Dairy Products and Processing

Abstract

The present invention relates to a heat-stable protein composition comprising whey protein and casein, and a method for producing such a heat-stable protein composition. The present invention further relates to a nutritional composition comprising the heat-stable protein composition, and a method for producing the nutritional composition.

Description

[Technical Field]

[0001] The present invention relates to a heat-stable protein composition comprising whey protein and casein, and a method for producing such a heat-stable protein composition. The present invention also relates to a nutritional composition comprising the heat-stable protein composition, a method for producing such a nutritional composition, and a method for providing nutrition to a subject in need thereof.

[0002] The thermostable protein compositions described herein are resistant to sedimentation and heat-induced coagulation or gelation. The protein particles contained in the compositions exhibit minimal particle size change after heat treatment and / or storage. Thus, the compositions described herein are particularly useful in high-protein and shelf-stable beverages that are heat-treated and stored for extended periods of time. [Background technology]

[0003] High protein foods and beverages can be made using ingredients with high protein content, the desirable properties of which include heat stability to allow subsequent heat treatment, such as retort or ultra-high temperature (UHT) processing, to ensure product safety and extended shelf life, and suspendability with minimal or substantially no settling, particularly in beverage applications.

[0004] Micronized whey protein concentrate (WPC) ingredients are known and used in the food industry to increase the protein content in a variety of applications, including cultured and ready-to-drink beverages, bars, and foods containing set or stirred yogurt.

[0005] Whey protein micronization is an advanced technology for producing whey protein particles. The size of whey protein particles is important in achieving the desired mouthfeel. Particles between 0.1 and 3 μm provide a creamy mouthfeel, whereas particles greater than 3 μm result in a chalky or even gritty sensation, and particles with a size of 0.1 μm result in a watery mouthfeel. In fact, particle sizes in the range below 0.1 μm are known to contribute to an oily mouthfeel, which is undesirable if this is perceived as the primary sensory characteristic.

[0006] The main mechanism of whey protein particle formation involves two steps: first, whey proteins unfold (denaturation) during heating, and second, the unfolded protein molecules aggregate primarily through disulfide bonds and hydrophobic interactions. Processing conditions, such as temperature, heating time, pH, and shear stress, determine the reaction rate and the physical and chemical properties of the particles.

[0007] Conventional micronization processes utilize shear or turbulence to limit the size of whey protein particles. However, when high concentrations of whey protein are used to produce protein particles, the micronization / aggregation reaction occurs very quickly due to the high molecular (protein) density and high collision efficiency. While these techniques are effective in preventing the formation of very large particles / aggregates, the average particle size of protein particles produced by conventional micronization processes is between 1 and 10 micrometers, with most particles exceeding 1 micrometer. This particle size range is still prone to sedimentation or settling in liquid applications, especially in low-viscosity liquid applications, throughout the product's shelf life. The presence of larger particles can also result in an undesirable grainy texture. Existing compositions can also exhibit poor thermal stability, especially at high protein concentrations, which can make them unsuitable for high-protein UHT- or retort-processed food / beverage products. For example, existing compositions are prone to particle size increase, viscosity increase, or gelation after UHT or retort processing. Furthermore, reducing the particle size of proteins while retaining all unfolded protein forms in stable microparticles has not been achieved.

[0008] Various methods for preparing particulate denatured whey protein compositions are known in the art. For example, McCarthy (US 5,350,590) prepared "loosely bound" aggregates containing whey protein and casein, generally having a volume average particle size of about 3.0 to about 15 microns. McCarthy reported that these aggregates were relatively loosely formed and "easily broken down to smaller, more organically efficient particle sizes by conventional dairy processing, e.g., homogenization alone."

[0009] There remains a need for protein compositions with suitable organoleptic properties (i.e., lack of a gritty or chalky mouthfeel) that are stable when exposed to a secondary heat treatment, and in particular (especially in the case of UHT or retort food / beverage products) compositions that do not exhibit substantial gelling and / or substantial changes in particle size distribution when exposed to a secondary heat treatment, and / or compositions that exhibit low or no sedimentation in low viscosity beverages.

[0010] It is an object of the present invention to provide improved or alternative thermostable protein compositions, methods for their preparation, nutritional compositions containing them, and / or methods for preparing nutritional compositions, or at least to provide the public with a useful choice.

