Method for producing dense whey protein nanogels, whey protein nanogels or nanogel compositions produced and food products

Abstract

The present invention relates to the production of special whey protein nanogels by denaturation of whey proteins, and also to the resulting whey protein nanogels and their use in food products such as beverages. The whey protein nanogel of the present invention provides an extremely low viscosity contribution when used in beverages and liquid products at high concentrations, which clearly shows the uniqueness of the whey protein nanogel of the present invention.

Description

[0001] Related Applications

[0002] This application is a divisional application of a Chinese invention patent application filed on December 29, 2020, with application number 2020800939051 and titled “Method for producing dense whey protein nanogel, the resulting whey protein nanogel or nanogel composition, and food containing such whey protein nanogel or nanogel composition”. Technical Field

[0003] The present invention relates to the production of special whey protein nanogels by denaturing whey protein, and further relates to the produced whey protein nanogels, and their use in food such as beverages. The whey protein nanogels of the present invention provide extremely low viscosity contributions when used in beverages and liquid products at high concentrations, which clearly demonstrates the uniqueness of the whey protein nanogels of the present invention. Background Art

[0004] The formation of micro- or nanoparticles of denatured whey proteins has been described previously and is known as a way to modify the functionality of whey proteins.

[0005] US 6,605,311 B2 discloses insoluble, denatured, heat-stable protein particles having an average diameter in the hydrated state of 0.1-3 micrometers, which can be dispersed in aqueous solutions and used in food and beverage products.

[0006] WO2007 / 110421A2 discloses the preparation of nano-sized whey protein micelles by thermal denaturation of whey protein. Example 11 describes the formation of a whey protein micelle concentrate having a protein content of 20% w / w (weight / weight). The concentrate is described as having a "creamy, semi-solid texture" on page 28 of Part 2 of WO2007 / 110421A2, which clearly indicates a high viscosity.

[0007] CN105542195 A discloses nanogels formed by conjugation of polysaccharides to whey proteins and subsequent heat treatment, and also discloses the use of these nanogels in food products. Summary of the invention

[0008] The present inventors have discovered that special process conditions can produce nanogels of denatured whey protein in solutions having surprisingly high concentrations of protein, in particular having surprisingly high concentrations of native beta-lactoglobulin (BLG), the major protein in whey and the driving force for thermal aggregation of whey protein. The ability to form nanogels at high protein concentrations is very advantageous because it reduces the energy consumption per kilogram of protein in the process, since less solution needs to be heated and subsequently cooled per kilogram of protein. In addition, the cost of transporting the nanogel product in liquid form is reduced, since less water needs to be transported, and the cost of converting the nanogel suspension to a powder is reduced, since less water needs to be removed.

[0009] Therefore, one aspect of the present invention relates to a method for producing a whey protein nanogel composition, the method comprising:

[0010] a) providing a whey protein solution, the solution having:

[0011] - a natural BLG content in an amount of at least 3% w / w,

[0012] - a pH value in the range of 5.8-7.5, preferably 5.8-6.5,

[0013] - a weight ratio of total calcium to natural BLG of at most 0.010, and

[0014] - The following total concentrations of monovalent metal cations:

[0015] - if the whey protein solution has a native BLG content of less than 10% w / w, up to 25 mM, or

[0016] - at most 20 mM if the native BLG content of the whey protein solution is at least 10% w / w,

[0017] b) heating the whey protein solution to a temperature of at least 68° C. for a period of time sufficient to form a suspension of whey protein nanogels,

[0018] c) optionally, concentrating the suspension of whey protein nanogels to obtain a concentrated suspension of whey protein nanogels,

[0019] d) optionally, drying the dryer feed comprising the whey protein nanogel originating from step b) or c).

[0020] Another aspect of the present invention relates to a method for producing a whey protein nanogel composition, the method comprising:

[0021] a) providing a whey protein solution, the solution having:

[0022] - a natural BLG content of at least 3% w / w,

[0023] - pH value in the range of 5.8-7.5, preferably 5.8-6.5,

[0024] -The weight ratio between total calcium and natural BLG is:

[0025] - at most 0.0041 x pH - 0.0209, and

[0026] - at least 0.0037 x pH - 0.0234 but greater than 0, and

[0027] - The following total concentrations of monovalent metal cations:

[0028] - if the whey protein solution has a native BLG content of less than 10% w / w, up to 25 mM, or

[0029] - at most 20 mM if the native BLG content of the whey protein solution is at least 10% w / w,

[0030] b) heating the whey protein solution to a temperature of at least 68° C. for a period of time sufficient to form a suspension of whey protein nanogels,

[0031] c) optionally, concentrating the suspension of whey protein nanogels to obtain a concentrated suspension of whey protein nanogels,

[0032] d) optionally, drying the dryer feed comprising the whey protein nanogel originating from step b) or c).

[0033] In addition to the above advantages, the inventors have observed that the nanogels of the present invention provide surprisingly low viscosity when present in liquids at high protein concentrations, such as 20% w / w protein or even 30% w / w, while being stable to heat treatment, even sterilizing heat treatment, at such protein concentrations.

[0034] Denatured whey protein particles of the prior art provide much higher viscosities at increasing protein concentrations and may not even be able to reach protein concentrations of 20% w / w in liquid solutions.

[0035] Therefore, another aspect of the present invention relates to a whey protein nanogel composition comprising whey protein nanogel in an amount of at least 30% relative to total protein and a total amount of protein of at least 30% w / w relative to total solids.

[0036] In some preferred embodiments of the present invention, the whey protein nanogel composition is a whey protein nanogel composition in powder form, comprising:

[0037] - whey protein nanogel in an amount of at least 90% w / w relative to the total protein,

[0038] - soluble whey protein aggregates in an amount of up to 5% w / w relative to the total protein,

[0039] - total BLG in an amount of at least 90% w / w relative to total protein, and

[0040] - A total amount of protein of at least 30% relative to total solids.

[0041] The low viscosity and extraordinary thermal stability of the nanogel of the present invention make it very suitable for high-protein beverages and enable the production of high-protein, heat-sterilized, low-viscosity beverage products, which are therefore very comfortable to drink.

[0042] Therefore, another aspect of the invention relates to a packaged heat treated beverage having a pH in the range of 3-8 and comprising whey protein nanogel in an amount of at least 1% w / w.Another aspect relates to a process for producing a packaged heat treated beverage.

[0043] Therefore, another aspect of the present invention relates to a process for producing an acidic thickened food, comprising the following steps:

[0044] - preparing a liquid food base having a pH of at least 5.7, said liquid food base comprising sufficient whey protein nanogel composition to provide protein in an amount of 4-20% w / w, said whey protein nanogel composition preferably being as defined herein, comprising:

[0045] - soluble whey protein aggregates in an amount of 15-70% w / w relative to the total protein,

[0046] - whey protein nanogel in an amount of at least 30-85% w / w relative to the total protein,

[0047] - heat treating said liquid food base at a temperature of at least 70°C for a period sufficient to at least pasteurize said liquid food base,

[0048] - optionally, homogenizing the heat-treated liquid food base,

[0049] - acidifying the heat-treated liquid food base to a pH of at most 5.4,

[0050] - optionally, homogenizing the acidified food base,

[0051] The acidic thickened food product is a mixture of an acidified food base or an acidified liquid food base and other ingredients such as, for example, sweeteners and / or flavorings.

[0052] Another aspect relates to an acidic, thickened food product obtainable by the process.

[0053] A further aspect of the present invention relates to the use of a whey protein nanogel composition as defined herein and / or a plurality of whey protein nanogels as defined herein for one or more of the following:

[0054] - As food ingredients,

[0055] - as a food ingredient for the production of an aseptic beverage containing at least 10% protein and even more preferably at least 21% protein,

[0056] - as a food ingredient for producing an acidic thickened food having a pH of less than 5.5 and having a viscosity of 100 % at 20°C and 300 s -1 The viscosity measured at a shear rate of greater than 200 cP, and

[0057] - As whitening agent, for example in coffee whitener.

[0058] Another aspect of the present invention relates to the use of whey protein nanogel and / or whey protein nanogel composition as a protein source for reducing the astringency and / or sourness of a heat-treated protein beverage having a pH of 3.0-5.0, most preferably 3.5-4.6.

[0059] - preferably, wherein the whey protein nanogel comprises at least 50% w / w of the total protein of the heat-processed beverage, more preferably at least 70% w / w, even more preferably at least 80% w / w, and most preferably at least 90% w / w of the total protein of the heat-processed beverage; and

[0060] - Preferably, wherein the heat treated beverage contains total protein in an amount of 2-35% w / w, more preferably 4-30% w / w, even more preferably 6-25% w / w and more preferably 8-20% w / w. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] Figure 1 Shown is the distribution of whey protein nanogels and soluble aggregates relative to total protein after heat treatment of whey protein solutions containing 4-16% native BLG protein at pH 5.9 at 90°C for 14 minutes.

[0062] Figure 2 Transmission electron microscopy images of whey protein nanogel suspensions prepared with A: 4% w / w native BLG; B: 8% w / w native BLG; C: 12% w / w native BLG and D: 16% w / w native BLG are shown. The scale bar for all images is 500 nm.

[0063] Figure 3Shown is the viscosity development during ultrafiltration (UF) concentration of a 14% w / w whey protein nanogel suspension (solid dots) and a native BLG solution (crosses) to approximately 30% w / w total protein.

[0064] Figure 4 Shown is the development of storage modulus (gel strength) of whey protein nanogel formulations containing soluble whey protein aggregates ranging from 1.9% (sample D) to 59.2% (sample A) relative to total protein. All samples were diluted to 4% total protein before determining the storage modulus. DETAILED DESCRIPTION

[0065] BLG is the most predominant protein in bovine whey and milk serum and exists in a variety of genetic variants, with the major variants in bovine milk labeled A and B. BLG is a lipocalin that can bind many hydrophobic molecules, suggesting a role in their transport. BLG has also been shown to be able to bind iron via siderophores and may play a role in combating pathogens. A homolog of BLG is lacking in human breast milk.

[0066] Bovine BLG is a relatively small protein of approximately 162 amino acid residues and a molecular weight of approximately 18.3-18.4 kDa. Under physiological conditions, it is primarily a dimer, but dissociates into monomers below about pH 3, retaining its native state, as determined using nuclear magnetic resonance spectroscopy. In contrast, BLG also occurs in aggregated forms of tetramers, octamers, and other polymers under various natural conditions.

[0067] In the context of the present invention, the term "native BLG" relates to undenatured BLG molecules, such as found in, for example, raw milk or whey or low-heat BLG isolates prepared, for example, by BLG crystallization, chromatography or filtration. Thus, native BLG in crystalline form is still native BLG. The amount of native BLG was quantified according to Assay 6.

[0068] In the context of the present invention, the term "total BLG" relates to the sum of native, denatured and aggregated BLG. The amount of total BLG was quantified according to assay 17.

[0069] In the context of the present invention, the term "crystalline" relates to a solid material whose constituents (eg atoms, molecules or ions) are arranged in a highly ordered microscopic structure, forming a lattice extending in all directions.

[0070] In the context of the present invention, the term "BLG crystals" relates to protein crystals that primarily contain non-aggregated and preferably native BLG arranged in a highly ordered microstructure, forming a lattice extending in all directions. BLG crystals can be monolithic or polycrystalline, and can be complete crystals, crystal fragments or a combination thereof. Crystal fragments are formed, for example, when complete crystals are subjected to mechanical shearing during processing. Crystal fragments also have a highly ordered crystal microstructure, but may lack the uniform surface and / or edges or corners of complete crystals. In both cases, BLG crystals or crystal fragments can be visually identified using an optical microscope, as well as a clear, compact and coherent structure. BLG crystals or crystal fragments are usually at least partially transparent. In addition, it is known that protein crystals are birefringent, and this optical property can be used to identify unknown particles with a crystalline structure.

[0071] In the context of the present invention, the term "edible composition" relates to a composition which is safe for human consumption and use as a food ingredient and does not contain problematic amounts of toxic ingredients such as toluene or other unwanted organic solvents. The whey protein nanogel composition and food of the present invention are preferably edible foods.

[0072] In the context of the present invention, the term "ALA" or "alpha-lactalbumin" relates to alpha-lactalbumin from mammalian species, e.g. in native and / or glycosylated form, 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 e.g. aggregated ALA is referred to. The total amount of ALA is determined according to Example 1.31. The term "aggregated ALA" relates to ALA that is typically at least partially unfolded and further aggregated with other denatured ALA molecules and / or other denatured whey proteins, typically by hydrophobic interactions and / or covalent bonds.

[0073] In the context of the present invention, the term "casein macropeptide" or "CMP" relates to a hydrophilic peptide, residues 106-169, derived from the hydrolysis of "κ-CN" or "κ-casein" from mammalian species, e.g. in native and / or glycosylated form, and includes naturally occurring genetic variants of aspartic proteases such as chymosin.

[0074] In the context of the present invention, the term "BLG isolate" refers to a composition comprising BLG in an amount of at least 85% w / w relative to total protein. 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.

[0075] The term "whey" relates to the liquid phase remaining after the precipitation and removal of casein in milk. Casein precipitation can be accomplished, for example, by acidifying the milk and / or using rennet. There are several types of whey: such as "sweet whey", which is a whey product produced by rennet-based precipitation of casein, and "acid whey" or "sour whey", which is a whey product produced by acid-based precipitation of casein. Acid-based precipitation of casein can be accomplished, for example, by adding edible acids or by bacterial cultures.

[0076] The term "milk serum" refers to the liquid remaining after casein and milk fat globules have been removed from milk, for example by microfiltration or macroporous ultrafiltration. Milk serum is also considered a type of whey and is sometimes referred to as "ideal whey."

[0077] The term "milk serum protein" or "serum protein" relates to proteins present in milk serum.

[0078] In the context of the present invention, the term "whey protein" relates to proteins found in whey or milk heap. The whey protein may be a subset of the protein species found in whey or milk heap, even a single whey protein species, or it may be a complete set of protein species found in whey or / and milk heap.

[0079] In the context of the present invention, the terms "liquid" and "solution" encompass both liquid compositions that do not contain particulate matter and liquid compositions that contain a combination of liquid and solid and / or semi-solid particles (e.g., protein crystals or other protein particles). However, "liquids" and "solutions" are preferably pumpable.

[0080] In the context of the present invention, the terms "whey protein concentrate" (WPC) and "serum protein concentrate" (SPC) relate to dry or aqueous compositions containing a total amount of protein of 20-89% w / w relative to total solids.

[0081] WPC or SPC preferably contains:

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

[0083] BLG at 15-70% w / w of total protein,

[0084] 8-50% w / w ALA relative to total protein, and

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

[0086] Alternatively, but preferably, the WPC or SPC may contain:

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

[0088] BLG with 15-90% w / w of total protein,

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

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

[0091] Preferably, the WPC or SPC contains:

[0092] 20-89% w / w protein relative to total solids,

[0093] BLG at 15-80% w / w of total protein,

[0094] 4-50% w / w ALA relative to total protein, and

[0095] 0-40% w / w CMP relative to protein.

[0096] More preferably, the WPC or SPC contains:

[0097] 70-89% w / w protein relative to total solids,

[0098] BLG with 30-90% w / w of total protein,

[0099] 4-35% w / w ALA relative to total protein, and

[0100] 0-25% w / w CMP relative to protein.

[0101] SPC generally contains no CMP or only trace amounts of CMP.

[0102] The terms "whey protein isolate" (WPI) and "serum protein isolate" (SPI) relate to dry or aqueous compositions containing a total amount of protein of 90-100% w / w relative to total solids.

[0103] WPI or SPI preferably contains:

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

[0105] BLG at 15-70% w / w of total protein,

[0106] 8-50% w / w ALA relative to total protein, and

[0107] 0-40% w / w CMP relative to total protein.

[0108] Alternatively, but preferably, the WPI or SPI may contain:

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

[0110] BLG with 30-95% w / w of total protein,

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

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

[0113] More preferably, the WPI or SPI may contain:

[0114] 90-100% w / w protein relative to total solids,

[0115] BLG with 30-90% w / w of total protein,

[0116] 4-35% w / w ALA relative to total protein, and

[0117] 0-25% w / w CMP relative to total protein.

[0118] SPI usually contains no CMP or only trace amounts of CMP.

[0119] In the context of the present invention, the term "whey protein nanogel" refers to nanometer-sized (usually about 150-1000nm) particles of denatured whey protein, which are usually spherical or nearly spherical in shape. Whey protein nanogels are also referred to as whey protein micelles, and are discussed, for example, in WO2007 / 110421A2, and their micellar properties are questionable. The amount of whey protein nanogel is quantified according to analysis 3. Protein nanogels have an opaque milky appearance when suspended, and are therefore very suitable for use in opaque beverages.

[0120] In the context of the present invention, the term "soluble whey protein aggregates" refers to small aggregates of denatured whey proteins, which can form strong gels (much stronger than native whey proteins) during acidification to pH 4.6 and which generally have linear, worm-like, branched or chain-like shapes and are generally submicron-sized. Soluble whey protein aggregates are well known to those skilled in the art, for example, described in WO2007 / 110421A2, where they are referred to as linear aggregates. The amount of soluble whey protein aggregates is quantified according to analysis 3. Soluble whey protein aggregates will generally form clear solutions when dissolved in water, and are therefore very suitable for clear beverages.

[0121] The terms “consists essentially of” and “consisting essentially of” mean that the claim or feature in question covers the specified materials or steps, as well as those that do not materially affect the basic and novel characteristics of the claimed invention.

[0122] In the context of the present invention, the phrase "Y and / or X" means "Y" or "X" or "Y and X". 1 、n 2 ,...,n i-1 and / or i " means "n 1 " or "n 2 "or...or" i-1 " or "n i " or quantity: n 1 、n 2 ,...,n i-1 and n i any combination of .

[0123] In the context of the present invention, the term "dry" or "dried" means that the composition or product in question contains at most 10% w / w, preferably at most 6% w / w, and more preferably even less water.

[0124] In the context of the present invention, the weight percent (% w / w) of a component of a composition, product or material refers to the weight percent of that component relative to the weight of the particular composition, product or material, unless another reference (e.g. total solids or total protein) is specifically mentioned.

[0125] In the context of the present invention, the term "weight ratio" between component X and component Y refers to the weight ratio calculated by m X / m Y The value obtained, where m X is the amount (weight) of component X and mY is the amount of component Y (by weight).

[0126] In the context of the present invention, the term "at least pasteurized" relates to a heat treatment having a microbial killing effect equal to or higher than a heat treatment at 70°C for 10 seconds. The reference for determining the bactericidal effect is Escherichia coli O157:H7.

[0127] In the context of the present invention, the term "sterile" means that the sterile composition or product in question does not contain any living microorganisms and therefore no microbial growth occurs during storage at room temperature. A composition that has been sterilized is sterile.

[0128] When a liquid, such as a beverage, is sterilized and aseptically packaged in a sterile container, it typically has a shelf life of at least six months at room temperature. The sterilization process kills spores and microorganisms that could cause the liquid to spoil.

[0129] Therefore, one aspect of the present invention relates to a method for producing a whey protein nanogel composition, the method comprising:

[0130] a) providing a whey protein solution, the solution having:

[0131] - a natural BLG content in an amount of at least 3% w / w,

[0132] - pH value in the range of 5.8-7.5, preferably 5.8-6.5,

[0133] - a weight ratio of total calcium to natural BLG of at most 0.010, and

[0134] - The following total concentrations of monovalent metal cations:

[0135] - if the whey protein solution has a native BLG content of less than 10% w / w, a maximum of 25

[0136] mM, or

[0137] - at most 20 mM if the native BLG content of the whey protein solution is at least 10% w / w,

[0138] b) heating the whey protein solution to a temperature of at least 68° C. for a period of time sufficient to form a suspension of whey protein nanogels,

[0139] c) optionally, concentrating the suspension of whey protein nanogels to obtain a concentrated suspension of whey protein nanogels,

[0140] d) optionally, drying the dryer feed comprising the whey protein nanogel originating from step b) or c).

[0141] Another aspect of the present invention relates to a method for producing a whey protein nanogel composition, the method comprising:

[0142] a) providing a whey protein solution, the solution having:

[0143] - a natural BLG content in an amount of at least 3% w / w,

[0144] - pH value in the range of 5.8-7.5, preferably 5.8-6.5,

[0145] -The weight ratio between total calcium and natural BLG is:

[0146] - at most 0.0041 x pH - 0.0209, and

[0147] - at least 0.0037 x pH - 0.0234 but greater than 0, and

[0148] - The following total concentrations of monovalent metal cations:

[0149] - if the whey protein solution has a native BLG content of less than 10% w / w, at most 25 mM, or

[0150] - at most 20 mM if the native BLG content of the whey protein solution is at least 10% w / w,

[0151] b) heating the whey protein solution to a temperature of at least 68° C. for a period of time sufficient to form a suspension of whey protein nanogels,

[0152] c) optionally, concentrating the suspension of whey protein nanogels to obtain a concentrated suspension of whey protein nanogels,

[0153] d) optionally, drying the dryer feed comprising the whey protein nanogel originating from step b) or c).

[0154] In some preferred embodiments of the present invention, the method provides a whey protein nanogel composition in powder form and comprises steps a), b), c) and d).

[0155] In other preferred embodiments of the present invention, the method provides a whey protein nanogel composition in powder form and comprises steps a), b) and d), but does not comprise step c).

[0156] In a further preferred embodiment of the present invention, the method provides a whey protein nanogel composition in liquid form and comprises steps a), b) and c), but does not comprise step d).

[0157] In a further preferred embodiment of the present invention, the method provides a whey protein nanogel composition in liquid form and comprises steps a) and b), but does not comprise steps c) and d).

[0158] As described above, step a) provides a whey protein solution containing native BLG and optionally other whey proteins. The whey protein solution is an aqueous solution and preferably contains water in an amount of at least 50% w / w. The whey protein solution is preferably edible and suitable as a food. In addition to dissolved protein, the whey protein solution may contain suspended particles.

[0159] The whey protein solution has a native BLG content of at least 3% w / w.

[0160] In some preferred embodiments of the invention, the whey protein solution comprises native BLG in an amount of at least 4% w / w, more preferably at least 6% w / w, even more preferably at least 8% w / w, and most preferably at least 10% w / w.

[0161] In other preferred embodiments of the invention the whey protein solution comprises native BLG in an amount of at least 11% w / w, more preferably at least 16% w / w, even more preferably at least 18% w / w and most preferably at least 20% w / w.

[0162] In a further preferred embodiment of the invention the whey protein solution comprises native BLG in an amount of at least 21% w / w, more preferably at least 23% w / w, even more preferably at least 25% w / w and most preferably at least 27% w / w.

[0163] Preferably, the whey protein solution comprises native BLG in an amount of 3-30% w / w, more preferably in an amount of 4-28% w / w, even more preferably in an amount of 6-26% w / w, and most preferably in an amount of 8-24% w / w.

[0164] In some preferred embodiments of the present invention, the whey protein solution comprises native BLG in an amount of 11-30% w / w, more preferably in an amount of 12-28% w / w, even more preferably in an amount of 14-26% w / w, and most preferably in an amount of 16-24% w / w.

[0165] In other preferred embodiments of the present invention, the whey protein solution comprises native BLG in an amount of 10-24% w / w, more preferably in an amount of 12-22% w / w, even more preferably in an amount of 14-20% w / w and most preferably in an amount of 16-18% w / w.

[0166] In a further preferred embodiment of the present invention the whey protein solution comprises native BLG in an amount of 21-32% w / w, more preferably in an amount of 22-31% w / w, even more preferably in an amount of 23-30% w / w and most preferably in an amount of 24-29% w / w.

[0167] In some preferred embodiments of the invention, the whey protein solution comprises total protein in an amount of at least 4% w / w, more preferably at least 6% w / w, even more preferably at least 8% w / w and most preferably at least 10% w / w.

[0168] Preferably, the whey protein solution comprises total protein in an amount of 1-30% w / w, more preferably 4-28% w / w, even more preferably 6-26% w / w, and most preferably 8-24% w / w.

[0169] In some preferred embodiments of the present invention, the whey protein solution comprises total protein in an amount of 11-30% w / w, more preferably 12-28% w / w, even more preferably 14-26% w / w, and most preferably 16-24% w / w.

[0170] In other preferred embodiments of the invention, the whey protein solution comprises total protein in an amount of 10-24% w / w, more preferably in an amount of 12-22% w / w, even more preferably in an amount of 14-20% w / w and most preferably in an amount of 16-18% w / w.

[0171] In a further preferred embodiment of the present invention the whey protein solution comprises total protein in an amount of 21-32% w / w, more preferably 22-31% w / w, even more preferably 23-30% w / w and most preferably 24-29% w / w.

[0172] In some preferred embodiments of the present invention, the whey protein solution comprises total protein in an amount of at least 30% w / w relative to the total solids, more preferably in an amount of at least 60% w / w relative to the total solids, even more preferably in an amount of at least 70% w / w relative to the total solids, and most preferably in an amount of at least 80% w / w relative to the total solids.

[0173] In other preferred embodiments of the present invention, the whey protein solution comprises total protein in an amount of at least 85% w / w relative to the total solids, more preferably in an amount of at least 90% w / w relative to the total solids, even more preferably in an amount of at least 92% w / w relative to the total solids, and most preferably in an amount of at least 95% w / w relative to the total solids.

[0174] In some preferred embodiments of the present invention, the whey protein solution comprises natural BLG in an amount of at least 50% w / w relative to the total protein, more preferably at least 60% w / w relative to the total protein, even more preferably at least 70% w / w relative to the total protein, and most preferably at least 80% w / w relative to the total protein.

[0175] In other preferred embodiments of the present invention, the whey protein solution comprises natural BLG in an amount of at least 85% w / w relative to the total protein, more preferably at least 90% w / w relative to the total protein, even more preferably at least 92% w / w relative to the total protein, and most preferably at least 95% w / w relative to the total protein. It is generally preferred that the whey protein solution comprises natural BLG in an amount of at least 97% w / w relative to the total protein.

[0176] In a further preferred embodiment of the invention, the whey protein solution comprises native BLG in an amount in the range of 50-80% w / w relative to the total protein, more preferably in the range of 52-75% w / w relative to the total protein, even more preferably in the range of 54-70% w / w relative to the total protein, and most preferably in the range of 55-65% w / w relative to the total protein. This is useful, for example, when highly demineralized WPI is used as the protein source of the whey protein solution.

[0177] The whey protein solution may contain other proteins than native BLG, and typically contains other whey proteins. In some preferred embodiments of the invention, the whey protein solution contains only proteins derived from mammalian whey.

[0178] In some preferred embodiments of the present invention, the whey protein solution has a degree of protein denaturation of at most 20% w / w relative to the total protein, more preferably at most 10% w / w relative to the total protein, even more preferably at most 5% w / w relative to the total protein, and most preferably at most 2% w / w relative to the total protein.

[0179] Even lower denaturation may be preferred, and in some preferred embodiments of the invention the whey protein solution has a degree of protein denaturation of at most 1% w / w relative to the total protein, more preferably at most 0.5% w / w relative to the total protein, even more preferably at most 0.2% w / w relative to the total protein, and most preferably at most 0.1% w / w relative to the total protein.

