordingly, in some preferred embodiments of the present invention, lactoferrin is present in a weight percentage of total protein, which is up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, most preferably up to 6% of its weight percentage of total protein in a standard whey protein concentrate from sweet whey. Even lower concentrations of lactoferrin may be preferred. Accordingly, in a further preferred embodiment of the present invention, lactoferrin is present in a weight percentage of total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of its weight percentage of total protein in a standard whey protein concentrate from sweet whey.

[0469] Similarly, in some preferred embodiments of the present invention, lactoperoxidase is present in a weight percentage of total protein, which is up to 25%, preferably up to 20%, more preferably up to 15%, even more preferably up to 10%, most preferably up to 6% of its weight percentage of total protein in a standard whey protein concentrate from sweet whey. Even lower concentrations of lactoperoxidase may be preferred. Accordingly, in a further preferred embodiment of the present invention, lactoperoxidase is present in a weight percentage of total protein, which is up to 4%, preferably up to 3%, more preferably up to 2%, and even more preferably up to 1% of its weight percentage of total protein in a standard whey protein concentrate from sweet whey.

[0470] Lactoferrin and lactoperoxidase are quantified according to Example 1.29.

[0471] In some preferred embodiments of the present invention, the BLG isolate powder has a moisture content of up to 10% w / w, preferably up to 7% w / w, more preferably up to 6% w / w, even more preferably up to 4% w / w, and most preferably up to 2% w / w.

[0472] In some preferred embodiments of the present invention, the BLG isolate powder contains an amount of carbohydrates up to 60% w / w, preferably up to 50% w / w, more preferably up to 20% w / w, even more preferably up to 10% w / w, even more preferably up to 1% w / w, and most preferably up to 0.1%. The BLG isolate powder may contain, for example, carbohydrates, for example, lactose, oligosaccharides and / or hydrolysis products of lactose (i.e., glucose and galactose), sucrose, and / or maltodextrin.

[0473] In some preferred embodiments of the present invention, the BLG isolate powder contains an amount of lipid 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.

[0474] The inventors have discovered that it may be advantageous to control the mineral content to achieve some of the desired properties of BLG isolated powder.

[0475] In some preferred embodiments of the present invention, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolate powder is up to 10 mmol / g protein. Preferably, the sum of the amounts of Na, K, Mg, and Ca in the BLG isolate powder is up to 6 mmol / g protein, more preferably up to 4 mmol / g protein, and even more preferably up to 2 mmol / g protein.

[0476] 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.

[0477] In another preferred embodiment of the present invention, the sum of the amounts of Mg and Ca in the BLG isolated powder is up to 5 mmol / g protein. Preferably, the sum of the amounts of Mg and Ca in the BLG isolated powder is up to 3 mmol / g protein, more preferably up to 1.0 mmol / g protein, and even more preferably up to 0.5 mmol / g protein.

[0478] In another preferred embodiment of the present invention, the sum of the amounts of Mg and Ca in the BLG isolated powder is at most 0.3 mmol / g protein. Preferably, the sum of the amounts of Mg and Ca in the BLG isolated powder is at most 0.2 mmol / g protein, more preferably at most 0.1 mmol / g protein, even more preferably at most 0.03 mmol / g protein, and most preferably at most 0.01 mmol / g protein.

[0479] The inventors have discovered that a low phosphorus / low potassium variant of BLG isolate powder, which is particularly useful for patients with kidney disease, can be used. To make such a product, the BLG isolate powder must have equally low phosphorus and potassium content.

[0480] Accordingly, in some preferred embodiments of the present invention, the BLG isolate powder has a total phosphorus content of up to 100 mg of phosphorus per 100 g of protein. Preferably, the BLG isolate powder has a total phosphorus content of up to 80 mg of phosphorus per 100 g of protein. More preferably, the BLG isolate powder has a total phosphorus content of up to 50 mg of phosphorus per 100 g of protein. Even more preferably, the BLG isolate powder has a total phosphorus content of up to 20 mg of phosphorus per 100 g of protein. The BLG isolate powder has a total phosphorus content of up to 5 mg of phosphorus per 100 g of protein.

[0481] 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.

[0482] The phosphorus content is related to the total amount of elemental phosphorus in the composition and is determined according to Example 1.19. Similarly, the potassium content is related to the total amount of elemental potassium in the composition and is determined according to Example 1.19.

[0483] In some preferred embodiments of the present invention, the BLG isolate powder contains up to 100 mg phosphorus and up to 700 mg potassium per 100 g of protein, preferably up to 80 mg phosphorus and up to 600 mg potassium per 100 g of protein, more preferably up to 60 mg phosphorus and up to 500 mg potassium per 100 g of protein, more preferably up to 50 mg phosphorus and up to 400 mg potassium per 100 g of protein, or more preferably up to 20 mg phosphorus and up to 200 mg potassium per 100 g of protein, or even more preferably up to 10 mg phosphorus and up to 50 mg potassium per 100 g of protein. In some preferred embodiments of the present invention, the BLG isolate powder contains up to 100 mg of phosphorus and up to 340 mg of potassium per 100 g of protein.

[0484] The low phosphorus and / or low potassium composition according to the present invention can be used as a food ingredient for the production of food for a group of patients with impaired kidney function.

[0485] The inventors have found that for some applications, for example, acidic foods and particularly acidic beverages, it is particularly advantageous to have an acidic BLG isolated powder having a pH of up to 4.9, more preferably up to 4.3. This applies particularly to high-protein, clear acidic beverages.

[0486] In the context of the present invention, a transparent liquid has a turbidity of up to 200 NTU as measured according to Example 1.7.

[0487] Accordingly, in some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of 2 to 4.9. Preferably, the BLG isolate powder has a pH in the range of 2.5 to 4.7, more preferably 2.8 to 4.3, even more preferably 3.2 to 4.0, and most preferably 3.4 to 3.9. Alternatively, but also preferably, the BLG isolate powder may have a pH in the range of 3.6 to 4.3.

[0488] The inventors have discovered that having pH-neutral BLG isolated powder is particularly advantageous for some applications, for example, pH-neutral foods and especially pH-neutral beverages. This applies particularly to high-protein transparent or opaque pH-neutral beverages.

[0489] Accordingly, in some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5. Preferably, the powder has a pH in the range of 6.1 to 8.5, more preferably 6.2 to 8.0, even more preferably 6.3 to 7.7, and most preferably 6.5 to 7.5.

[0490] 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.

[0491] Advantageously, the BLG isolate powder used in the present invention has a concentration of at least 0.20 g / cm³ 3 , preferably at least 0.30 g / cm³ 3 , more preferably at least 0.40 g / cm³ 3 , more preferably at least 0.45 g / cm³ 3 , more preferably at least 0.50 g / cm³ 3, most preferably at least 0.6 g / cm³ 3 It can have a bulk density.

[0492] Low-density powders, such as freeze-dried BLG isolates, are fluffy and easily enter the air at the production site during use. This is problematic because freeze-dried powders increase the risk of cross-contamination with other foods, and dusty environments are known to cause hygiene issues. In extreme cases, dusty environments also increase the risk of dust explosions.

[0493] The high-density variant of the present invention is easy to handle and has a low tendency to enter the surrounding air.

[0494] An additional advantage of the high-density variant of the present invention is that it takes up less space during transport and thus increases the weight of the BLG isolate powder that can be transported in a single volume unit.

[0495] However, the advantage of the high-density variant of the present invention is that it is suitable for other powdered food ingredients, for example, powdered sugar (approximately 0.56 g / cm³). 3 bulk density), granulated sugar (approximately 0.71 g / cm³) 3 bulk density), powdered citric acid (approximately 0.77 g / cm³) 3 When used in a powder mixture with bulk density, the tendency to separate is small.

[0496] The BLG isolated powder of the present invention has a content of 0.2 to 1.0 g / cm³ 3 Range, preferably 0.30 to 0.9 g / cm³ 3 Range, more preferably 0.40 to 0.8 g / cm³ 3 Range, more preferably 0.45 to 0.75 g / cm³ 3 Range, more preferably 0.50 to 0.75 g / cm³ 3 Range, most preferably 0.6 to 0.75 g / cm³ 3 It can have a bulk density within a range.

[0497] The bulk density of the powder is measured according to Example 1.17.

[0498] The inventors have discovered that it is advantageous to maintain the natural form of BLG, and have observed indications that when BLG is used in acidic beverages, increased unfolding of BLG causes an increased level of dry texture.

[0499] The intrinsic tryptophan fluorescence emission ratio (I330 / I350) measures the degree of unfolding of BLG, and the inventors found that less dryness is observed at a high intrinsic tryptophan fluorescence emission ratio associated with less or no unfolding of BLG. The intrinsic tryptophan fluorescence emission ratio (I330 / I350) is measured according to Example 1.1.

[0500] 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.

[0501] In some preferred embodiments of the present invention, the BLG isolate powder has an intrinsic tryptophan fluorescence emission ratio (I330 / I350) of at least 1.12, preferably at least 1.13, more preferably at least 1.15, even more preferably at least 1.17, most preferably at least 1.19.

[0502] If the BLG isolated powder contains a significant amount of non-protein material, it is desirable to isolate the protein fraction before measuring the intrinsic tryptophan fluorescence emission ratio. Accordingly, in some preferred embodiments of the present invention, the protein fraction of the BLG isolated powder has an intrinsic tryptophan fluorescence emission ratio of at least 1.11.

[0503] In some preferred embodiments of the present invention, the protein fraction of the BLG isolate powder has an intrinsic tryptophan fluorescence emission ratio (I330 / I350) of at least 1.12, preferably at least 1.13, more preferably at least 1.15, even more preferably at least 1.17, most preferably at least 1.19.

[0504] The protein fraction can be separated from the BLG isolate powder by, for example, dissolving the BLG isolate powder in deionized water and dialyzing or ultrafiltration-based volume filtration of the solution using a protein-retaining filter. If the BLG isolate powder contains interfering levels of lipids, such lipids can be removed, for example, by microfiltration. Both lipids and small molecules can be removed from the protein fraction by combining the microfiltration and ultrafiltration / volume filtration steps.

[0505] It is often desirable that a significant amount of BLG in the BLG isolate powder is non-aggregated BLG. Preferably, at least 50% of the BLG is non-aggregated BLG. More preferably, at least 80% of the BLG is non-aggregated BLG. Even more preferably, at least 90% of the BLG is non-aggregated BLG. Most preferably, at least 95% of the BLG is non-aggregated BLG. Even more preferably, approximately 100% of the BLG in the BLG isolate powder is non-aggregated BLG.

[0506] 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%.

[0507] However, for example, if an opaque beverage is required, it may also be desirable for the BLG isolate powder to have a significant level of protein denaturation. Accordingly, in another preferred embodiment of the present invention, the BLG isolate powder has a degree of protein denaturation of at least 11%, preferably at least 20%, more preferably at least 40%, even more preferably at least 50%, even more preferably at least 75%, and most preferably at least 90%.

[0508] If the BLG isolated powder has a significant level of protein denaturation, it is often desirable to maintain a low level of insoluble protein material, that is, precipitated protein material that precipitates into the beverage during storage. The level of insoluble material is measured according to Example 1.10.

[0509] In some preferred embodiments of the present invention, the BLG isolated powder comprises up to 20% w / w insoluble protein material, preferably up to 10% w / w insoluble protein material, more preferably up to 5% w / w insoluble protein material, even more preferably up to 3% w / w insoluble protein material, and most preferably up to 1% w / w insoluble protein material. It may be more preferable that the BLG isolated powder does not contain any insoluble protein material at all.

[0510] In some preferred embodiments of the present invention, the BLG isolate powder has a crystallinity of BLG of up to 19%, preferably up to 10%, more preferably up to 5%, and most preferably 0%.

[0511] In another preferred embodiment of the present invention, the BLG isolate powder has a crystallinity of at least 20%, preferably at least 40%, more preferably at least 60%, and most preferably at least 80% of BLG. These embodiments contain a significant amount of dried BLG crystals and provide the advantage of having a protein source existing in a dense solid form.