[0011] Other objects of the present invention may become apparent from the following description, which is given by way of example only.

[0012] External sources of information, including patent specifications and other literature, are referenced herein, typically to provide a context for discussing features of the present invention. In any jurisdiction, unless otherwise stated, citation of such sources shall not be construed as an acknowledgement that such sources are prior art or form part of the common general knowledge in the field. Summary of the Invention

[0013] In a first aspect, the present invention provides a heat-stable protein composition comprising whey protein and casein, The composition comprises β-lactoglobulin and casein in a weight ratio of about 1.6:1 to about 5:1, the whey protein comprises at least about 65% (w / w) denaturable whey protein; at least about 40% (w / w) of the total β-lactoglobulin in the composition is covalently aggregated; the composition comprises protein particles; At least 40% by volume of the protein particles have a particle size of less than 1 μm; and At least a portion of the protein particles comprise co-aggregates of denatured whey protein and casein.

[0014] In a second aspect, the present invention provides a method for preparing a thermostable protein composition, the method comprising: a. An aqueous composition having a pH of 5.5 to 6.8, i. a total whey protein content of at least about 18 g / 100 g aqueous composition; ii. a whey protein content that is at least about 9g / 100g aqueous composition; and iii. a total protein content that is at least about 20 g / 100 g aqueous composition; and iv. providing an aqueous composition comprising β-lactoglobulin and casein in a weight ratio of about 1.6:1 to about 5:1; and b. heat-treating the aqueous composition to at least about 70°C for a time sufficient to cause protein denaturation, wherein the heat-treating comprises heating the aqueous composition under high shear stress to provide a thermostable protein composition.

[0015] In a third aspect, the present invention provides a method for preparing a thermostable protein composition, the method comprising: a. contacting a whey protein source with an oxidizing agent in combination with a catalyst, such as an enzymatic or chemical catalyst, preferably a peroxidase enzyme; b. An aqueous composition having a pH of 5.5 to 6.8 obtained by contacting a whey protein source with a casein protein source, i. a total whey protein content of at least about 14 g / 100 g aqueous composition; ii. a β-lactoglobulin content that is at least about 4 g / 100 g aqueous composition; iii. a total protein content that is at least about 15 g / 100 g aqueous composition; and iv. providing an aqueous composition comprising β-lactoglobulin and casein in a weight ratio of about 1.6:1 to about 5:1; and c. heat-treating the aqueous composition to at least about 70°C for a time sufficient to cause protein denaturation, wherein the heat-treating comprises heating the aqueous composition under high shear stress to provide a thermostable protein composition.

[0016] In a fourth aspect, the present invention provides a method for preparing a thermostable protein composition, the method comprising: a. An aqueous composition having a pH of 5.5 to 6.8, i. a total whey protein content of at least about 14 g / 100 g aqueous composition; ii. a β-lactoglobulin content that is at least about 4 g / 100 g aqueous composition; iii. a total protein content that is at least about 15 g / 100 g aqueous composition; and iv. providing an aqueous composition comprising β-lactoglobulin and casein in a weight ratio of about 1.6:1 to about 5:1; b. contacting the aqueous composition with an oxidizing agent in combination with a catalyst, such as an enzymatic or chemical catalyst, preferably a peroxidase enzyme; and c. heat-treating the aqueous composition to at least about 70°C for a time sufficient to cause protein denaturation, wherein the heat-treating comprises heating the aqueous composition under high shear stress to provide a thermostable protein composition.

[0017] In a fifth aspect, the present invention provides a thermostable protein composition produced by the method of the second, third or fourth aspect.

[0018] In a sixth aspect, the present invention provides a nutritional composition comprising the thermostable protein composition of the first or fifth aspect.

[0019] In a seventh aspect, the present invention provides a liquid composition comprising the thermostable protein composition of the first or fifth aspect.

[0020] In an eighth aspect, the present invention provides a sterilized and / or pasteurized, shelf-stable liquid nutritional composition comprising a thermostable protein composition of the first or fifth aspect.

[0021] In a ninth aspect, the present invention provides a food product comprising the thermostable protein composition of the first or fifth aspect.

[0022] In a further aspect, the present invention provides the use of the thermostable protein composition of the first or fifth aspect in the preparation of a nutritional composition.