[0180] The extent of protein denaturation was measured according to Assay 3.

[0181] In some preferred embodiments of the present invention, the whey protein solution has a total solid content of 3-50% w / w, more preferably 4-40% w / w, even more preferably 6-35% w / w, and most preferably 8-30% w / w.

[0182] In other preferred embodiments of the present invention, the whey protein solution has a total solids content in an amount of 10-50% w / w, more preferably in an amount of 12-40% w / w, even more preferably in an amount of 14-35% w / w and most preferably in an amount of 16-30% w / w.

[0183] In some preferred embodiments of the present invention, the whey protein solution has a water content in an amount of 50-97% w / w, more preferably in an amount of 60-96% w / w, even more preferably in an amount of 65-94% w / w, and most preferably in an amount of 70-92% w / w.

[0184] The non-solid part of the whey protein solution is preferably water.

[0185] The pH of the whey protein solution is in the range of 5.8-7.5.

[0186] In some preferred embodiments of the present invention, the pH of the whey protein solution is in the range of 5.8-6.5, more preferably 5.9-6.3, and most preferably 5.9-6.2.

[0187] In some preferred embodiments of the present invention, the pH of the whey protein solution is in the range of 5.9-6.5, more preferably 6.0-6.4, and most preferably 6.1-6.3.

[0188] In a further preferred embodiment of the present invention, the pH of the whey protein solution is in the range of 6.5-7.5, more preferably 6.6-7.2, even more preferably 6.7-7.0.

[0189] pH adjustment preferably uses a method that does not increase the amount of Na + and K + The alkalizing agent with a monovalent metal ion content of 1.5 wt % is preferably Ca(OH) 2 Alternatively, but also preferably, the alkalizing agent used may comprise a combination of one or more bases containing monovalent metal ions and one or more bases containing divalent metal ions. For example, if only Ca(OH) 2 Such combinations are useful if adjusting the pH provides an excessively high weight ratio between calcium and native BLG, and the resulting monovalent metal ion concentration does exceed the threshold required for efficient nanogel formation.

[0190] In some particularly preferred embodiments of the present invention, the weight ratio between the total amount of calcium in the whey protein solution and the natural BLG is:

[0191] - at most 0.0041 x pH - 0.0209, and

[0192] - At least 0.0037 x pH - 0.0234 but greater than 0.

[0193] The term "greater than zero" means that the weight ratio between the total amount of calcium and native BLG will always be greater than absolute zero, as a whey protein solution will always contain trace amounts of calcium, even though it may be difficult to detect. However, the weight ratio can be very close to zero.

[0194] It is further preferred that the total weight of other divalent metals of the whey protein solution is less than the total weight of calcium, more preferably at most 50% of the total weight of calcium, and most preferably at most 20% of the total weight of calcium.

[0195] In some preferred embodiments of the present invention, the weight ratio between the total amount of calcium in the whey protein solution and the natural BLG is:

[0196] - at most 0.0041 x pH - 0.0220, and

[0197] - At least 0.0037 x pH - 0.0222 but greater than 0.

[0198] In other preferred embodiments of the present invention, the weight ratio between the total amount of calcium in the whey protein solution and the natural BLG is:

[0199] - at most 0.0040 x pH - 0.0221, and

[0200] - At least 0.0039 x pH - 0.0221 but greater than 0.

[0201] In some preferred embodiments of the present invention, the weight ratio between the total amount of calcium of the whey protein solution and the native BLG is at most 0.0070, more preferably at most 0.0065, even more preferably at most 0.0060, and most preferably at most 0.0050.

[0202] In other preferred embodiments of the present invention, the weight ratio between the total amount of calcium of the whey protein solution and the native BLG is at most 0.0045, more preferably at most 0.0040, even more preferably at most 0.0035, and most preferably at most 0.0030.

[0203] Even lower weight ratios are generally preferred, and in some preferred embodiments of the invention the whey protein solution has a weight ratio between the total amount of calcium and native BLG of at most 0.0025, more preferably at most 0.0020, even more preferably at most 0.0015, and most preferably at most 0.0005.

[0204] In some preferred embodiments of the present invention, the weight ratio between the total amount of calcium of the whey protein solution and the native BLG is in the range of 0.0005-0.0065, more preferably in the range of 0.0010-0.0050, even more preferably in the range of 0.0015-0.0040, and most preferably in the range of 0.0015-0.0030, such as preferably in the range of 0.0017-0.0027.

[0205] In addition, the total concentration of monovalent metal cations in the whey protein solution is

[0206] - if the whey protein solution has a native BLG content of less than 10% w / w, at most 25 mM, or

[0207] - at most 20 mM if the native BLG content of the whey protein solution is at least 10% w / w.

[0208] In some preferred embodiments of the present invention, the native BLG content of the whey protein solution is less than 10% w / w and the total concentration of monovalent metal cations of the whey protein solution is at most 24 mM, more preferably at most 22 mM, even more preferably at most 21 mM, and most preferably at most 20 mM.

[0209] In some preferred embodiments of the invention, regardless of the content of native BLG, the total concentration of monovalent metal cations of the whey protein solution is at most 19 mM, more preferably at most 17 mM, even more preferably at most 15 mM, and most preferably at most 10 mM. Even lower contents of monovalent metal cations may be preferred, and in some preferred embodiments of the invention, the total concentration of monovalent metal cations of the whey protein solution is at most 8 mM, more preferably at most 6 mM, even more preferably at most 4 mM, and most preferably at most 2 mM.

[0210] The method of the present invention makes it possible to produce a whey protein nanogel composition having a very high yield of whey protein nanogel and a low content of soluble whey protein aggregates and microparticles, which are usually undesirable. Therefore, the method of the present invention provides an improved and more cost-effective way to produce a whey protein nanogel composition having a high purity whey protein nanogel, and enables the production of novel whey protein nanogel compositions that are not available in the prior art.

[0211] The whey protein solution may comprise other macronutrients besides protein, such as for example carbohydrates and / or lipids.

[0212] However, it is generally preferred that the whey protein solution contains carbohydrates in an amount of at most 15% w / w relative to the total solids, more preferably at most 5% w / w relative to the total solids, even more preferably at most 1% w / w relative to the total solids, and most preferably at most 0.1% w / w relative to the total solids.

[0213] It is also generally preferred that the whey protein solution contains lipids in an amount of at most 8% w / w relative to the total solids, more preferably at most 2% w / w relative to the total solids, even more preferably at most 0.5% w / w relative to the total solids, and most preferably at most 0.1% w / w relative to the total solids.

[0214] The whey protein solution may contain other macronutrients and ingredients besides protein. The embodiments and preferences described below in the context of packaged heat-processed beverages relating to macronutrients and additional ingredients are equally applicable to the whey protein solution and we refer to these embodiments here rather than repeating them.

[0215] In some particularly preferred embodiments of the invention, the whey protein solution has the same chemical composition as the packaged heat-processed beverage, except that the protein is in its native form.

[0216] In other particularly preferred embodiments of the present invention, the whey protein solution has the same chemical composition as the packaged heat-treated beverage.

[0217] The protein source of the whey protein solution can be any source that provides natural BLG and optionally also provides minerals. The whey protein source can be, for example, a whey protein isolate, a milk protein isolate, a milk protein concentrate or a combination thereof. Particularly preferred are whey protein isolates and / or milk protein isolates. The whey protein source preferably has a very low mineral content.

[0218] A currently preferred whey protein source is a BLG isolate obtainable by crystallization and recovery of BLG crystals as outlined in WO 2018 / 115520 A1, i.e. by crystallizing BLG in a salinization mode from a pH 5-6 aqueous solution containing whey protein. The protein source may be, for example, a wet crystal slurry obtained according to the process of WO 2018 / 115520 A1 or a spray-dried crystal slurry obtained according to WO 2018 / 115520 A1. Alternatively, but also preferably, the protein source may be a BLG isolate according to PCT / EP2019 / 066998, and preferably obtainable by the process outlined in PCT / EP2019 / 066998. WO 2018 / 115520 A1 and PCT / EP2019 / 066998 are incorporated herein by reference for all purposes.

[0219] Alternatively, but also preferably, the protein source may be a BLG isolate obtainable according to de Jongh et al. (Mild Isolation Procedure Discloses New Protein Structural Properties of beta-Lactoglobulin, J Dairy Sci., Vol. 84(3), 2001, pp. 562-571) and subsequently further demineralized by dialysis.

[0220] Alternatively, but also preferably, the protein source may be commercially available high quality WPI which is demineralized by dialysis prior to use.

[0221] The whey protein used in the whey protein solution is whey protein from mammalian milk, such as, for example, from the milk of cows, goats, mares, sheep, camels and / or buffaloes. Bovine whey protein is particularly preferred.

[0222] The proteins of the various compositions described herein are preferably edible proteins and may contain other edible proteins in addition to BLG. It is particularly preferred that the proteins of the present invention are derived from mammalian milk, for example, proteins provided by isolating and / or modifying one or more protein species from mammalian milk. Most preferably, the proteins of the present invention are whey proteins or protein materials derived from whey proteins, preferably by heat denaturation. However, it is contemplated and generally preferred that foods containing the whey protein nanogel or whey protein nanogel composition of the present invention may contain non-milk proteins in addition to protein species from milk.

[0223] Ideally, a whey protein solution is provided by dissolving a whey protein source in demineralized water and optionally adjusting the pH. In addition, it may be necessary to vary the mineral composition to obtain a useful whey protein solution as described herein.

[0224] Step b) involves heating the whey protein solution to a temperature of at least 68°C for a period sufficient to form a suspension of whey protein nanogels. Step b) may also involve cooling the suspension to terminate the heat treatment unless the suspension is to be used warm, for example for direct spray drying or for the production of food products.

[0225] If step b) involves cooling, the suspension is typically cooled to a temperature of at most 50°C, more preferably at most 40°C, even more preferably at most 20°C and more preferably at most 10°C.

[0226] In some preferred embodiments of the present invention, the heating of step b) heats the whey protein solution to a temperature of at least 70° C., more preferably at least 75° C., even more preferably at least 80° C. and most preferably at least 85° C. Even higher temperatures have proven useful, and in some preferred embodiments of the present invention, the heating of step b) heats the whey protein solution to a temperature of at least 90° C., more preferably at least 95° C., even more preferably at least 100° C. and most preferably at least 120° C.

[0227] In some preferred embodiments of the present invention, the heating of step b) heats the whey protein solution to a temperature in the range of 68-160°C, more preferably 75-150°C, even more preferably 80-120°C and most preferably 85-100°C.

[0228] It is especially preferred that the heating of step b) heats the whey protein solution to a temperature in the range of 80-95°C, more preferably 82-92°C, even more preferably 84-90°C and most preferably 85-89°C.

[0229] The duration of the heat treatment of step b) should be sufficient to form a large amount of nanogels. The duration is preferably at least 500 milliseconds, and is usually longer. This duration is preferably selected to denature a large amount of native BLG of the whey protein solution.

[0230] In some preferred embodiments of the present invention, step b) heating is performed for a duration sufficient to denature at least 50% w / w of native BLG, more preferably at least 80% w / w of native BLG, even more preferably at least 90% of native BLG, and most preferably at least 95% w / w of native BLG. Even higher levels of BLG denaturation may be preferred, and in some preferred embodiments of the present invention, step b) heating is performed for a duration sufficient to denature at least 96% w / w of native BLG, more preferably at least 97% w / w of native BLG, even more preferably at least 98% of native BLG, and most preferably at least 99% w / w of native BLG.

[0231] The present inventors have found that a high level of BLG denaturation results in a better thermal stability of the resulting whey protein nanogel composition.

[0232] The duration of the heat treatment depends on the temperature, but is generally between 1 minute and 1 hour, preferably between 4-50 minutes, more preferably between 6-45 minutes, even more preferably between 8-40 minutes, and most preferably between 10-30 minutes.

[0233] Surprisingly, despite the use of relatively high protein concentrations in the whey protein solution, the inventive method allows the production of whey protein nanogel compositions at high protein concentrations using only limited mechanical shearing or even without mechanical shearing. In some preferred embodiments of the present invention, the thermal treatment of step b) does not involve mechanical shearing, such as scraped surface heat exchangers or high pressure homogenization. However, the present invention does not exclude mechanical shearing, and in other preferred embodiments of the present invention, the thermal treatment of step b) does involve mechanical shearing.

[0234] The compositional features described in the context of the whey protein solution of step a) apply equally to the suspension of whey protein nanogels obtained in step b), except that a considerable amount of the native proteins have been denatured and thus converted into e.g. whey protein nanogels.

[0235] Step c) is optional, but in some embodiments is preferred.

[0236] Therefore, in some preferred embodiments of the present invention, the method comprises a step c) which involves concentrating the suspension of whey protein nanogels to obtain a concentrated suspension of whey protein nanogels.

[0237] However, in other preferred embodiments of the present invention, the process does not comprise step c), and thus the suspension is not concentrated.

[0238] In the context of the present invention, the term "suspension of concentrated whey protein nanogels" does not involve a drying step, like for example spray drying but removes at least water to increase the concentration of at least the whey protein nanogels.

[0239] The suspension of whey protein nanogel is preferably concentrated by one or more of the following methods: microfiltration, ultrafiltration, nanofiltration, reverse osmosis and / or evaporation. Ultrafiltration, nanofiltration or reverse osmosis are particularly preferred because they preserve the protein content of the suspension.

[0240] In some embodiments of the invention, the concentration of step c) increases the content of whey protein nanogel in the suspension by at least 25%, more preferably by at least 50%, even more preferably by at least 75%, and most preferably by at least 100%. Even higher levels of concentration may be required, and in some preferred embodiments of the invention, the concentration of step c) increases the content of whey protein nanogel in the suspension by at least 200%, more preferably by at least 300%, even more preferably by at least 500%, and most preferably by at least 700%.

[0241] In some embodiments of the present invention, the concentration in step c) increases the content of whey protein nanogel in the suspension by 25-900%, more preferably by 100-850%, even more preferably by 200-825%, and most preferably by 300-800%.

[0242] In some preferred embodiments of the invention, the concentration of step c) provides a concentrated suspension of whey protein nanogels comprising at least 21% w / w, more preferably at least 25%, even more preferably at least 28% w / w, and most preferably at least 30% w / w of total protein.

[0243] Therefore, in some preferred embodiments of the present invention, the concentrated suspension of whey protein nanogels comprises at least 21% w / w, more preferably at least 25%, even more preferably at least 28% w / w, and most preferably at least 30% w / w of total protein.

[0244] In other preferred embodiments of the invention, the concentration of step c) provides a concentrated suspension of whey protein nanogels comprising 21-35% w / w, more preferably 25-33%, even more preferably 28-32% w / w, and most preferably 29-31% w / w of total protein.

[0245] Therefore, in other preferred embodiments of the present invention, the concentrated suspension of whey protein nanogels comprises 21-35% w / w, more preferably 25-33%, even more preferably 28-32% w / w, and most preferably 29-31% w / w of total protein.

[0246] In some preferred embodiments of the present invention, the concentration of step c) provides a concentrated suspension of whey protein nanogel comprising whey protein nanogel in an amount of at least 21% w / w, more preferably at least 25%, even more preferably at least 28% w / w, and most preferably at least 30% w / w.

[0247] Therefore, in some preferred embodiments of the present invention, the concentrated suspension of whey protein nanogel comprises whey protein nanogel in an amount of at least 21% w / w, more preferably at least 25%, even more preferably at least 28% w / w, and most preferably at least 30% w / w.

[0248] In other preferred embodiments of the present invention, the concentration of step c) provides a concentrated suspension of whey protein nanogel comprising whey protein nanogel in an amount of 21-35% w / w, more preferably 25-33%, even more preferably 28-32% w / w, and most preferably 29-31% w / w.

[0249] Therefore, in other preferred embodiments of the present invention, the concentrated suspension of whey protein nanogel comprises whey protein nanogel in an amount of 21-35% w / w, more preferably 25-33%, even more preferably 28-32% w / w, and most preferably 29-31% w / w.

[0250] The present inventors have found that it is preferred to control the content of soluble whey protein aggregates of the suspension and concentrated suspension and keep them to a minimum.

[0251] Therefore, in some preferred embodiments of the present invention, the concentrated suspension of whey protein nanogel contains soluble whey protein aggregates in an amount of at most 8% w / w, more preferably at most 6%, even more preferably at most 3% w / w, and most preferably at most 1% w / w. Even lower amounts of soluble whey protein aggregates are also feasible, and in some preferred embodiments of the present invention, the concentrated suspension of whey protein nanogel contains soluble whey protein aggregates in an amount of at most 1% w / w, more preferably at most 0.5%, even more preferably at most 0.3% w / w, and most preferably at most 0.1% w / w.

[0252] In some preferred embodiments of the present invention, the concentrated suspension of whey protein nanogel comprises whey protein nanogel in an amount of at least 60% w / w relative to the total protein, more preferably at least 70% w / w relative to the total protein, even more preferably at least 80% w / w, and most preferably at least 90% w / w.

[0253] Higher amounts of whey protein nanogels are both feasible and generally preferred. Thus, in some preferred embodiments of the present invention, the concentrated suspension of whey protein nanogels comprises whey protein nanogels in an amount of at least 92% w / w relative to total protein, more preferably at least 94% w / w relative to total protein, even more preferably at least 96% w / w, and most preferably at least 98% w / w.

[0254] The compositional features described in the context of the whey protein solution of step a) are equally applicable to the concentrated suspension of whey protein nanogels obtained in step c), except that a considerable amount of native proteins have been denatured and thus converted into, for example, whey protein nanogels. In addition, due to the concentration, the content of total protein is increased.

[0255] If the pH of the whey protein nanogel composition should be different from the pH used during the formation of the whey protein nanogel, the method preferably comprises one or more steps of adjusting the pH of one or more streams containing the whey protein nanogel obtained by step b) or after step b), the stream is for example a suspension of the whey protein nanogel, a concentrated suspension of the whey protein nanogel and / or a dryer feed. One or more pH adjustments are sufficient to provide a whey protein nanogel composition with a desired pH and preferably with one of the preferred ranges mentioned in the context of the whey protein nanogel composition. One or more pH adjustments preferably use one or more suitable acids or bases, preferably selected from acids or bases mentioned herein.

[0256] Step d) is optional, but in some embodiments is preferred.

[0257] In some preferred embodiments of the present invention, the process does not comprise step d) and no drying step is performed.

[0258] However, in other preferred embodiments of the present invention, the method comprises a step d) which involves drying the dryer feed comprising the whey protein nanogel originating from step b) or c). Step d) is particularly preferred because it provides a whey protein nanogel composition in the form of a powder which is easy to transport and has better storage stability than an aqueous suspension.

[0259] In the context of the present invention the term "dryer feed" relates to the liquid feed fed to the dryer for converting the feed into a powder. The dryer feed of the present invention contains the whey protein nanogel produced in step b).

[0260] In some preferred embodiments of the present invention, the dryer feed comprises solids of the whey protein nanogel suspension of step b) and / or solids of the whey protein nanogel concentrated suspension of step c).

[0261] In other preferred embodiments of the present invention, the solids of the dryer feed comprise the solids of the whey protein nanogel suspension of step b) and / or the solids of the whey protein nanogel concentrated suspension of step c).

[0262] In some preferred embodiments of the present invention, the dryer feed is the whey protein nanogel suspension of step b) and / or the whey protein nanogel concentrated suspension of step c).

[0263] Preferably, the dryer feed comprises a total amount of protein of at least 21% w / w, more preferably at least 25%, even more preferably at least 28% w / w and most preferably at least 30% w / w.

[0264] In some preferred embodiments of the invention, the dryer feed comprises a total amount of protein of 21-35% w / w, more preferably 25-33% w / w, even more preferably 28-32% w / w and most preferably 29-31% w / w.

[0265] In other preferred embodiments of the present invention the dryer feed comprises a total amount of protein of 15-32% w / w, more preferably 17-31 %, even more preferably 18-30% w / w and most preferably 19-29% w / w.

[0266] Preferably, the dryer feed comprises whey protein nanogel in an amount of at least 21% w / w, more preferably at least 25%, even more preferably at least 28% w / w and most preferably at least 30% w / w.

[0267] In some preferred embodiments of the invention, the dryer feed comprises whey protein nanogel in an amount of 21-35% w / w, more preferably 25-33%, even more preferably 28-32% w / w, and most preferably 29-31% w / w.

[0268] In other preferred embodiments of the present invention, the dryer feed comprises whey protein nanogel in an amount of 10-35% w / w, more preferably 12-30%, even more preferably 14-25% w / w, and most preferably 15-20% w / w.

[0269] The inventors have seen evidence that drying a dryer feed comprising whey protein nanogel in an amount of 10-25% w / w provides a dried whey protein nanogel composition with improved dispersibility when reconstituted in water. Therefore, in some preferred embodiments of the invention, the dryer feed comprises whey protein nanogel in an amount of 10-25% w / w, more preferably 12-25%, even more preferably 14-20% w / w and most preferably 15-20% w / w.

[0270] It is preferred that the dryer feed comprises soluble whey protein aggregates in an amount of at most 8% w / w, more preferably at most 6%, even more preferably at most 3% w / w and most preferably at most 1% w / w.

[0271] In some preferred embodiments of the invention, the dryer feed comprises whey protein nanogel in an amount of at least 60% w / w relative to the total protein, more preferably at least 70% w / w relative to the total protein, even more preferably at least 80% w / w, and most preferably at least 90% w / w.

[0272] Higher amounts of whey protein nanogels are both feasible and generally preferred. Thus, in some preferred embodiments of the invention, the dryer feed comprises whey protein nanogels in an amount of at least 92% w / w relative to total protein, more preferably at least 94% w / w relative to total protein, even more preferably at least 96% w / w, and most preferably at least 98% w / w.

[0273] In some preferred embodiments of the present invention, the dryer feed comprises:

[0274] - a total BLG content in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein,

[0275] - a total protein content in an amount of at least 80% w / w, most preferably at least 90% w / w relative to the total solids,

[0276] - a total solids content in the range of 21-40% w / w, most preferably 24-35% w / w,

[0277] - whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein, and

[0278] - Soluble whey protein aggregates in an amount of at most 9% w / w, and most preferably at most 5% w / w, relative to total protein.

[0279] In a further preferred embodiment of the present invention, the dryer feed comprises:

[0280] - a total BLG content in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein,

[0281] - a total protein content in an amount of at least 80% w / w, most preferably at least 90% w / w relative to the total solids,

[0282] - a total solids content in the range of 8-25% w / w, most preferably 12-22% w / w,

[0283] - whey protein nanogel in an amount of 30-85% w / w, most preferably 35-70% w / w relative to total protein, and

[0284] - Soluble whey protein aggregates in an amount of 15-70% w / w, and most preferably 30-65% w / w relative to total protein.

[0285] In a further preferred embodiment of the present invention, the dryer feed comprises:

[0286] - a total BLG content in an amount of at least 50% w / w, most preferably at least 60% w / w relative to the total protein,

[0287] - a total protein content in an amount of at least 80% w / w, most preferably at least 90% w / w relative to the total solids,

[0288] - a total solids content in the range of 8-25% w / w, most preferably 12-22% w / w,

[0289] - 30-85% w / w, most preferably 35-70% w / w whey protein nanogel relative to total protein, and

[0290] - 15-70% w / w, and most preferably 30-65% w / w, of soluble whey protein aggregates relative to total protein.

[0291] It is particularly preferred that the dryer feed contains carbohydrates in an amount of at most 15% w / w relative to the total solids, more preferably at most 5% w / w relative to the total solids, even more preferably at most 1% w / w relative to the total solids, and most preferably at most 0.1% w / w relative to the total solids.

[0292] In some preferred embodiments of the present invention, the dryer feed contains carbohydrates in an amount in the range of 1-15% w / w, more preferably 5-15% w / w and most preferably 8-15% w / w relative to the total solids. The inventors have seen evidence that such embodiments can be used for whey protein nanogel compositions in powder form, as they appear to aid in the reconstitution and hydration of the powder. The carbohydrates preferably comprise or even consist of digestible carbohydrates such as sucrose, lactose, glucose, galactose and / or maltodextrin.

[0293] The carbohydrates may be present in the whey protein solution prior to the heat treatment in step b).However, in some preferred embodiments of the invention at least some of the carbohydrates of the dryer feed are added after the heat treatment in step b).

[0294] It is further particularly preferred that the dryer feed contains lipids in an amount of at most 8% w / w relative to the total solids, more preferably at most 2% w / w relative to the total solids, even more preferably at most 0.5% w / w relative to the total solids, and most preferably at most 0.1% w / w relative to the total solids.

[0295] In some preferred embodiments of the present invention, the method is a method for producing a whey protein nanogel composition in powder form, the method comprising:

[0296] a) providing a whey protein solution, the solution having:

[0297] - natural BLG content in an amount of 8-30% w / w, most preferably 12-24% w / w,

[0298] - a native BLG content in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein,

[0299] - a total protein content in an amount of at least 80% w / w, most preferably at least 90% w / w relative to the total solids,

[0300] - a pH value in the range of 5.8-7.5, most preferably 5.8-6.5,

[0301] -The weight ratio between total calcium and natural BLG is:

[0302] - at most 0.0041 x pH - 0.0209, and

[0303] - at least 0.0037 x pH - 0.0234 but greater than 0, and

[0304] - The following total concentrations of monovalent metal cations:

[0305] - if the native BLG content of the whey protein solution is less than 10% w / w, then at most 20 mM, and most preferably at most 15 mM, or

[0306] - if the native BLG content of the whey protein solution is at least 10% w / w, then at most 15 mM, and most preferably at most 10 mM,

[0307] b) heating the whey protein solution to 80-100°C, most preferably at least 85-95°C, for a time sufficient to form a suspension of whey protein nanogels, the time being sufficient to denature at least 90%, most preferably at least 95%, of the native BLG,

[0308] c) optionally, concentrating the suspension of whey protein nanogels to obtain a concentrated suspension of whey protein nanogels,

[0309] d) drying a dryer feed comprising the whey protein nanogel originating from step b) or c) by spray drying, preferably the dryer feed is the suspension of step b) or the concentrated suspension of step c).