[0512] The inventors have discovered that the thermal stability of the BLG isolated powder at pH 3.9 is an excellent indicator of its usefulness for clear high-protein beverages. Thermal stability at pH 3.9 is measured according to Example 1.2.

[0513] It is particularly desirable that the BLG isolated powder has a thermal stability of up to 200 NTU at pH 3.9, preferably up to 100 NTU, more preferably up to 60 NTU, even more preferably up to 40 NTU, and most preferably up to 20 NTU. Even better thermal stability is possible, and the BLG isolated powder preferably has a thermal stability of up to 10 NTU at pH 3.9, preferably up to 8 NTU, more preferably up to 4 NTU, and even more preferably up to 2 NTU.

[0514] It is desirable to keep the microbial content of BLG isolated powder to a minimum. However, since the process for reducing microorganisms tends to result in protein unfolding and denaturation, it is a difficult task to obtain both a high degree of protein naturalness and a low microbial content. The present invention enables obtaining a very low microbial content while maintaining a high level of BLG naturalness.

[0515] Accordingly, in some preferred embodiments of the present invention, the BLG isolate powder contains up to 15,000 colony-forming units (CFU) / g. Preferably, the BLG isolate powder contains up to 10,000 CFU / g. More preferably, the BLG isolate powder contains up to 5,000 CFU / g. Even more preferably, the BLG isolate powder contains up to 1,000 CFU / g. Even more preferably, the BLG isolate powder contains up to 300 CFU / g. Most preferably, the BLG isolate powder contains up to 100 CFU / g, for example, up to 10 CFU / g. In a particularly preferred embodiment, the powder is sterile. Sterile BLG isolate powder can be produced by combining several physical microbiome reduction processes, such as microfiltration and heat treatment at an acidic pH, while producing the BLG isolate powder.

[0516] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of i) 2 to 4.9, ii) 6.1 to 8.5, or iii) 5.0 to 6.0, and comprises:

[0517] - Total protein in an amount of at least 30% w / w, preferably at least 80% w / w, more preferably at least 90% w / w,

[0518] - At least 85% w / w, preferably at least 90% w / w, of beta-lactoglobulin (BLG) relative to total protein,

[0519] - Up to 6% w / w of water,

[0520] - Up to 2% w / w, preferably up to 0.5% w / w of lipids;

[0521] The above BLG isolated powder has the following:

[0522] - At least 1.11 intrinsic tryptophan fluorescence emission ratio (I330 / I350),

[0523] - Up to 10% protein denaturation, and

[0524] - Thermal stability of up to 200 NTU at pH 3.9.

[0525] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of i) 2 to 4.9 or ii) 6.1 to 8.5 and comprises:

[0526] - Total protein in an amount of at least 30% w / w, preferably at least 80% w / w, more preferably at least 90% w / w,

[0527] - At least 85% w / w, preferably at least 90% w / w, and more preferably at least 94% w / w of beta-lactoglobulin (BLG) relative to total protein,

[0528] - Up to 6% w / w of water,

[0529] - Up to 2% w / w, preferably up to 0.5% w / w of lipids;

[0530] The above BLG isolated powder has the following:

[0531] - At least 1.11 intrinsic tryptophan fluorescence emission ratio (I330 / I350),

[0532] - Protein denaturation of up to 10%, preferably up to 5%, and

[0533] - Thermal stability of up to 70 NTU at pH 3.9, preferably up to 50 NTU, more preferably up to 40 NTU.

[0534] In some preferred embodiments of the present invention, the BLG isolate powder has a pH in the range of i) 2 to 4.9 or ii) 6.1 to 8.5 and comprises:

[0535] - At least 30% w / w of total protein,

[0536] - At least 85% w / w, preferably at least 90% w / w, of beta-lactoglobulin (BLG) relative to total protein,

[0537] - Up to 6% w / w of water,

[0538] The above BLG isolated powder has the following:

[0539] - At least 0.2 g / cm³ 3 The bulk density of,

[0540] - At least 1.11 intrinsic tryptophan fluorescence emission ratio (I330 / I350),

[0541] - Up to 10% protein denaturation, and

[0542] - Thermal stability of up to 200 NTU at pH 3.9.

[0543] In another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 2 to 4.9 and comprises:

[0544] - at least 80% w / w, preferably at least 90% w / w, more preferably at least 94% w / w of total protein,

[0545] - At least 85% w / w, preferably at least 90% w / w, of beta-lactoglobulin (BLG) relative to total protein, more preferably at least 94% w / w relative to total protein,

[0546] - Up to 6% w / w of water,

[0547] - Up to 2% w / w, preferably up to 0.5% w / w of lipids;

[0548] The above BLG isolated powder has the following:

[0549] - At least 0.2 g / cm³ 3 , preferably at least 0.3 g / cm³ 3 , more preferably at least 0.4 g / cm³ 3 The bulk density of,

[0550] - At least 1.11 intrinsic tryptophan fluorescence emission ratio (I330 / I350),

[0551] - Protein denaturation of up to 10%, preferably up to 5%, more preferably up to 2%, and

[0552] - Thermal stability of up to 50 NTU at pH 3.9, preferably up to 30 NTU, more preferably up to 10 NTU.

[0553] In another preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5 and comprises:

[0554] - at least 80% w / w, preferably at least 90% w / w, more preferably at least 94% w / w of total protein,

[0555] - At least 85% w / w, preferably at least 90% w / w, of beta-lactoglobulin (BLG) relative to total protein, more preferably at least 94% w / w relative to total protein,

[0556] - Up to 6% w / w of water,

[0557] - Up to 2% w / w, preferably up to 0.5% w / w of lipids;

[0558] The above BLG isolated powder has the following:

[0559] - At least 0.2 g / cm³ 3 , preferably at least 0.3 g / cm³ 3 , more preferably at least 0.4 g / cm³ 3 The bulk density of,

[0560] - Protein denaturation of up to 10%, preferably up to 5%, more preferably up to 2%, and

[0561] - Thermal stability of up to 50 NTU at pH 3.9, preferably up to 30 NTU, more preferably up to 10 NTU.

[0562] In a further preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 6.1 to 8.5 and comprises:

[0563] - at least 80% w / w, preferably at least 90% w / w, more preferably at least 94% w / w of total protein,

[0564] - At least 85% w / w, preferably at least 90% w / w, of beta-lactoglobulin (BLG) relative to total protein, more preferably at least 94% w / w relative to total protein,

[0565] - Up to 6% w / w of water,

[0566] - Up to 2% w / w, preferably up to 0.5% w / w of lipids;

[0567] The above BLG isolated powder has the following:

[0568] - At least 0.2 g / cm³ 3 , preferably at least 0.3 g / cm³ 3 , more preferably at least 0.4 g / cm³ 3 The bulk density of,

[0569] - Protein denaturation of up to 10%, preferably up to 5%, more preferably up to 2%, and

[0570] - Thermal stability of up to 50 NTU at pH 3.9, preferably up to 30 NTU, more preferably up to 10 NTU.

[0571] In a further preferred embodiment of the present invention, the BLG isolate powder has a pH in the range of 5.0 to 6.0 and comprises:

[0572] - at least 80% w / w, preferably at least 90% w / w, more preferably at least 94% w / w of total protein,

[0573] - At least 85% w / w, preferably at least 90% w / w, of beta-lactoglobulin (BLG) relative to total protein, more preferably at least 94% w / w relative to total protein,

[0574] - Up to 6% w / w of water,

[0575] - Up to 2% w / w, preferably up to 0.5% w / w of lipids;

[0576] The above BLG isolated powder has the following:

[0577] - At least 0.2 g / cm³ 3 , preferably at least 0.3 g / cm³ 3 , more preferably at least 0.4 g / cm³ 3 The bulk density of,

[0578] - Protein denaturation of up to 10%, preferably up to 5%, more preferably up to 2%,

[0579] - Thermal stability of up to 50 NTU at pH 3.9, preferably up to 30 NTU, more preferably up to 10 NTU, and

[0580] - Preferably, BLG crystallinity of less than 10%.

[0581] BLG isolate powder containing at least 85% w / w of BLG relative to total protein is typically provided by a method comprising the following steps:

[0582] a) A step of providing a liquid BLG isolate having the following:

[0583] i) pH in the range of 2 to 4.9,

[0584] ii) pH in the range of 6.1 to 8.5, or

[0585] iii) pH in the range of 5.0 to 6.0

[0586] The above liquid BLG isolate contains an amount of BLG of at least 85% w / w relative to total protein, step,

[0587] b) Optionally, a step of physically reducing microorganisms in the liquid BLG isolate,

[0588] c) A step of drying the liquid BLG isolated material, preferably spray-drying.

[0589] BLG isolates are preferably prepared from mammalian milk, preferably ruminant milk, e.g., milk of dairy cows, sheep, goats, buffalo, camels, llamas, mares, and / or deer. Proteins derived from cow's milk are particularly preferred. Thus, BLG is preferably bovine BLG.

[0590] Liquid BLG isolates can be provided in a number of different ways.

[0591] Typically, the provision of a liquid BLG isolate comprises or even consists of the step of isolating BLG from a whey protein feed to provide a BLG-rich composition by one or more of the following methods:

[0592] - Crystallization or precipitation of BLG by salt treatment,

[0593] - Crystallization or precipitation of BLG by salting out,

[0594] - Ion exchange chromatography, and

[0595] - Separation of whey proteins by ultrafiltration

[0596] A particularly preferred method for providing a BLG-rich composition is by crystallization of BLG, preferably by salting or alternatively by salting out.

[0597] The whey protein feedstock is preferably WPC, WPI, SPC, SPI, or a combination thereof.

[0598] The term "whey protein feedstock" relates to a BLG-rich composition and subsequently a composition from which a liquid BLG isolate is derived.

[0599] In some embodiments of the present invention, the preparation of the BLG-rich composition comprises or even consists of high salt BLG crystallization in the pH range of 3.6 to 4.0 in accordance with US No. 2,790,790 A1.

[0600] In another embodiment of the present invention, the preparation of the BLG-rich composition comprises or is even made by the method described by de Jongh et al. (Mild Isolation Procedure Discloses New Protein Structural Properties of β-Lactoglobulin, J Dairy Sci., vol. 84(3), 2001, pages 562-571) or Vyas et al. (Scale-Up of Native β-Lactoglobulin Affinity Separation Process, J. Dairy Sci. 85:1639-1645, 2002).

[0601] However, in a particularly preferred embodiment of the present invention, the BLG-rich composition is prepared by crystallization at pH 5 to 6 under salt-soluble conditions as described in PCT application No. PCT / EP2017 / 084553, incorporated herein by reference for all purposes.

[0602] In some preferred embodiments of the present invention, the BLG-rich composition is an edible BLG composition according to PCT / EP2017 / 084553 containing at least 90% BLG with respect to total protein and preferably containing BLG crystals.

[0603] If the BLG-rich composition isolated from a whey protein feed does not yet possess the necessary characteristics to be used as a liquid BLG isolate, it may be processed in one or more steps selected from the following group as part of providing a liquid BLG isolate:

[0604] - Desalination,

[0605] - Mineral addition,

[0606] - Dilution,

[0607] - Concentration,

[0608] - Physical microbial reduction, and

[0609] - pH adjustment.

[0610] Non-limiting examples of desalination include, for example, dialysis, gel filtration, UF / dialysis filtration, NF / dialysis filtration, and ion exchange chromatography.

[0611] Non-limiting examples of mineral addition include the addition of salts permitted in water-soluble foods, e.g., salts of Na, K, Ca, and / or Mg. Such salts may be, e.g., phosphate salts, chloride salts, or salts of edible acids, e.g., citrate salts or lactate salts. Minerals may be added in solid, suspended, or dissolved form.

[0612] 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.

[0613] Non-limiting examples of concentration include, for example, evaporation, reverse osmosis, nanofiltration, ultrafiltration, and combinations thereof.

[0614] If it is necessary to increase the protein concentration relative to the total solids, it is desirable to use a concentration step such as ultrafiltration or alternatively dialysis. If it is not necessary to increase the protein concentration relative to the total solids, methods such as evaporation, nanofiltration, and / or reverse osmosis may be useful.