[0023] In a further aspect, the present invention provides a method for providing nutrition to a subject in need thereof, the method comprising administering to the subject the nutritional composition, liquid composition, sterilized and / or pasteurized shelf-stable liquid nutritional composition, and / or food product of any one of aspects 6 to 9.

[0024] In a further aspect, the present invention provides a method for preparing a nutritional composition, the method comprising: a. a thermostable protein composition of the first or fifth aspect; b. contacting with one or more additional components.

[0025] In a further aspect, the present invention provides a method for preparing a liquid composition, the method comprising: a. a thermostable protein composition of the first or fifth aspect; b. contacting with one or more additional components.

[0026] The following embodiments and preferences may relate to any of the above aspects, either alone or in any two or more combinations.

[0027] In some embodiments, the casein comprises, consists essentially of, or consists of non-micellar casein. In some embodiments, the composition comprises β-lactoglobulin and non-micellar casein in a weight ratio of about 1.6:1 to about 5:1. In some embodiments, at least a portion of the protein particles comprise co-aggregates of denatured whey protein and non-micellar casein. In some embodiments, the composition comprises β-lactoglobulin and non-micellar casein in a weight ratio of about 1.6:1 to about 5:1, and at least a portion of the protein particles comprise co-aggregates of denatured whey protein and non-micellar casein.

[0028] In some embodiments, at least about 40% by volume of the protein particles have a particle size of less than 1 μm, e.g., at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% by volume have a particle size of less than 1 μm, with useful ranges ranging from any of these values. may be selected (e.g., about 40% by volume to about 100% by volume, about 40% by volume to about 95% by volume, about 40% by volume to about 90% by volume, about 40% by volume to about 85% by volume, about 40% by volume to about 80% by volume, about 45% by volume to about 100% by volume, about 45% by volume to about 95% by volume, about 45% by volume to about 90% by volume, about 45% by volume to about 85% by volume, about 45% by volume to about 80% by volume, about 50% by volume to about 100% by volume, about 50% by volume to about 95% by volume, about 50% by volume to about 90% by volume, about 50% by volume to about 85% by volume, or about 50% by volume to about 80% by volume).

[0029] In some embodiments, at least about 40% by volume of the protein particles have a particle size of less than 1 μm, e.g., at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% by volume have a particle size of less than 1 μm, with useful ranges ranging from any of these values. may be selected (e.g., about 40% by volume to about 100% by volume, about 40% by volume to about 95% by volume, about 40% by volume to about 90% by volume, about 40% by volume to about 85% by volume, about 40% by volume to about 80% by volume, about 45% by volume to about 100% by volume, about 45% by volume to about 95% by volume, about 45% by volume to about 90% by volume, about 45% by volume to about 85% by volume, about 45% by volume to about 80% by volume, about 50% by volume to about 100% by volume, about 50% by volume to about 95% by volume, about 50% by volume to about 90% by volume, about 50% by volume to about 85% by volume, or about 50% by volume to about 80% by volume).

[0030] In some embodiments, less than about 55% by volume of the protein particles have a particle size of 1 to 5 μm, e.g., less than about 50%, less than about 45%, less than about 40%, less than about 35%, less than about 30%, or less than about 25% by volume have a particle size of 1 to 5 μm, and useful ranges can be selected from between any of these values ​​(e.g., 25% to 55%, 25% to 50%, 25% to 45%, 25% to 40%, 25% to 35%, 25% to 30%, 30% to 55%, 35% to 55%, 40% to 55%, or 45% to 55% by volume).

[0031] In some embodiments, less than about 5% by volume of the protein particles have a particle size of at least 5 μm, e.g., less than about 4%, less than about 3%, less than about 2%, less than about 1%, or about 0% by volume have a particle size of at least 5 μm, and useful ranges can be selected between any of these values ​​(e.g., 0% to 5%, 0% to 4%, 0% to 3%, 0% to 2%, or 0% to 1% by volume).

[0032] In some embodiments, at least about 65% (w / w) of the denaturable whey protein is denatured, e.g., at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% (w / w) denatured, and useful ranges can be selected between any of these values ​​(e.g., 65%-100%, 65%-95%, 65%-90%, 70%-100%, 70%-95%, 70%-90%, 75%-100%, 75%-95%, 75%-90%, 80%-100%, 80%-95%, 80%-90%, 85%-100%, 85%-95%, 85%-90%, 90%-100%, or 90%-95%).