[0310] In some preferred embodiments of the present invention, the method is a method for producing a whey protein nanogel composition in powder form, the method comprising:

[0311] a) providing a whey protein solution, the solution having:

[0312] - natural BLG content in an amount of 8-30% w / w, most preferably 12-24% w / w,

[0313] - a native BLG content of at least 90% w / w, most preferably at least 95% w / w relative to the total protein,

[0314] - a total protein content in an amount of at least 80% w / w, most preferably at least 90% w / w relative to the total solids,

[0315] - a pH value in the range of 5.8-7.5, most preferably 5.8-6.5,

[0316] -The weight ratio between total calcium and natural BLG is:

[0317] - at most 0.0041 x pH - 0.0209, and

[0318] - at least 0.0037 x pH - 0.0234 but greater than 0, and

[0319] - The following total concentrations of monovalent metal cations:

[0320] - if the native BLG content of the whey protein solution is less than 10% w / w, then at most 20 mM, and most preferably at most 15 mM, or

[0321] - if the native BLG content of the whey protein solution is at least 10% w / w, then at most 15 mM, and most preferably at most 10 mM,

[0322] b) heating the whey protein solution to 80-100°C, most preferably at least 85-95°C, for a time sufficient to form a suspension of whey protein nanogels, which time is sufficient to denature at least 90%, most preferably at least 95% of the native BLG,

[0323] c) if the concentration of the whey protein nanogel in the suspension of step b) is lower than the desired whey protein nanogel concentration of step d), concentrating the suspension of whey protein nanogel to obtain a whey protein nanogel concentrated suspension having the desired whey protein nanogel concentration,

[0324] d) drying a dryer feed by spray drying, the dryer feed comprising the whey protein nanogel originating from step b) or c), preferably, the dryer feed is the suspension of step b) or the concentrated suspension of step c), the dryer feed comprising:

[0325] - a total BLG content in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein,

[0326] - a total protein content in an amount of at least 80% w / w, most preferably at least 90% w / w relative to the total solids,

[0327] - a total solids content in the range of 21-40% w / w, most preferably 24-35% w / w,

[0328] - whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein, and

[0329] - Soluble whey protein aggregates in an amount of at most 9% w / w, and most preferably at most 5% w / w, relative to total protein.

[0330] In some preferred embodiments of the present invention, the method is a method for producing a whey protein nanogel composition in powder form, the method comprising:

[0331] a) providing a whey protein solution, the solution having:

[0332] - natural BLG content in an amount of 16-30% w / w, most preferably 18-24% w / w,

[0333] - a native BLG content of at least 90% w / w, most preferably at least 95% w / w relative to the total protein,

[0334] - a total protein content in an amount of at least 80% w / w, most preferably at least 90% w / w relative to the total solids,

[0335] - a pH value in the range of 5.8-7.5, most preferably 5.8-6.5,

[0336] -The weight ratio between total calcium and natural BLG is:

[0337] - at most 0.0041 x pH - 0.0209, and

[0338] - at least 0.0037 x pH - 0.0234 but greater than 0, and

[0339] - The following total concentrations of monovalent metal cations:

[0340] - if the native BLG content of the whey protein solution is less than 10% w / w, then at most 20 mM, and most preferably at most 15 mM, or

[0341] - if the native BLG content of the whey protein solution is at least 10% w / w, then at most 15 mM, and most preferably at most 10 mM,

[0342] b) heating the whey protein solution to 80-100°C, most preferably at least 85-95°C, for a time sufficient to form a suspension of whey protein nanogels, which time is sufficient to denature at least 90%, most preferably at least 95% of the native BLG,

[0343] d) drying a dryer feed comprising the whey protein nanogel originating from step b) or c) by spray drying, the dryer feed being the suspension of step b).

[0344] In some preferred embodiments of the present invention, the method is a method for producing a whey protein nanogel composition in powder form, the method comprising:

[0345] a) providing a whey protein solution, the solution having:

[0346] - natural BLG content in an amount of 3-10% w / w, most preferably 4-8% w / w,

[0347] - a native BLG content in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein,

[0348] - a total protein content in an amount of at least 80% w / w, most preferably at least 90% w / w relative to the total solids,

[0349] - pH value in the range of 5.8-7.5, most preferably 5.8-6.5,

[0350] -The weight ratio between total calcium and natural BLG is:

[0351] - at most 0.0041 x pH - 0.0209, and

[0352] - at least 0.0037 x pH - 0.0234 but greater than 0, and

[0353] - The following total concentrations of monovalent metal cations:

[0354] - at most 15 mM, and most preferably at most 10 mM,

[0355] b) heating the whey protein solution to 80-100°C, most preferably at least 85-95°C, for a time sufficient to form a suspension of whey protein nanogels, which time is sufficient to denature at least 90%, most preferably at least 95% of the native BLG,

[0356] c) concentrating the suspension of whey protein nanogels to obtain a concentrated suspension of whey protein nanogels having the desired whey protein nanogel concentration in step d),

[0357] d) drying a dryer feed comprising the whey protein nanogel originating from step b) or c) by spray drying, the dryer feed being the concentrated suspension of step c), the dryer feed comprising:

[0358] - a total BLG content in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein,

[0359] - a total protein content in an amount of at least 80% w / w, most preferably at least 90% w / w relative to the total solids,

[0360] - a total solids content in the range of 21-40% w / w, most preferably 24-35% w / w,

[0361] - whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein, and

[0362] - Up to 9% w / w, and most preferably up to 5% w / w, soluble whey protein aggregates relative to total protein.

[0363] It is particularly preferred that the whey protein nanogel composition contains carbohydrates in an amount of at most 15% w / w relative to the total solids, more preferably at most 5% w / w relative to the total solids, even more preferably at most 1% w / w relative to the total solids, and most preferably at most 0.1% w / w relative to the total solids.

[0364] In some preferred embodiments of the invention, the whey protein nanogel composition contains carbohydrates in an amount of 1-15% w / w relative to the total solids, more preferably 5-15% w / w relative to the total solids and most preferably 8-15% w / w. The inventors have seen evidence that such embodiments can be used for whey protein nanogel compositions in powder form, as they appear to aid in the reconstitution and hydration of the powder. The carbohydrates preferably comprise or even consist of digestible carbohydrates such as sucrose, lactose, glucose, galactose and / or maltodextrin.

[0365] It is further particularly preferred that the whey protein nanogel composition contains lipid in an amount of at most 8% w / w relative to the total solids, more preferably at most 2% w / w relative to the total solids, even more preferably at most 0.5% w / w relative to the total solids, and most preferably at most 0.1% w / w relative to the total solids.

[0366] In some preferred embodiments of the present invention, the method is a method for producing a whey protein nanogel composition in powder form, the method comprising:

[0367] a) providing a whey protein solution, the solution having:

[0368] - natural BLG content in an amount of 8-20% w / w, most preferably 12-18% w / w,

[0369] - natural protein in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein

[0370] BLG content,

[0371] - a total protein content in an amount of at least 80% w / w, most preferably at least 90% w / w relative to the total solids,

[0372] - pH value in the range of 5.8-7.5, most preferably 5.9-6.5,

[0373] -The weight ratio between total calcium and natural BLG is:

[0374] - at most 0.0041 x pH - 0.0209, and

[0375] - at least 0.0037 x pH - 0.0234 but greater than 0, and

[0376] - The following total concentrations of monovalent metal cations:

[0377] - if the whey protein solution has a native BLG content of less than 10% w / w,

[0378] 18-25 mM, and most preferably 20-25 mM, or

[0379] - if the whey protein solution has a native BLG content of at least 10% w / w

[0380] 15-20mM, most preferably 17-20mM,

[0381] b) heating the whey protein solution to 80-100°C, most preferably at least 85-95°C, for a time sufficient to form a suspension of whey protein nanogels, which time is sufficient to denature at least 90%, most preferably at least 95% of the native BLG,

[0382] c) optionally, concentrating the suspension of whey protein nanogels to obtain a concentrated suspension of whey protein nanogels,

[0383] d) drying a dryer feed by spray drying, the dryer feed comprising the whey protein nanogel originating from step b) or c), preferably the dryer feed is the suspension of step b) or the concentrated suspension of step c), preferably the dryer feed comprises:

[0384] - a total BLG content in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein,

[0385] - a total protein content in an amount of at least 80% w / w, most preferably at least 90% w / w relative to the total solids,

[0386] - a total solids content in the range of 8-25% w / w, most preferably 12-22% w / w,

[0387] - whey protein nanogel in an amount of 30-85% w / w, most preferably 35-70% w / w relative to total protein, and

[0388] - Soluble whey protein aggregates in an amount of 15-70% w / w, and most preferably 30-65% w / w relative to total protein.

[0389] Yet another aspect of the present invention relates to a whey protein nanogel composition comprising whey protein nanogel in an amount of at least 30% relative to total protein and a total amount of protein of at least 30% w / w relative to total solids. The whey protein nanogel composition is preferably obtainable by the process described herein.

[0390] In some preferred embodiments of the present invention, the whey protein nanogel composition comprises whey protein nanogel in an amount of at least 60% w / w relative to the total protein, more preferably at least 70% w / w relative to the total protein, even more preferably at least 80% w / w, and most preferably at least 90% w / w.

[0391] Even higher contents of whey protein nanogel are generally preferred, and in other preferred embodiments of the present invention, the whey protein nanogel composition comprises whey protein nanogel in an amount of at least 92% w / w relative to the total protein, more preferably at least 94% w / w relative to the total protein, even more preferably at least 96% w / w, and most preferably at least 98% w / w.

[0392] It may even be preferred that the whey protein nanogel composition contains whey protein nanogel in an amount of about 100% w / w relative to the total protein.

[0393] In some preferred embodiments of the present invention, the whey protein nanogel composition comprises soluble whey protein aggregates in an amount of at most 30% w / w relative to the total protein, more preferably at most 20% w / w relative to the total protein, even more preferably at most 10% w / w, and most preferably at most 5% w / w.

[0394] Even lower contents of soluble whey protein aggregates are generally preferred, and the whey protein nanogel composition preferably comprises soluble whey protein aggregates in an amount of at most 3% w / w relative to the total protein, more preferably at most 2% w / w relative to the total protein, even more preferably at most 1% w / w, and most preferably at most 0.5% w / w.

[0395] In other preferred embodiments of the present invention, the whey protein nanogel composition comprises soluble whey protein aggregates in an amount of at most 30% w / w relative to the total weight of the composition, more preferably at most 20% w / w relative to the total weight of the composition, even more preferably at most 10% w / w, and most preferably at most 5% w / w.

[0396] Even lower contents of soluble whey protein aggregates are generally preferred, and the whey protein nanogel composition preferably comprises soluble whey protein aggregates in an amount of at most 3% w / w relative to the total weight of the composition, more preferably at most 2% w / w relative to the total weight of the composition, even more preferably at most 1% w / w relative to the total weight of the composition, and most preferably at most 0.5% w / w relative to the total weight of the composition.

[0397] The whey protein nanogel composition having a low content of soluble whey protein aggregates is particularly suitable for high protein acidic beverages.

[0398] The inventors have seen evidence that such whey protein nanogel compositions with increased whey protein nanogel content appear to undergo slower gastrointestinal digestion and form less gel than soluble whey protein aggregates in the acidic environment of the stomach. Therefore, the whey protein nanogels appear to contribute less to structural development and satiety than soluble whey protein aggregates. This makes these whey protein nanogels suitable for clinical nutrition in high-energy drinks of any pH that should not promote satiety.

[0399] The inventors have observed that whey protein nanogel compositions with significant contents of soluble whey protein aggregates are stable to heat treatment and further produce gels upon acidification. This makes such whey protein nanogel compositions very suitable for use in acidic thickened foods produced by pasteurizing a liquid food base and subsequently acidifying the heat treated food base. For example, this is suitable for use in place of carbohydrate based hydrocolloids such as starch, gums or pectins.

[0400] In some preferred embodiments of the present invention, the whey protein nanogel composition comprises:

[0401] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0402] - whey protein nanogel in an amount of 30-85% w / w, most preferably 35-70% w / w relative to total protein, and

[0403] - Soluble whey protein aggregates in an amount of 15-70% w / w, and most preferably 30-65% w / w relative to total protein.

[0404] In other preferred embodiments of the present invention, the whey protein nanogel composition comprises:

[0405] - a total amount of BLG of at least 50% w / w, most preferably at least 60% w / w relative to total protein,

[0406] - whey protein nanogel in an amount of 30-85% w / w, most preferably 35-70% w / w relative to total protein, and

[0407] - Soluble whey protein aggregates in an amount of 15-70% w / w, and most preferably 30-65% w / w relative to total protein.

[0408] The inventors have seen that these whey protein nanogel compositions with increased soluble whey protein aggregate content appear to undergo faster gastrointestinal digestion and are thought to form gels in the acidic environment of the stomach and promote structural development and satiety. This makes these whey protein nanogel compositions suitable for beverages that can promote improved satiety and / or rapid digestion of protein.

[0409] The whey protein nanogel composition containing soluble whey protein aggregates in an amount of 15-70% w / w and most preferably 30-65% w / w relative to the total protein can be used as a food ingredient for producing a thickened food having a pH value of less than 5.5 and having a viscosity of 100 μg / ml at 20°C and 300 s -1 The viscosity measured under shear rate is greater than 200 cP.

[0410] It is generally preferred that the whey protein nanogel composition of the present invention contains a small amount of larger whey protein particles, which may cause precipitation in the beverage, or even a sandy impression. Therefore, in some preferred embodiments of the present invention, the whey protein nanogel composition comprises whey protein microparticles in an amount of at most 10% w / w relative to the total protein, more preferably at most 5% w / w relative to the total protein, even more preferably at most 3% w / w, and most preferably at most 1% w / w. The content of whey protein microparticles was determined according to Assay 3.

[0411] The whey protein nanogel composition of this embodiment typically contains less than 4% w / w whey protein particles, if any.

[0412] In some preferred embodiments of the present invention, the whey protein nanogel composition comprises total protein in an amount of at least 30% w / w relative to the total solids, more preferably at least 60% w / w relative to the total solids, even more preferably at least 70% w / w relative to the total solids, and most preferably at least 80% w / w relative to the total solids.

[0413] In other preferred embodiments of the present invention, the whey protein nanogel composition comprises total protein in an amount of at least 85% w / w relative to the total solids, more preferably at least 90% w / w relative to the total solids, even more preferably at least 92% w / w relative to the total solids, and most preferably at least 95% w / w relative to the total solids.

[0414] In some preferred embodiments of the present invention, the whey protein nanogel composition comprises total protein in an amount of at least 30% w / w, more preferably at least 60% w / w, even more preferably at least 70% w / w, and most preferably at least 80% w / w.

[0415] In other preferred embodiments of the present invention, the whey protein nanogel composition comprises total protein in an amount of at least 85% w / w, more preferably at least 90% w / w, even more preferably at least 92% w / w, and most preferably at least 95% w / w.

[0416] In some preferred embodiments of the present invention, the whey protein nanogel composition comprises total BLG in an amount of at least 50% w / w relative to total protein, more preferably at least 60% w / w relative to total protein, even more preferably at least 70% w / w relative to total protein, and most preferably at least 80% w / w relative to total protein.

[0417] In other preferred embodiments of the present invention, the whey protein nanogel composition comprises total BLG in an amount of at least 85% w / w relative to total protein, more preferably at least 90% w / w relative to total protein, even more preferably at least 92% w / w relative to total protein, and most preferably at least 95% w / w relative to total protein. It is generally preferred that the whey protein nanogel composition comprises total BLG in an amount of at least 97% w / w relative to total protein.

[0418] In a further preferred embodiment of the present invention, the whey protein nanogel composition comprises total BLG in an amount in the range of 50-80% w / w relative to total protein, more preferably in the range of 52-75% w / w relative to total protein, even more preferably in the range of 54-70% w / w relative to total protein, and most preferably in the range of 55-65% w / w relative to total protein.

[0419] In the context of the present invention, the terms "amount of total BLG" and "total amount of BLG" are used interchangeably and both relate to the total amount of BLG of the composition in question, including native and denatured BLG.

[0420] In some preferred embodiments of the present invention, the whey protein nanogel composition is liquid and comprises total solids in an amount of 1-50% w / w, more preferably in an amount of 10-45% w / w, even more preferably in an amount of 15-40% w / w, and most preferably in an amount of 20-35% w / w.

[0421] Preferably, if the whey protein nanogel composition is liquid, it comprises water in an amount of 50-99% w / w, more preferably in an amount of 55-90% w / w, even more preferably in an amount of 60-85% w / w, and most preferably in an amount of 65-80% w / w.

[0422] In case of a liquid whey protein nanogel composition, it is preferred that soluble whey protein aggregates are present in an amount of at most 8% w / w, more preferably at most 6%, even more preferably at most 3% w / w, and most preferably at most 1% w / w.

[0423] In some preferred embodiments of the present invention, the whey protein nanogel composition is a powder and comprises total solids in an amount of 90-99% w / w, more preferably in an amount of 92-99% w / w, even more preferably in an amount of 94-99% w / w, and most preferably in an amount of 95-99% w / w.

[0424] Preferably, if the whey protein nanogel composition is a powder, it comprises water in an amount of 1-10% w / w, more preferably 1-8% w / w, even more preferably 1-6% w / w, and most preferably 1-5% w / w.

[0425] In some preferred embodiments of the present invention, the whey protein nanogel composition is a paste and comprises total solids in an amount of 51-89% w / w, more preferably in an amount of 55-90% w / w, even more preferably in an amount of 60-85% w / w, and most preferably in an amount of 65-80% w / w.

[0426] Preferably, if the whey protein nanogel composition is a powder, it comprises water in an amount of 11-49% w / w, more preferably in an amount of 13-45% w / w, even more preferably in an amount of 15-40% w / w, and most preferably in an amount of 20-35% w / w.

[0427] Although a wide range of pH is possible, the whey protein nanogel composition preferably has a pH in the range of 3-8, more preferably 4-7, even more preferably 5-7, and most preferably 6-7.

[0428] If the whey protein nanogel composition contains a large amount of soluble whey protein aggregates, it is preferred that the pH is in the range of 5.7-8, and most preferably in the range of 6.0-7.5.

[0429] The present inventors have found that the pH neutral variant of the whey protein nanogel composition is particularly suitable for pH neutral food applications, where the pH of the final food, such as a beverage, is close to the pH of the whey protein nanogel composition. This reduces the risk of unwanted protein aggregation during protein processing, particularly during pH adjustment.

[0430] In some preferred embodiments of the present invention, the pH range of the whey protein nanogel composition is 6-8, most preferably 6-7. In other preferred embodiments of the present invention, the pH range of the whey protein nanogel composition is 7-8.

[0431] In a further preferred embodiment of the present invention, the pH range of the whey protein nanogel composition is 6.5-8.0, and more preferably 6.6-7.5, even more preferably 6.8-7.5, and most preferably 6.9-7.5.

[0432] Similar to the case of the pH neutral variant of the whey protein nanogel composition, the inventors have found that the acidic variant of the whey protein nanogel composition is particularly suitable for acidic food applications, where the pH of the final food, such as a beverage, is close to the pH of the whey protein nanogel composition; because it reduces the risk of unwanted protein aggregation occurring during protein processing, particularly during pH adjustment.

[0433] In some preferred embodiments of the present invention, the pH range of the whey protein nanogel composition is 3-5, most preferably 3-4.

[0434] In other preferred embodiments of the present invention, the pH range of the whey protein nanogel composition is 4-5.

[0435] In a further preferred embodiment of the present invention, the pH range of the whey protein nanogel composition is 3-5.5, more preferably 3.0-5.0, even more preferably 3.0-4.7, and most preferably 3.5-4.5.

[0436] The nanogels of the present invention typically have a particle size in the nanometer range. In some preferred embodiments of the present invention, the particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-1000 nm, more preferably 170-900 nm, even more preferably 200-800 nm, and most preferably 250-700 nm as measured by dynamic light scattering.

[0437] In some preferred embodiments of the present invention, the particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-800 nm, more preferably 250-700 nm, even more preferably 300-600 nm and most preferably 350-500 nm.

[0438] If the nanogels are provided as powdered whey protein nanogel compositions, the measurement of particle size and polydispersity index requires reconstitution and hydration of the whey protein nanogels in water prior to measuring particle size, and preferably subjecting the reconstituted whey protein nanogels to homogenization to ensure that they are hydrated.

[0439] In some preferred embodiments of the present invention, the particles of the whey protein nanogel composition have a polydispersity index of at most 0.3, more preferably at most 0.2, even more preferably at most 0.1, and most preferably at most 0.05.

[0440] If the whey protein nanogel composition is prepared, for example, by directly drying the suspension of the whey protein nanogel of step b) or the concentrated suspension of step c), it contains the same or slightly less minerals calculated on total solids than the whey protein solution.

[0441] Therefore, in some preferred embodiments of the present invention, the whey protein nanogel composition has a weight ratio between the total amount of calcium and native BLG of at most 0.0070, more preferably at most 0.0065, even more preferably at most 0.0060, and most preferably at most 0.0050.

[0442] In other preferred embodiments of the present invention, the whey protein nanogel composition has a weight ratio between the total amount of calcium and the total BLG of at most 0.0045, more preferably at most 0.0040, even more preferably at most 0.0035 and most preferably 0.0030.

[0443] Even lower weight ratios are generally preferred, and in some preferred embodiments of the invention the whey protein nanogel composition has a weight ratio between total calcium and total BLG of at most 0.0025, more preferably at most 0.0020, even more preferably at most 0.0015, and most preferably 0.0010.

[0444] In other preferred embodiments of the present invention, the weight ratio between the total amount of calcium and the total BLG of the whey protein nanogel composition is in the range of 0.0010-0.0030, and most preferably in the range of 0.0017-0.0027.

[0445] The whey protein nanogel composition may preferably have a total concentration of monovalent metal cations of at most 20 mM, more preferably at most 15 mM, even more preferably at most 10 mM, and more preferably at most 5 mM when standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water.

[0446] In some preferred embodiments of the present invention, the whey protein nanogel composition has a total content of monovalent metal ions of at most 0.5% w / w relative to the total solids, more preferably at most 0.3% w / w, even more preferably at most 0.1% w / w relative to the total solids, and most preferably at most 0.01% w / w relative to the total solids.

[0447] Although the present method limits the weight ratio between calcium and native BLG during the heat treatment in step b), it is possible to enrich the resulting whey protein nanogel suspension with calcium and other minerals. This may be advantageous, for example, if the whey protein nanogel composition is used to enrich the mineral content of a food.

[0448] Therefore, in some preferred embodiments of the present invention, the weight ratio between the total amount of calcium and the total BLG of the whey protein nanogel composition is at least 0.005, more preferably at least 0.01, even more preferably at least 0.02, and most preferably at least 0.03.

[0449] In other preferred embodiments of the present invention, the weight ratio of the total amount of calcium to the total BLG of the whey protein nanogel composition is 0.005-0.050, more preferably 0.010-0.045, even more preferably 0.015-0.040, and most preferably 0.020-0.035.

[0450] In some preferred embodiments of the present invention, the whey protein nanogel composition has a total concentration of monovalent metal cations greater than 20 mM, more preferably at least 25 mM, even more preferably at least 50 mM, and more preferably at least 100 mM when standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water.

[0451] In other preferred embodiments of the present invention, the whey protein nanogel composition has a total concentration of monovalent metal cations of 20-400 mM, more preferably at least 40-350 mM, even more preferably 50-300 mM, and most preferably 60-250 mM when standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water.

[0452] The whey protein nanogel composition may comprise other macronutrients besides protein, such as carbohydrates and / or lipids.

[0453] However, it is generally preferred that the whey protein nanogel composition contains carbohydrates in an amount of at most 15% w / w relative to the total solids, more preferably at most 5% w / w relative to the total solids, even more preferably at most 1% w / w relative to the total solids, and most preferably at most 0.1% w / w relative to the total solids.

[0454] In some preferred embodiments of the present invention, the whey protein nanogel composition contains carbohydrates in an amount of 1-15% w / w relative to the total solids, more preferably 5-15% w / w relative to the total solids and most preferably 8-15% w / w. The inventors have seen evidence that such embodiments are suitable for whey protein nanogel compositions in powder form, as they appear to aid in the reconstitution and hydration of the powder. The carbohydrates preferably comprise or even consist of digestible carbohydrates such as sucrose, lactose, glucose, galactose and / or maltodextrin.

[0455] Furthermore, it is generally preferred that the whey protein nanogel composition contains lipid in an amount of at most 8% w / w relative to the total solids, more preferably at most 2% w / w relative to the total solids, even more preferably at most 0.5% w / w relative to the total solids, and most preferably at most 0.1% w / w relative to the total solids.

[0456] The whey protein nanogel composition may contain other macronutrients and ingredients in addition to protein. The embodiments and preferences described below in the context of packaged heat-processed beverages relating to macronutrients and additional ingredients are equally applicable to the whey protein nanogel composition and we refer to these embodiments here rather than repeating them.

[0457] In some particularly preferred embodiments of the present invention, the whey protein nanogel composition has the same chemical composition as the packaged heat-processed beverage, except that the protein is in native form.

[0458] In other particularly preferred embodiments of the present invention, the whey protein nanogel composition has the same chemical composition as the packaged heat-processed beverage.

[0459] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form, and the whey protein nanogel composition comprises:

[0460] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0461] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0462] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0463] - whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein, and

[0464] - soluble whey protein aggregates in an amount of up to 9% w / w, and most preferably up to 5% w / w, relative to the total protein,

[0465] And one or more of the following:

[0466] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0467] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 150 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 100 cP at a shear rate of

[0468] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0469] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form, and the whey protein nanogel composition comprises:

[0470] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0471] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0472] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0473] - at least 70% w / w, most preferably at least 80% w / w whey protein nanogel relative to total protein, and

[0474] - soluble whey protein aggregates in an amount of at most 9% w / w, and more preferably at most 5% w / w, relative to the total protein,

[0475] And one or more of the following:

[0476] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0477] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 150 cP, and most preferably at 20 ° C and 300 s-1 has a viscosity of at most 100 cP at a shear rate of

[0478] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0479] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form, and the whey protein nanogel composition comprises:

[0480] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0481] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0482] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0483] - at least 80% w / w, most preferably at least 85% w / w whey protein nanogel relative to total protein, and

[0484] - up to 20% w / w, and most preferably up to 15% w / w, of soluble whey protein aggregates relative to total protein,

[0485] And one or more of the following:

[0486] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0487] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 300 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 200 cP at a shear rate of

[0488] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0489] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form, and the whey protein nanogel composition comprises:

[0490] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0491] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0492] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0493] - whey protein nanogel in an amount of at least 70% w / w, most preferably at least 80% w / w relative to the total protein, and

[0494] - soluble whey protein aggregates in an amount of up to 9% w / w, and most preferably up to 5% w / w, relative to the total protein,

[0495] And one or more of the following:

[0496] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0497] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 300 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 200 cP at a shear rate of

[0498] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0499] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form, and the whey protein nanogel composition comprises:

[0500] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0501] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0502] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0503] - whey protein nanogel in an amount of 30-85% w / w, most preferably 35-70% w / w relative to total protein, and

[0504] - soluble whey protein aggregates in an amount of 15-70% w / w, and most preferably 30-65% w / w, relative to the total protein,

[0505] And one or more of the following:

[0506] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering, and

[0507] iii) the whey protein nanogel composition particles in hydrated form have a polydispersity index of at most 0.2.

[0508] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form, and the whey protein nanogel composition comprises:

[0509] - at least 92% w / w, most preferably at least 94% w / w, total solids relative to the total weight of the composition,

[0510] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0511] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0512] - whey protein nanogel in an amount of 30-59% w / w, most preferably 35-50% w / w relative to total protein, and

[0513] - soluble whey protein aggregates in an amount of 21-50% w / w, and most preferably 30-45% w / w, relative to the total protein,

[0514] And one or more of the following:

[0515] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering, and

[0516] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0517] The powders of the invention are preferably dried by spray drying.