[0615] Non-limiting examples of physical microbial reduction include, for example, heat treatment, bacterial filtration, UV radiation, high-pressure treatment, pulsed electric field treatment, and ultrasound. These methods are well known to those skilled in the art.

[0616] Non-limiting examples of pH adjustment include, for example, the addition of a base and / or acid, preferably a base and / or acid acceptable to food. It is particularly desirable to use an acid and / or base capable of chelating divalent metal cations. Examples of such acids and / or bases are citric acid, citrate salts, EDTA, lactic acid, lactate salts, phosphoric acid, phosphate salts, and combinations thereof.

[0617] Next, there are a number of preferred embodiments providing a liquid BLG isolate of step a) from a BLG-rich composition. In this context, the process step mentioned is applied to a BLG-containing product stream following the BLG-rich composition.

[0618] The present invention has been described above with reference to specific embodiments. However, other embodiments other than those described above are equally possible within the scope of the present invention. Different features and steps of various embodiments and aspects of the present invention may be combined in ways different from those described herein, unless otherwise noted.

[0619] Examples

[0620] Example 1: Analysis Method

[0621] Example 1.1: Determination of protein naturalness by intrinsic tryptophan fluorescence

[0622] Tryptophan (Trp) fluorescence spectroscopy is a well-described tool for monitoring protein folding and unfolding. Trp residues embedded within natural proteins typically exhibit the highest fluorescence emission around 330 nm compared to when they are located at higher solvent exposure sites, such as in unfolded proteins. In unfolded proteins, the wavelength for Trp fluorescence emission typically shifts to higher wavelengths and is often measured around 350 nm. Here, the inventors utilize this shift to monitor thermally induced unfolding by calculating the ratio between fluorescence emission at 330 nm and 350 nm to investigate the effect of heating temperature.

[0623] The analysis includes the following steps:

[0624] The beverage composition was diluted to 0.6 mg / ml in MQ water.

[0625] ㆍ Transfer 300 μl of the sample to a white 96-well plate to prevent air bubbles, or transfer 3 mL to a 10 mm quartz cuvette.

[0626] The tryptophan fluorescence emission intensity between 310 nm and 400 nm was recorded from the top by excitation at 295 using a 5 nm slit.

[0627] ㆍ Samples were measured at 22°C using a Cary Eclipse fluorescence spectrophotometer equipped with a plate reader accessory (G9810A) or a single cuvette holder.

[0628] The luminescence intensity ratio was calculated by dividing the fluorescence luminescence intensity measured at 330 nm by the luminescence intensity at 350 nm (R = I330 / I350) and used as a measure of protein nativity.

[0629] o At least R in 1.11 describes the dominant natural BLG form, and

[0630] o R less than 1.11 reports at least partial unfolding and aggregation.

[0631] Example 1.2: Thermal stability at pH 3.9

[0632] Thermal stability at pH 3.9:

[0633] Thermal stability at pH 3.9 is a measure of the ability of a protein composition to remain clear during long-term pasteurization at pH 3.9.

[0634] Thermal stability at pH 3.9 is determined by the steps of: mixing a sample of powder or liquid to be tested with water (or alternatively, if it is a dilute liquid, concentrating it by low-temperature evaporation) to form an aqueous solution containing 6.0% w / w protein with a pH of 3.9, and adjusting the pH to 3.9 with the minimum required amount of 0.1 M NaOH or 0.1 M HCl.

[0635] After leaving the pH-adjusted mixture for 30 minutes, transfer 25 mL of the mixture to a 30 mL thin-walled glass test tube. Immerse the tube in a water bath at 75.0°C and heat it to 75.0°C for 300 seconds. Immediately after heating, transfer the glass test tube to an ice bath to cool it to 1 to 5°C, and measure the turbidity of the heat-treated sample according to Example 1.7.

[0636] Example 1.3: Determination of Protein Denaturation Degree of Whey Protein Composition

[0637] It is known that denatured whey protein has lower solubility at pH 4.6 than at pH values ​​below or above pH 4.6, and therefore, the degree of denaturation of a whey protein composition is determined by measuring the amount of soluble protein at pH 4.6 relative to the total amount of protein at the pH at which the protein in solution is stable.

[0638] 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:

[0639] - A first aqueous solution containing 5.0% (w / w) total protein and having a pH of 7.0 or 3.0, and

[0640] - A second aqueous solution containing 5.0% (w / w) total protein and having a pH of 4.6.

[0641] pH adjustment is performed using 3% (w / w) NaOH (aqueous) or 5% (w / w) HCl (aqueous).

[0642] Total protein content of the first aqueous solution (PpH 7.0 또는 3.0 ) is determined according to Example 1.5.

[0643] The second aqueous solution is stored at room temperature for 2 hours, and then centrifuged at 3000 g for 5 minutes. A sample of the supernatant is recovered and analyzed according to Example 1.5 to obtain the supernatant (S pH4.6 Provides the protein concentration of ).

[0644] The degree of protein denaturation, D, of the whey protein composition is calculated as follows:

[0645] D = ((P pH 7.0 또는 3.0 -S pH 4.6 ) / P pH 7.0 또는 3.0 )*100%

[0646] Example 1.4 Determination of protein denaturation using reverse-phase UPLC analysis (using pH 4.6 acid precipitation).

[0647] BLG samples (e.g., non-heated reference and heated BLG beverage compositions) were diluted to 2% in MQ water. 5 mL of protein solution, 10 mL of Milli-Q, 4 mL of 10% acetic acid, and 6 mL of 1.0 M NaOAc were mixed and stirred for 20 minutes to allow precipitation aggregation of the denatured protein at a pH of around 4.6. The solution was filtered through a 0.22 μm filter to remove aggregates and non-natural proteins.

[0648] All samples were diluted to the same degree by adding polished water.

[0649] For each sample, the same volume was loaded into a UPLC system equipped with a UPLC column (protein BEH C4; 300 Å; 1.7 μm; 150 x 2.1 mm) and detected at 214 nm.

[0650] The sample was executed using the following conditions:

[0651] Buffer A: Milli-Q water, 0.1% w / w TFA

[0652] Buffer B: HPLC-grade acetonitrile, 0.1% w / w TFA

[0653] Flow: 0.4 ml / min

[0654] Gradient: 0 to 6.00 min 24 to 45% B; 6.00 to 6.50 min 45 to 90% B; 6.50 to 7.00 min 90% B; 7.00 to 7.50 min 90 to 24% B and 7.50 to 10.00 min 24% B.

[0655] The concentration of natural BLG in the sample (5-level calibration curve) was determined using the area of ​​the BLG peak relative to the protein standard (Sigma L0130).

[0656] The sample was further diluted, and reinjected when it fell outside the linear range.

[0657] Example 1.5: Determination of Total Protein

[0658] The total protein content (net protein) of the sample is determined by the following:

[0659] 1) ISO 8968-1 / 2|IDF 020-1 / 2- Determination of Total Nitrogen in Samples Based on Milk - Determination of Nitrogen Content - Part 1 / 2: Determination of Nitrogen Content Using the Kjeldahl Method

[0660] 2) ISO 8968-4|IDF 020-4- Determination of Non-Protein Nitrogen of Samples According to Milk - Determination of Nitrogen Content - Part 4: Determination of Non-Protein Nitrogen Content.

[0661] 3) Total amount of protein (m 총 질소 - m 비-단백질-질소 Calculated as )*6.38.

[0662] Example 1.6: Determination of non-aggregated BLG, ALA, and CMP

[0663] The content of non-aggregated alpha-lactalbumin (ALA), beta-lactoglobulin (BLG), and caseinomacropeptide (CMP), respectively, was analyzed by HPLC at 0.4 mL / min. 25 μL of filtered samples were injected into two TSKgel3000PWxl (7.8 mm 30 cm, Tosohass, Japan) columns connected in series with an attached pre-column PWxl (6 mm x 4 cm, Tosohass, Japan) equilibrated in an eluent (consisting of 465 g Milli-Q water, 417.3 g acetonitrile, and 1 mL trifluoroacetic acid), and a UV detector was used at 210 nm.

[0664] Natural alpha-lactalbumin (C 알파 ), beta-lactoglobulin (C 베타 ), and caseinomacropeptide (C CMP The quantitative determination of the content of ) was performed by comparing the peak area obtained for the corresponding standard protein with the peak area of ​​the sample.

[0665] The total amount of additional protein (non-BLG protein) was determined by subtracting the amount of BLG from the amount of total protein (determined according to Example 1.5).

[0666] Example 1.7: Determination of Turbidity

[0667] Turbidity is the cloudiness or haziness of a fluid caused by a large number of particles that are generally invisible to the naked eye, similar to smoke in the air.

[0668] Turbidity is measured in NTU (nephelometric turbidity units).

[0669] 20 mL of beverage / sample was added to an NTU glass and placed in a Turbiquant® 3000 IR turbidimeter. NTU values ​​were measured after stabilization and repeated twice.

[0670] Example 1.8: Determination of Viscosity

[0671] The viscosity of the beverage formulation was measured using a rheometer (Anton Paar, Physica MCR301).

[0672] 3.8 mL of the sample was added to cup DG26.7. The sample was equilibrated to 22°C, and then 50 seconds -1 Pre-shear for 30 seconds, followed by 30 seconds of equilibrium time and 1 s -1 and 200s -1 and 1s -1 A shear rate sweep was performed between them.

[0673] Viscosity is 100s unless otherwise specified. -1 It is expressed in units of centipoise (cP) at the shear rate. The higher the measured cP value, the higher the viscosity.

[0674] Alternatively, viscosity was estimated using Viscoman (manufacturer: Gilson) and was approximately 300s -1 It was reported at a shear rate of .

[0675] Example 1.9: Determination of color

[0676] Color was measured using a Chroma Meter (Konica Minolta, CR-400). A 15 g sample was placed in a small Petri dish (55 x 14.2 mm, VWR Cat# 391-0895) to prevent bubble formation. The protein content of the sample was standardized to a protein of 6.0 w / w % or less.

[0677] The colorimeter was calibrated with a white calibration plate (No. 19033177). The illuminant was set to D65 and the observer was set to 2 degrees. The hue (CIELAB color space, a*-, b*-, L*- values) was measured as the average of three individual readings at different locations on the Petri dish with the lid covering the suspension.

[0678] Demineralized water standards have the following values:

[0679] L* 39.97±0.3

[0680] a* 0.00±0.06

[0681] b* -0.22±0.09

[0682] The measurement was converted into a delta / difference value based on the demineralized water measurement.

[0683] Delta L* = L 6.0 w / w % 단백질로 표준화된 샘플 * - L 탈염수 *, measured at room temperature

[0684] Delta a* = a 6.0 w / w % 단백질로 표준화된 샘플 * - a 탈염수 *, measured at room temperature

[0685] Delta b* = b 6.0 w / w % 단백질로 표준화된 샘플 * - b 탈염수 *, measured at room temperature

[0686] The sample is standardized to less than 6.0 w / w % protein.

[0687] The L*a*b* color space (also referred to as the CIELAB space) is one of the uniform color spaces defined by the International Commission on Illumination (CIE) in 1976 and was used to quantitatively report brightness and hue (ISO 11664-4:2008(E) / CIE S 014- 4 / E:2007).

[0688] In this space, L* represents brightness (values ​​from 0 to 100), the darkest black at L* = 0, and the brightest white at L* = 100.

[0689] Color channels a* and b* represent true neutral gray values ​​at a* = 0 and b* = 0. The a* axis represents the green-red component, where green is in the negative direction and red is in the positive direction. The b* axis represents the blue-yellow component, where blue is in the negative direction and yellow is in the positive direction.

[0690] Example 1.10 Beverage Stability Test / Insoluble Protein Material

[0691] When less than 15% of the total protein precipitates from a heated sample during centrifugation at 3000 g for 5 minutes, the whey protein beverage composition was considered stable.

[0692] Approximately 20 g of the sample was added to a centrifuge tube and centrifuged at 3000 g for 5 minutes.

[0693] Protein recovery was quantified using Kjeldahl analysis of the protein before centrifugation and the supernatant after centrifugation. Refer to Example 1.5.