[0033] In some embodiments, at least about 40% (w / w) of the total amount of β-lactoglobulin in the composition is covalently aggregated, for example, at least about 45%, 50%, 55%, 60%, 65%, 70%, 75%, or at least about 80% is covalently aggregated, and useful ranges can be selected between any of these values ​​(e.g., 40%-80%, 50%-80%, 60%-80%, 70%-80%, 40%-70%, 50%-70%, 60%-70%, or 55%-65%).

[0034] In some embodiments, the composition comprises a weight ratio of total whey protein to casein of at least about 3:1, e.g., at least about 4:1, at least about 4.1:1, at least about 4.2:1, at least about 4.3:1, at least about 4.4:1, at least about 4.5:1, at least about 5:1, at least about 55:1, at least about 6:1, at least about 7:1, at least about 8:1, at least about 9:1, or about 10:1, with useful ranges ranging from any of these values. can be selected (e.g., 3:1 to 10:1, 3:1 to 9:1, 3:1 to 8:1, 3:1 to 7:1, 3:1 to 6:1, 3:1 to 5:1, 4:1 to 10:1, 4:1 to 9:1, 4:1 to 8:1, 4:1 to 7:1, 4:1 to 6:1, 4:1 to 5:1, 4.5:1 to 10:1, 4.5:1 to 9:1, 4.5:1 to 8:1, 4.5:1 to 7:1, 4.5:1 to 6:1, 4.5:1 to 5:1, 5:1 to 10:1, 5:1 to 9:1, 5:1 to 8:1, 5:1 to 7:1, or 5:1 to 6:1).

[0035] In some embodiments, at least about 70% w / w / w of the total protein in the composition is whey protein, e.g., at least about 75% w / w, at least about 80% w / w, at least about 85% w / w, or at least about 90% w / w is whey protein, and useful ranges can be selected between any of these values ​​(e.g., 50% w / w to 90% w / w, 60% w / w to 90% w / w, 70% w / w to 90% w / w, 75% w / w to 90% w / w, or 80% w / w to 90% w / w).

[0036] In some embodiments, at least about 35% w / w of the total protein in the composition is β-lactoglobulin, e.g., at least about 36% w / w, at least about 37% w / w, at least about 38% w / w, at least about 39% w / w, at least about 40% w / w, at least about 42% w / w, at least about 44% w / w, at least about 46% w / w, at least about 48% w / w, at least about 50% w / w, at least about 52% w / w, at least about 54% w / w, at least about 56% w / w, at least about 58% w / w. / w, or at least about 60% w / w, is β-lactoglobulin, and useful ranges can be selected between any of these values ​​(e.g., 35% w / w to 60% w / w, 35% w / w to 56% w / w, 35% w / w to 52% w / w, 35% w / w to 50% w / w, 38% w / w to 60% w / w, 38% w / w to 56% w / w, 38% w / w to 50% w / w, 40% w / w to 60% w / w, 40% w / w to 56% w / w, 40% w / w to 54% w / w, 40% w / w to 52% w / w, or 40% w / w to 50% w / w).

[0037] In some embodiments, less than about 30% w / w of the total protein in the composition is casein, e.g., less than about 25%, less than about 20%, less than about 15%, or less than about 10% is casein, and useful ranges can be selected between any of these values ​​(e.g., 10%-30%, 10%-25%, 10%-20%, 10%-15%, 15%-30%, 15%-25%, or 15%-20%).

[0038] In some embodiments, about 80% to about 90% w / w of the total protein in the composition is whey protein and about 10% to about 20% w / w is casein.