[0518] In some preferred embodiments of the present invention, the whey protein nanogel composition is in liquid form, and the whey protein nanogel composition comprises:

[0519] - total protein in an amount of 21-35% w / w, most preferably 24-32% w / w, relative to the total weight of the whey protein nanogel composition,

[0520] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0521] - at least 90% w / w, most preferably at least 95% w / w, total BLG relative to total protein,

[0522] - whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein, and

[0523] - soluble whey protein aggregates in an amount of up to 9% w / w, and most preferably in an amount of up to 5% w / w relative to the total protein,

[0524] And one or more of the following:

[0525] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering, and

[0526] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 has a viscosity of at most 200 cP at a shear rate of

[0527] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0528] In some preferred embodiments of the present invention, when standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, the whey protein nanogel composition in liquid form is stable at 20°C and 300s -1 The shear rate is at most 200 cP, more preferably at 20°C and 300 s -1 The shear rate has a maximum of 100 cP at a shear rate of 20°C and 300 s -1 The shear rate is at most 50 cP, and most preferably at 20 ° C and 300 s -1 The viscosity of the composite is at most 20 cP at a shear rate of 1.5 %.

[0529] Although other methods can be used to prepare the whey protein nanogel composition of the present invention, it is preferred that the whey protein nanogel composition is obtainable by the method described herein.

[0530] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 6-8, most preferably 7-8, the whey protein nanogel composition comprising:

[0531] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0532] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0533] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0534] - whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein, and

[0535] - soluble whey protein aggregates in an amount of up to 9% w / w, and most preferably up to 5% w / w, relative to the total protein,

[0536] And one or more of the following:

[0537] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0538] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 150 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 100 cP at a shear rate of

[0539] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0540] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 6-8, most preferably 7-8, the whey protein nanogel composition comprising:

[0541] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0542] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0543] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0544] - whey protein nanogel in an amount of at least 70% w / w, most preferably at least 80% w / w relative to the total protein, and

[0545] - soluble whey protein aggregates in an amount of at most 9% w / w, and more preferably at most 5% w / w, relative to the total protein,

[0546] And one or more of the following:

[0547] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0548] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 150 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 100 cP at a shear rate of

[0549] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0550] In other preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 6-8, most preferably 7-8, said whey protein nanogel composition comprising:

[0551] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0552] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0553] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0554] - whey protein nanogel in an amount of at least 50% w / w, most preferably at least 60% w / w relative to the total protein, and

[0555] - soluble whey protein aggregates in an amount of at most 9% w / w, and more preferably at most 5% w / w, relative to the amount of total protein,

[0556] And one or more of the following:

[0557] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0558] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 150 cP, and most preferably at 20 ° C and 300 s -1has a viscosity of at most 100 cP at a shear rate of

[0559] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0560] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 6-8, most preferably 7-8, the whey protein nanogel composition comprising:

[0561] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0562] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0563] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0564] - whey protein nanogel in an amount of at least 80% w / w, most preferably at least 85% w / w relative to the total protein, and

[0565] - soluble whey protein aggregates in an amount of up to 20% w / w, and most preferably up to 15% w / w relative to the total protein,

[0566] And one or more of the following:

[0567] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0568] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 300 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 200 cP at a shear rate of

[0569] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0570] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 6-8, most preferably 7-8, the whey protein nanogel composition comprising:

[0571] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0572] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0573] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0574] - whey protein nanogel in an amount of at least 70% w / w, most preferably at least 80% w / w relative to the total protein, and

[0575] - soluble whey protein aggregates in an amount of up to 9% w / w, and most preferably up to 5% w / w, relative to the total protein,

[0576] And one or more of the following:

[0577] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0578] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 300 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 200 cP at a shear rate of

[0579] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0580] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 6-8, most preferably 7-8, the whey protein nanogel composition comprising:

[0581] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0582] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0583] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0584] - whey protein nanogel in an amount of at least 50% w / w, most preferably at least 60% w / w relative to the total protein, and

[0585] - soluble whey protein aggregates in an amount of up to 9% w / w, and most preferably up to 5% w / w, relative to the total protein,

[0586] And one or more of the following:

[0587] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0588] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 300 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 200 cP at a shear rate of

[0589] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0590] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 6-8, most preferably 7-8, the whey protein nanogel composition comprising:

[0591] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0592] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0593] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0594] - whey protein nanogel in an amount of 30-85% w / w, most preferably 35-70% w / w relative to total protein, and

[0595] - soluble whey protein aggregates in an amount of 15-70% w / w, and most preferably 30-65% w / w, relative to the total protein,

[0596] And one or more of the following:

[0597] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering, and

[0598] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0599] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 6-8, most preferably 7-8, the whey protein nanogel composition comprising:

[0600] - at least 92% w / w, most preferably at least 94% w / w, total solids relative to the total weight of the composition,

[0601] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0602] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0603] - whey protein nanogel in an amount of 30-59% w / w, most preferably 35-50% w / w relative to total protein, and

[0604] - soluble whey protein aggregates in an amount of 21-50% w / w, and most preferably 30-45% w / w, relative to the total protein,

[0605] And one or more of the following:

[0606] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering, and

[0607] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0608] The powders of the invention are preferably dried by spray drying.

[0609] In some preferred embodiments of the present invention, the whey protein nanogel composition is in liquid form and has a pH in the range of 6-8, most preferably 7-8, the whey protein nanogel composition comprising:

[0610] - total protein in an amount of 21-35% w / w, most preferably 24-32% w / w, relative to the total weight of the whey protein nanogel composition,

[0611] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0612] - at least 90% w / w, most preferably at least 95% w / w, total BLG relative to total protein,

[0613] - whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein, and

[0614] - soluble whey protein aggregates in an amount of up to 9% w / w, and most preferably in an amount of up to 5% w / w, relative to the total protein,

[0615] And one or more of the following:

[0616] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering, and

[0617] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 has a viscosity of at most 200 cP at a shear rate of

[0618] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0619] In some preferred embodiments of the present invention, the whey protein nanogel composition is in liquid form and has a pH in the range of 6-8, most preferably 7-8, the whey protein nanogel composition comprising:

[0620] - total protein in an amount of 21-35% w / w, most preferably 24-32% w / w, relative to the total weight of the whey protein nanogel composition,

[0621] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0622] - at least 50% w / w, most preferably at least 55% w / w, total BLG relative to total protein,

[0623] - whey protein nanogel in an amount of at least 50% w / w, most preferably at least 55% w / w relative to the total protein, and

[0624] - soluble whey protein aggregates in an amount of up to 9% w / w, and most preferably up to 5% w / w, relative to the total protein,

[0625] And one or more of the following:

[0626] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering, and

[0627] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 has a viscosity of at most 200 cP at a shear rate of

[0628] iii) the whey protein nanogel composition particles in hydrated form have a polydispersity index of at most 0.2.

[0629] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 3-5, most preferably 3-4, the whey protein nanogel composition comprising:

[0630] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0631] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0632] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0633] - whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein, and

[0634] - soluble whey protein aggregates in an amount of up to 9% w / w, and most preferably up to 5% w / w, relative to the total protein,

[0635] And one or more of the following:

[0636] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0637] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 150 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 100 cP at a shear rate of

[0638] iii) the whey protein nanogel composition particles in hydrated form have a polydispersity index of at most 0.2.

[0639] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 3-5, most preferably 3-4, the whey protein nanogel composition comprising:

[0640] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0641] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0642] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0643] - whey protein nanogel in an amount of at least 70% w / w, most preferably at least 80% w / w relative to the total protein, and

[0644] - soluble whey protein aggregates in an amount of at most 9% w / w, and more preferably at most 5% w / w, relative to the total protein,

[0645] And one or more of the following:

[0646] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0647] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 150 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 100 cP at a shear rate of

[0648] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0649] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 3-5, most preferably 3-4, the whey protein nanogel composition comprising:

[0650] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0651] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0652] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0653] - whey protein nanogel in an amount of at least 50% w / w, most preferably at least 60% w / w relative to the total protein, and

[0654] - soluble whey protein aggregates in an amount of at most 9% w / w, and more preferably at most 5% w / w, relative to the amount of total protein,

[0655] And one or more of the following:

[0656] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0657] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 150 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 100 cP at a shear rate of

[0658] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0659] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 3-5, most preferably 3-4, the whey protein nanogel composition comprising:

[0660] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0661] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0662] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0663] - whey protein nanogel in an amount of at least 80% w / w, most preferably at least 85% w / w relative to the total protein, and

[0664] - soluble whey protein aggregates in an amount of up to 20% w / w, and most preferably up to 15% w / w relative to the total protein,

[0665] And one or more of the following:

[0666] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0667] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 300 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 200 cP at a shear rate of

[0668] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0669] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 3-5, most preferably 3-4, the whey protein nanogel composition comprising:

[0670] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0671] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0672] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0673] - whey protein nanogel in an amount of at least 70% w / w, most preferably at least 80% w / w relative to the total protein, and

[0674] - up to 9% w / w, and most preferably up to 5% w / w, of soluble whey protein aggregates relative to total protein,

[0675] And one or more of the following:

[0676] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0677] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 300 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 200 cP at a shear rate of

[0678] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0679] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 3-5, most preferably 3-4, the whey protein nanogel composition comprising:

[0680] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0681] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0682] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0683] - whey protein nanogel in an amount of at least 50% w / w, most preferably at least 60% w / w relative to the total protein, and

[0684] - soluble whey protein aggregates in an amount of up to 9% w / w, and most preferably up to 5% w / w, relative to the total protein,

[0685] And one or more of the following:

[0686] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering,

[0687] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 The shear rate is at most 300 cP, and most preferably at 20 ° C and 300 s -1 has a viscosity of at most 200 cP at a shear rate of

[0688] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0689] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form and has a pH in the range of 3-5, most preferably 3-4, the whey protein nanogel composition comprising:

[0690] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0691] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0692] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0693] - 30-85% w / w, most preferably 35-70% w / w whey protein nanogel relative to total protein, and

[0694] - soluble whey protein aggregates in an amount of 15-70% w / w, and most preferably 30-65% w / w, relative to the total protein,

[0695] And one or more of the following:

[0696] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering, and

[0697] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0698] In some preferred embodiments of the present invention, the whey protein nanogel composition is in powder form, having a pH in the range of 3-5, most preferably 3-4, and the whey protein nanogel composition comprises:

[0699] - total solids in an amount of at least 92% w / w, most preferably at least 94% w / w relative to the total weight of the composition,

[0700] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0701] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0702] - whey protein nanogel in an amount of 30-59% w / w, most preferably 35-50% w / w relative to total protein, and

[0703] - soluble whey protein aggregates in an amount of 21-50% w / w, and most preferably 30-45% w / w, relative to the total protein,

[0704] And one or more of the following:

[0705] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering, and

[0706] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0707] The powders of the invention are preferably dried by spray drying.

[0708] In some preferred embodiments of the present invention, the whey protein nanogel composition is in liquid form and has a pH in the range of 3-5, most preferably 3-4, the whey protein nanogel composition comprising:

[0709] - total protein in an amount of 21-35% w / w, most preferably 24-32% w / w, relative to the total weight of the whey protein nanogel composition,

[0710] - total protein in an amount of at least 80% w / w, most preferably at least 90% w / w relative to total solids,

[0711] - at least 90% w / w, most preferably at least 95% w / w, total BLG relative to total protein,

[0712] - whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein, and

[0713] - soluble whey protein aggregates in an amount of up to 9% w / w, and most preferably up to 5% w / w, relative to the total protein,

[0714] And one or more of the following:

[0715] i) particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-900 nm as measured by dynamic light scattering, and

[0716] ii) When standardized to 20% w / w total protein by vacuum evaporation or addition of milli-Q water, at 20°C and 300 s -1 has a viscosity of at most 200 cP at a shear rate of

[0717] iii) the particles of the hydrated form of the whey protein nanogel composition have a polydispersity index of at most 0.2.

[0718] The whey protein nanogel composition is preferably prepared by heat denaturing whey protein at a pH in the range of 5.8-7.5 or in one of the preferred sub-ranges described in the context of the whey protein solution in step a).

[0719] Yet another aspect of the present invention relates to whey protein nanogels obtainable by the methods described herein. Individual whey protein nanogels can be isolated, for example, by microfiltration, differential centrifugation and density gradient centrifugation.

[0720] The whey protein nanogels of the present invention in various hydrated forms preferably have a z-average diameter of 150-1000 nm, more preferably 170-900 nm, even more preferably 200-800 nm and most preferably 250-700 nm.

[0721] In some preferred embodiments of the present invention, the various hydrated forms of the whey protein nanogels of the present invention have a z-average diameter of 150-800 nm, more preferably 250-700 nm, even more preferably 300-600 nm and most preferably 350-500 nm.

[0722] In some preferred embodiments of the present invention, when the whey protein nanogel of the present invention is present in demineralized water in an amount of 20.0% w / w, the whey protein nanogel is heated to 20°C and 300 s -1 Provide up to 200 cP at a shear rate of 20°C and 300 s -1 Provide up to 100 cP at a shear rate of 20°C and 300 s -1 Provide up to 50 cP at a shear rate of 1.5 s, and most preferably at 20°C and 300 s -1 Provides a viscosity of up to 40 cP at a shear rate of 1.5 %.

[0723] In some preferred embodiments of the present invention, the various hydrated forms of the whey protein nanogels of the present invention have a polydispersity index of at most 0.3, more preferably at most 0.2 and most preferably at most 0.1.

[0724] Yet another aspect of the present invention relates to a food product comprising a whey protein nanogel composition as defined herein or a plurality of whey protein nanogels as defined herein.

[0725] The food product preferably contains at least one or more additional ingredients in addition to the whey protein nanogel composition or the whey protein nanogel and / or has been converted into a product which is no longer a whey protein nanogel composition.

[0726] The food is preferably a beverage, an acidic thickened food, or a solid food.

[0727] The inventors have found that the whey protein nanogel and whey protein nanogel composition of the present invention are particularly suitable for use in beverages, and preferably for use in packaged heat-processed beverages having a high protein content.

[0728] Thus, one particular aspect of the present invention relates to a packaged heat-processed beverage comprising at least a whey protein nanogel.

[0729] In some preferred embodiments of the present invention, the packaged heat-treated beverage has a pH in the range of 3-8 and comprises whey protein nanogel in an amount of at least 1% w / w.

[0730] It is particularly preferred that the whey protein nanogel for the beverage is provided by a whey protein nanogel composition as defined herein, or is a whey protein nanogel as defined herein.It is further particularly preferred that the whey protein nanogel for the beverage is obtainable by the method as described herein.

[0731] In some preferred embodiments of the invention, the packaged heat treated beverage comprises total protein in an amount ranging from 1-32% w / w, more preferably 5-31% w / w, even more preferably 10-30% w / w, and most preferably 21-30% w / w.

[0732] In some preferred embodiments of the invention, the packaged heat treated beverage comprises total protein in an amount ranging from 17-32% w / w, more preferably 21-31% w / w, even more preferably 22-30% w / w, and most preferably 24-30% w / w.

[0733] In other preferred embodiments of the invention, the packaged heat treated beverage comprises total protein in an amount ranging from 8-32% w / w, more preferably 9-24% w / w, even more preferably 10-22% w / w, and most preferably 11-20% w / w.

[0734] In some preferred embodiments of the present invention, the packaged heat-treated beverage comprises total BLG in an amount of at least 50% w / w relative to total protein, more preferably at least 60% w / w relative to total protein, even more preferably at least 70% w / w relative to total protein, and most preferably at least 80% w / w relative to total protein.

[0735] In other preferred embodiments of the present invention, the packaged heat-treated beverage comprises total BLG in an amount of at least 85% w / w relative to total protein, more preferably at least 90% w / w relative to total protein, even more preferably at least 92% w / w relative to total protein, and most preferably at least 95% w / w relative to total protein. It is generally preferred that the whey protein nanogel composition comprises total BLG in an amount of at least 97% w / w relative to total protein.

[0736] In a further preferred embodiment of the present invention, the packaged heat-treated beverage comprises total BLG in an amount in the range of 50-80% w / w relative to total protein, more preferably in the range of 52-75% w / w relative to total protein, even more preferably in the range of 54-70% w / w relative to total protein, and most preferably in the range of 55-65% w / w relative to total protein.

[0737] In some preferred embodiments of the invention, the packaged heat-treated beverage comprises whey protein nanogel in an amount of at least 5% w / w, more preferably at least 8% w / w, even more preferably at least 10% w / w, and most preferably up to at least 11% w / w.

[0738] Preferably, the packaged heat treated beverage may comprise whey protein nanogel in an amount of 5-32% w / w, more preferably 8-24% w / w, even more preferably 10-22% w / w, and most preferably up to 11-20% w / w.

[0739] In some preferred embodiments of the invention, the packaged heat-treated beverage comprises whey protein nanogel in an amount of at least 21% w / w, more preferably at least 22% w / w, even more preferably at least 25% w / w, and most preferably at least 28% w / w.

[0740] Preferably, the packaged heat treated beverage comprises whey protein nanogel in an amount of 21-32% w / w, more preferably 22-31% w / w, even more preferably 23-30% w / w, and most preferably 24-30% w / w.

[0741] In some preferred embodiments of the invention, the packaged heat-treated beverage comprises whey protein nanogel in an amount of at least 50% w / w relative to the total protein, more preferably at least 70% w / w relative to the total protein, even more preferably at least 80% w / w, and most preferably at least 90% w / w. Even higher amounts of whey protein nanogel are generally preferred, and in other preferred embodiments of the invention, the packaged heat-treated beverage comprises whey protein nanogel in an amount of at least 92% w / w relative to the total protein, more preferably at least 94% w / w relative to the total protein, even more preferably at least 96% w / w, and most preferably at least 98% w / w.

[0742] The degree of BLG denaturation of the beverage is preferably very high. In some preferred embodiments of the present invention, the packaged heat-treated beverage has a degree of BLG denaturation relative to the total protein of at least 70%, more preferably at least 80%, even more preferably at least 85%, and most preferably at least 90%. Even higher degrees of BLG denaturation are generally preferred, and in other preferred embodiments of the present invention, the packaged heat-treated beverage has a degree of BLG denaturation of at least 92%, more preferably at least 94%, even more preferably at least 96%, and most preferably at least 98%.

[0743] In some preferred embodiments of the present invention, the packaged heat-treated beverage comprises soluble whey protein aggregates in an amount of at most 30% w / w relative to the total protein, more preferably at most 20% w / w relative to the total protein, even more preferably at most 10% w / w, and most preferably at most 5% w / w.

[0744] Even lower levels of soluble whey protein aggregates are generally preferred and the packaged heat-treated beverage preferably comprises soluble whey protein aggregates in an amount of at most 3% w / w relative to the total protein, more preferably at most 2% w / w relative to the total protein, even more preferably at most 1% w / w and most preferably at most 0.5% w / w.

[0745] In other preferred embodiments of the present invention, the packaged heat-treated beverage comprises soluble whey protein aggregates in an amount of at most 8% w / w relative to the total weight of the beverage, more preferably at most 5% w / w relative to the total weight of the beverage, even more preferably at most 2% w / w, and most preferably at most 0.5% w / w.

[0746] It is generally preferred that the packaged heat-treated beverage of the present invention contains a small amount of larger whey protein particles which may cause sedimentation in the beverage or even a sandy impression. Thus, in some preferred embodiments of the present invention, the packaged heat-treated beverage comprises whey protein microparticles in an amount of at most 10% w / w relative to the total protein, more preferably at most 5% w / w relative to the total protein, even more preferably at most 3% w / w, and most preferably at most 1% w / w. The content of whey protein microparticles is determined according to Assay 3.

[0747] In some preferred embodiments of the present invention, the packaged heat-treated beverage comprises:

[0748] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0749] - whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein, and

[0750] - Soluble whey protein aggregates in an amount of at most 9% w / w, and most preferably at most 5% w / w, relative to total protein.

[0751] In other preferred embodiments of the present invention, the packaged heat-treated beverage comprises:

[0752] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0753] - whey protein nanogel in an amount of at least 85% w / w, most preferably at least 89% w / w relative to the total protein, and

[0754] - Soluble whey protein aggregates in an amount of at most 15% w / w, and most preferably at most 11% w / w, relative to total protein.

[0755] In some preferred embodiments of the present invention, the packaged heat-treated beverage has a total solids content in an amount of 3-50% w / w, more preferably 4-40% w / w, even more preferably 6-35% w / w, and most preferably 8-30% w / w.

[0756] In other preferred embodiments of the present invention, the packaged heat-treated beverage has a total solids content in an amount of 10-50% w / w, more preferably in an amount of 12-40% w / w, even more preferably in an amount of 14-35% w / w, and most preferably in an amount of 16-30% w / w.

[0757] In some preferred embodiments of the present invention, the packaged heat treated beverage has a water content in the amount of 50-97% w / w, more preferably in the amount of 60-96% w / w, even more preferably in the amount of 65-94% w / w, and most preferably in the amount of 70-92% w / w.

[0758] The part of the packaged heat-treated beverage which is not a solid is preferably water.

[0759] The pH of a beverage can range from acidic to slightly alkaline.

[0760] In some preferred embodiments of the present invention, the pH of the packaged heat-treated beverage is in the range of 5.5-8.0, more preferably 6.0-7.5, even more preferably 6.2-7.3, and most preferably 6.3-7.2.

[0761] In other preferred embodiments of the present invention, the pH of the packaged heat-treated beverage is in the range of 6.0-7.5, more preferably 6.2-7.5, and most preferably 6.3-7.5.

[0762] In a further preferred embodiment of the present invention the pH of the packaged heat treated beverage is in the range of 6.0-8.0, more preferably 6.6-7.7, even more preferably 6.7-7.6, and most preferably 6.8-7.5.

[0763] In other preferred embodiments of the present invention, the pH of the packaged heat treated beverage is in the range of 3.0-5.4, more preferably 3.5-5.0, even more preferably 3.7-4.8, and most preferably 4.0-4.6.

[0764] In a further preferred embodiment of the present invention the pH of the packaged heat treated beverage is in the range of 3.0-5.4, more preferably 3.0-5.0, even more preferably 3.5-5.0, and most preferably 3.5-4.6.

[0765] In a still further preferred embodiment of the present invention the pH of the packaged heat treated beverage is in the range of 3.0-5.4, more preferably 3.1-5.0, even more preferably 3.2-4.6, and most preferably 3.5-4.0.

[0766] The inventors have found that the viscosity contribution of soluble whey protein aggregates becomes less significant below pH 4.0. Therefore, in some preferred embodiments of the present invention, the pH of the packaged heat-treated beverage is in the range of 3.0-4.0, more preferably 3.0-3.8, and most preferably 3.0-3.6. Preferably, the pH of the packaged heat-treated beverage may be in the range of 3.0-4.0, more preferably 3.0-3.8, and most preferably 3.0-3.6; and the whey protein nanogel may account for 50-95% w / w of the total protein of the packaged heat-treated beverage, more preferably 55-85% w / w, even more preferably 60-75% w / w; and the soluble whey protein aggregates may account for 5-50% w / w of the total protein of the packaged heat-treated beverage, more preferably 15-45% w / w, and most preferably 25-30% w / w.

[0767] The inventors have also found that a high content of whey protein nanogel relative to total protein allows the production of low viscosity, high protein beverages with a pH range of 4.0-5.0, most preferably 4.2-4.8. Therefore, in some preferred embodiments of the present invention, the packaged heat-treated beverage has a pH in the range of 4.0-5.0, and most preferably 4.2-4.8. Preferably, the packaged heat-treated beverage may have a pH in the range of 4.0-5.0, most preferably 4.2-4.8, and the whey protein nanogel may account for at least 80% w / w of the total protein of the packaged heat-treated beverage, most preferably at least 90% w / w.

[0768] The inventors have observed that heat-treated acidic protein beverages containing the whey protein nanogels of the invention surprisingly have less astringency than similar beverages containing other types of denatured whey proteins. This is documented, for example, in Example 8. This is advantageous, for example, for beverages that have been subjected to a heat treatment for protein denaturation, for example at temperatures above 100°C.

[0769] One advantage of the packaged heat-treated beverage of the present invention is that it has a surprisingly low viscosity compared to the amount of protein it contains.

[0770] In some preferred embodiments of the present invention, the packaged heat-treated beverage is heated at 20°C and 300s -1 The shear rate is at most 200 cP, more preferably at 20°C and 300 s -1 The shear rate has a maximum of 100 cP at a shear rate of 20°C and 300 s -1 The shear rate is at most 50 cP, and most preferably at 20 ° C and 300 s -1 The viscosity of the composite is at most 20 cP at a shear rate of 1.5 %.

[0771] Even lower viscosities are possible and often desired. Therefore, in other preferred embodiments of the present invention, the packaged heat treated beverage is heated at 20°C and 300 s -1 The shear rate is at most 15 cP, more preferably at 20 ° C and 300 s -1 The shear rate is at most 10 cP, even more preferably at 20 ° C and 300 s -1 The shear rate is at most 8 cP, and most preferably at 20 ° C and 300 s -1 The viscosity of the composite is at most 5 cP at a shear rate of 1.5 %.

[0772] It is particularly preferred that the packaged heat treated beverage is aseptic.

[0773] In some preferred embodiments of the present invention, the packaged heat-treated beverage has a shelf life of at least 6 months, more preferably at least 1 year, even more preferably at least 2 years at ambient temperature.

[0774] The packaged heat-treated beverage of the present invention may contain other macronutrients besides protein, such as for example carbohydrates and / or lipids.

[0775] In some embodiments of the present invention, the packaged heat-treated beverage further comprises carbohydrates.The total carbohydrate content in the heat-treated beverage of the present invention depends on the intended use of the heat-treated beverage.

[0776] The carbohydrates of the packaged heat-treated beverage are preferably provided by one or more carbohydrate sources.

[0777] Useful carbohydrate sources can be selected from the group consisting of: sucrose, maltose, dextrose, galactose, maltodextrin, corn syrup solids, sucralose (sucromalt), glucose polymers, corn syrup, modified starch, resistant starch, rice derived carbohydrates, isomaltulose, white sugar, glucose, fructose, lactose, high fructose corn syrup, honey, sugar alcohols, oligofructose, soy fiber, corn fiber, guar gum, konjac flour, polydextrose, fiber sol and combinations thereof. In some embodiments of the present invention, the packaged heat-treated beverage comprises non-digestible carbohydrates such as fructans, and the fructans comprise inulin or oligofructose.

[0778] In some preferred embodiments of the present invention, the packaged heat-treated beverage contains carbohydrates in the range between 0 and 95% of the total energy content of the beverage, more preferably in the range between 10 and 85% of the total energy content of the beverage, even more preferably in the range between 20 and 75% of the total energy content of the beverage, and most preferably in the range between 30 and 60% of the total energy content of the beverage.

[0779] Even lower carbohydrate contents are generally preferred, thus in some preferred embodiments of the invention preferably in the range between 0 and 30% of the total energy content of the beverage, more preferably in the range between 0 and 20% of the total energy content of the beverage, even more preferably in the range between 0 and 10% of the total energy content of the beverage.

[0780] In some preferred embodiments of the invention, the beverage is particularly useful as a sports drink and comprises a total amount of carbohydrates, e.g. at most 75%, more preferably at most 40E%, even more preferably at most 10E%, and most preferably at most 5E%, of the total energy content (E) of the beverage.