[0694] Protein loss is calculated as follows:

[0695]

[0696] Additionally, this parameter is sometimes referred to as the level of insoluble protein material and can be used to analyze both liquid and powder samples. If the sample is a powder, 10 g of powder is suspended in 90 g of deionized water and hydrated at 22°C for 1 hour under gentle stirring. Approximately 20 g of the sample (e.g., liquid sample or suspended powder sample) was added to a centrifuge tube and centrifuged at 3000 g for 5 minutes. Before centrifugation (P 총 ) Protein and after centrifugation (P 3000xg Protein recovery according to Example 1.5 was quantified using Kjeldahl analysis of the supernatant.

[0697] The amount of insoluble protein material is calculated as follows:

[0698]

[0699] Example 1.11: Sensory Evaluation

[0700] The heat-treated beverage formulations underwent descriptive sensory evaluation. The beverage formulations were heat-treated using a plate heat exchanger.

[0701] Mix 1 volume of sample with 1 volume of water, compare with non-heated whey protein isolate, and also use lactic acid and citric acid to form an attribute list prior to the final tasting session:

[0702] Category Attributes:

[0703] Flavored whey, acidic (sour dairy)

[0704] Basic taste: acid, bitter

[0705] Flavoring whey, citric acid, lactic acid

[0706] Dry texture, astringent taste

[0707] Participants' mouths were cleaned between each sample using crackers, white tea, melon, and water.

[0708] A 15 mL test sample was placed in a small cup at ambient temperature (20 to 25℃).

[0709] Test samples were provided three times to 10 individuals in each of three different blocks in a random order.

[0710] The properties (see table above) were evaluated on a 15 cm scale with 0 = low strength and 15 = high strength.

[0711] Statistical analysis was performed in 'Panelcheck' software using a 3-way ANOVA test for multiple replications. Samples were fixed and panels were randomized. Significant differences between samples were evaluated using Bonferroni correction, which implies the minimum significant difference value (paired comparison of groups related to letters).

[0712] Example 1.12: Determination of transparency by imaging

[0713] A photograph of the beverage formulation was taken by placing a sample in a turbidity NTU measuring vial that came into contact with a piece of paper containing the text 'lorem ipsum'. The vial was photographed using a smartphone, and the inventors evaluated whether the text could be clearly observed through the vial.

[0714] Example 1.13: Determination of Ash Content

[0715] The ash content of food is determined according to NMKL 173:2005 "Ashes in food, weight measurement".

[0716] Example 1.14: Determination of Conductivity

[0717] The "conductivity" (sometimes referred to as "specific conductivity") of an aqueous solution is a measure of the solution's ability to conduct electricity. Conductivity can be determined, for example, by measuring the AC resistance of the solution between two electrodes, and the results are typically provided in millisiemens per centimeter (mS / cm). Conductivity can be measured, for example, according to the EPA (the US Environmental Protection Agency) Method No. 120.1.

[0718] Conductivity values ​​mentioned herein have been normalized to 25°C unless otherwise specified.

[0719] Conductivity is measured using a conductivity meter (WTW Cond 3210 with Tetracon 325 electrodes).

[0720] The system is calibrated as described in the manual before use. The electrode is thoroughly rinsed in the same type of medium where the measurement is to be performed to avoid local dilution. The electrode is lowered into the medium so that the area where the measurement takes place is completely submerged. Then, the electrode is shaken to remove any trapped air. Then, the electrode is held in place until a stable value is obtained and can be recorded from the display.

[0721] Example 1.15: Determination of total solid content of the solution

[0722] The total solid content of the solution is [NMKL 110 2 nd It can be determined according to [Edition, 2005 (Total solids (Water) - Gravimetric determination in milk and milk products)]. NMKL is an abbreviation for "Nordic Food Methodology Committee (Nordisk Metodikkomite for Naeringsmidler)".

[0723] The water content of the solution can be calculated as 100% minus the total solid content (% w / w).

[0724] Example 1.16: Determination of pH

[0725] All pH values ​​are measured using a pH glass electrode and normalized to 25°C.

[0726] The pH glass electrode (with temperature compensation) is carefully rinsed and compensated before use.

[0727] If the sample is in liquid form, the pH is measured directly in the liquid solution at 25°C.

[0728] If the sample is a powder, dissolve 10 grams of powder in 90 ml of deionized water at room temperature while stirring vigorously. Then, the pH of the solution is measured at 25°C.

[0729] Example 1.17: Determination of loose density and bulk density

[0730] The density of a dry powder is defined as the relationship between the weight and volume of the powder analyzed using a special Stampf volumetric meter (i.e., a measuring cylinder) under specified conditions. Density is typically expressed in g / ml or kg / L.

[0731] In this method, a sample of the dried powder is placed in a measuring cylinder. After a specified number of taps, the volume of the product is read and the density is calculated.

[0732] Three types of density can be defined by this method:

[0733] ■ Poured density, calculated by dividing the mass by the volume of the powder after transferring to the specified measuring cylinder

[0734] ■ Loose density, calculated by dividing the mass by the volume of the powder after 100 taps according to the conditions specified in this standard.

[0735] ■ Bulk density, calculated by dividing the mass by the volume of the powder after 625 taps according to the conditions specified in this standard.

[0736] The above method uses a special measuring cylinder, 250 ml, graduated from 0 to 250 ml, with a weight of 190 ± 15 g (J. Engelsmann AG 67059 Ludwigshafen / Rh) and a Stampf volumetric measuring instrument, e.g., J. Engelsmann A. G.

[0737] The loose density and bulk density of the dried product are determined by the following procedure.

[0738] Preprocessing:

[0739] The sample to be measured is stored at room temperature.

[0740] Next, rotate and turn the container repeatedly to thoroughly mix the sample (to prevent particle crushing). The container is not filled more than 2 / 3 full.

[0741] procedure:

[0742] Weigh 100.0 ± 0.1 grams of powder and transfer it to a measuring cylinder. Read the volume V0 in ml units.

[0743] If 100 g of powder does not fit into the cylinder, the amount must be reduced to 50 or 25 g.

[0744] Secure the measuring cylinder to the Stampf volumetric meter and tap it 100 times. Flatten the surface with a spatula and volume V 100 It is read in ml units.

[0745] Change the number of taps to 625 (including 100 taps). After tapping, flatten the surface, and volume V 625 It is read in ml units.

[0746] Density calculation:

[0747] Calculate the loose density and bulk density in g / ml according to the following formula:

[0748] Bulk density = M / V

[0749] Here, M represents the weighed sample in grams, and V represents the volume after 625 taps in ml.

[0750] Example 1.18: Determination of moisture content of powder

[0751] The moisture content of food is determined according to ISO 5537:2004 (milk powder - determination of moisture content (reference method)). NMKL is an abbreviation for the "Nordic Food Methodology Committee (Nordisk Metodikkomite for Naeringsmidler)".

[0752] Example 1.19: Determination of amounts of calcium, magnesium, sodium, potassium, and phosphorus (ICP-MS method)

[0753] The total amount of calcium, magnesium, sodium, potassium, and phosphorus is determined using a procedure to first decompose the sample using microwave digestion, and then the total amount of mineral(s) is determined using an ICP device.

[0754] device:

[0755] The microwave is from the manufacturer (Anton Paar) and the ICP is the Optima 2000DV from the manufacturer (PerkinElmer Inc.).

[0756] ingredient:

[0757] 1 M HNO3

[0758] Yttrium in 2% HNO3

[0759] Standards suitable for calcium, magnesium, sodium, potassium, and phosphorus in 5% HNO3

[0760] Preprocessing:

[0761] Weigh a certain amount of powder and transfer it to a microwave digestion tube. Add 5 mL of 1 M HNO3. Dilute the sample in the microwave according to the microwave instructions. Place the digested tube into a fume cupboard, remove the lid, and allow the volatile fumes to evaporate.

[0762] Measurement Procedure:

[0763] Transfer the pretreated sample to DigiTUBE using a known amount of Milli-Q water. Add 2% HNO3 yttrium solution to the digestion tube (approx. 0.25 mL per 50 mL diluted sample) and dilute to a known volume using Milli-Q water. Analyze the sample in ICP using the procedure described by the manufacturer.

[0764] A blind sample is prepared by using Milli-Q water to dilute a mixture of 10 mL of 1 M HNO3 and 0.5 mL of yttrium solution in 2% HNO3 to a final volume of 100 mL.

[0765] Prepare at least three standard samples with concentrations in parentheses, with the expected sample concentrations.

[0766] Example 1.20: Determination of Furosine Value:

[0767] The furosine value is determined as described in the literature ["Maillard Reaction Evaluation by Furosine Determination During Infant Cereal Processing", Guerra-Hernandez et al, Journal of Cereal Science 29 (1999) 171-176], and the total amount of protein is determined according to Example 1.5. The furosine value is reported in mg of furosine units per 100 g of protein.

[0768] Example 1.21: Determination of the degree of crystallinity of BLG in liquid

[0769] The following method is used to determine the degree of crystallization of BLG in a liquid having a pH in the range of 5 to 6.

[0770] a) Transfer a 10 mL sample of the liquid to a Maxi-Spin filter with a 0.45 micron pore size CA membrane.

[0771] b) Immediately spin the filter at 1500 g for 5 minutes. Maintain the centrifuge at 2°C.

[0772] c) Add 2 mL of cold Milli-Q water (2°C) to the retentate side of the spin filter, immediately centrifuge the filter at 1500 g for 5 minutes while keeping the centrifuge cooled to 2°C, collect the permeate (permeate A), measure the volume, and determine the BLG concentration by HPLC using the method summarized in Example 1.31.

[0773] d) Add 4 mL of 2 M NaCl to the retaining surface of the filter, shake rapidly, and let the mixture sit at 25°C for 15 minutes.

[0774] e) Immediately rotate the filter at 1500 g for 5 minutes and collect the permeate (permeate B).

[0775] f) Determine the total weight of BLG in permeate A and permeate B using the method summarized in Example 1.31, and convert the result into the total weight of BLG instead of weight percentage. The weight of BLG in permeate A is m 투과액 A It is referred to as, The weight of BLG in permeate B is m 투과액 B It is referred to as.

[0776] g) For BLG, the degree of crystallinity of the liquid is determined as follows:

[0777] Degree of crystallization = m 투과액 B / (m 투과액A + m 투과액 B )*100%

[0778] Example 1.22: Determination of the degree of crystallinity of BLG in dry powder

[0779] This method is used to determine the degree of crystallinity of BLG in dry powder.

[0780] a) Mix 5.0 grams of powder sample with 20.0 grams of cold Milli-Q water (2°C) and let it stand for 5 minutes at 2°C.

[0781] b) Transfer a sample of the liquid to a Maxi-Spin filter with a 0.45 micron CA membrane.

[0782] c) Immediately spin the filter at 1500 g for 5 minutes. Maintain the centrifuge at 2°C.

[0783] d) Add 2 mL of cold Milli-Q water (2°C) to the retention side of the spin filter, immediately spin the filter at 1500 g for 5 minutes, collect the permeate (Permeate A), measure its volume, determine the BLG concentration via HPLC using the method summarized in Example 1.6, and convert the result to a result relative to the total weight of BLG instead of a weight percentage. The weight of BLG in Permeate A is m 투과액 A It is referred to as.

[0784] f) Next, the degree of crystallinity of BLG in the powder is calculated using the following formula:

[0785]

[0786] Here, m BLG 총 is the total amount of BLG in the powder sample of step a).

[0787] If the total amount of BLG in the powder sample is unknown, it can be determined by suspending another 5 g powder sample (from the same powder source) in 20.0 g of Milli-Q water, adding aqueous NaOH to adjust the pH to 7.0, leaving the mixture at 25°C for 1 hour under stirring, and finally determining the total amount of BLG in the powder sample using Example 1.31.

[0788] Example 1.23: Determination of UF permeate conductivity

[0789] Transfer 15 mL of the sample to an Amicon Ultra-15 centrifuge filter device with a 3 kDa cutoff (3000 NMWL) and centrifuge at 4000 g for 20 to 30 minutes, or until a sufficient volume of UF permeate for conductivity measurement accumulates at the bottom of the filter device. Conductivity is measured immediately after centrifugation. Sample handling and centrifugation are performed at the temperature of the sample source.