[0039] In some embodiments, the composition comprises a weight ratio of β-lactoglobulin to casein of at least about 1.6:1, at least about 1.8:1, at least about 2:1, at least about 2.2:1, at least about 2.4:1, at least about 2.6:1, at least about 2.8:1, at least about 3:1, at least about 3.2:1, at least about 3.4:1, at least about 3.6:1, at least about 3.8:1, at least about 4:1, at least about 4.2:1, at least about 4.4:1, at least about 4.6:1, at least about 4.8:1, or at least about 5:1, with useful ranges being within these values. It can be selected from any of the following (for example, 1.6:1 to 5:1, 1.6:1 to 4:1, 1.6:1 to 3.5:1, 1.6:1 to 3:1, 1.8:1 to 5:1, 1.8:1 to 4:1, 1.8:1 to 3.5:1, 1.8:1 to 3:1, 2:1 to 5:1, 2:1 to 4:1, 2:1 to 3.5:1, 2:1 to 3:1, 2.2:1~5:1, 2.2:1~4:1, 2.2:1~3.5:1, 2.2:1~3:1, 2.4:1~5:1, 2.4:1~4:1, 2.4:1~3.5:1, 2.4:1~3:1, 2.6:1~5:1, 2.6:1~4:1, 2.6:1~3.5:1, or 2.6:1~3:1).

[0040] In some embodiments, an aqueous composition comprising a sufficient amount of a thermostable protein composition to provide a 15% w / w total protein content and subjected to a secondary heat treatment at 120°C for 4 minutes contains at least about 40% by volume of protein particles having a particle size of 0.1 to 1 μm, e.g., at least about 41%, at least about 42%, at least about 43%, at least about 44%, at least about 45%, at least about 46%, at least about 47%, at least about 48%, at least about 49%, at least about 50%, at least about 51%, at least about 52%, at least about 53%, at least about 54%, at least about 55%, at least about 56%, at least about 57%, at least about 58%, at least about 59%, at least about 60%, at least about 61%, at least about 62%, at least about 63%, at least about 64%, at least about 65%, at least about 66%, at least about 67%, at least about 68%, at least about 69%, at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81% by volume of protein particles having a particle size of 0.1 to 1 μm. 8% by volume, at least about 49% by volume, at least about 50% by volume, at least about 51% by volume, at least about 52% by volume, or at least about 53% by volume comprise protein particles having a particle size of 0.1 to 1 μm, and useful ranges can be selected between any of these values ​​(e.g., 40% by volume to 53% by volume, 40% by volume to 52% by volume, 40% by volume to 51% by volume, 40% by volume to 50% by volume, 41% by volume to 53% by volume, 41% by volume to 52% by volume, 41% by volume to 51 ... 0% by volume, 42% to 53% by volume, 42% to 52% by volume, 42% to 51% by volume, 42% to 50% by volume, 43% to 53% by volume, 43% to 52% by volume, 43% to 51% by volume, 43% to 50% by volume, 44% to 53% by volume, 44% to 52% by volume, 44% to 51% by volume, 44% to 50% by volume, 45% to 53% by volume, 45% to 52% by volume, 45% to 51% by volume, 45% to 50% by volume, 46% to 53% by volume, 46% by volume ~52% by volume, 46% by volume to 51% by volume, 46% by volume to 50% by volume, 47% by volume to 53% by volume, 47% by volume to 52% by volume, 47% by volume to 51% by volume, 47% by volume to 50% by volume, 48% by volume to 53% by volume, 48% by volume to 52% by volume, 48% by volume to 51% by volume, 48% by volume to 50% by volume, 49% by volume to 53% by volume, 49% by volume to 52% by volume, 49% by volume to 51% by volume, 49% by volume to 50% by volume, 50% by volume to 53% by volume, 50% by volume to 52% by volume, or 50% by volume to 51% by volume.

[0041] In some embodiments, an aqueous composition comprising a sufficient amount of a thermostable protein composition to provide a 15% w / w total protein content and subjected to a secondary heat treatment at 120°C for 4 minutes contains less than about 60% by volume of protein particles having a particle size of 1-5 μm, e.g., less than about 58%, less than about 56%, less than about 55%, less than about 54%, less than about 52%, less than about 50%, less than about 48%, less than about 46%, less than about 44%, less than about 42%, or less than about 40% by volume of protein particles having a particle size of 1-5 μm. For protein particles, useful ranges can be selected from between any of these values ​​(e.g., 40% to 60% by volume, 40% to 58% by volume, 40% to 56% by volume, 40% to 54% by volume, 44% to 60% by volume, 44% to 58% by volume, 44% to 56% by volume, 44% to 54% by volume, 46% to 60% by volume, 46% to 58% by volume, 46% to 56% by volume, 46% to 54% by volume, 48% to 60% by volume, 48% to 58% by volume, 48% to 56% by volume, or 48% to 54% by volume).