[0781] In some preferred embodiments of the invention, the packaged heat-treated beverage is particularly useful as a nutritional supplement for the nutritionally incomplete and comprises a total amount of carbohydrates, for example in the range between 70-95%, preferably between 80-90E%, of the total energy content (E) of the beverage.

[0782] In some preferred embodiments of the present invention, the packaged heat treated beverage comprises a total amount of carbohydrates in the range between 30-60% of the total energy content of the beverage, and most preferably in the range between 35-50E%.Such beverages are particularly suitable for nutritionally complete beverages.

[0783] In some embodiments of the present invention, the packaged heat-treated beverage further comprises at least one additional ingredient selected from the group consisting of vitamins, flavorings, minerals, sweeteners, antioxidants, food acids, lipids, carbohydrates, prebiotics, probiotics and non-whey proteins, and combinations thereof.

[0784] Additional ingredients may be used to adjust the nutritional contribution as well as the taste and flavor profile of the beverage.

[0785] In one embodiment of the invention, the beverage comprises at least one high intensity sweetener (HIS). The at least one HIS is preferably selected from the group consisting of aspartame, cyclamate, sucralose, acesulfame salt, neotame, saccharin, stevia extract, steviol glycosides, such as rebaudioside A, or a combination thereof.

[0786] In some embodiments of the invention, it is particularly preferred that the sweetener comprises or even consists of one or more high intensity sweeteners.

[0787] HIS is found in both natural and artificial sweeteners and its sweetness intensity is typically at least 10 times that of sucrose.

[0788] If used, the total amount of HIS of the beverage is typically in the range of 0.001-2% w / w. Preferably, the total amount of HIS is in the range of 0.005-1% w / w. Most preferably, the total amount of HIS is in the range of 0.01-0.5% w / w.

[0789] The choice of sweetener may depend on the beverage to be produced, for example, high intensity sweeteners (such as aspartame, acesulfame K or sucralose) may be used in beverages that do not require sweeteners to provide energy, while for beverages with a natural character, natural sweeteners (such as steviol glycosides, sorbitol or sucrose) may be used.

[0790] In addition it is preferred that sweetener comprises one or more polyol sweeteners or even consists of one or more polyol sweeteners.The limiting examples of useful polyol sweeteners are maltitol, mannitol, lactitol, sorbitol, inositol, xylitol, threitol, galactitol or its combination.If used, the total amount of the polyol sweetener of beverage is usually in the scope of 1-20%w / w.More preferably, the total amount of the polyol sweetener of beverage is in the scope of 2-15%w / w.Even more preferably, the total amount of polyol sweetener can be in the scope of 4-10%w / w.

[0791] In some preferred embodiments of the present invention, the packaged heat-treated beverage comprises:

[0792] - at most 1% w / w, more preferably at most 0.5% w / w, and most preferably at most 0.1% w / w of total carbohydrates, and

[0793] - Total amount of HIS in the range of 0.001-2% w / w, more preferably in the range of 0.005-1% w / w, and most preferably in the range of 0.01-0.5% w / w.

[0794] In some embodiments of the present invention, the packaged heat-treated beverage further comprises lipids.The total lipid content in the heat-treated beverage of the present invention depends on the intended use of the heat-treated beverage.

[0795] In some preferred embodiments of the present invention, the packaged heat-treated beverage has a lipid content in the range between 0 and 50% of the total energy content of the beverage, or preferably in the range between 0 and 40% of the total energy content of the beverage, or preferably in the range between 0 and 30% of the total energy content of the beverage, or preferably in the range between 0 and 20% of the total energy content of the beverage, or preferably in the range between 0 and 10% of the total energy content of the beverage, or preferably in the range between 0 and 5% of the total energy content of the beverage.

[0796] In some preferred embodiments of the invention, the beverage is particularly useful as a sports drink and comprises, for example, at most 10 E%, preferably at most 1 E% of total lipids.

[0797] In some preferred embodiments of the invention, the packaged heat-treated beverage is particularly useful as a nutritional supplement for the nutritionally incomplete and comprises a total amount of lipids, for example, of at most 10%, preferably at most 1E%, of the total energy content of the beverage.

[0798] In some preferred embodiments of the present invention, the beverage, such as a sports drink, comprises:

[0799] a total amount of protein in the range of 1-30% w / w relative to the weight of the beverage, more preferably 4-24% w / w relative to the weight of the beverage, even more preferably 8-20% w / w relative to the weight of the beverage, and most preferably 10-20% w / w relative to the weight of the beverage,

[0800] - at most 75%, more preferably at most 40E%, even more preferably at most 10E%, and most preferably at most 5E% of the total energy content (E) of the beverage in terms of total carbohydrates, and

[0801] - at most 10 E%, more preferably at most 6 E%, even more preferably at most 3 E%, and most preferably at most 1 E% of the total amount of lipids.

[0802] In other preferred embodiments of the invention, the beverage, for example in the form of a low carbohydrate sports drink, comprises:

[0803] a total amount of protein in the range of 1-30% w / w relative to the weight of the beverage, more preferably 4-24% w / w relative to the weight of the beverage, even more preferably 8-20% w / w relative to the weight of the beverage, and most preferably 10-20% w / w relative to the weight of the beverage,

[0804] - at most 10 E%, more preferably at most 6 E%, even more preferably at most 3 E%, and most preferably at most 1 E% of the total amount of carbohydrates,

[0805] - at most 5E%, more preferably at most 4E%, even more preferably at most 3E%, and most preferably at most 1E% of the total amount of lipids, and

[0806] - Total amount of HIS in the range of 0.001-2% w / w, more preferably in the range of 0.005-1% w / w, and most preferably in the range of 0.01-0.5% w / w.

[0807] In other preferred embodiments of the present invention, the packaged heat-treated beverage, for example in the form of a nutritionally complete beverage, comprises:

[0808] a total amount of protein in the range of 1-30% w / w relative to the weight of the beverage, more preferably 4-24% w / w relative to the weight of the beverage, even more preferably 8-20% w / w relative to the weight of the beverage, and most preferably 10-20% w / w relative to the weight of the beverage,

[0809] - a total amount of carbohydrates in the range between 30-60% and most preferably in the range between 35-50E% of the total energy content of the beverage, and

[0810] - Total amount of lipids in the range between 20-50%, more preferably 25-45 E%, and most preferably 30-40 E% of the total energy content.

[0811] In some preferred embodiments of the invention, the packaged heat-treated beverage has a pH in the range of 3.0-5.4, most preferably 3.5-4.6, and comprises:

[0812] - a total amount of protein in the range of 8-32% w / w relative to the weight of the beverage, and most preferably in the range of 10-22% w / w relative to the weight of the beverage,

[0813] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0814] - a whey protein nanogel in an amount of at least 85% w / w, most preferably at least 89% w / w, relative to the total protein, and

[0815] - Soluble whey protein aggregates in an amount of at most 15% w / w, and most preferably at most 11% w / w, relative to the total protein.

[0816] In other preferred embodiments of the present invention, the packaged heat treated beverage has a pH in the range of 3.0-5.4, most preferably in the range of 3.5-4.6, and comprises:

[0817] - a total amount of protein in the range of 8-32% w / w relative to the weight of the beverage, and most preferably in the range of 10-22% w / w relative to the weight of the beverage,

[0818] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0819] - a whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w, relative to the total protein, and

[0820] - Soluble whey protein aggregates in an amount of up to 10% w / w, and most preferably up to 5% w / w, relative to the total protein.

[0821] In some preferred embodiments of the invention, the packaged heat-treated beverage has a pH in the range of 3.0-5.4, most preferably 3.5-4.6, and comprises:

[0822] - a total amount of protein in the range of 8-32% w / w relative to the weight of the beverage, and most preferably in the range of 10-22% w / w relative to the weight of the beverage,

[0823] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0824] - a whey protein nanogel in an amount of at least 50% w / w, most preferably at least 60% w / w, relative to the total protein, and

[0825] - Soluble whey protein aggregates in an amount of at most 15% w / w, and most preferably at most 11% w / w, relative to the total protein.

[0826] In other preferred embodiments of the present invention, the packaged heat treated beverage has a pH in the range of 3.0-5.4, most preferably in the range of 3.5-4.6, and comprises:

[0827] - a total amount of protein in the range of 8-32% w / w relative to the weight of the beverage, and most preferably in the range of 10-22% w / w relative to the weight of the beverage,

[0828] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0829] - a whey protein nanogel in an amount of at least 50% w / w, most preferably at least 60% w / w, relative to the total protein, and

[0830] - Soluble whey protein aggregates in an amount of up to 10% w / w, and most preferably up to 5% w / w, relative to the total protein.

[0831] In some preferred embodiments of the invention, the packaged heat-treated beverage has a pH in the range of 6.2-7.5, most preferably 6.8-7.5, and comprises:

[0832] - a total amount of protein in the range of 8-32% w / w relative to the weight of the beverage, and most preferably in the range of 10-22% w / w relative to the weight of the beverage,

[0833] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0834] - a whey protein nanogel in an amount of at least 85% w / w, most preferably at least 89% w / w, relative to the total protein, and

[0835] - Soluble whey protein aggregates in an amount of at most 15% w / w, and most preferably at most 11% w / w, relative to the total protein.

[0836] In other preferred embodiments of the present invention, the packaged heat treated beverage has a pH in the range of 6.2-7.5, most preferably in the range of 6.8-7.5, and comprises:

[0837] - a total amount of protein in the range of 8-32% w / w relative to the weight of the beverage, and most preferably in the range of 10-22% w / w relative to the weight of the beverage,

[0838] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0839] - a whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w, relative to the total protein, and

[0840] - Soluble whey protein aggregates in an amount of up to 10% w / w, and most preferably up to 5% w / w, relative to the total protein.

[0841] In some preferred embodiments of the invention, the packaged heat-treated beverage has a pH in the range of 6.2-7.5, most preferably 6.8-7.5, and comprises:

[0842] - a total amount of protein in the range of 8-32% w / w relative to the weight of the beverage, and most preferably in the range of 10-22% w / w relative to the weight of the beverage,

[0843] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0844] - a whey protein nanogel in an amount of at least 50% w / w, most preferably at least 60% w / w, relative to the total protein, and

[0845] - Soluble whey protein aggregates in an amount of at most 15% w / w, and most preferably at most 11% w / w, relative to the total protein.

[0846] In other preferred embodiments of the present invention, the packaged heat treated beverage has a pH in the range of 6.2-7.5, most preferably in the range of 6.8-7.5, and comprises:

[0847] - a total amount of protein in the range of 8-32% w / w relative to the weight of the beverage, and most preferably in the range of 10-22% w / w relative to the weight of the beverage,

[0848] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0849] - a whey protein nanogel in an amount of at least 50% w / w, most preferably at least 60% w / w, relative to the total protein, and

[0850] - Soluble whey protein aggregates in an amount of up to 10% w / w, and most preferably up to 5% w / w, relative to the total protein.

[0851] In some preferred embodiments of the present invention, the beverage, such as a sports drink, comprises:

[0852] - at most 75%, more preferably at most 40E%, even more preferably at most 10E%, and most preferably at most 5E% of the total energy content (E) of the beverage in terms of total carbohydrates, and

[0853] - at most 10 E%, more preferably at most 6 E%, even more preferably at most 3 E%, and most preferably at most 1 E% of the total amount of lipids.

[0854] In other preferred embodiments of the present invention, the packaged heat-treated beverage, for example in the form of a nutritionally complete beverage, comprises:

[0855] - a total amount of carbohydrates in the range between 30-60% and most preferably in the range between 35-50E% of the total energy content of the beverage, and

[0856] - Total amount of lipids in the range between 20-50%, more preferably 25-45 E%, and most preferably 30-40 E% of the total energy content.

[0857] One aspect of the present invention relates to a process for producing a packaged heat-treated beverage having a pH in the range of 3-8, comprising the following steps:

[0858] 1) Providing a liquid solution having a pH in the range of 3-8 and comprising:

[0859] - 1 to 32% by weight of total protein, of which at least 50 w / w % of the protein is beta-lactoglobulin (BLG),

[0860] -optionally, sweeteners and / or flavoring agents,

[0861] 2) packaging the liquid solution,

[0862] Wherein the liquid solution of step a) and / or the packaged liquid solution of step b) is subjected to a heat treatment comprising at least pasteurization.

[0863] Thus, a heat treated beverage is a heat treated liquid solution.

[0864] In some particularly preferred embodiments of the present invention, the liquid solution is a whey protein solution according to step a) of the method for producing a whey protein nanogel composition as described above and the heat treatment of the method for producing a packaged heat-treated beverage is step b) of the method as described above). The heat treatment is preferably sterilization, so that it is possible to package the sterile liquid whey protein nanogel composition as the heat-treated liquid solution in step 2) of the process for producing a packaged heat-treated beverage.

[0865] In other particularly preferred embodiments of the present invention, the liquid solution is:

[0866] - the nanogel suspension obtained in step b) of the above method for producing a whey protein nanogel composition, and / or

[0867] - a concentrated suspension of nanogel obtained from step c),

[0868] And the heat treatment of the production process of packaged heat-treated beverages sterilizes the liquid solution or the packaged liquid solution.

[0869] In some preferred embodiments, the liquid solution has a pH in the range of 3-8 and comprises whey protein nanogel in an amount of at least 1% w / w.

[0870] It is particularly preferred that the whey protein nanogel of liquid solution is provided by a whey protein nanogel composition, preferably in powder form, as defined herein, and / or is a whey protein nanogel as defined herein. It is further particularly preferred that the whey protein nanogel of liquid solution can be obtained by the method described herein. The provision of the liquid solution generally involves reconstitution of the whey protein nanogel composition in powder form in water, and if necessary, homogenization of the reconstructed whey protein nanogel composition until sufficient dispersion is obtained. Preferably, the particle size of the whey protein nanogel of liquid solution is substantially the same as its particle size before the drying step d).

[0871] In some preferred embodiments of the invention, the liquid solution comprises total protein in an amount ranging from 1-32% w / w, more preferably 5-31% w / w, even more preferably 10-30% w / w, and most preferably 21-30% w / w.

[0872] In some preferred embodiments of the invention, the liquid solution comprises total protein in an amount ranging from 17-32% w / w, more preferably 21-31% w / w, even more preferably 22-30% w / w, and most preferably 24-30% w / w.

[0873] In other preferred embodiments of the invention, the liquid solution comprises total protein in an amount ranging from 8-32% w / w, more preferably 9-24% w / w, even more preferably 10-22% w / w, and most preferably 11-20% w / w.

[0874] In some preferred embodiments of the present invention, the liquid solution comprises total BLG in an amount of at least 50% w / w relative to total protein, more preferably at least 60% w / w relative to total protein, even more preferably at least 70% w / w relative to total protein, and most preferably at least 80% w / w relative to total protein.

[0875] In other preferred embodiments of the present invention, the liquid solution comprises total BLG in an amount of at least 85% w / w relative to total protein, more preferably at least 90% w / w relative to total protein, even more preferably at least 92% w / w relative to total protein, and most preferably at least 95% w / w relative to total protein. It is generally preferred that the whey protein nanogel composition comprises total BLG in an amount of at least 97% w / w relative to total protein.

[0876] In a further preferred embodiment of the present invention, the liquid solution comprises total BLG in an amount in the range of 50-80% w / w relative to total protein, more preferably in the range of 52-75% w / w relative to total protein, even more preferably in the range of 54-70% w / w relative to total protein, and most preferably in the range of 55-65% w / w relative to total protein.

[0877] In some preferred embodiments of the invention, the liquid solution comprises whey protein nanogel in an amount of at least 5% w / w, more preferably at least 8% w / w, even more preferably at least 10% w / w, and most preferably up to at least 11% w / w.

[0878] Preferably, the liquid solution may comprise whey protein nanogel in an amount of 5-32% w / w, more preferably 8-24% w / w, even more preferably 10-22% w / w, and most preferably up to 11-20% w / w.

[0879] In some preferred embodiments of the invention, the liquid solution comprises whey protein nanogel in an amount of at least 21% w / w, more preferably at least 22% w / w, even more preferably at least 25% w / w, and most preferably at least 28% w / w.

[0880] Preferably, the liquid solution comprises whey protein nanogel in an amount of 21-32% w / w, more preferably 22-31% w / w, even more preferably 23-30% w / w, and most preferably 24-30% w / w.

[0881] In some preferred embodiments of the invention, the liquid solution comprises whey protein nanogel in an amount of at least 50% w / w relative to the total protein, more preferably at least 70% w / w relative to the total protein, even more preferably at least 80% w / w, and most preferably at least 90% w / w. Even higher amounts of whey protein nanogel are generally preferred, and in other preferred embodiments of the invention, the liquid solution comprises whey protein nanogel in an amount of at least 92% w / w relative to the total protein, more preferably at least 94% w / w relative to the total protein, even more preferably at least 96% w / w, and most preferably at least 98% w / w.

[0882] The degree of BLG denaturation of the liquid solution is generally very high, especially when the protein of the liquid solution is mainly in the form of whey protein nanogels and optionally soluble whey protein aggregates. In some preferred embodiments of the present invention, the degree of BLG denaturation of the liquid solution is at least 70%, more preferably at least 80%, even more preferably at least 85%, and most preferably at least 90% relative to the total protein. Even higher degrees of BLG denaturation are generally preferred, and in other preferred embodiments of the present invention, the liquid solution has a degree of BLG denaturation of at least 92%, more preferably at least 94%, even more preferably at least 96%, and most preferably at least 98%.

[0883] In some preferred embodiments of the present invention, the liquid solution comprises soluble whey protein aggregates in an amount of at most 30% w / w relative to the total protein, more preferably at most 20% w / w relative to the total protein, even more preferably at most 10% w / w, and most preferably at most 5% w / w.

[0884] Even lower contents of soluble whey protein aggregates are generally preferred, and the liquid solution preferably comprises soluble whey protein aggregates in an amount of at most 3% w / w relative to the total protein, more preferably at most 2% w / w relative to the total protein, even more preferably at most 1% w / w, and most preferably at most 0.5% w / w.

[0885] In other preferred embodiments of the present invention, the liquid solution comprises soluble whey protein aggregates in an amount of at most 8% w / w relative to the total weight of the liquid solution, more preferably at most 5% w / w relative to the total weight of the liquid solution, even more preferably at most 2% w / w and most preferably at most 0.5% w / w.

[0886] In some preferred embodiments of the present invention, the liquid solution comprises:

[0887] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0888] - whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein, and

[0889] - Soluble whey protein aggregates in an amount of up to 9% w / w, and most preferably up to 5% w / w relative to total protein.

[0890] It is generally preferred that the liquid solution contains a small amount of larger whey protein particles, which may cause sedimentation in the beverage, or even a sandy impression. Therefore, in some preferred embodiments of the present invention, the liquid solution comprises whey protein microparticles in an amount of at most 10% w / w relative to the total protein, more preferably at most 5% w / w relative to the total protein, even more preferably at most 3% w / w, and most preferably at most 1% w / w.

[0891] The pH of a liquid solution can range from acidic to slightly alkaline.

[0892] Nearly pH neutral liquid solutions are particularly preferred for producing near pH neutral beverages.In some preferred embodiments of the present invention, the pH of the liquid solution is in the range of 5.5-8.0, more preferably 6.0-7.5, even more preferably 6.2-7.3, and most preferably 6.3-7.2.

[0893] In other preferred embodiments of the present invention, the pH of the packaged heat-treated beverage is in the range of 6.0-7.5, more preferably 6.2-7.5, and most preferably 6.3-7.5.

[0894] In a further preferred embodiment of the present invention, the pH of the liquid solution is in the range of 6.0-8.0, more preferably 6.6-7.7, even more preferably 6.7-7.6, and most preferably 6.8-7.5.

[0895] The acidic liquid solution is particularly preferably used for producing acidic beverages.In some preferred embodiments of the present invention, the pH of the liquid solution is in the range of 3.0-5.4, more preferably 3.5-5.0, even more preferably 3.7-4.8, and most preferably 4.0-4.6.

[0896] In a further preferred embodiment of the present invention, the pH of the liquid solution is in the range of 3.0-5.4, more preferably 3.0-5.0, even more preferably 3.5-5.0, and most preferably 3.5-4.6.

[0897] In a further preferred embodiment of the present invention, the pH of the liquid solution is in the range of 3.0-5.4, more preferably 3.1-5.0, even more preferably 3.2-4.6, and most preferably 3.3-3.8.

[0898] The inventors have found that the viscosity contribution of soluble whey protein aggregates becomes less significant below pH 4.0. Therefore, in some preferred embodiments of the present invention, the pH of the liquid solution is in the range of 3.0-4.0, more preferably 3.0-3.8, and most preferably 3.0-3.6. Preferably, the pH of the liquid solution may be in the range of 3.0-4.0, more preferably 3.0-3.8, and most preferably 3.0-3.6; and the whey protein nanogel may account for 50-95% w / w of the total protein of the liquid solution, more preferably 55-85% w / w, even more preferably 60-75% w / w; and the soluble whey protein aggregates may account for 5-50% w / w of the total protein of the liquid solution, more preferably 15-45% w / w, and most preferably 25-30% w / w.

[0899] The inventors have also found that a high content of whey protein nanogel relative to total protein allows the production of a low viscosity, high protein beverage with a pH range of 4.0-5.0, most preferably 4.2-4.8. Therefore, in some preferred embodiments of the present invention, the pH of the liquid solution is in the range of 4.0-5.0, and most preferably 4.2-4.8. Preferably, the pH of the liquid solution may be in the range of 4.0-5.0, and most preferably 4.2-4.8, and the whey protein nanogel may account for at least 80% w / w of the total protein of the liquid solution, most preferably at least 90% w / w.

[0900] Generally any suitable food acid or food base can be used to adjust the pH of the liquid solution. Those skilled in the art will recognize suitable means for adjusting pH. Suitable food bases include sodium carbonate or potassium carbonate, sodium bicarbonate or potassium bicarbonate, or ammonium hydroxide. Alternatively, KOH or NaOH can be used, for example, to adjust the pH value. Suitable food acids include, for example, citric acid, hydrochloric acid, malic acid or tartaric acid or phosphoric acid.

[0901] In some preferred embodiments of the present invention, the liquid solution is heated at 20°C and 300s -1 The shear rate is at most 200 cP, more preferably at 20°C and 300 s -1 The shear rate has a maximum of 100 cP at a shear rate of 20°C and 300 s -1 The shear rate has a maximum of 50 cP, and most preferably at 20 ° C and 300 s -1 The viscosity of the composite is at most 20 cP at a shear rate of 1.5 %.

[0902] Even lower viscosities are possible and often desired. Therefore, in other preferred embodiments of the present invention, the liquid solution is at 20°C and 300 s -1 The shear rate is at most 15 cP, more preferably at 20 ° C and 300 s -1At most 10 cP at 20 ° C and 300 s -1 The shear rate is at most 8 cP, and most preferably at 20 ° C and 300 s -1 The viscosity of the composite is at most 5 cP at a shear rate of 1.5 %.

[0903] It is particularly preferred that the liquid solution is sterile.

[0904] The liquid solution of the present invention may contain other macronutrients and ingredients in addition to protein. The embodiments and preferences described in the context of packaged heat-treated beverages and relating to macronutrients and additional ingredients are equally applicable to the liquid solution, and we refer to these embodiments rather than repeating them here.

[0905] In some particularly preferred embodiments of the present invention, the liquid solution has the same chemical composition as the packaged heat-treated beverage.

[0906] In other particularly preferred embodiments of the present invention, the liquid solution has the same chemical composition as the packaged heat-treated beverage, except for less protein denaturation.

[0907] It is particularly preferred that the provision of a liquid solution comprises mixing a whey protein nanogel composition in powder form as defined herein with water and optionally one or more additional ingredients.

[0908] It is preferred that the whey protein nanogel composition in powder form comprises at least 50% w / w of the whey protein nanogel of the liquid solution, more preferably at least 80% w / w, even more preferably at least 90% w / w, and most preferably all of the whey protein nanogel of the liquid solution.

[0909] It is further preferred that the whey protein nanogel composition in powder form comprises at least 50% w / w of the protein of the liquid solution, more preferably at least 80% w / w, even more preferably at least 90% w / w, and most preferably all of the protein of the liquid solution.

[0910] In some preferred embodiments of the present invention, the liquid solution has a pH in the range of 3.0-5.4, most preferably 3.5-4.6, and comprises:

[0911] - a total amount of protein in the range of 8-32% w / w relative to the weight of the liquid solution, and most preferably in the range of 10-22% w / w relative to the weight of the liquid solution,

[0912] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0913] - whey protein nanogel in an amount of at least 85% w / w, most preferably at least 89% w / w relative to the total protein, and

[0914] - Soluble whey protein aggregates in an amount of at most 15% w / w, and most preferably at most 11% w / w, relative to total protein.

[0915] In other preferred embodiments of the present invention, the liquid solution has a pH in the range of 3.0-5.4, most preferably 3.5-4.6, and comprises:

[0916] - a total amount of protein in the range of 8-32% w / w relative to the weight of the liquid solution, and most preferably in the range of 10-22% w / w relative to the weight of the liquid solution,

[0917] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0918] - whey protein nanogel in an amount of at least 90% w / w, most preferably at least 95% w / w relative to the total protein, and

[0919] - Soluble whey protein aggregates in an amount of up to 10% w / w, and most preferably up to 5% w / w, relative to the total protein.

[0920] In some preferred embodiments of the present invention, the liquid solution has a pH in the range of 3.0-5.4, most preferably 3.5-4.6, and comprises:

[0921] - a total amount of protein in the range of 8-32% w / w relative to the weight of the liquid solution, and most preferably in the range of 10-22% w / w relative to the weight of the liquid solution,

[0922] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0923] - whey protein nanogel in an amount of at least 50% w / w, most preferably at least 60% w / w relative to the total protein, and

[0924] - Up to 15% w / w, and most preferably up to 11% w / w, soluble whey protein aggregates relative to total protein.

[0925] In other preferred embodiments of the present invention, the liquid solution has a pH in the range of 3.0-5.4, most preferably 3.5-4.6, and comprises:

[0926] - a total amount of protein in the range of 8-32% w / w relative to the weight of the liquid solution, and most preferably in the range of 10-22% w / w relative to the weight of the liquid solution,

[0927] - a total amount of BLG of at least 50% w / w, most preferably at least 55% w / w relative to total protein,

[0928] - at least 50% w / w, most preferably at least 60% w / w whey protein nanogel relative to total protein, and

[0929] - Soluble whey protein aggregates in an amount of up to 10% w / w, and most preferably up to 5% w / w, relative to the total protein.

[0930] In some preferred embodiments of the present invention, the liquid solution has a pH in the range of 6.2-7.5, most preferably 6.8-7.5, and comprises:

[0931] - a total amount of protein in the range of 8-32% w / w relative to the weight of the liquid solution, and most preferably in the range of 10-22% w / w relative to the weight of the liquid solution,

[0932] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0933] - at least 85% w / w, most preferably at least 89% w / w whey protein nanogel relative to total protein, and

[0934] - Up to 15% w / w, and most preferably up to 11% w / w, soluble whey protein aggregates relative to total protein.