[0790] Example 1.24: Detection of dried BLG crystals in powder

[0791] The presence of dried BLG crystals in the powder can be confirmed in the following way:

[0792] A sample of the powder to be analyzed was resuspended, gently mixed in deionized water at a temperature of 4°C in a weight ratio of 2 parts water to 1 part powder, and rehydrated at 4°C for 1 hour.

[0793] The rehydrated sample is examined under a microscope to confirm the presence of crystals, and preferably, birefringence is detected using plane polarization.

[0794] To prove the existence of a crystal structure, it is desirable to isolate the crystalline material and process it by X-ray crystallography, and also to verify whether the crystal lattice (space group and unit cell dimensions) matches the crystal lattice of the BLG crystal.

[0795] Analyze the chemical composition of the separated crystalline material to verify whether its solid is mainly composed of BLG.

[0796] Example 1.25: Determination of the total amount of lactose

[0797] The total amount of lactose is determined according to: ISO 5765-2:2002 (IDF 79-2: 2002) "Dried milk, dried ice-mixes and processed cheese - Determination of lactose content - Part 2: Enzymatic method utilizing the galactose moiety of the lactose".

[0798] Example 1.26: Determination of Total Amount of Carbohydrates:

[0799] The amount of carbohydrates is determined using the Sigma Aldrich Total Carbohydrate Test Kit (Cat MAK104-1KT), in which carbohydrates are hydrolyzed and converted into furfural and hydroxyfurfural, which are converted into chromagen monitored by a spectrophotometer at 490 nm.

[0800] Example 1.27: Determination of Total Amount of Lipids

[0801] The amount of lipids is determined according to ISO 1211:2010 (Determination of fat content - Rose-Gottlieb weight method).

[0802] Example 1.28: Determination of Brix

[0803] Brix measurements were performed using a PAL-α digital handheld refractometer (Atago) calibrated for polished water (water filtered by reverse osmosis to obtain a conductivity of up to 0.05 mS / cm).

[0804] Approximately 500 μl of sample was transferred to the prism surface of the instrument and measurement was started. The measured value was read and recorded.

[0805] The Brix of the whey protein solution is proportional to the content of total solids (TS), and TS (% w / w) is approximately Brix * 0.85.

[0806] Example 1.29 Determination of Lactoferrin and Lactoperoxidase

[0807] The concentration of lactoferrin is determined by ELISA immunoassay as summarized in the literature [Soyeurt 2012 (Soyeurt et al; Mid-infrared prediction of lactoferrin content in bovine milk: potential indicator of mastitis; Animal (2012), 6:11, pp 1830-1838)].

[0808] The concentration of lactoperoxidase is determined using a commercially available bovine lactoperoxidase kit.

[0809] Example 1.30: Determination of the number of colony-forming units

[0810] The determination of the number of colony-forming units per gram of sample is performed according to: ISO 4833-1:2013(E): Microbiology of food and animal feed - Horizontal method for microbial counting - Colony counting technique at 30°C.

[0811] Example 1.31: Determination of the total amount of BLG, ALA, and CMP

[0812] This procedure is a liquid chromatography (HPLC) method for the quantitative analysis of proteins, such as ALA, BLG, and CMP, and optionally other protein species of the composition. Unlike the method of Example 1.6, this method also measures proteins present in aggregated form and thus provides a measurement of the total amount of protein species in the composition.

[0813] The separation mode is size exclusion chromatography (SEC), and the method uses 6 M guanidine HCl buffer as both the sample solvent and the HPLC mobile phase. Mercaptoethanol is used as a reducing agent to reduce disulfides (SS) in proteins or protein aggregates to produce unfolded monomer structures.

[0814] Sample preparation is easily achieved by dissolving 10 mg of protein equivalent in the mobile phase.

[0815] Two TSK-GEL G3000SWXL (7.7 mm x 30.0 cm) columns (GPC columns) and a guard column are arranged in series to achieve proper separation of major proteins in the raw material.

[0816] The eluted analyte is detected and quantified by UV detection (280 nm).

[0817] Equipment / Materials:

[0818] 1. HPLC Pump 515 with Manual Seal Cleaning Function (Waters)

[0819] 2. HPLC Pump Controller Module II (Waters)

[0820] 3. Autosampler 717 (Waters)

[0821] 4. Dual Absorbance Detector 2487 (Waters)

[0822] 5. Computer software capable of generating quantitative reports (Empower 3, Waters)

[0823] 6. Analysis columns: 2 TSK-GEL G3000SWXL (7.8 x 300 mm, P / N: 08541).

[0824] Guard Column: TSK-Guard Column SWxL (6.0 x 40 mm, P / N: 08543).

[0825] 7. Ultrasonic Bath (Branson 5200)

[0826] 8. 25 mm syringe filter with 0.2 μm cellulose acetate membrane (514-0060, VWR)

[0827] procedure:

[0828] Lee Dong-sang:

[0829] A. Stock buffer solution.

[0830] 1. Weigh 56.6 g of Na2HPO4, 3.5 g of NaH2PO4, and 2.9 g of EDTA into a 1000 mL beaker. Dissolve them in 800 mL of water.

[0831] 2. Measure the pH and, if necessary, adjust it to 7.5 ± 0.1 using HCl (to decrease pH) or NaOH (to increase pH).

[0832] 3. Transfer to a 1000 mL volumetric flask and dilute the volume with water.

[0833] B. 6 M guanidine HCl mobile phase.

[0834] 1. Weigh 1146 g of guanidine HCl into a 2000 mL beaker and add 200 mL of stock buffer solution (A).

[0835] 2. Dilute this solution with water to about 1600 mL while mixing using a magnetic stirring bar (50°C).

[0836] 3. Adjust the pH to 7.5 ± 0.1 using NaOH.

[0837] 4. Transfer to a 2000 mL volumetric flask and dilute the volume with water.

[0838] 5. It is filtered using a solvent filtration device equipped with a 0.22 μm membrane filter.

[0839] Correction standard.

[0840] The calibration standards for each protein to be quantified are prepared in the following manner:

[0841] 1. Accurately weigh about 25 mg of protein reference standard (to the right of 0.01 mg) into a 10 mL volumetric flask and dissolve it in 10 mL of water.

[0842] This is a standard protein stock solution (S1) of protein.

[0843] 2. Pipette 200 μl of S1 into a 20 mL volumetric flask and dilute the volume with the mobile phase.

[0844] This is the low working standard solution WS1.

[0845] 3. Pipette 500 μL of S1 into a 10 mL volumetric flask and dilute the volume with the mobile phase.

[0846] This is standard solution WS2.

[0847] 4. Pipette 500 μL of S1 into a 5 mL volumetric flask and dilute the volume with the mobile phase. .

[0848] This is standard solution WS3.

[0849] 5. Pipette 750 μL of S1 into a 5 mL volumetric flask and dilute the volume with the mobile phase.

[0850] This is standard solution WS4.

[0851] 6. Pipette 1.0 mL of S1 into a 5 mL volumetric flask and dilute the volume with the mobile phase.

[0852] This is a high working standard solution WS5.

[0853] 7. Using a graduated disposable pipette, transfer 1.5 mL of WS1 to 5 into a separate vial.

[0854] Add 10 μL of 2-mercaptoethanol to each vial and cap. Vortex the solution for 10 seconds.

[0855] Maintain the standard at ambient temperature for about 1 hour.

[0856] 8. Filter the standard using a 0.22 μm cellulose acetate syringe filter.

[0857] The purity of the protein is measured using Kjeldahl (N x 6.38) and HPLC as the area % of the standard solution WS5.

[0858] protein (mg) = "Standard protein weight" (mg) x P1 x P2

[0859] P1 = P% (Kildahl)

[0860] P2 = Protein Area % (HPLC)

[0861] Sample manufacturing

[0862] 1. Weigh 25 mg of the protein equivalent of the original sample into a 25 mL volumetric flask.

[0863] 2. Add approximately 20 mL of mobile phase and dissolve the sample for about 30 minutes.

[0864] 3. Add the mobile phase to the volume, and add 167 μL of 2-mercaptoethanol to 25 mL of the sample solution.

[0865] 4. After sonicating for about 30 minutes, keep the sample at ambient temperature for about 1½ hours.

[0866] 5. Mix the solution and filter it using a 0.22 μl cellulose acetate syringe filter.

[0867] HPLC System / Column

[0868] Column equilibration

[0869] 1. Connect the GPC guard column and two GPC analysis columns in series.

[0870] New columns are typically shipped with phosphate-salt buffer.

[0871] 2. Gradually flow water through the new column at a rate of 0.1 mL / min to 0.5 mL / min over 30 to 60 minutes.

[0872] Continue flushing for about 1 hour.

[0873] 3. Gradually decrease the flow rate from 0.5 mL / min to 0.1 mL / min and replace the mobile phase in the reservoir.

[0874] 4. To prevent pressure shock and maintain at 0.5 mL / min, gradually increase the pump flow rate from 0.1 mL / min to 0.5 mL / min within 30 to 60 minutes.

[0875] 5. Inject 10 samples to saturate the column and wait until the peak elutes.

[0876] This will help with column conditioning.

[0877] This step is performed without having to wait for each injection to be completed before the next injection.

[0878] 6. Equilibrate with the mobile phase for at least 1 hour.

[0879] Calculation of results

[0880] The quantitative determination of the content of proteins to be quantified, e.g., alpha-lactalbumin, beta-lactoglobulin, and caseinomacropeptide, is performed by comparing the peak area obtained for the corresponding standard protein with the peak area of ​​the sample. The results are reported as g of the specific protein per 100 g of the original sample, or as a weight percentage of the specific protein relative to the weight of the original sample.

[0881] Example 2: Crystallization of beta-lactoglobulin from crude whey protein concentrate

[0882] Protocol:

[0883] Lactose-depleted UF retained from sweet whey from a standard cheese production process, filtered through a 1.2-micron filter, was used as a feed for the BLG crystallization process. The sweet whey feed was conditioned in an ultrafiltration setup using a Koch HFK-328 type membrane with 46 mil spacers and a feed pressure of 1.5 to 3.0 bar, using a feed concentration of 21% TS (Total Solids) ± 5 and polished water (water filtered by reverse osmosis to obtain a conductivity of up to 0.05 mS / cm) as the rectification medium. During ultrafiltration, the temperatures of the feed and retained were The temperature was approximately 12°C. Subsequently, HCl was added to adjust the pH to approximately 5.40. Volume filtration was continued over a period of 20 minutes until the conductivity drop of the retention was less than 0.03 mS / cm. Subsequently, the retention was concentrated to approximately 30% TS (approximately 23.1% total protein relative to the total weight of the concentrated retention). A sample of the concentrated retention was centrifuged at 3000 g for 5 minutes, but no visible pellet formed. The supernatant was analyzed by HPLC. The composition of the feed is shown in Table 1.

[0884] The concentrated retained material was seeded with 0.5 g / L pure BLG crystalline material obtained from spontaneous BLG crystallization (as described in Example 3 in relation to feed 2). The seeding material was prepared by washing the BLG crystal slurry five times in milliQ water and collecting the BLG crystals after each wash. After washing, the BLG crystals were freeze-dried, ground using a mortar and pestle, and then passed through a 200-micron sieve. Thus, the crystallization seed had a particle size of less than 200 microns.

[0885] The concentrated retention was transferred to a 300 L crystallization tank, where it was cooled to approximately 4°C and maintained at this temperature overnight with gentle stirring. The following morning, samples of the cooled concentrated retention were transferred to test tubes and examined both visually and microscopically. Rapidly precipitating crystals had clearly formed overnight. Laboratory samples of the mixture containing both the crystals and the mother liquor were further cooled to 0°C in an ice water bath. The mother liquor and crystals were separated by centrifugation at 3000 g for 5 minutes, and samples of the supernatant and pellet were taken for HPLC analysis. The crystals were washed once with cold polished water, and the pellet was centrifuged again before freeze-drying.

[0886] Table 1: Concentrations of selected components in feed standardized to 95% w / w total solids

[0887]

[0888] Quantitative determination of BLG relative yield by HPLC:

[0889] All samples were diluted to the same degree by adding polished water. The samples were filtered through a 0.22 micron filter. For each sample, an equal volume was loaded into an HPLC system equipped with a Phenomenex Jupiter® 5 μm C4 300 Å, LC column 250 x 4.6 mm (Part Number: 00G-4167-E0) and detected at 214 nm.

[0890] The sample was executed using the following conditions:

[0891] Buffer A: MilliQ water, 0.1% w / w TFA

[0892] Buffer B: HPLC-grade acetonitrile, 0.085% w / w TFA

[0893] Flow rate: 1 ml / min

[0894] Gradient: 0 to 30 min 82 to 55% A and 18 to 45% B; 30 to 32 min 55 to 10% A and 45 to 90% B; 32.5 to 37.5 min 10% A and 90% B; 38 to 48 min 10 to 82% A and 90 to 18% B.

[0895] Data Processing:

[0896] Since all samples were processed in the same manner, the inventors can obtain the relative yield by directly comparing the areas of the BLG peaks. Since the crystals contain only BLG and all samples were processed in the same manner, the concentration of alpha-lactalbumin (ALA) and, therefore, the area of ​​ALA must be the same in all samples. Accordingly, the area of ​​ALA before and after crystallization is used as a correction factor (cf) when calculating the relative yield.

[0897]

[0898] Relative yield is calculated using the following formula:

[0899]

[0900] result:

[0901] From the chromatogram obtained before and after the crystallization of BLG from sweet whey, it is evident that the concentration of BLG has decreased significantly, and using the previously described yield calculation, the yield of removed BLG was determined to be 64.5% w / w.

[0902] The crystal slurry was examined under a microscope. The sample contained hexagonal crystals, many of which were significantly larger than 200 microns, indicating that the observed crystals were not only seeding crystals. The crystals were easily crushed when pressed with a needle, confirming that they were protein crystals.

[0903] The chromatogram of the washed crystal product shows that BLG occupies 98.9% of the total area of ​​the chromatogram. The purity of the BLG product can be further increased by additional washing.

[0904] conclusion:

[0905] This example demonstrates that BLG can be selectively crystallized from crude whey protein concentrate containing more than 48% non-BLG protein relative to total protein, and that the obtained BLG crystallized isolate is of very high purity. These findings present a new approach for industrial milk protein separation, wherein BLG is preferably separated from other protein components in a mild manner, avoiding prolonged exposure to high temperatures and problematic chemicals.

[0906] Example 3: Crystallization of BLG in three types of whey protein solutions

[0907] Protocol:

[0908] Three different types of whey protein-containing feedstocks were tested as feedstocks for crystallization using the same experimental and analytical setup as in Example 2. However, seeding was not used in the test performed with feedstock 2. Feedstocks 1 and 2 were based on sweet whey and fat-reduced via a Synder FR membrane prior to processing as described in Example 2. Feedstock 3 was derived from acid whey.

[0909] The compositions of the three feeds can be seen in Tables 2, 3, and 4 below. Feed 3 was crystallized at 21% TS (total protein of 13.3% w / w relative to the total weight of the feed), which was a lower concentration than the other two (total protein of 26.3% w / w in Feed 1 and 25.0% w / w in Feed 2).

[0910] The slurry of crystallized feed 1 was centrifuged at 1500 g for 5 minutes in a Maxi-spin filter with a 0.45 micron CA membrane. Subsequently, 2 volumes of MilliQ water were added to the filter cake before centrifugation again. The resulting filter cake was analyzed by HPLC. The pellet from feed 2 was washed with 2 volumes of MilliQ water and centrifuged again under standard conditions before analysis by HPLC. The pellet from feed 3 was analyzed without washing.

[0911] Crystals made from feed 2 were diluted with 10% TS, and crystallization was reversed by adjusting the pH to pH 7 using 1 M NaOH. Crystallization was reversed by adding NaCl to the crystal slurry from feed 2, 36% TS.

[0912] Table 2: Concentrations of selected components of Feed 1 (whey protein concentrate based on sweet whey). BDL = Below detection limit in wet samples

[0913]

[0914] Table 3: Concentrations of selected components in Feed 2 (ALA-reduced whey protein concentrate based on sweet whey). BDL = Below the detection limit in non-standardized wet samples.

[0915]

[0916] Table 4: Concentrations of selected components of Feed 3 (whey protein concentrate based on acid whey).

[0917]

[0918] result:

[0919] Supply 1:

[0920] From the protein composition of the feed and the chromatogram of the mother liquor, it is evident that a large portion of the BLG was recovered as crystals by the process. The yield of isolated BLG (calculated as described in Example 2) is approximately 65% ​​of the total amount of BLG in the feed.

[0921] From micrographs of the sample taken during the early stages of crystallization and those taken when crystallization was complete, it is evident that BLG crystals are relatively brittle. Some of the crystals appear to break during stirring, transforming from hexagonal or rhombus shapes into crystal fragments that, while still appearing very dense and well-defined, possess a more irregular shape.

[0922] From the chromatogram of the BLG crystals separated and washed in the spin filter, it can be seen that the purity is very high and the removal of other whey proteins is very efficient.

[0923] Supply 2:

[0924] From the protein composition of feed 2 and the chromatogram of the obtained mother liquor, it is evident that a large portion of BLG was removed and the calculated yield was 82% of the total amount of BLG in feed 2.

[0925] By studying feed 2 before and after crystallization, it can be seen that during crystallization, the feed changes from a transparent liquid (the stirring magnet was visible) to a milky white opaque liquid.

[0926] Microscopic images of BLG crystals show a hexagonal shape, although most of the crystals are broken.

[0927] The chromatogram of the isolated pellet of BLG crystals after washing with 2 volumes of MilliQ water clearly shows that the crystals contain BLG with very high purity.

[0928] In both cases, increasing conductivity (by adding NaCl) or changing pH (by adding NaOH to adjust pH to 7) so that the environment no longer favors a crystalline structure shows that the milky white suspension turns into a clear liquid as the BLG crystals dissolve.

[0929] The mineral composition of the crystalline preparation obtained from feedstock 2 is provided in Table 5. The inventors note that the ratio of phosphorus to protein is very low, which means the crystalline preparation is suitable as a protein source for patients with kidney disease.

[0930] Table 5: Concentration of selected components in the crystalline formulation obtained from Feed 2

[0931]

[0932] Supply 3:

[0933] From the protein composition of feed 3 and the chromatogram of the resulting mother liquor, it is evident that a large portion of BLG is isolated (calculated yield of 70.3% of the total amount of BLG in the feed). If the protein content had been higher before crystallization, the yield obtained would have been even higher.

[0934] Microscopic images of BLG crystals isolated from feed 3 (substantially CMP-free) show that the crystals have a rectangular shape, as opposed to a hexagonal shape. The rectangular crystals appeared more solid than the hexagonal crystals. In the case of the chromatogram of the crystal pellet isolated without washing; the chromatogram clearly shows that the crystals are BLG crystals despite the rectangular shape rather than the hexagonal shape.

[0935] Table 6: Concentration of selected components of the crystalline formulation obtained from Feed 3

[0936]

[0937] The crystalline preparation derived from feed 3 contained 45 mg P per 100 g of protein. The inventors note that the ratio of phosphorus to protein is very low, which makes the crystalline preparation suitable as a protein source for patients with kidney disease.

[0938] conclusion:

[0939] All three feedstocks were suitable for the BLG crystallization process. The BLG crystals were easily dissolved by adding salt or increasing the pH or temperature. The new method enables the production of BLG formulations with very low phosphorus content, making them suitable as a protein source for patients with kidney disease.

[0940] Example 4: Preparation of spray-dried BLG crystals and determination of bulk density

[0941] A portion of the BLG crystals produced in Example 3 (using feed 2) was separated in a decanter centrifuge at 1200 g, 5180 RPM, 110 RPM Diff., 64 mil spacers (mil means 1 / 1000 inch), and a flow rate of 25 to 30 L / h. Subsequently, the BLG crystal phase was mixed with polished water in a 1:1 ratio and then separated again in a decanter centrifuge using the same settings. Subsequently, the BLG crystal phase was mixed with polished water to prepare a slurry containing approximately 25% dry matter and a crystallinity of approximately 80 of BLG, and then dried in a pilot plant spray dryer with an inlet temperature of 180°C and an outlet temperature of 85°C without any preheating. The temperature of the liquid stream was 10 to 12°C until spray-drying. The generated powder sampled at the outlet had a moisture content of 4.37% w / w.

[0942] The degree of crystallization of BLG in the slurry was approximately 90%.

[0943] The inventors also successfully separated a slurry of BLG crystals and mother liquor in a decanter centrifuge at 350 g, 2750 RPM, 150 RPM Diff., with 64 mil spacers and a flow rate of 75 L / h. Subsequently, the BLG crystal phase was mixed with polished water in a 1:2 ratio. Then, to make a thinner slurry, the BLG crystal phase was mixed with polished water and then dried in a pilot plant spray dryer using the same parameters as described above.

[0944] Next, the bulk density of the spray-dried powder was measured according to Example 1.17 and compared to the bulk density of standard WPI dried in the same equipment. Standard WPI was found to have a bulk density of 0.39 g / mL (based on 625 stamping), which is the upper limit of the normal range for WPI powder. However, the spray-dried BLG crystalline formulation had a bulk density of 0.68 g / mL, which was more than 75% higher than the bulk density of standard WPI. This is truly remarkable and offers numerous logistical and application-related advantages.

[0945] Table 7: Concentrations of selected components in the spray-dried BLG crystalline formulation of Example 7. BDL = Below detection limit

[0946]

[0947] Afterward, samples of the spray-dried BLG crystal preparations were resuspended in cold deionized water, and the BLG crystals were still clearly visible under a microscope. The addition of citric acid or NaCl dissolved the BLG crystals and transformed the opaque crystal suspension into a clear liquid.

[0948] The inventors have observed indications that extended heating during the drying step reduces the amount of BLG in crystalline form. Therefore, it is desirable that the heat exposure of the BLG crystalline formulation be as low as possible.

[0949] conclusion:

[0950] This embodiment demonstrates that a slurry containing BLG crystals can be spray-dried, and if heating is controlled during the drying step, the BLG crystals still exist as a resuspended spray-dried powder.

[0951] The inventors also discovered that the bulk density of whey protein powder containing BLG crystals is significantly higher than that obtained by normal spray-drying of a dissolved protein stream. Since the high-density powder requires less packaging material per kg of powder, more cost-effective packaging and logistics of the powder are possible, and more powder (mass) can be transported by a given container or truck.

[0952] High-density powder also appears easy to handle during manufacturing and use, and seems fluffy and less dusty.

[0953] Example 5: Low-phosphorus protein drink

[0954] Six low-phosphorus instant beverage powders were prepared using the purified BLG product from Example 3 (crystalline formulation obtained from feed 3). All dry ingredients were blended to obtain the instant beverage powder, which was then mixed with deionized water to obtain 10 kg of each sample and hydrated at 10°C for 1 hour.

[0955] Table 8: Composition of 6 beverage samples.

[0956]

[0957] The turbidity of the six sample subsamples was measured using a Turbiquant® 3000 IR turbidimeter, and the viscosity was measured using a vicoman (manufacturer: Gilson). The results are shown in the table below.

[0958] Table 9: Measured viscosity and turbidity of 6 beverage samples.

[0959]

[0960] Figure 3 shows a photograph of test tubes containing subsamples of six low-phosphorus beverage samples. From left to right, the subsamples were samples A, B, C, D, E, and F. Visual inspection of the test tubes confirmed turbidity measurements and documented that all beverage samples were clear, and samples C and D (pH 3.0) in particular were very clear. The low viscosity demonstrates that the beverage samples were easily drinkable.