[0935] In other preferred embodiments of the present invention, the liquid solution has a pH in the range of 6.2-7.5, most preferably 6.8-7.5, and comprises:

[0936] - a total amount of protein in the range of 8-32% w / w relative to the weight of the liquid solution, and most preferably in the range of 10-22% w / w relative to the weight of the liquid solution,

[0937] - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein,

[0938] - at least 90% w / w, most preferably at least 95% w / w whey protein nanogel relative to total protein, and

[0939] - Soluble whey protein aggregates in an amount of up to 10% w / w, and most preferably up to 5% w / w, relative to the total protein.

[0940] In some preferred embodiments of the present invention, the liquid solution, for example, for preparing a sports drink, comprises:

[0941] - at most 75%, more preferably at most 40E%, even more preferably at most 10E%, and most preferably at most 5E% of the total energy content (E) of the liquid solution, and

[0942] - at most 10 E%, more preferably at most 6 E%, even more preferably at most 3 E%, and most preferably at most 1 E% of the total amount of lipids.

[0943] In other preferred embodiments of the present invention, the liquid solution, for example for preparing a nutritionally complete beverage, comprises:

[0944] - a total amount of carbohydrates in the range between 30-60% and most preferably in the range between 35-50E% of the total energy content of the liquid solution, and

[0945] - Total amount of lipids in the range of 20-50%, more preferably between 25-45 E%, and most preferably 30-40 E% of the total energy content.

[0946] The packaging of step 2) can be any suitable packaging technology, and any suitable container can be used to package the liquid solution.

[0947] However, in a preferred embodiment of the present invention, the packaging of step 2) is aseptic packaging, i.e. the liquid solution is packaged under aseptic conditions. For example, an aseptic filling system can be used for aseptic packaging, preferably the liquid solution is filled into one or more aseptic containers.

[0948] Aseptic filling and sealing is particularly preferred if the liquid solution is already sterile or has a very low microbial content prior to filling.

[0949] Examples of useful containers are eg bottles, cartons, bricks and / or bags.

[0950] In some preferred embodiments of the method of the present invention, the liquid solution of step 1) is subjected to a heat treatment including at least pasteurization and then packaged in step 2).

[0951] In another embodiment of the process of the invention, the packaging liquid solution of step 2) is subjected to a heat treatment comprising at least pasteurization.

[0952] In some preferred embodiments, thermal treatment involves heating the liquid solution to a temperature in the range of 70-80°C.

[0953] In some preferred embodiments of the present invention, the temperature of the heat treatment is in the range of 70-80°C, preferably in the range of 70-79°C, more preferably in the range of 71-78°C, even more preferably in the range of 72-77°C, and most preferably in the range of 73-76°C, such as about 75°C.

[0954] Preferably, the duration of the heat treatment is from 1 second to 60 minutes when carried out in a temperature range of 70-80°C. The longest exposure time is most suitable for the lowest temperature in the temperature range, and vice versa.

[0955] In other preferred embodiments, the temperature of the heat treatment is 70°C for at least 60 minutes or preferably 75°C for at least 45 minutes or preferably 80°C for at least 30 minutes or preferably 85°C for at least 22 minutes or preferably 90°C for at least 10 minutes.

[0956] In a particularly preferred embodiment of the invention, the heat treatment provides 70-78°C for 1 second to 30 minutes, more preferably 71-77°C for 1 minute to 25 minutes, and even more preferably 72-76°C for 2 minutes to 20 minutes.

[0957] In some preferred embodiments of the present invention, the process of heat treatment involves heating to a temperature of 85°C-95°C for 1 to 30 minutes.

[0958] In some embodiments, higher temperatures may also be preferred, particularly if it is desired to expand and optionally aggregate the BLG. For example, the temperature of the heat treatment may be at least 81°C, preferably at least 91°C, preferably at least 95°C, more preferably at least 100°C, even more preferably at least 120°C, and most preferably at least 140°C.

[0959] In other preferred embodiments of the present invention, the temperature of the heat treatment is preferably at least 95° C., more preferably at least 100° C., even more preferably at least 120° C., and most preferably at least 140° C. For example, the temperature of the heat treatment is preferably in the range of 95-160° C., more preferably in the range of 100-155° C., even more preferably in the range of 120-153° C., and most preferably in the range of 140-152° C.

[0960] In some preferred embodiments of the invention, the heat treatment is heat sterilization and preferably involves a temperature in the range of 120 to 155°C for a time sufficient to obtain sterility, typically 0.3 seconds to 10 minutes at 140 to 155°C, and more preferably 4-30 seconds.

[0961] The heat treatment may, for example, involve a temperature in the range of 90-130°C and a duration in the range of 5 seconds-10 minutes. The heat treatment may, for example, involve heating to a temperature in the range of 90-95°C for 1-10 minutes, such as approximately 120°C for approximately 20 seconds. Alternatively, the heat treatment may involve heating to a temperature in the range of 115-125°C for 5-30 seconds, such as approximately 120°C for approximately 20 seconds.

[0962] Alternatively, but particularly preferably, the heat treatment is a sterilising ultra high temperature (UHT) type treatment, which typically involves a temperature in the range 135-146°C and a duration in the range 2-10 seconds.

[0963] Alternatively, but also preferably, the heat treatment may involve a temperature in the range of 145-180°C and a duration in the range of 0.01-2 seconds, more preferably a temperature in the range of 150-180°C and a duration in the range of 0.01-0.3 seconds.

[0964] The implementation of heat treatment may involve the use of equipment such as plate or tube heat exchangers, scraped surface heat exchangers or retort systems. Alternatively, for heat treatments above 95°C, it is particularly preferred that direct heating based on steam can be used, for example using direct steam injection, direct steam injection or spray cooking. In addition, this direct heating based on steam is preferably used in combination with flash cooling. Suitable examples for implementing spray cooking can be found in WO2009113858A1, which is incorporated herein for all purposes. Suitable examples for implementing direct steam injection and direct steam injection can be found in WO2009113858A1 and WO 2010 / 085957 A3, which are incorporated herein for all purposes. General aspects of high temperature treatment can be found, for example, in "Thermal technologies in food processing" ISBN 185573558 X, which is incorporated herein by reference for all purposes.

[0965] In some preferred embodiments of the present invention, the heat treatment involves or even consists of a retort heat treatment, preferably at a temperature of at least 80° C., more preferably at a temperature of at least 95° C., even more preferably at least 100° C., and most preferably at least 120° C. As shown in Example 10, the inventors have demonstrated that the beverage according to the present invention can be subjected to a retort-type heat treatment at 150° C. for 10 minutes.

[0966] In other preferred embodiments of the present invention, the heat treatment involves or even consists of steam injection or spray cooking, preferably at a temperature of at least 100°C, more preferably at a temperature of at least 120°C, even more preferably at least 130°C, and most preferably at least 140°C.

[0967] In some preferred embodiments of the invention, pasteurization is combined with physical microbial reduction.

[0968] Useful examples of physical microbial reduction involve one or more of bacterial filtration, ultraviolet radiation, high pressure treatment, pulsed electric field treatment, and ultrasound.

[0969] In some preferred embodiments of the invention, the heat treatment is a sterilization heat treatment, thereby producing a sterile liquid solution and thus a sterile beverage. This sterilization can be obtained, for example, by combining bacterial filtration and pasteurization or by heat treatment at least 100°C for a time sufficient to obtain sterilization.

[0970] In some particularly preferred embodiments of the present invention, the heat treatment involves heating the liquid solution to a temperature in the range of 100-160°C for a time sufficient to sterilize the liquid solution. This preferably involves heating the liquid solution to a temperature in the range of 120 to 155°C for a time sufficient to obtain sterility, typically 0.3 seconds to 10 minutes, more preferably heating to 140 to 155°C for 0.3-30 seconds.

[0971] It is beneficial to cool the liquid solution after the heat treatment. According to a preferred embodiment of the process of the present invention, after the heat treatment, the heat treated liquid solution is cooled in an optional step to preferably 0 to 50°C, preferably 0 to 25°C or preferably 0 to 20°C or preferably 0 to 15°C, preferably 0 to 10°C or preferably 4 to 8°C or preferably 2 to 5°C or preferably 1 to 5°C.

[0972] If the liquid solution has been at least pasteurized, it is preferably cooled to 0 to 15°C, more preferably to 1 to 5°C, after the heat treatment.

[0973] Cooling can be performed before the filling step or after the filling step.

[0974] In some preferred embodiments of the present invention, a packaged heat-treated beverage is obtainable by the above-described process.

[0975] Alternatively, the food may be an acidic thickened food. The inventors have observed that whey protein nanogel compositions containing a large amount of soluble whey protein aggregates in addition to the whey protein nanogel are both heat stable and capable of increasing viscosity upon acidification. Such whey protein nanogel compositions are suitable for use in liquid food bases that are pasteurized or sterilized at near-neutral pH and subsequently acidified chemically or by bacterial acidification.

[0976] In the context of the present invention, a "thickened food" is a food that has been heated to 20°C and 300 s -1 The viscosity of the thickened food may be a thick but still pourable liquid or a viscous gel.

[0977] In the context of the present invention, an "acidic food" has a pH of at most 5.6 at 25°C.

[0978] Therefore, a specific aspect of the present invention relates to a process for producing an acidic thickened food, comprising the following steps:

[0979] - preparing a liquid food base having a pH of at least 5.7, said liquid food base comprising sufficient whey protein nanogel composition to provide protein in an amount of 4-20% w / w, said whey protein nanogel composition, preferably as defined herein, comprising:

[0980] - soluble whey protein aggregates in an amount of 15-70% w / w, more preferably 20-50% w / w relative to the total protein,

[0981] - whey protein nanogel in an amount of 30-85%, more preferably 50-80% w / w relative to the total protein,

[0982] - heat treating said liquid food base at a temperature of at least 70°C for a period sufficient to at least pasteurize said liquid food base,

[0983] - optionally, homogenizing the heat-treated liquid food base,

[0984] - acidifying the heat-treated liquid food base to a pH of at most 5.4,

[0985] - optionally, homogenizing the acidified food base,

[0986] The acidic thickened food product is a mixture of an acidified food base or an acidified liquid food base and other ingredients such as, for example, a sweetener and / or a flavoring agent.

[0987] Another particular aspect of the present invention relates to an acidic thickened food product obtainable by the above process, preferably in the form of a pourable viscous liquid or a non-pourable gel.

[0988] A further aspect of the present invention relates to the use of a whey protein nanogel composition as defined herein and / or a plurality of whey protein nanogels as defined herein for one or more of the following:

[0989] - As food ingredients,

[0990] - as a food ingredient for the production of a sterile beverage containing at least 10% w / w protein, even more preferably at least 21% w / w protein,

[0991] - as a food ingredient for the production of a thickened food having a pH of less than 5.5 and a viscosity of 100 μg / ml at 20°C and 300 s -1 A viscosity greater than 200 cP measured at a shear rate of

[0992] - As a whitening agent, for example in coffee whiteners.

[0993] Another aspect of the present invention relates to the use of whey protein nanogel and / or whey protein nanogel composition as a protein source for reducing the astringency and / or sourness of a heat-treated protein beverage having a pH of 3.0-5.0, most preferably 3.5-4.6,

[0994] - preferably wherein the whey protein nanogel comprises at least 50% w / w of the total protein of the heat processed beverage, more preferably at least 70% w / w, even more preferably at least 80% w / w, and most preferably at least 90% w / w of the total protein of the heat processed beverage; and

[0995] - Preferably wherein the heat treated beverage contains total protein in an amount of 2-35% w / w, more preferably 4-30% w / w, even more preferably 6-25% w / w and most preferably 8-20% w / w.

[0996] Preferably, the whey protein nanogel of the present invention contains a total amount of BLG of at least 60% w / w, more preferably at least 70% w / w, even more preferably at least 80% w / w and most preferably at least 90% w / w.

[0997] The whey protein nanogel and / or whey protein nanogel composition for the above use is preferably a whey protein nanogel and / or a whey protein nanogel composition as defined herein, and is preferably obtainable by the method of the present invention.

[0998] The present invention has been described above with reference to specific embodiments. However, within the scope of the present invention, other embodiments other than those described above are equally possible. Unless otherwise indicated, the different features and steps of the various embodiments and aspects of the present invention may be combined in other ways than those described herein.

[0999] Example

[1000] Analytical methods

[1001] Analysis 1: Determination of aggregate morphology by transmission electron microscopy

[1002] The samples were fixed in 2% glutaraldehyde phosphate buffer (pH 7.2) at room temperature in a 1:1 volume ratio for 40 minutes. Thereafter, the fixed samples were diluted in water to a protein concentration of approximately 0.2%. 4 μl of the diluted sample was placed on a discharged polyvinyl acetate methyl vinyl ester (formvar) / carbon film grid (300 mesh) for 1 minute and then blotted with filter paper. Staining was performed by adding 4 μl of phosphotungstic acid (1%, pH 7.0) for 1 minute and then blotted. The grid was washed once with MilliQ water and dried, then inserted into a Philips CM-100 electron microscope operating at 100 kV. Images were taken at least five different locations on the grid.

[1003] Analysis 2: Aggregate size determination by dynamic light scattering

[1004] The hydrodynamic diameter of whey protein nanogels was measured using a Malvern Nanosizer S instrument. The whey protein nanogel formulation diluted to a protein concentration of 0.05% or less in milliQ water was placed in a 2.5 mL 12.5 x 12.5 x 45 mm disposable cuvette with the arrow marking pointing in the direction of the instrument laser. After balancing for 30 seconds with automatic adjustment of the attenuation factor and measurement duration determined by the software, the samples were measured at 20°C.

[1005] The average Z-average hydrodynamic diameter (in nanometers) of five measurements provided by the software is given.

[1006] Analysis 3: Quantification of the amount of whey protein nanogels, soluble whey protein aggregates, microparticles and residual protein.

[1007] As described below, the amount of whey protein nanogel, whey protein microparticles, soluble whey protein aggregates and residual protein was determined by isolating the sample to be analyzed (diluted to 4% protein (18.2 MΩ) in milliQ water and adjusted to pH 7.0). The protein content of each fraction was determined as described in Assay 7.

[1008] The total amount of protein (P) of the initial subsample was measured by filtering a 5.0 mL subsample of the 4% protein solution through a 2.0 μm syringe filter and measuring the total amount of protein (P 总 ) and the total protein content of the filtrate (P 滤液 The amount of whey protein microparticles relative to total protein is calculated as follows:

[1009]

[1010] Denatured whey protein is known to be less soluble at pH 4.6 than at pHs below or above pH 4.6, so the amount of non-denatured (native) whey protein is determined by measuring the amount of protein that does not precipitate upon centrifugation at pH 4.6 relative to the total amount of protein at the same pH.

[1011] Therefore, a sample solution containing 4.0% (w / w) total protein and a pH of 4.6 was prepared by adjusting the pH using 5% (w / w) HCl (aq). The solution was stored at room temperature for 2 hours and then centrifuged at 50.000 g for 1 hour. The total protein content (P pH4.6,supernatant after 50.000g ) and protein mass before centrifugation (P pH4.6,离心前 ). The amount of non-denatured protein relative to total protein is calculated as follows:

[1012]

[1013] The fraction of soluble whey protein aggregates was determined by measuring the amount of protein remaining in the supernatant after centrifugation of a pH 7.0 solution of the sample (measuring soluble whey protein aggregates and non-denatured whey protein) and subtracting the amount of protein remaining in the supernatant after centrifugation of a pH 4.6 solution (measuring non-denatured whey protein only).

[1014] Therefore, a subsample of the 4.0% protein solution (pH 7.0) was stored at room temperature for 2 hours and then centrifuged at 50.000 g for 1 hour. The supernatant (P pH7.0，50.000g后上清液 ) and the subsample before centrifugation (P pH7.0,离心前 ) of the total amount of protein.

[1015] The amount of soluble whey protein aggregates relative to total protein was determined as:

[1016]

[1017] The amount of whey protein nanogel relative to total protein was determined by subtracting the relative amounts of microparticles, non-denatured protein and soluble whey protein aggregates from the total amount of protein content (100%):

[1018] Nanogel (%) = 100% - microparticles (%) - soluble whey protein aggregates (%) - non-denatured protein (%)

[1019] The degree of protein denaturation was determined as 100% - residual protein (%).

[1020] Analysis 4: Viscosity of Liquid Samples

[1021] The viscosity was measured using a handheld capillary viscometer (Viscoman, Gilson) at 20 °C and at 300 s. -1 The shear rate is reported.

[1022] Analysis 5: Quantification of gel formation during acidification

[1023] Rheometer (Anton Paar Physica MCR301) is used to evaluate the gel formation and viscosity development of samples containing whey protein nanogel during acidification. The whey protein nanogel samples are diluted to a total protein content of 4w / w% using demineralized water and equilibrated at 42°C for 10 minutes. GDL (D-glucono δ-lactone) is added to the stirred solution from powder form to a final concentration of 1w / w% and allowed to dissolve for 1 minute under stirring.

[1024] 20 mL of solution was added to a cup (CC27-SS) in a rheometer pre-equilibrated to 42°C, and the storage and loss moduli were measured at 0.1 Hz and 0.5% strain for 60 minutes. The pH was recorded throughout the experiment using a WTW Multi3410 pH recorder calibrated with a standard solution at 42°C.

[1025] Analysis 6: Determination of native BLG, native ALA and CMP and determination of the degree of BLG denaturation

[1026] Protein samples (unheated and heated) 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 precipitate and aggregate the denatured protein at about pH 4.6. The solution was filtered through a 0.22 μm filter to remove coagulation and non-native proteins.

[1027] All samples were diluted to the same extent by adding polished water.

[1028] For each sample, the same volume was loaded onto a UPLC column (Protein BEH C4; 1.7 μm; 150x2.1 mm) on a UPLC system and detected at 214 nm.

[1029] The samples were run using the following conditions:

[1030] Buffer A: Milli-Q water, 0.1% w / w TFA

[1031] Buffer B: HPLC grade acetonitrile, 0.1% w / w TFA

[1032] Flow rate: 0.4ml / min

[1033] Gradient: 0-6.00 min 24-45% B; 6.00-6.50 min 45-90% B; 6.50-7.00 min 90% B; 7.00-7.50 min 90-24% B and 7.50-10.00 min 24% B.

[1034] The BLG peak area relative to a protein standard (Sigma L0130) was used to determine the concentration of native BLG in the samples (5-level calibration curve)

[1035] If outside the linear range, further dilute the sample and re-inject.

[1036] The degree of BLG denaturation (D) provided by heat treatment was calculated as:

[1037] D=((BLG 天然，未加热 -BLG 天然，加热 ) / BLG 天然，未加热 )×100%.

[1038] Analysis 6 was also used to determine the samples' native BLG, native ALA and CMP content.

[1039] Analysis 7: Determination of total protein

[1040] The total protein content (true protein) of the samples was determined by:

[1041] 1) Determine the total nitrogen of the sample according to ISO 8968-1 / 2 | IDF 020-1 / 2 - Milk - Determination of nitrogen content - Part 1 / 2: Nitrogen content using the Kjeldahl method.

[1042] 2) Determine the non-protein nitrogen content of the sample in accordance with ISO 8968-4 | IDF 020-4 - Milk - Determination of nitrogen content - Part 4: Determination of non-protein nitrogen content.

[1043] 3) Calculate the total amount of protein (m 总氮 -m 非蛋白质氮 )×6.38.

[1044] Analysis 8: Determination of total lipid content

[1045] According to ISO 1211:2010 (Determination of fat content - -Gottlieb gravimetric method) to determine the amount of lipids.

[1046] Analysis 9: Determination of total lactose

[1047] The total amount of lactose was determined according to ISO 5765-2:2002 (IDF 79-2:2002) "Milk powder, dry ice mix and processed cheese - Determination of lactose content - Part 2: Enzymatic method using the galactose portion of lactose".

[1048] Analysis 10: Determination of Brix

[1049] Brix measurements were made using an Atago PAL-α digital handheld refractometer calibrated for polished water (water filtered by reverse osmosis to obtain a conductivity of at most 0.05 mS / cm).

[1050] Transfer approximately 500 μl of sample to the prism surface of the instrument and start the measurement. Read and record the measurement.

[1051] The Brix of a whey protein solution is directly proportional to the total solids (TS) content, and TS (measured in % w / w) is approximately Brix x 0.85.

[1052] Analysis 11: Determination of ash content

[1053] The ash content of foods is determined according to NMKL 173:2005 "Gravimetric determination of ash in foods".

[1054] Analysis 12: Determination of total solids and water content

[1055] The total solids of the product can be determined according to NMKL 110 2nd edition, 2005 (Total solids (water) - Gravimetric determination in milk and milk products). NMKL is the "Nordisk Method for ” abbreviation.

[1056] The water content of the product can be calculated as 100% minus the relative amount of total solids (% w / w).

[1057] Analysis 13: Measurement of pH

[1058] All pH values ​​were measured at 25 °C using a pH glass electrode.

[1059] The pH glass electrode (with temperature compensation) was rinsed carefully before use and calibrated before use.

[1060] When the sample was in liquid form, the pH was measured directly in the liquid solution at 25°C.

[1061] When the sample was a powder, 10 g of the powder was dissolved in 90 ml of demineralized water at room temperature while stirring vigorously. The pH of the solution was then measured at 25°C.

[1062] Analysis 14: Determination of the amount of calcium, magnesium, sodium, potassium and phosphorus (ICP-MS method)

[1063] The total amounts of calcium, magnesium, sodium, potassium and phosphorus were determined using the following procedure: the samples were first decomposed using microwave digestion and then the total amount of minerals was determined using an ICP device.

[1064] equipment:

[1065] The microwave was from Anton Paar and the ICP was an Optima 2000DV from PerkinElmer Inc.

[1066] Material:

[1067] 1M HNO 3

[1068] 2% HNO3 Yttrium

[1069] 5% HNO 3 Applicable standards for calcium, magnesium, sodium, potassium and phosphorus in pretreatment:

[1070] Weigh a certain amount of powder and transfer the powder into a microwave digestion tube.

[1071] Add 5 mL 1M HNO 3 Digest samples in a microwave according to the microwave instructions.

[1072] Place the digested tubes in a fume hood with the lid removed to allow the volatile fumes to evaporate.

[1073] Measurement procedure:

[1074] Transfer the pre-treated samples to the DigiTUBE using a known amount of Milli-Q water. 3 The yttrium solution in 1% 2% NH 2 O was added to the digestion tube (approximately 0.25 mL per 50 mL diluted sample) and diluted to a known volume with Milli-Q water. The samples were analyzed on ICP using the procedure described by the manufacturer.

[1075] Prepare 10 mL of 1 M HNO by using Milli-Q water 3 and 0.5 mL of yttrium in 2% HNO 3 A blind sample was prepared by diluting the mixture of solutions in to a final volume of 100 mL.

[1076] Prepare at least 3 standards with concentrations covering the expected sample concentrations.

[1077] Analysis 16: Determination of thermal coagulation time

[1078] The thermal stability of whey proteins is particularly important for high protein beverages in the neutral pH range where ultra-high temperature processing is required to ensure extended shelf life of the beverage composition.

[1079] Therefore, the thermosetting time analysis was developed as a measure of thermal stability.

[1080] 1.0 mL of the whey protein nanogel formulation was transferred to a 2 mL 32 x 11.5 mm injection vial (Mikrolab, part number ML33003V) and crimp sealed. The sample was then heated by incubation in an aluminum block preheated to 150°C in a Mikrolab ultra high temperature heating unit.

[1081] The aluminum block is drilled with holes that precisely match the dimensions of the vials, ensuring efficient heat transfer. A temperature increase of 20-100°C is achieved in 40 seconds, and a temperature of 140°C is reached in 132 seconds.

[1082] Aliquots of individual samples were incubated (min:sec) 0:45, 1:00, 1:20, 1:47, 2:22, 3:10, 4:13, 5:38, 7:30 or 10:00, where the temperature in the sample was measured to exceed 140°C after 132 seconds (2:12) to simulate ultra-high temperature conditions. The samples were immediately transferred to cold water after heating to stop further reaction. The samples were inverted and their ability to flow to the bottom was recorded at each heating time. The heat coagulation time (HCT) was determined as the first heating time at which the sample gelled and therefore did not flow to the bottom of the vial when inverted.

[1083] The temperature was recorded by placing a thermocouple thermometer connected to an Omega Hh802 digital thermometer into a vial containing 1 mL of oil to avoid boiling and evaporation of the sample at high temperatures.

[1084] Analysis 17: Determination of total BLG and ALA

[1085] The total amount of BLG and the total amount of ALA in the sample, including BLG and / or ALA in polymerized form, were determined according to Example 1.31 of PCT application number PCT / EP2019 / 067039.

[1086] Example 1. Preparation of dense whey protein nanogel solution

[1087] The inventors previously discovered by chance that whey protein nanogels can be produced in solutions with surprisingly high protein contents. The purpose of this example was to further explore the possibility of making whey protein nanogels at high protein concentrations and to characterize the obtained whey protein nanogel compositions.

[1088] Materials and methods:

[1089] Table 1 Characteristics of exemplary whey protein powders used to prepare whey protein nanogels.

[1090]

[1091]

[1092] An exemplary whey protein solution was prepared by dissolving BLG isolate powder produced as described in WO 2018 / 115520, Example 7. The characteristics of the BLG isolate powder are described in Table 1.

[1093] Mineral content was determined using assay 14, fat, lactose, ash, total solids, and total protein were determined by assays 8, 9, 10, 11, 12, and 7, respectively. Protein composition was determined using assay 6.

[1094] The powders were weighed to produce final whey protein compositions with protein contents of 4, 8, 10, 12 and 16%, taking into account protein / total solids. 90% milli-Q water was added to the samples and the pH was adjusted to 5.9 (or pH 6.0 at 10% protein) using 3M NaOH.

[1095] The remaining water was added and the final pH verified. 15 mL of each sample in Duran GL18 threaded tubes (1.8 mm wall thickness, 16 mm outer diameter, catalog number 28625320) equipped with PBT screw caps (Duran) were heat treated at 90° C. for 14 minutes (or 10 minutes at 10% protein) in a water bath and then immediately cooled in an ice water bath.

[1096] The whey protein nanogels were characterized by dynamic light scattering (Z-average hydrodynamic diameter), transmission electron microscopy, and the fraction of soluble whey protein aggregates and whey protein nanogels as described in analyses 1, 2, and 3, respectively.

[1097] result:

[1098] The whey protein nanogel formulation was formed by heat treating BLG at 90°C for 14 minutes. Surprisingly, it was found that the whey nanogel formulation remained liquid even when heated at high protein concentrations up to at least 16% protein, as shown in Table 2.

[1099] It is generally found that heat treatment at, for example, 90°C for 10 minutes results in 95-97% denaturation, with only 3-5% non-denatured protein present after heat treatment (by assay 6).

[1100] The whey nanogel preparations were characterized by z-average diameters of 196-287 nm, and the fractions of soluble whey protein aggregates and nanogels varied from 6% to 20% and 94% to 80%, respectively, with increasing protein concentration (see Figure 1 ).

[1101] Figure 2 Transmission electron microscopy images of highly uniform spherical nanogels are shown, and the individual sizes of these nanogels are in good agreement with the dynamic light scattering measurements (Table 2). During the TEM sample preparation, staining and drying can affect the particles and cause them to aggregate together, such as Figure 2 shown.

[1102] Although non-spherical aggregates were visually abundant in the TEM images of the 12–16% whey protein nanogel formulations, the mass fraction of spherical aggregates accounted for >80% due to their compact structure.

[1103] It was further found that the fraction of soluble aggregates increased with increasing protein concentration, which was consistent with the appearance of smaller stranded aggregates in TEM images, indicating that increasing protein concentration and / or increasing Na and K contents led to an increase in the fraction of soluble aggregates.

[1104] Table 2 Characteristics of 4-16% whey protein nanogel formulations. nd = not determined

[1105] protein 4％ 8％ 10％ 12％ 16％ pH 5.91 5.92 6.0 5.93 5.92 Ca:BLG w:w ratio 0.0016 0.0016 0.0016 0.0016 0.0016 Na+K(mM) 3.8 7.6 9.5 11.4 15.2 Z-average diameter, nm 211±1 211±3 196 222±3 287±3 Nanogel 94％ 90％ nd 88％ 80％ Soluble aggregates 6％ 10％ nd 12％ 20％

[1106] in conclusion:

[1107] When using a whey protein solution characterized by low calcium (0.0016Ca:BLG w:w) and monovalent salt, a whey protein nanogel preparation can be prepared even at a concentration of at least 16% protein. The major portion (≥80%) of the whey protein nanogel preparation thus produced is composed of spherical nanogels with a size of 211-287 nm, with a small fraction of soluble whey protein aggregates.

[1108] The fraction of soluble whey protein aggregates increases with increasing protein and / or monovalent salt (Na+K) content.

[1109] Example 2: Whey protein nanogel formulation with low content of soluble aggregates

[1110] The purpose of the whey protein nanogel preparation of the present invention is to demonstrate how the presence of monovalent and divalent cations during the heat treatment of a whey protein composition affects the distribution between the nanogel and the soluble whey protein aggregates, and ultimately to enable the heat treatment of a whey protein solution at a high protein concentration without gelation to form a whey protein nanogel.

[1111] Materials and methods:

[1112] Whey protein nanogel preparations were prepared from the BLG powders having the compositions described in Table 1 used in Example 1 by weighing the BLG powders to achieve 3, 4, 5 and 16% BLG, respectively, all with a calcium content of 0.0016 g / g BLG.

[1113] The powders were dispersed in MilliQ water to the desired concentration and dissolved by adjusting the pH to 6.0 using 3M NaOH, while recording the amount of mineral added. 0-50 mM NaCl was added to the formulations from 1 M stock solutions. The total molar concentration of Na+K was calculated by summing up the inherent content of Na+K in the powders using their respective molecular weights, and the amount of Na added as NaCl and the resulting total amount of Na+K are presented in Table 3.

[1114] 1 mL of each sample in 2 mL clear crimp seal vials (Agilent Technologies, Germany) was heated in an aluminum block preheated to 90° C. for 10 min.

[1115] The size of the whey protein nanogels was assessed by light scattering using Assay 2, by assessing the flow behavior of the sample toward the bottom when the sample was inverted (gel / liquid) and by measuring the sample viscosity using a Viscoman as described in Assay 4.

[1116] Samples AE were prepared from the same BLG powder to prepare a 12% BLG composition with an inherent Ca content of 0.0016 g / g BLG. BLG powder was weighed, dispersed in MilliQ water and dissolved by adding 3M NaOH to reach pH 5.9 (Sample A).

[1117] Samples BE were prepared by further titration with 3 M NaOH to reach pH 6.0-6.1-6.3 and 6.5, respectively.

[1118] 15 mL of each sample (AE) was heated in a Duran glass tube at 95° C. for 15 minutes by immersion in a water bath and immediately cooled in an ice water bath after heating.

[1119] Sample FL was prepared from the same BLG powder to prepare a 14% BLG solution with a calcium content of 0.0021 g / g BLG. The BLG powder was weighed, dispersed in MilliQ water and diluted with 5% Ca(OH) 2 The slurry was dissolved by titration (with stirring to ensure homogeneity) to a Ca content of 0.0021 g / g BLG and a pH of 5.97 (Sample F).

[1120] Sample GK was prepared from sample F by adjusting to pH 6.1-6.7 (see table) with 3M NaOH while recording the amount of NaOH added.

[1121] 15 mL of each sample FL was taken and heated at 95° C. for 14 minutes in a water bath to prepare whey protein nanogel, and then cooled immediately after heating.

[1122] Sample set M was prepared as described for sample FL, except that after the initial pH adjustment to 5.97, 1 M CaCl 2 7.5 mM CaCl 2 to achieve a calcium content of 0.0044 g / g BLG. The pH was adjusted to 5.9-6.5 using 3M NaOH and the sample was heated as described for sample FL.

[1123] Sample set N was prepared as described for sample set M, but with the addition of 14.9 mM CaCl 2 to achieve a Ca content of 0.0065 g / g BLG.The pH of an aliquot of the sample was adjusted to within the range of 5.9-6.7 using 3M NaOH and heated as described for sample FL.

[1124] By using 5% Ca(OH) 2 The slurry was adjusted to pH 6.04-5.94-5.89 to reach a Ca content of 0.0022, 0.0024 and 0.0023 g / g BLG, respectively, and the BLG powder was dissolved to reach 14-16-20% BLG to prepare additional whey protein nanogel formulations OPQ. The pH was then adjusted using 3M NaOH to reach pH 6.11, 6.10 and 6.08, respectively. The whey protein nanogel formulation OPQ with a low content of soluble whey protein aggregates was then prepared by heating 15 mL of the sample in a Duran glass tube immersed in a 95°C water bath for 14 minutes.

[1125] The viscosity of sample OPQ was determined according to Analysis 4.

[1126] Samples R and S were prepared as described for sample F and further treated with 5% Ca(OH) 2 Titrated to pH 6.2 and 6.4 respectively and heated as described for sample FL. As described in Table 4, the calcium content of the samples was 0.0028 and 0.0030 g / g BLG.

[1127] The Z-average size and distribution between nanogels and soluble whey protein aggregates in the whey protein nanogel formulations were characterized by assays 2 and 3, respectively. Viscosity was determined by assay 4 and the mineral content was as described in assay 14.

[1128] result:

[1129] 1.0 mL of a whey protein composition characterized by an inherent calcium content of 0.0016 g / g BLG was adjusted to pH 6.0 with 0-50 mM added NaCl and heat treated in an aluminum block at 90° C. for 10 minutes. The amount of NaCl added and the resulting total amount of Na+K after adjustment can be found in Table 3 (3%, 4% and 5% whey protein solutions) and Table 4 (16% whey protein solution).

[1130] Having demonstrated that whey protein nanogel formulations can be formed even at native BLGs as high as at least 16% (Example 1), the inventors found that it was very surprising that there was an apparent threshold concentration at about 20 mM total Na+K above which whey protein solutions formed gels upon heat treatment. At lower protein concentrations (3-5%), whey protein nanogel formulations can be formed at total Na+K of up to at least 23-25 ​​mM.

[1131] Heat treatment of monovalent minerals above this approximate content resulted in sample gelation, regardless of whether the protein dose was 3%, 4%, 5% (Table 3) or even 16% protein (Table 4). In contrast, all samples containing <20 mM Na+K (16%) or <25 mM (3-5%) remained liquid, thus clearly demonstrating the feasibility of preparing whey protein nanogel formulations at low monovalent mineral contents.

[1132] Table 4 further shows that even a slight increase in total Na+K from 15.2 to 19.5 increased viscosity from 24 cP to >100 cP when heat treating the 16% whey protein solution.

[1133] Table 3 Effect of added NaCl content on flow behavior when samples are inverted after heat treatment at 90°C for 14 minutes to form whey protein nanogels with a protein content of 3-5% and a calcium content of 0.0016 g / g BLG at pH 6.0. The flow behavior of the sample was evaluated as liquid when the sample flowed to the bottom when the vial was inverted and did not stay as it was (i.e., gelled state).

[1134]

[1135]

[1136] Table 4 Effect of Na+K content on viscosity and aggregate size of 16% whey protein nanogel formulations after heat treatment at 90°C for 10 minutes at pH 6.0. nd = not determined.

[1137]

[1138] To minimize the addition of monovalent minerals, try using Ca(OH) 2 The pH of the 14% whey protein composition was adjusted by using a slurry so that divalent cations were added during titration instead of monovalent cations.

[1139] Therefore, a 14% whey protein composition was prepared by dissolving BLG using Ca(OH) 2The pH was adjusted to pH 5.97 to bring the Ca content to 0.0021 g / g BLG. However, as shown in Table 6 (sample FL), heat treatment at 95°C for 10 minutes resulted in gelation of the sample.

[1140] Therefore, the pH was further increased by using 3M NaOH to keep the calcium content constant at 0.0021 g / g BLG while increasing the electrostatic repulsion between proteins. This unexpectedly allowed the formation of a whey protein nanogel formulation of 14% protein by heat treatment at 95°C for 10 minutes at a pH of up to pH 6.1 to as high as at least 6.7. As shown in Table 6, in this pH range, total Na+K remained ≤20 mM.

[1141] Surprisingly, the fraction of soluble aggregates in heated whey protein nanogel preparations remained very low (≤ 2%) at pH 6.1-6.3 with no signs of precipitation in the samples despite the high protein concentrations used, making the preparation of nanogels in this pH range particularly beneficial.

[1142] More surprisingly, it was found that by (1) using Ca(OH) 2 A calcium content of approximately 0.0022-0.0024 (sample OPQ) g / g BLG achieved by two-step pH adjustment to achieve the desired calcium content and (2) increasing the pH to approximately 6.1 to ensure total Na+K remained well below 20 mM allowed heat treatment up to protein concentrations of at least 20% BLG, as shown in Table 6.

[1143] However, if calcium levels are not increased by adding CaCl 2 , Ca(OH) 2 or other sources, the 12% whey protein nanogel preparations (samples AD, 0.0016 g / g BLG) heat-treated at pH 5.9-6.1 shown in Table 6 showed that the fraction of soluble whey protein was significantly higher and increased with increasing pH, eventually leading to gelation at pH ≥ 6.3.

[1144] Table 6 (sample group MN) further shows that if CaCl is added 2 Increasing the calcium content in the whey protein composition from 0.0043 to 0.0065 g / g BLG in the pH range between 5.9-6.7 resulted in gelation upon heat treatment.

[1145] Surprisingly, even by using only Ca(OH) 2To increase the pH, liquid whey protein nanogel samples R and S were also prepared by heat treatment at 94°C for 14 min at pH 6.2-6.48, respectively. Nanogel formation was feasible at 0.0028-0.0030 g Ca / g BLG and 3.7 mM total Na+K (see Table 6).

[1146] Table 5. Raw materials and process characteristics used to prepare whey protein nanogels.

[1147] sample FL M N protein 14％ 14％ 14％ <![CDATA[pH setpoint 1 Ca(OH) 2 > 5.97 5.97 5.97 <![CDATA[Added CaCl 2 , mM]]> - 7.5 14.9 pH during heat treatment 5.97-6.67 5.9-6.48 5.87-6.66 Calcium / BLG (g / g) 0.0021 0.0043 0.0064 Monovalent salt 3.3-12.1 3.26-11.3 3.26-15.9 <![CDATA[ Heat Treatment ]]> 95℃ 14min 95℃ 14min 95℃ 14min

[1148] Table 6 Characteristics of heat-treated whey protein nanogel formulations

[1149]

[1150] in conclusion:

[1151] By controlling the amount of calcium per BLG and pH while keeping the monovalent mineral content low (≤20 mM total Na+K), whey protein nanogel formulations containing up to at least 98% nanogels were successfully prepared. 2 Combined with NaOH to adjust pH to achieve proper mineral levels, appropriate calcium content and total Na+K levels are achieved.

[1152] Aggregate sizes at ≥90% nanogel content were observed in the range of 276-563nm. It was observed that nanogel formation requires a higher pH at higher calcium levels to avoid gelation, which can also result in gelation due to the formation of large amounts of soluble whey protein aggregates. Therefore, an intermediate pH tailored to a specific calcium content is required to achieve optimal performance.

[1153] Example 3: Whey protein nanogel prepared at high temperature

[1154] Whey protein solutions containing 4-16% BLG were heat treated at temperatures ranging from 85 to 150°C to evaluate the opportunity of producing whey protein nanogels over a wide temperature range.

[1155] Materials and methods:

[1156] A whey protein solution was prepared by dissolving BLG as described in Example 1 by adjusting to pH 5.9 with 3 M NaOH.

[1157] Sample group A was heat treated at 85°C for 22 minutes. 120 μl of each 4-16% pH 5.9 BLG solution was transferred to a 96-well PCR plate (VWR, catalog number 732-2387) and sealed with adhesive aluminum foil (VWR catalog number 60941-126). The plate was heated in an EscoHealthcare Swiftmax MaxPro thermal cycler, the program was set to balance the sample at 20°C for 5 minutes, then heated to the target temperature at 4°C / second and kept at the target temperature of 85°C for 22 minutes, and then the sample was cooled to 4°C.

[1158] Sample Group B was the same as Sample Group A, but was heat treated at a higher temperature of 95°C for 14 minutes to produce a whey protein nanogel formulation.

[1159] Sample set C consisted of 12% BLG solutions prepared as described above and pH adjusted using 3M NaOH in the range of 5.9-6.0 and 6.1. As described in Example 1, 10 mL of 12% BLG solutions were heated at 95°C for 10 minutes at each pH in a water bath.

[1160] Sample group D is composed of 12% BLG solution of pH 5.9-6.0 prepared as described for sample group C. However, sample group D is heat treated by incubation at 150°C for 0-10 minutes. 1.0mL sample is transferred to 2mL 32x11.5 mm injection bottle (Mikrolab, part number ML33003V) and crimped and sealed. The sample is then heated by incubation in an aluminum block preheated to 150°C in the Mikrolab ultra-high temperature heating unit. Holes precisely matching the size of the vial are drilled on the aluminum block to ensure effective heat transfer. Aliquots of individual samples are usually incubated (minutes: seconds) 0:45, 1:00, 1:20, 1:47, 2:22, 3:10, 4:13, 5:38 or 7:30, and the temperature in the sample is measured to exceed 140°C after 132 seconds to simulate ultra-high temperature conditions. The sample is transferred to cold water immediately after heating to stop further reaction.

[1161] Sample E was prepared as a 14% BLG solution as described in Example 2 for sample set FL and the pH was adjusted to 6.2 using 3 M NaOH.

[1162] The Z-average size of the whey protein nanogel formulations was determined as described in Assay 2. Mineral content was determined using Assay 14. Thermoagulation time and distribution between nanogels and soluble whey protein aggregates were determined using Assays 16 and 3, respectively.

[1163] result:

[1164] Table 7 highlights that whey protein nanogels characterized by a total Na and K concentration of 3.8 to 16 mM and a Z-average diameter of about 180-390 nm can be prepared by heating up to 4 to at least 12% protein solution having a calcium content of 0.0016 g / g BLG and heat treatment at about pH 6 in the temperature range of up to 85°C to at least 95°C.

[1165] Furthermore, when a 14% native whey protein composition characterized by 0.0022 g calcium / g BLG and 6.9 mM Na+K in total was heat treated by incubation in a 150°C heat block for 0-7:30 minutes, it was surprisingly found that the resulting nanogel formulation surprisingly remained liquid and flowed freely to the bottom of the HPLC vial when inverted, even after heat treatment at 150°C for at least 5:38 to 7:30 minutes at pH 6.0 and 5.9, respectively.

[1166] Table 7 Characteristics of heat-treated whey protein nanogel formulations.

[1167]

[1168]

[1169] Table 8 shows that nanoparticulation is feasible at high protein concentrations even under ultra-high temperature conditions when mineral conditions and pH are appropriate by incubating 12-14% whey protein compositions at 150°C for heat treatment at pH 5.9-6.2.

[1170]

[1171]

[1172] in conclusion:

[1173] It was found that whey protein nanogel formulations could be prepared over a surprisingly wide temperature range (eg 85°C / 22 min or 150°C / 7:30 min), and at very high BLG concentrations (up to 4% to at least 16% protein).

[1174] Since 140°C was reached after 2 minutes 12 seconds of heating in a 150°C heat block, with a surprisingly long effective hold time of more than 4 minutes 30 seconds, high heat loads could be applied to 12-14% of the sample while still producing whey protein nanogels in liquid form.

[1175] While it is generally believed that heating high protein concentrations under UHT conditions without shearing will lead to gelation and equipment clogging, it was surprisingly found that treating native BLG at 12-14% under UHT conditions did not lead to sample gelation, even in the absence of applied shear forces. This is particularly surprising because BLG is considered one of the least heat stable whey proteins.

[1176] The size of the obtained nanogels was about 184-389 nm, which was consistent with the previous examples and Figure 2 The aggregate sizes observed by TEM are very consistent.

[1177] Example 4: Heat-stable whey protein nanogel formulation

[1178] This study aimed to demonstrate that protein nanogel formulations are amenable to ultrahigh temperature processing, making them well suited for high-protein beverage applications at neutral pH.

[1179] Materials and methods:

[1180] The 14-16-20% whey protein nanogel formulation is as described in Example 2, sample OPQ.

[1181] The 14-16% and 20% nanogel formulations were subjected to a secondary heat treatment at ultra-high temperature to evaluate thermal stability defined by the first heating time when the sample coagulated. This time is called the thermocoagulation time and is measured according to assay 16.

[1182] In addition, 14% sample is concentrated to 24% protein by following ultrafiltration: by placing 150mL whey protein nanogel preparation in 200mL Amicon stirring cell and it is concentrated, and this stirring cell is equipped with the Koch HFK 328 membrane of nominal molecular weight cutoff being 5kDa.Apply 3 bar (bar) N2 gas to pressurize the cell, thereby allow to concentrate sample by forcing liquid through ultrafiltration membrane.Measure the Brix of retentate and permeate, and calculate the retentate protein content as described in analysis 10.During the concentration process, temporarily release pressure and remove the top of the cell, so that by Brix (analysis 10) measure protein concentration and as described in analysis 4 measure viscosity.

[1183] The thermosetting time of the 24% concentrate was determined according to assay 16.

[1184] The Z-average diameter and viscosity of the whey protein nanogel formulations were determined by assays 2 and 4, respectively. Assay 3 was used to determine the distribution between whey protein nanogels and soluble whey protein aggregates.

[1185] result:

[1186] Table 9 summarizes the characteristics of whey protein nanogel formulations prepared by heat treatment at high protein concentrations.

[1187] The non-heated solution was characterized by 0.0022-0.0024 g Ca / g BLG and low levels of monovalent ions as indicated by less than 11 mM total Na+K.

[1188] The whey protein nanogel preparations were characterized by surprisingly low viscosities of 2.9-4.7 cP despite high protein content (14-20%, respectively), and the Z-average diameter of the aggregates was found to be in the range of about 350-560 nm. The content of nanogels relative to total protein was found in the samples at very high levels (>98%).

[1189] As can be seen in Table 9, it was surprisingly found that the nanogel formulations were particularly heat stable, as all samples at 14%, 16%, 20% protein and even the 14% whey protein nanogel formulation concentrated to 24% remained liquid after 2 minutes 20 seconds of heating and were therefore resistant to ultra-high temperatures, and required even further heating at 150°C for at least 7 minutes 30 seconds at 14-16% protein or 3 minutes 10 seconds at 20% and 24% protein to force gelation, see Table 9.

[1190] Table 9 Characteristics of whey protein nanogel formulations prepared by heat treatment at 90°C for 10 minutes at 14-20% protein concentration.

[1191]

[1192]

[1193] in conclusion:

[1194] Heat stable whey protein nanogel formulations with diameters of about 353-562 nm and low viscosity can be prepared at high protein concentrations by heat treatment at pH 14-20% protein, 0.0022-0.0024 g calcium / g BLG, and less than 10.6 mM Na+K.

[1195] The samples were characterized by a high ratio of whey protein nanogels (>98%) and were found to be remarkably resistant to heat treatment as they remained liquid even after heat treatment above 140°C, with even the 14% sample concentrated to 24% protein by ultrafiltration resisting heating to ultrahigh temperatures.

[1196] Example 5: Low viscosity whey protein preparation

[1197] To produce whey protein nanogel formulations with low content of soluble aggregates to reduce the protein contribution of soluble whey protein aggregates and to evaluate the feasibility of preparing highly concentrated whey protein nanogel formulations.

[1198] High protein at low viscosity is a strong advantage in industrial processing, where the ability to achieve higher protein concentrations directly increases the capacity of the spray drying equipment. The development of low viscosity is further needed in production because the transmembrane flux (such as in the case of ultrafiltration) is inversely proportional to the viscosity of the liquid to be filtered.

[1199] Furthermore, low viscosity is required in high protein whey protein drinks and high protein yogurts.

[1200] Materials and methods:

[1201] By dispersing the powder into MilliQ, 5% Ca(OH) 2 The slurry was adjusted to pH 6.0 for dissolution and 200 mL of 14% BLG composition was prepared using 3M NaOH to adjust the final pH to 6.2. The sample was dispensed into several 15 mL aliquots, immersed in a 95°C water bath for 14 minutes, immediately cooled after heating, and combined into a single sample.

[1202] The samples were concentrated by ultrafiltration (UF) while tracking protein concentration and viscosity at appropriate time points.

[1203] The viscosity of the whey protein nanogel formulation was measured using a rheometer (Anton Paar, Physica MCR301). 19.6 mL of sample was added to a cup CC27-SS (SN33864) and the sample was equilibrated to 8°C. A 1 s -1 With 1000s -1 The viscosity was scanned at a shear rate of 300s -1 The shear rate is expressed in centipoise (cP).

[1204] Ultrafiltration concentration was performed by placing 150 mL of the 14% sample in a 200 mL Amicon stirred cell equipped with a Kock HFK328 membrane. 3 bar nitrogen was applied to pressurize the cell and concentrate the sample. The Brix of the permeate was always measured as 0.0, thus equal to water, indicating that protein did not pass through the membrane. During the concentration process, the pressure was temporarily released so that the protein concentration could be measured according to the Brix as described in Analysis 10 and the viscosity was as described in Analysis 4. The sample was removed after concentration to 24% protein and left overnight at 4°C.

[1205] result:

[1206] Table 10 describes the characteristics of a nanogel formulation containing 95.5% nanogel, which was prepared by heat treatment at 95°C for 14 minutes at 14% protein in the presence of a calcium content of 0.0022 g / g BLG and Na and K combined to a concentration of 6.9 mM. Although such heat treatment of concentrated whey protein formulations would normally result in gelation and / or require high shear using, for example, a scraped heat exchanger to avoid gelation during heating, the inventors have found that the heat treated whey protein formulations maintain a very low viscosity, similar to the viscosity of a native whey protein solution within experimental error, as shown in Table 10.

[1207] The whey protein nanogel preparations were further concentrated by ultrafiltration while monitoring the viscosity development with protein content, e.g. Figure 3 Surprisingly, it was found that the nanogel formulations could be concentrated up to at least 29.5% protein and, contrary to expectations, the viscosity even closely followed that of the non-heated whey protein solution up to at least 29.5% protein.

[1208] Table 10 Characteristics of whey protein nanogel formulations used to study flow behavior

[1209]

[1210]

[1211] in conclusion:

[1212] The combination of high protein (14%), low Na+K and 0.0022 g calcium / g BLG with heat treatment at 95°C for 14 min resulted in a stable whey protein nanogel formulation characterized by 95.5% nanogel relative to total protein and a Z-average size of 305.5 ± 2.8 nm.

[1213] It was found that the viscosity of the concentrated whey protein nanogel formulations was consistently low and approximately equal to the viscosity of native BLG solutions, even when concentrated to high protein levels up to at least 29.5%.

[1214] This allows for a concentration of the UHT temperature resistant whey protein nanogel (24% in the previous example), making it particularly useful at neutral pH where UHT treatment is a prerequisite for, for example, medical beverages requiring high protein for several patient groups.

[1215] Example 6: Low viscosity acidified product

[1216] Nanogel formulations containing small amounts of soluble aggregates were produced to demonstrate that low viscosity acidified products could be prepared.

[1217] method:

[1218] Samples AC were prepared from BLG powder to prepare a 12% BLG composition with an inherent calcium content of 0.0016 g / g BLG. BLG powder was weighed, dispersed in MilliQ water and dissolved by adding 3M NaOH to reach pH 5.9 (Sample A).

[1219] Sample BC was prepared by further titration with 3 M NaOH to reach pH 6.0 and 6.2, respectively.

[1220] Sample D was prepared from BLG powder to prepare a 16% BLG whey protein nanogel formulation with a particularly low acid gellable protein content. The protein was weighed, dispersed in MilliQ water and diluted with 5% Ca(OH) 2 The slurry was titrated to pH 5.94. The pH was then adjusted to 6.0 using 3M NaOH.

[1221] 3 x 18 mL of each sample AD was heated in a Duran glass tube at 95 °C for 15 min by immersion in a water bath and immediately cooled in an ice water bath after heating.

[1222] Weigh 15 grams each of Samples AD and add 30 grams of MilliQ (Samples AC) or 45 mL of MilliQ (Sample D) to dilute the samples to 4% protein. The diluted samples were assessed for gel formation after acidification according to Assay 5.

[1223] The mineral composition of the whey protein solution was determined using assay 14.

[1224] The distribution between whey protein nanogels and soluble whey protein aggregates was determined by assay 3. The structure formation during acidification was determined using assay 5.

[1225] result:

[1226] 12-16% whey protein nanogel formulations with different contents of soluble aggregates were successfully prepared as shown in Table 11, including the 16% sample, which had a very low soluble aggregate content of 1.9%.

[1227] Figure 4It was shown that acidification of the samples in the rheometer (all diluted to 4% protein before analysis) resulted in a substantial increase in the storage modulus (gel strength) of the samples containing a large amount of soluble aggregates (e.g., a maximum of 1257 was observed for sample A at about 18 minutes (pH 4.4)), while a decrease in the maximum gel strength was observed as the fraction of soluble whey protein aggregates decreased. For example, a storage modulus of 7.1 Pa was the maximum value measured during acidification of a 16% sample (D) containing 1.9% acid gelable protein, which makes this whey protein nanogel formulation particularly suitable for acidified / fermented products requiring a low degree of protein gelation, such as acidified whey protein beverages and high-protein yogurt.

[1228] Table 11 BLG whey protein nanogel preparation

[1229]

[1230]

[1231] in conclusion:

[1232] Whey protein nanogel formulations produced at high protein concentrations under conditions that favor the formation of small amounts of soluble aggregates are particularly suitable for low-viscosity acidified products because the viscosity remains low even after acidification.

[1233] Example 7: Preparation of whey protein nanogel using a mixture of WPI and BLG

[1234] Whey protein nanogel formulations were produced by mixing BLG compositions with WPI to demonstrate that mineral contributions from WPI sources prevented the formation of whey protein nanogel formulations at high protein concentrations at ≥20-25 mM Na+K and / or excessively high Ca:BLG ratios.