[0961] All ingredients used to manufacture the beverage were low in phosphorus and did not contain unnecessary minerals. Therefore, the resulting beverage had a phosphorus content of approximately 45 mg P per 100 g of protein and generally very low mineral content. Thus, the six liquid foods prepared from the instant beverage powder were suitable for use as instant protein drinks for patients with kidney disease.

[0962] Example 6: Crystal separation by dynamic cross-flow filtration

[0963] Lactose-depleted UF retained from sweet whey from a standard cheese production process, filtered through a 1.2-micron filter, was used as a feed for the crystallization process. The sweet whey feed was conditioned in an ultrafiltration setup using a Koch HFK-328 type membrane with 46 mil spacers and a feed pressure of 1.5 to 3.0 bar, using a feed concentration of 10% TS (Total Solids) ± 5 and polished water (water filtered by reverse osmosis to obtain a conductivity of up to 0.05 mS / cm) as the rectification filtration medium. During ultrafiltration, the temperatures of the feed and retained were The temperature was approximately 12°C. Subsequently, HCl was added to adjust the pH to approximately 5.60. Volume filtration was continued until the conductivity of the retained was less than 1.30 mS / cm. Then, the feed was heated to 25°C before the retained was concentrated to approximately 27% TS (approximately 21% total protein relative to the total weight of the concentrated retained). The conductivity of the permeate was 0.33 mS / cm at the end of the concentration. A sample of the concentrated retained was centrifuged at 3000 g for 5 minutes, but no visible pellet formed.

[0964] The concentrated retention was transferred to a 300 L crystallization tank, where it was cooled to approximately 6°C and maintained at this temperature overnight with gentle stirring. The next morning, the retention had crystallized. The mother liquor and crystals were separated by centrifugation at 3000 g for 5 minutes, and samples of the supernatant and pellet were taken for HPLC analysis. From this process, the yield of BLG was calculated to be 67%.

[0965] Crystalline slurry from a 300 L tank was used as the feed for an Andritz DCF 152S system using a single disc membrane with a pore size of 500 nm. Filtration was performed at 8°C, the rotation speed was 32 Hz, and the membrane penetration pressure was 0.4 bar. Unlike larger devices where retained matter is continuously removed, the system operates as a dead-end filtration system where retained matter accumulates in the filtration chamber. Filtration was performed in a stable manner for more than 40 minutes, at which point the solids accumulated in the filtration chamber began to affect the filtration.

[0966] The amount of crystal mass increased significantly during DFC operation.

[0967] Conclusion: DCF provides a stable and efficient means of separating crystals from ML. If necessary, a washing solution can be added to the DCF.

[0968] Example 7: Protein denaturation of different whey protein products

[0969] The protein denaturation degree of four isolated BLG samples was compared with that of a commercial product. The isolated BLG samples are suitable for manufacturing the instant beverage powder of the present invention. The samples are listed below.

[0970]

[0971] Samples B through E were manufactured in the following manner:

[0972] A crystal slurry was prepared as described in Example 6 and separated as described in Example 4. A portion of the separated BLG slurry was taken out and divided into four parts.

[0973] Sample B: The first portion of the separated BLG crystal slurry was redissolved without any drying by adjusting the pH of the BLG crystal slurry to 7.01 using 3% NaOH, and then the sample was diluted to Brix 6 to make an approximately 5% protein solution.

[0974] Sample C: A second portion of the isolated BLG crystal slurry was freeze-dried. Subsequently, the powder was resuspended in polished water, the pH was adjusted to 7.09 using 3% NaOH, and then the sample was diluted to Brix 6 to make an approximately 5% protein solution.

[0975] Sample D: A third portion of the isolated BLG crystal slurry was redissolved using 3% NaOH to adjust the pH to 7.0 and then freeze-dried. Subsequently, the freeze-dried powder was resuspended in polished water, and the pH was measured to be 7.07. Then, the sample was diluted to Brix 6 to prepare an approximately 5% protein solution.

[0976] Sample E: A fourth portion of the separated BLG crystal slurry was processed as described in Example 4 and spray-dried. Subsequently, the powder was resuspended in polished water, and the pH was adjusted to 7.04 using 3% NaOH. Then, the sample was diluted to Brix 6 to prepare an approximately 5% protein solution.

[0977] The degree of protein denaturation of each sample was determined according to Example 1.3, and the results are presented in the table below.

[0978] Table: Comparison of protein denaturation levels of a commercially available WPI product (Bipro) and four BLG products that can be used in the instant beverage powder of the present invention.

[0979]

[0980] conclusion:

[0981] Regardless of the drying method, BLG isolates have a surprisingly low degree of denaturation; it is only 1 / 10 of that found in commercially available WPI used for comparison. It is particularly surprising that the spray-dried BLG crystal slurry product still has the lowest degree of denaturation among all products.

[0982] Example 8: Production of spray-dried, acidic BLG isolate powder

[0983] whey protein supply

[0984] Lactose-depleted UF retained from sweet whey from a standard cheese production process was filtered through a 1.2 micron filter and fat-reduced through a Synder FR membrane before being used as a feed for the BLG crystallization process. The chemical composition of the feed can be seen in Table 10. The inventors note that all weight percentages of specific proteins, such as BLG and ALA, mentioned in this example relate to the weight percentage of non-aggregated protein relative to total protein.

[0985] conditioning

[0986] Sweet whey feed was filtered using polished water (water filtered by reverse osmosis to obtain a conductivity of up to 0.05 mS / cm) as the rectified filtration medium at a feed concentration of 21% Total Solids (TS) ± 5, with a 46 mil spacer and a feed pressure of 1.5 to 3.0 bar using a Koch HFK-328 type membrane (70 m 2 It was conditioned at an ultrafiltration setting at 20°C using a membrane. Subsequently, HCl was added to adjust the pH to approximately 5.5. Volumetric filtration was continued over a period of 20 minutes until the conductivity drop of the retained material was less than 0.1 mS / cm. Subsequently, the permeate flow rate was 1.43 L / h / m² 2 The retention was concentrated until it was less than [amount]. The first sample of the concentrated retention was taken and centrifuged at 3000 g for 5 minutes. The supernatant of the first sample was used to determine the BLG yield.

[0987] crystallization

[0988] The concentrated retained material was transferred to a 300 L crystallization tank and seeded with pure BLG crystalline material made from rehydrated and spray-dried BLG crystals. Afterwards, the seeded whey protein solution was cooled from 20°C to approximately 6°C over approximately 10 hours to allow BLG crystals to form and grow.

[0989] After cooling, a sample of the crystal-containing whey protein solution (second sample) was taken, and BLG crystals were separated by centrifugation at 3000 g for 5 minutes. The supernatant and crystal pellet from the second sample were analyzed by HPLC as described below. The crystallization yield was calculated as summarized below and was determined to be 57%.

[0990] Table 10: Chemical composition of the feed

[0991]

[0992] Determination of BLG Yield Using HPLC:

[0993] The supernatants of the first and second samples were diluted to the same degree by adding polished water, and the diluted supernatants were filtered through a 0.22 μm filter. For each filtered and diluted supernatant, an equal volume was loaded into an HPLC system equipped with a Phenomenex Jupiter® 5 μm C4 300 Å, LC column 250 x 4.6 mm, Ea., and detected at 214 nm.

[0994] The sample was executed using the following conditions:

[0995] Buffer A: MilliQ water, 0.1% w / w TFA

[0996] Buffer B: HPLC-grade acetonitrile, 0.085% w / w TFA

[0997] Flow: 1 mL / min

[0998] Column temperature: 40℃

[0999] Gradient: 0 to 30 min 82 to 55% A and 18 to 45% B; 30 to 32 min 55 to 10% A and 45 to 90% B; 32.5 to 37.5 min 10% A and 90% B; 38 to 48 min 10 to 82% A and 90 to 18% B.

[1000] Data Processing:

[1001] Since both supernatants were treated in the same manner, the relative yield can be calculated by directly comparing the areas of the BLG peaks. Since the crystals contain only BLG and all samples were treated in the same manner, the concentration of alpha-lactalbumin (ALA) and, consequently, the area of ​​ALA must be the same in all samples. Therefore, the area of ​​ALA before and after crystallization is used as a correction factor (cf) when calculating the relative yield.

[1002]

[1003] The relative yield is calculated by the following formula:

[1004]

[1005] Acid dissolution of BLC crystals

[1006] The remaining material from the crystallization tank was separated using a decanter at 350 g, 2750 RPM, 150 RPM Diff., 64 spacers, and a feed flow rate of 75 L / h before separating the feed mixed with polished water in a 1:2 ratio. Subsequently, the BLG crystal / solid phase from the decanter was mixed with polished water to form a thinner slurry before adding phosphoric acid to rapidly dissolve the crystals and lower the pH to approximately 3.0.

[1007] After dissolving the BLG crystals, the pure BLG protein liquid was concentrated to 15 Brix at the same UF setting used to prepare the feed for crystallization, and the pH was adjusted to a final pH of approximately 3.8. Subsequently, the liquid BLG isolate was heated to 75°C for 5 minutes and then cooled to 10°C. Heat treatment was found to reduce the microbial load from 137,000 CFU / g before heat treatment to <1,000 CFU / g after heat treatment. Heat treatment did not cause any protein denaturation, and the intrinsic tryptophan fluorescence ratio (330 nm / 350 nm) was determined to be 1.20, which indicated the natural form of the BLG molecule.

[1008] BLG was dried in a pilot plant spray dryer with an inlet temperature of 180°C and an outlet temperature of 75°C. The resulting powder sampled at the outlet had a moisture content of approximately 4% w / w, and the chemical composition of the powder is shown in Table 11. A sample of the dried powder was dissolved, the degree of protein denaturation was determined to be 1.5%, and the intrinsic tryptophan fluorescence emission ratio (I330 / I350) was measured as 1.20.

[1009] Table 11: Composition of BLG isolated powder (BDL = below detection limit)

[1010]

[1011] The bulk density (625 tab) of the spray-dried powder is 0.2 to 0.3 g / cm³ 3 It was estimated as.

[1012] Conclusion: By using the process described above, the inventors were able to produce a high-purity BLG product that can be heat-treated without substantial protein denaturation or protein unfolding during treatment. Heat treatment significantly reduced bacterial levels without damaging the protein product.

[1013] The inventors have observed indications that higher bulk density can be obtained by increasing the protein content prior to spray-drying. Additionally, the inventors have observed that a lower degree of denaturation is obtained when the inlet and / or outlet temperatures used for spray-drying are reduced.

[1014] Example 9: Production of Spray-Dried pH-Neutral BLG Isolate Powder

[1015] When using the same protocol and experimental setup as in Example 2, the lactose-reduced whey protein isolates shown in Table 12 were conditioned and used as a feed for crystallization. The yield of crystallization was calculated to be 68%.

[1016] The inventors note that all weight percentages of specific proteins, such as BLG and ALA, mentioned in the embodiments are related to the weight percentage of non-aggregated proteins relative to the total protein.

[1017] Table 12: Composition of the supply

[1018]

[1019] The remaining material from the crystallization tank was separated in a decanter at a feed rate of 350 g, 2750 RPM, 150 RPM, 64 spacers, and 75 L / h, before separating the feed mixed with polished water in a 1:2 ratio. Subsequently, the BLG crystal / solid phase from the decanter was mixed with polished water to form a thinner slurry before adding 0.1 M potassium hydroxide to rapidly dissolve the crystals and adjust the pH to approximately 7.

[1020] After dissolving the crystals, the pure BLG protein liquid was concentrated to 15 Brix at the same UF setting used to prepare the whey protein solution for crystallization, and the pH was adjusted to a final pH of 7.0. The BLG was dried in a pilot plant spray dryer with an inlet temperature of 180°C and an outlet temperature of 75°C. The resulting powder sampled at the outlet had a moisture content of approximately 4% w / w. The composition of the powder is shown in Table 13. After drying, a portion of the powder was dissolved in deionized water, and the protein denaturation was determined to be 9.0%, with an intrinsic tryptophan fluorescence ratio (330 nm / 350 nm) of 1.16.

[1021] Table 13: Chemical composition of BLG isolated powder

[1022]

[1023] The bulk density (625 tab) of the spray-dried powder is 0.2 to 0.3 g / cm³ 3 It was estimated as.