[1235] method:

[1236] A 12% whey protein composition (A) was prepared by dispersing BLG isolate powder (see Example 1) in MilliQ water and dissolving by adjusting the pH to 6.0 using 3M NaOH. A 12% whey protein composition (B) was prepared by further titrating to pH 6.4 using 3M NaOH.

[1237] The third whey protein composition (C) was thawed (frozen), diluted to 12% in MilliQ water, and adjusted to pH 6.0 using 3M NaOH. The protein composition of composition C is provided in Table 12.

[1238] 12% whey protein solutions consisting of 100% 12% BLG A and mixtures of 80:20, 75:25, 66.7:33.3 and 50:50 of 12% BLG A solution and 12% WPI solution were prepared by mixing appropriate volumes of BLG A and WPI.

[1239] Additionally, a 66.7:33.3 mixture of BLG B and WPI was prepared to give a whey protein composition with a pH of 6.2.

[1240] All samples were heated at 95°C for 14 minutes and immediately cooled after heating.

[1241] Assess the condition of the sample by inverting the glass tube.

[1242] The distribution between soluble aggregates and nanogels was assessed by using the Brix measured before / after centrifugation at 50.000 xg for 1 hour.

[1243] Table 12 Composition of WPI solution

[1244]

[1245]

[1246] The mineral content was determined using assay 14.

[1247] The distribution between whey protein nanogels and soluble whey protein aggregates was determined by Assay 3.

[1248] result:

[1249] Table 13 shows the whey protein nanogel formulations prepared by mixing BLG with WPI.

[1250] All nanogel formulations had low total Na+K contents, and when the calcium content was 0.0032 g / g BLG, the formulations remained liquid after heating at pH 6.0, while higher calcium contents resulted in extensive formation of gel pieces or gelation of the entire sample.

[1251] Surprisingly, it was found that increasing the pH to 6.2 reduced gelation and allowed the formation of a 12% whey protein nanogel formulation, even when the calcium content was 0.0037 g / g BLG and more ALA and CMP were present in the sample.

[1252] Table 13 Whey protein nanogel preparations prepared from BLG and WPI

[1253]

[1254] The non-denatured proteins found in the whey protein nanogel formulations consisted mainly of CMP.

[1255] in conclusion:

[1256] When the Na and K contents are low enough and the pH is increased to compensate for the higher calcium levels, whey protein nanogel formulations can be prepared at higher protein concentrations, in the presence of other proteins besides BLG, and at elevated calcium levels.

[1257] Example 8: Use of nanogel for preparing low-astringency acidic beverages

[1258] The inventors have found that whey protein in nanogel form has less astringency than other types of denatured whey. Astringency is not a taste, but a mouth sensation, and feels like cheek muscle contraction and increased saliva secretion. For example, eating foods such as unripe fruits, red wine, and heat-sterilized acidic protein drinks may produce astringency. In this embodiment, the inventors studied the low astringency of whey protein nanogels.

[1259] method

[1260] The BLG nanogel formulation was prepared as described in Example 2 "OPQ" with the following modifications:

[1261] Using 5% Ca(OH) 2 Slurry 10% BLG characterized by the properties of Example 1, Table 1 was adjusted to a Ca:BLG ratio of 0.0021 w:w and further adjusted to 6.0 using 10% NaOH.

[1262] A pilot-scale scraped heat exchanger was used for thermal treatment at 93 °C for 10 min, followed by cooling to 10 °C. Finally, the nanogel formulation was concentrated to 20.4% protein using a spiral wound reverse osmosis membrane (Alfa-laval RO98pHt-65-263364) at 20 bar pressure on a MMS SW40 RO pilot unit and spray dried on an anhydrous Microspray MS 750 to prepare nanogel powder.

[1263] 7.3 kg of whey protein nanogel powder having the characteristics described in Table 14 below was dispersed in 30.8 kg of demineralized water in a scanima mixer to prepare an 18% BLG nanogel formulation and allowed to hydrate overnight at 10°C with gentle stirring.

[1264] Disperse homogeneously at 200 bar, then adjust the pH using 9% HCl and dilute in demineralized water to prepare a 10% BLG nanogel solution at pH 3.5.

[1265] The nanogel solution was sterilized using an OMVE HT320 pilot plant ultra-high temperature system equipped with a plate heat exchanger for thermal treatment at 120°C for 20 seconds and introduced into 100 mL screw cap bottles.

[1266] 1.1 kg of acidic BLG powder having the characteristics described in Table 14 prepared according to Example 2 of PCT application WO 2020 / 002426 was dissolved in 8.9 kg of demineralized water to produce a 10% feed solution. The pH was 3.51 and no further adjustment was required for use. The BLG solution was sterilized using an OMVE HT320 pilot plant ultra-high temperature system equipped with a plate heat exchanger for heat treatment at 120°C for 20 seconds and introduced into 100 mL screw cap bottles.

[1267] The viscosity of the beverage formulations was measured as described in Analysis 4. The perceived astringency level was assessed as described in PCT Application WO 2020 / 002435, with the modification that non-diluted samples were assessed in this example.

[1268] As described in PCT application WO 2020 / 002435, astringency is assessed on a scale of 0-15, with 0 indicating no intensity and 15 indicating high intensity.

[1269] Brightness and turbidity were determined according to Example 1.9 and Example 1.7 of PCT application WO 2020 / 002435. Mineral, total solids, ash and protein contents of the raw material, nanogel and beverage composition were determined according to the current assays 14, 12, 11 and 7, respectively.

[1270] result

[1271] A 10% nanogel formulation characterized by a z-average hydrodynamic diameter of 458 nm (Assay 2), a viscosity of 5.3 cP (Assay 4), 89% nanogel and 11% soluble aggregates was prepared, concentrated and dried to prepare a nanogel powder. The nanogel powder was rehydrated and homogenized to facilitate its use in beverage compositions.

[1272] As shown in Table 15 below, a 10% whey protein nanogel beverage composition was prepared at pH 3.5 by heat treatment at 120°C / 20 seconds and compared with a 10% acidic BLG composition also heat treated at 120°C / 20 seconds. The protein fraction of the resulting ultra-high temperature treated 10% reference beverage consisted essentially of soluble aggregates and monomeric proteins (98%) and very little nanogel (2%).

[1273] Table 14 Characteristics of nanogel powder and acidic BLG powder.

[1274]

[1275] The nanogel beverage composition produced a beverage with a milky appearance, as evidenced by high turbidity and lightness (L*), while in contrast the acidic BLG reference beverage was clear, also indicated by the amount of soluble aggregates and monomeric protein (see Table 15).

[1276] The phosphorus content in the nanogel drink was below the detection limit, making this nanogel formulation particularly suitable for patients with chronic kidney disease.

[1277] The sensory evaluation confirmed the inventors' preliminary findings and clearly demonstrated that the astringency of the nanogel beverage composition was significantly reduced compared to the clear reference beverage (see Table 15). This effect is believed to be closely related to the low content of soluble aggregates and monomeric proteins in the milky nanogel beverage compared to the clear BLG reference beverage, and indicates that whey protein nanogels are particularly useful for acidic whey protein beverages processed by ultra-high temperature.

[1278] Table 15 Characteristics of nanogel beverage compositions and their processing.

[1279]

[1280] in conclusion

[1281] A 'milky' nanogel beverage with high turbidity and whiteness of a nutritional composition was prepared. The beverage contained extremely low levels of phosphorus, which is particularly important but not limited to making a high protein beverage for chronic kidney disease that cannot tolerate high levels of phosphorus.

[1282] Sensory testing showed that the UHT treated nanogel beverage composition had essentially no astringency and was much less astringent than the UHT treated BLG reference beverage.

[1283] Example 9: Use of nanogel in preparing acidic high-energy beverages

[1284] The feasibility of using nanogels to prepare high-energy nutritional compositions processed by ultra-high temperature under acidic conditions was evaluated.

[1285] method

[1286] 10.8 kg of nanogel powder prepared as described in Example 8 was dispersed in 68.5 kg of tap water at 65° C. Emulsifier, oil and dry ingredients were mixed into the liquid using a Brine Mixer Rotostat for 15 minutes.

[1287] The pH was adjusted to 3.5 using 10% phosphoric acid and the nutritional composition was homogenized at 150 bar. After homogenization, the nutritional composition was heat treated at 120° C. for 30 seconds on a tubular heat exchanger and cooled to 10° C. before being filled into 100 mL sterile bottles.

[1288] Viscosity was measured according to assay 4, and astringency and sourness were determined according to PCT application WO 2020 / 002435, but without dilution prior to the sensory testing.

[1289] result

[1290] The inventors have found that it is possible to prepare an acidic, high energy drink characterized by a surprisingly low viscosity of 11.3 cP. In contrast, it was found that replacing the protein in nanogel form with an equal protein mass of acidic, unfractionated whey protein isolate resulted in clogging and plugging of ultra high temperature equipment. A low viscosity, acidic, high energy nutritional composition comprising 10% protein, 4% fat and 16% carbohydrates was prepared to provide a high energy density of 140 kcal / 100 mL in a small package.

[1291] Tasting the nanogel composition revealed a surprisingly low level of astringency, scoring only 1 on a 0-15 (low-high) scale, compared to a score of 5 for the 10% BLG reference beverage evaluated in Example 8. In addition, despite the high protein concentration (10%) and the undiluted sample (e.g., compared to Figure 15 of PCT application WO 2020 / 002435), the sourness of the nanogel beverage composition at pH 3.5 was surprisingly only 2 on a 0-15 scale.

[1292] in conclusion

[1293] By combining the nanogel with a fat and carbohydrate source under acidic conditions, a low viscosity, high energy nutritional nanogel formulation was successfully prepared after UHT treatment with excellent sensory properties (low perceived astringency and sourness / acidity). Attempts to produce a reference product based on a similar but unfractionated native whey protein isolate resulted in clogging of the UHT apparatus due to uncontrolled aggregation upon heating.

[1294] Example 10: Use of nanogels for preparing high energy pH neutral beverages

[1295] The feasibility of using nanogels to prepare ultra-high temperature processed high energy liquid nutritional compositions at neutral pH was evaluated.

[1296] method

[1297] Whey protein nanogel powder was prepared as described in Example 8 and used to prepare 18% whey protein nanogel solution, which was hydrated overnight at 4°C and then homogenized at 200 bar. The homogenate was adjusted to pH 7.0 using 3M NaOH and preheated to 50°C in a water bath. 0.5 g / L Grindsted Citrem LR10, rapeseed oil, sucrose and the remaining water used to form the nutritional nanogel composition shown in Table 16 were mixed into the homogenate sample. The weight fractions relative to the final liquid nutritional composition are provided.

[1298] Finally, the nutritional composition was homogenized at 150 bar.

[1299] 1 mL of each nutritional composition was transferred to a crimp sealed HPLC vial and heated by incubating the vial in an aluminum block at 150° C. for 10 minutes. The heated samples were analyzed for their ability to flow when the sample was inverted and the viscosity was measured after the vial was uncapped according to Assay 4.

[1300] result

[1301] Nutritional nanogel compositions were prepared that included a source of carbohydrate (range 8-17 w:w % sucrose), a source of fat (rapeseed oil; range 4-9 w:w %), and a source of nanogel (range 10-15 w:w %). Surprisingly, even after prolonged extensive heating at 150°C for 10 minutes, all evaluated nanogel compositions remained liquid and free-flowing, regardless of the protein, fat, and carbohydrate content evaluated.

[1302] Even at the highest protein dose evaluated of 15%, the viscosity remained very low, and the nanogels could even be used to prepare sterilized nutritional compositions with an energy content of at least 200 kcal / 100 mL by combining 15% protein, 8% fat, and 17% carbohydrates, as evidenced by the liquid state and low measured viscosity after heating at 150 °C for 10 min.

[1303] Table 16 Composition and characteristics of sterilized nutritional compositions.

[1304]

[1305] in conclusion

[1306] A low viscosity, free-flowing, high energy nutritional nanogel composition can be prepared by heat treatment, which can resist heat treatment at a temperature of at least 150° C. for up to at least 10 minutes, and has an energy content of up to at least 200 kcal / 100 mL. Therefore, a high protein beverage based on the whey protein nanogel of the present invention can be sufficiently heat stable to withstand retort-type heat treatment even at high protein concentrations.

[1307] Example 11: Acidic and pH neutral whey protein nanogel powders and use for preparing high protein beverages.

[1308] Whey protein nanogel powders at acidic pH (pH 3.5) and neutral pH (pH 7.0) were prepared and used to prepare high-protein beverages.

[1309] method

[1310] The nanogel powder was produced essentially as described in Example 8 with the following modifications:

[1311] The nanogel preparation was concentrated to 16% protein and divided into three equal parts, two of which were adjusted to pH 3.5 and 7.0 with 4.5% HCl and 2% NaOH, respectively. The three feeds were spray dried separately to prepare nanogel powders.

[1312] 20% nanogel dispersion of each nanogel powder (pH 3.5, 6.6 and 7.0) was rehydrated in demineralized water and homogenized at 150 bar. The homogenate was diluted to 10-20% in demineralized water and pH was readjusted if necessary. 1 mL of sample was transferred to an HPLC injection bottle (Mikrolab, part number ML33003V) and crimped to seal. At 170°C, the sample was inserted into an aluminum block having a drilled hole that precisely matched the size of the vial to ensure effective heat transfer. The sample was kept at 170°C for 120 seconds, or kept at 90°C for up to 10 minutes, and cooled immediately.

[1313] The first indication regarding flow properties (liquid / gel) was determined by the ability of the vial to flow when inverted, and the viscosity was determined by assay 4.

[1314] The particle size after heating was determined using a Malvern Mastersizer 3000 equipped with a HydroLV liquid dispersion unit. The heated nanogel solution was dripped into a dispersion unit containing water under stirring at 1500 rpm until an obscuration of 8-10% was achieved. When calculating the particle size in the Mastersizer 3000 software version 3.62, spherical particles were assumed, using an RI of 1.4, an absorption index of 0.01, and a solvent (water) refractive index of =1.33 to estimate the particle size.

[1315] result

[1316] Nanogel formulations characterized by a particle size of 0.424 microns were successfully prepared, followed by pH adjustment, concentration and drying to produce pH 3.5, 6.6 and 7.0, resulting in nanogel powders of the following composition (% w / w of total powder):

[1317]

[1318] Nanogel formulations were used to evaluate the accessible protein range of heat-treated, low-viscosity but protein-rich nanogel beverages:

[1319] Table 17 Characteristics and process parameters of heat treated beverages.

[1320]

[1321] in conclusion

[1322] Powders at pH 3.5, 6.6 and 7.0 were successfully prepared and used to produce acidic and pH neutral high protein beverages.

[1323] A nutritious high protein beverage was prepared by heat treating a beverage containing the following nanogels under the following conditions:

[1324] Up to at least 18% at pH 7.0 by heating at 170°C for up to at least 120 seconds. Up to at least 20% at pH 7.0 by heating at 90°C for up to at least 10 minutes. Up to at least 20% at pH 6.6 by heating at 90°C for up to at least 10 minutes. Up to at least 14% at pH 3.5 by heating at 170°C for 120 seconds.

Claims

1. A method for producing a whey protein nanogel composition, the method comprising: a) providing a whey protein solution, the solution having: - a natural BLG content in an amount of at least 3% w / w, - a pH value in the range of 5.8-7.5, preferably in the range of 5.8-6.5, -The weight ratio between total calcium and natural BLG is: - up to 0.010, and - The following total concentrations of monovalent metal cations: - if the native BLG content of the whey protein solution is below 10% w / w, Then the total concentration is at most 25 mM, or - if the native BLG content of the whey protein solution is at least 10% w / w, the total concentration is at most 20 mM, b) heating the whey protein solution to a temperature of at least 68° C. for a period of time sufficient to form a suspension of whey protein nanogels, c) optionally, concentrating the suspension of whey protein nanogel to obtain a concentrated suspension of whey protein nanogel, d) optionally, drying the dryer feed comprising the whey protein nanogel originating from step b) or c).

2. The method according to claim 1, wherein: The whey protein solution comprises native BLG in an amount of 3-30% w / w, more preferably in an amount of 4-28% w / w, even more preferably in an amount of 6-26% w / w and most preferably in an amount of 8-24% w / w.

3. The method according to any one of the preceding claims, wherein the weight ratio between the total amount of calcium of the whey protein solution and native BLG is at most 0.0070, more preferably at most 0.0065, even more preferably at most 0.0060, and most preferably at most 0.0050.

4. The method according to any one of the preceding claims, wherein the weight ratio between the total amount of calcium of the whey protein solution and native BLG is at most 0.0045, more preferably at most 0.0040, even more preferably at most 0.0035, most preferably at most 0.0030.

5. The method according to any one of the preceding claims, wherein the weight ratio between the total amount of calcium of the whey protein solution and native BLG is at most 0.0025, more preferably at most 0.0020, even more preferably at most 0.0015, and most preferably at most 0.0005.

6. The method according to any one of the preceding claims, wherein the weight ratio between the total amount of calcium of the whey protein solution and native BLG is in the range of 0.0005-0.0065, more preferably in the range of 0.0010-0.0050, even more preferably in the range of 0.0015-0.0040, and most preferably in the range of 0.0015-0.0030, such as preferably in the range of 0.0017-0.0027.

7. A method according to any one of the preceding claims, wherein: The whey protein solution comprises native BLG in an amount of 11-30% w / w, more preferably in an amount of 12-28% w / w, even more preferably in an amount of 14-26% w / w and most preferably in an amount of 16-24% w / w.

8. A method according to any one of the preceding claims, wherein: The whey protein solution comprises native BLG in an amount of at least 50% w / w relative to the total protein, more preferably at least 60% w / w relative to the total protein, even more preferably at least 70% w / w relative to the total protein, and most preferably at least 80% w / w relative to the total protein.

9. A method according to any one of the preceding claims, wherein: The whey protein solution comprises native BLG in an amount of at least 85% w / w relative to the total protein, more preferably at least 90% w / w relative to the total protein, even more preferably at least 92% w / w relative to the total protein, and most preferably at least 95% w / w relative to the total protein.

10. A method according to any one of the preceding claims, wherein: The whey protein solution has a degree of protein denaturation of at most 20% w / w relative to the total protein, more preferably at most 10% w / w relative to the total protein, even more preferably at most 5% w / w relative to the total protein, and most preferably at most 2% w / w relative to the total protein.

11. A method according to any one of the preceding claims, wherein: The pH value of the whey protein solution is in the range of 5.8-6.5, more preferably 5.9-6.3, and even more preferably 5.9-6.

2.

12. A method according to any one of the preceding claims, wherein: The whey protein solution has a total concentration of monovalent metal cations of at most 19 mM, more preferably at most 17 mM, even more preferably at most 15 mM, and most preferably at most 10 mM.

13. A method according to any one of the preceding claims, wherein: The heating of step b) is performed for a duration sufficient to denature at least 50% of the native BLG on a weight / weight basis, more preferably at least 80% of the native BLG on a weight / weight basis, even more preferably at least 90% of the native BLG, and most preferably at least 95% of the native BLG on a weight / weight basis.

14. The method according to any one of the preceding claims, comprising step c) and wherein the concentrated suspension of whey protein nanogels comprises a total amount of protein of 21-35% w / w, more preferably 25-33%, even more preferably 28-32% w / w, and most preferably 29-31% w / w.

15. A method according to any one of the preceding claims, wherein: The concentrated suspension of whey protein nanogel comprises soluble whey protein aggregates in an amount of at most 8% w / w, more preferably at most 6%, even more preferably at most 3% w / w, and most preferably at most 1% w / w.

16. The process according to any one of the preceding claims, comprising step d) and wherein the dryer feed comprises a total amount of protein of 21-35% w / w, more preferably 25-33%, even more preferably 28-32% w / w, and most preferably 29-31% w / w.

17. A whey protein nanogel composition in powder or liquid form, comprising: - whey protein nanogel in an amount of at least 60% w / w relative to the total protein, - soluble whey protein aggregates in an amount of up to 10% and more preferably 5% w / w relative to the total protein, - total BLG in an amount of at least 50% w / w relative to total protein, and - A total amount of protein of at least 30% relative to total solids.

18. The whey protein nanogel composition according to claim 17, wherein the particles of the hydrated form of the whey protein nanogel composition have a z-average diameter of 150-1000 nm, more preferably 170-900 nm, even more preferably 200-800 nm, and most preferably 250-700 nm as measured by dynamic light scattering.

19. The whey protein nanogel composition according to claim 17 or 18, wherein the whey protein nanogel composition comprises soluble whey protein aggregates in an amount of at most 20% w / w, more preferably at most 10% w / w, and most preferably at most 5% w / w relative to total protein.

20. The whey protein nanogel composition according to claim 17 or 18, wherein the whey protein nanogel composition comprises soluble whey protein aggregates in an amount of at most 3% w / w, more preferably at most 2% w / w, even more preferably at most 1% w / w, and most preferably at most 0.5% w / w relative to total protein.

21. The whey protein nanogel composition according to any one of claims 17 to 20, wherein the whey protein nanogel composition comprises total BLG in an amount of at least 60% w / w relative to total protein, more preferably at least 70% w / w relative to total protein, and most preferably at least 80% w / w relative to total protein.

22. The whey protein nanogel composition according to any one of claims 17 to 21, wherein the whey protein nanogel composition in liquid form has a viscosity of 100 % w / w total protein at 20°C and 300 s -1 The viscosity of the present invention is at most 200 cP at a shear rate of 20°C and 300 s -1 The viscosity of the product is at most 100 cP at a shear rate of 20°C and 300 s -1 The viscosity is at most 50 cP at a shear rate of 1.5 s, and most preferably at 20°C and 300 s -1 The viscosity of the composite is at most 20 cP at a shear rate of 1.5 %.

23. The whey protein nanogel composition according to any one of claims 17 to 22 is used for one or more of the following: - As food ingredients, - as a food ingredient for the production of an aseptic beverage containing at least 10% protein and even more preferably at least 21% protein, and - As whitening agent, for example in coffee whitener.

24. A packaged heat treated beverage having a pH in the range of 3-8 and comprising whey protein nanogel in an amount of at least 1% w / w, preferably, wherein the packaged heat treated beverage is aseptic.

25. A packaged heat treated beverage according to claim 24, wherein the packaged heat treated beverage comprises total protein in an amount in the range of 1-32% w / w, more preferably 5-31% w / w, even more preferably 10-30% w / w, and most preferably 21-30% w / w.

26. A packaged heat treated beverage according to claim 24 or 25, wherein the packaged heat treated beverage comprises total protein in an amount in the range 8-32% w / w, more preferably 9-24% w / w, even more preferably 10-22% w / w, and most preferably 11-20% w / w.

27. A packaged heat-treated beverage according to any one of claims 24-26, wherein the packaged heat-treated beverage comprises total BLG in an amount of at least 50% w / w relative to total protein, more preferably at least 60% w / w relative to total protein, even more preferably at least 70% w / w relative to total protein, and most preferably at least 80% w / w relative to total protein.

28. A packaged heat-treated beverage according to any one of claims 24-27, wherein the packaged heat-treated beverage comprises at least 85% w / w total BLG relative to total protein, more preferably at least 90% w / w relative to total protein, even more preferably at least 92% w / w relative to total protein, and most preferably at least 95% w / w relative to total protein.

29. A packaged heat-treated beverage according to any one of claims 24 to 28, wherein the packaged heat-treated beverage comprises whey protein nanogel in an amount of at least 50% w / w, more preferably at least 70% w / w, even more preferably at least 80% w / w, and most preferably at least 90% w / w relative to total protein.

30. A packaged heat-treated beverage according to any one of claims 24 to 29, wherein the packaged heat-treated beverage comprises soluble whey protein aggregates in an amount of at least 30% w / w, more preferably at least 20% w / w, even more preferably at least 10% w / w, and most preferably at least 5% w / w relative to total protein.

31. A packaged heat-treated beverage according to any one of claims 24 to 30, wherein the packaged heat-treated beverage comprises whey protein microparticles in an amount of at most 10% w / w, more preferably at most 5% w / w, even more preferably at most 3% w / w, and most preferably at most 1% w / w relative to total protein.

32. A packaged heat-treated beverage according to any one of claims 24-31, wherein the packaged heat-treated beverage has a total solids content in an amount of 3-50% w / w, more preferably in an amount of 4-40% w / w, even more preferably in an amount of 6-35% w / w, and most preferably in an amount of 8-30% w / w.

33. A packaged heat-treated beverage according to any one of claims 24 to 32, comprising: - a total amount of BLG of at least 90% w / w, most preferably at least 95% w / w relative to total protein, - whey protein nanogel in an amount of at least 85% w / w, most preferably at least 89% w / w relative to the total protein, and - Soluble whey protein aggregates in an amount of at most 15% w / w, and most preferably at most 11% w / w, relative to total protein.

34. A packaged heat treated beverage according to any one of claims 24 to 33 having: - a pH value in the range of 5.5-8.0, more preferably in the range of 6.0-7.5, even more preferably in the range of 6.2-7.3, and most preferably in the range of 6.3-7.2, or - a pH value in the range of 3.0-5.4, more preferably in the range of 3.5-5.0, even more preferably in the range of 3.7-4.8, and most preferably in the range of 4.0-4.

6.

35. A packaged heat treated beverage according to any one of claims 24 to 34, wherein the packaged heat treated beverage comprises a total amount of carbohydrates in the range between 30-60% and most preferably in the range between 35-50E% of the total energy content of the beverage, and Preferably, the total amount of lipids is in the range of 20-50% of the total energy content, more preferably in the range of 25-45E%, and most preferably in the range of 30-40E%.

36. A packaged heat-treated beverage according to any one of claims 24-35, wherein the whey protein nanogel in hydrated form has a z-average diameter of 150-1000 nm, more preferably 170-900 nm, even more preferably 200-800 nm, and most preferably 250-700 nm.

37. A method for producing a packaged heat treated beverage having a pH in the range of 3-8 according to any one of claims 1-14, said method comprising the steps of: 1) Providing a liquid solution having a pH in the range of 3-8 and comprising: - 1 to 32% by weight of total protein, of which at least 50 w / w % of the protein is beta-lactoglobulin (BLG), - optionally, a sweetener and / or a flavoring agent, and wherein the liquid solution comprises whey protein nanogel in an amount of at least 50% w / w relative to the total protein, 2) packaging the liquid solution, The liquid solution of step 1) and / or the packaged liquid solution of step 2) is subjected to a heat treatment including at least pasteurization.

38. A beverage comprising: a total amount of protein in the range of 1-30% w / w relative to the weight of the beverage, more preferably 4-24% w / w relative to the weight of the beverage, even more preferably 8-20% w / w relative to the weight of the beverage, and most preferably 10-20% w / w relative to the weight of the beverage, - at most 75%, more preferably at most 40E%, even more preferably at most 10E%, and most preferably at most 5E% of the total energy content (E) of the beverage in terms of total carbohydrates, and - at most 10 E%, more preferably at most 6 E%, even more preferably at most 3 E%, and most preferably at most 1 E% of the total amount of lipids.

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