[1024] Conclusion: By using the process described above, the inventors can produce pH-neutral, high-purity BLG products with minimal or no protein denaturation during processing. The inventors have observed indications that higher bulk density can be obtained by increasing the protein content prior to spray-drying. Additionally, the inventors have observed that lower degrees of denaturation are obtained when the inlet and / or outlet temperatures used for spray-drying are reduced. The level of denaturation can also be reduced by decreasing the mineral content prior to spray-drying.

[1025] Example 10: Preparation of Coated BLG Isolated Powder

[1026] Coated BLG isolates were produced in a fluidized bed (DIOSNA, MINILAB XP No. 365-146l) using spray-dried BLG isolates produced as described in Example 4 or Example 9. The inlet temperature was 60°C. The powder temperature was between 40°C and 50°C during the process, and the airflow was 25 to 35 m / h. 3 The coating material was dissolved in 50 g of deionized water and slowly injected into a fluidized bed to be sprayed. For each batch, 500 g of spray-dried BLG isolate was used. After the coating material was added, drying was continued until the coated BLG isolate had a moisture content of 4 to 5%. Using this setup, BLG isolate coated with 25 g of citric acid and BLG isolate coated with 30 g of trisodium citrate were produced.

[1027] Test samples were prepared as shown in the table and analyzed for solubility as described below. In addition to the described 10% w / w solution, a 30% w / w solution of BLG isolated powder was prepared and tested. Test samples were further analyzed for wettability as described below. Additionally, test samples were sensorially evaluated by two trained testers according to the parameters set in Example 1.11.

[1028] Using the BLG isolated powder from Example 8, lecithin-coated powder was produced in the same fluidized bed as above. The inlet temperature at which 500 g of powder was added to the fluidized bed was 75°C, and the airflow was 25 m 3 / h. When the powder temperature reached 38℃, 50 mL of water was slowly added through a spray bottle. The powder temperature was raised to a maximum of 45℃, and 5 mL of lecithin was injected through a spray bottle. The powder was heated to 65℃ and dried until it contained less than 5% moisture.

[1029] Solubility Test: The solubility and readiness of the instant powder to be dissolved can be measured by this test. Add 10 grams of powder to 90 grams of deionized water (8°C) in a sealable, transparent test tube. Shake the mixture vigorously by hand for 30 seconds. Evaluate the mixture immediately, let it stand for 1 minute, and then evaluate the mixture again. The evaluation is performed by visual inspection of the following parameters: transparency of the liquid phase, foam formation, color, and the degree of powder dissolution.

[1030] result

[1031]

[1032]

[1033] Conclusion: The obtained BLG isolate powders possessed an excellent aggregation structure and dissolved easily in water. All coated BLG isolate powders (Samples 1, 3-5, and 7-8) performed equally well as or better than the standard coated WPI (Test Sample 2) in this test. Surprisingly, when testing the 30% version, both the coated and uncoated BLG isolates (Test Samples 1, 3-5, and 7-8) were easy to dissolve, suggesting the possibility of increasing protein concentrations above 30%. In the 30% solution (Sample 7-8), both the coated and uncoated BLG isolates dissolved easily compared to the standard WPI (Sample 9), which contained visible particles in the foam.

[1034] Wetness: This method is used to describe the wetness of a powder. Wetness is defined as the time it takes for the entire sample to become wet. 0.5 grams of powder are measured and placed on the surface of 100 g of deionized water (5°C) inside a cylindrical container with a diameter of 5 cm. After placing the powder on the surface of the water, the time until the powder dissolves or passes over the water surface is measured.

[1035] result

[1036]

[1037] Conclusion: The coated BLG isolate powders (Samples 10, 12, 14) were wetted in a manner similar to that of standard instantized WPI (Samples 11, 15), with the exception of the BLG isolate powder coated with citric acid (Sample 13). It was surprising that the uncoated BLG isolate powder was wetted even better than the standard WPI (Sample 6).

[1038] Example 11: Wetness of uncoated BLG isolate

[1039] In this embodiment, the wettability of the uncoated acidic BLG isolate and the uncoated neutral BLG isolate is compared with the wettability of the uncoated whey protein isolate (WPI). Wetness is defined as the time taken before the entire sample becomes wet. 0.5 grams of powder is measured and placed on the surface of 100 g of deionized water (10°C) in a cylindrical container with a diameter of 5 cm. The time from placing the powder on the surface of the water until the powder dissolves or passes over the water surface is measured.

[1040] result

[1041]

[1042] Conclusion: It was surprising that the uncoated BLG isolate powder (Samples 2, 3) wetted better than standard WPI (Sample 1).

[1043] Example 12: Preparation of instant beverage powder

[1044] Manufacture of instant powder used as a nutritional supplement.

[1045] 100 g of instant powder is prepared by blending the following ingredients: 91 g of whey protein concentrate containing at least 85% w / w of BLG prepared in Example 4 and 4 g of soy lecithin.

[1046] 100 grams of instant powder contains 360 kcal with the following energy distribution: 2 E% from lipids, 1 E% from carbohydrates, and 97 E% from protein. Foods prepared from instant powder can be used as protein supplements for the treatment of patients who are malnourished or at risk of malnutrition, for example, by mixing the instant powder with water to obtain a beverage or by adding the powder to a regular meal. 10 to 15 grams of instant powder are stirred with water at a temperature of 15 to 25°C. The prepared instant powder beverage has good taste, color, and viscosity.

[1047] Example 13: Preparation of instant beverage powder

[1048] Manufacture of nutritionally complete instant powder containing BLG.

[1049] 100 g of instant powder is prepared by blending 18.2 g of vegetable oil (a mixture consisting of palm kernel oil, coconut oil, rapeseed oil, and sunflower oil), 56.4 g of glucose syrup, 20 g of whey protein concentrate containing at least 90% w / w of BLG as prepared in Example 4, and about 3 g of soy lecithin.

[1050] Potassium citrate, sodium citrate, sodium chloride, magnesium hydrogen phosphate, potassium hydrogen phosphate, magnesium chloride, choline chloride, calcium carbonate, calcium phosphate, sodium L-ascorbate, aroma, iron sulfate, L-ascorbic acid, zinc sulfate, magnesium citrate, DL-alpha-tocopheryl acetate, manganese sulfate, nicotinamide, D-biotin, copper sulfate, calcium-D-pantothenate, pteroyl monoglutamic acid, sodium fluoride, DL-alpha-tocopherol, thiamine hydrochloride, pyridoxine hydrochloride, carotenoids, retinyl palmitate, riboflavin, cyanocobalamin, cholecalciferol, chromium chloride, sodium molybdenum, potassium iodide, sodium selenite, and phytomethadione are additionally added to obtain an instant powder having the following nutritional profile: 381 μg RE Vitamin A, 0.91 mg Carotenoids, 3.3 μg Vitamin D, 5.9 mg α-TE Vitamin E, 24 μg Vitamin K, 0.69 mg Vitamin B1, 0.74 mg Vitamin B2, 8.3 mg-NE Niacin, 2.5 mg Pantothenic Acid, 0.79 μg Vitamin B6, 123 μg Folic Acid, 0.98 μg Vitamin B12, 24 μg Biotin, 60 mg Vitamin C, 156 mg Choline, 1.17 gram Salt, 469 mg Sodium, 705 mg Potassium, 578 mg Chloride, 371 mg Calcium, 337 mg Phosphorus, 107 mg Magnesium, 7.4 mg Iron, 5.6 mg Zinc, 0.83 mg Copper, 1.5 mg Manganese, 0.5 g Fluorine, 48 μg Molybdenum, 26 μg selenium, 25 μg chromium, 61 μg iodide.

[1051] The instant powder contains 462 kcal / 100 grams with the following energy distribution: 35.6 E% from lipids, 48.7 E% from carbohydrates, and 15.7 E% from protein. Foods prepared from the instant powder can be used as nutritionally complete foods to treat patients who are malnourished or at risk of malnutrition, for example, by mixing the instant powder with water to obtain a beverage or using it for tube feeding. 22 grams of instant powder are stirred (boiled) in 85 ml of cold water to obtain a beverage containing 100 kcal. 33 grams of instant powder are stirred (boiled) in 78 ml of cold water to obtain a beverage containing 150 kcal. The prepared instant powder beverages have good taste, color, and viscosity.

Claims

Claim 1 An instant beverage powder comprising at least 30% w / w beta-lactoglobulin (BLG), wherein at least 91% w / w of the protein in the instant beverage powder consists of BLG, and further comprising at least one additional ingredient selected from the group consisting of vitamins, flavoring agents, coloring agents, minerals, sweeteners, antioxidants, edible acids, lipids, carbohydrates, prebiotics, probiotics, antifoaming agents, and non-whey proteins. Claim 2 The powder according to claim 1 further comprises one or more of the following: i. a sweetener, ii. a flavoring agent, iii. at least one edible acid, iv. the sum of the amounts of Na, K, Mg, and Ca in the instant beverage powder is at most 10 mmol / g protein, and the solution in which the powder is dissolved in deionized water at a concentration of 10% w / w has a pH in the range of 2 to 8. Claim 3 The powder according to claim 1 or 2, wherein the powder comprises at least 92% w / w BLG with respect to total protein, at least 93% w / w BLG with respect to total protein, at least 94% w / w BLG with respect to total protein, at least 95% w / w BLG with respect to total protein, at least 96% w / w BLG with respect to total protein, at least 97% w / w BLG with respect to total protein, at least 98% w / w BLG with respect to total protein, or at least 99% w / w BLG with respect to total protein. Claim 4 The powder according to claim 1 or 2, wherein the powder contains a moisture content of up to 6% w / w, up to 5% w / w, up to 4% w / w, up to 3% w / w, or up to 2% w / w. Claim 5 The powder according to claim 1 or 2, wherein the powder comprises i. up to 6% w / w water, ii. at least 15% w / w total protein with respect to total solids, and iii. at least 91% w / w BLG with respect to total protein, and the powder is a dry powder. Claim 6 The powder according to claim 1 or 2, wherein the powder has a bulk density of at least 0.30 g / mL, at least 0.4 g / mL, or 0.5 g / mL. Claim 7 A powder according to claim 1 or 2, wherein the sum of the amounts of Na, K, Mg, and Ca is at most 10 mmol / g protein. Claim 8 The powder according to claim 1 or 2, wherein the solution in which the powder is dissolved in deionized water at a concentration of 10% w / w has a pH in the range of 2 to 8 at room temperature. Claim 9 The powder according to claim 1 or 2, wherein the powder has a protein denaturation degree of up to 10%, up to 4%, up to 1%, up to 0.4%, or up to 0.1%. Claim 10 The powder according to claim 1 or 2, wherein the powder has a crystallinity of at least 20% or at least 40% of BLG. Claim 11 Powder for use in alleviating or preventing malnutrition in paragraph 1 or 2. Claim 12 Powder for use in alleviating or preventing kidney disease in paragraph 1 or 2. Claim 13 A kit comprising a powder according to claim 1 or 2, wherein the kit comprises i. a tool for measuring the powder, and ii. a container having a lid for opening and closing the container, wherein the container is for mixing the powder with a liquid to form a food, and wherein the container is suitable for drinking the food directly from the container. Claim 14 A method for manufacturing a liquid food comprising a liquid and an instant beverage powder according to claim 1 or 2, comprising: i. adding the powder according to claim 1 or 2 to the liquid; ii. optionally adding at least one additional ingredient; and iii. mixing the obtained powder and the liquid to form a homogeneous mixture. Claim 15 In paragraph 14, the method wherein the additional ingredient is selected from fruits or vegetables. Claim 16 delete Claim 17 delete Claim 18 delete Claim 19 delete Claim 20 delete Claim 21 delete Claim 22 delete Claim 23 delete Claim 24 delete Claim 25 delete Claim 26 delete Claim 27 delete Claim 28 delete Claim 29 delete Claim 30 delete Claim 31 delete Claim 32 delete Claim 33 delete Claim 34 delete Claim 35 delete Claim 36 delete Claim 37 delete Claim 38 delete

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