[0042] In some embodiments, an aqueous composition comprising a sufficient amount of a thermostable protein composition to provide a 15% w / w total protein content and subjected to a secondary heat treatment at 120° C. for 4 minutes contains less than about 10% by volume of protein particles having a particle size of at least 5 μm, e.g., less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, less than about 1%, or about 0% by volume of protein particles having a particle size of at least 5 μm, and useful ranges can be selected between any of these values ​​(e.g., about 0% to about 10%, about 0% to about 9%, about 0% to about 10%, and about 0% by volume). % to about 8 volume%, about 0 volume% to about 7 volume%, about 0 volume% to about 6 volume%, about 0 volume% to about 5 volume%, about 0 volume% to about 4 volume%, about 0 volume% to about 3 volume%, about 0 volume% to about 2 volume%, about 0 volume% to about 1 volume%, about 1 volume% to about 10 volume%, about 1 volume% to about 9 volume%, about 1 volume% to about 8 volume%, about 1 volume% to about 7 volume%, about 1 volume% to about 6 volume%, about 1 volume% to about 5 volume%, about 1 volume% to about 4 volume%, about 1 volume% to about 3 volume%, about 2 volume% to about 10 volume%, about 2 volume% to about 9 volume%, about 2 volume% to about 8 volume%, about 2 volume% to about 7 volume%, about 2 volume% to about 6 volume%, about 2 volume% to about 5 volume%, about 2 volume% to about 4 volume%, or about 2 volume% to about 3 volume%.

[0043] In some embodiments, an aqueous composition comprising a sufficient amount of a thermostable protein composition to provide a 15% w / w total protein content, and which has been subjected to a secondary heat treatment at 120°C for 4 minutes, comprises protein particles having a D[4,3] of less than about 5, e.g., less than about 4, less than about 3, less than about 2, less than about 1.8, less than about 1.6, less than about 1.4, less than about 1.2, less than about 1.0, or less than about 0.9, with useful ranges selected between any of these values ​​(e.g., from about 0.9 to about 5, from about 0.9 to about 4, from about 0.9 to about 3, from about 0.9 to about 2, from about 0.9 to about 1.8, from about 0.9 to about 1.6, from about 0.9 to about 1.4, from about 0.9 to about 1.2, or from about 0.9 to about 1.0).

[0044] In some embodiments, an aqueous composition comprising a thermostable protein composition in an amount sufficient to provide a 15% w / w total protein content and subjected to a second heat treatment at 120° C. for 4 minutes is -1 and the like, and a useful range can be selected between any of these values ​​(e.g., 5-100 mPa·s, 5-80 mPa·s, 5-70 mPa·s, 5-60 mPa·s, 5-50 mPa·s, 5-40 mPa·s, 5-80 mPa·s, 5-70 mPa·s, 5-60 mPa·s, 5-50 mPa·s, 5-40 mPa·s, 5-80 mPa·s, 5-70 mPa·s, 5-60 mPa·s, mPa·s, 5–50 mPa·s, 5–40 mPa·s, 5–30 mPa·s, 10–100 mPa·s, 10–80 mPa·s, 10–70 mPa·s, 10–60 mPa·s, 10–50 mPa·s, 10–40 mPa·s, or 10–30 mPa·s).

[0045] In some embodiments, the method comprises contacting a whey protein source with a casein protein source to provide the aqueous composition of step a).

[0046] In some embodiments, the whey protein source comprises or consists of whey protein concentrate (WPC), whey protein isolate (WPI), or a combination thereof.

[0047] In some embodiments, the casein protein source comprises, consists essentially of, or consists of non-micellar casein or casein-calcium-sodium / potassium-phosphate complex. In some embodiments, the casein protein source comprises or consists of caseinate, a β-casein-enriched fraction, a κ-casein-enriched fraction, a decalcified milk protein concentrate (MPC), a decalcified micellar casein concentrate (MCC), a total milk protein (TMP), or a combination of any two or more thereof. In some embodiments, the casein protein source comprises or consists of caseinate. In some embodiments, the casein comprises or consists of a decalcified milk protein concentrate (MPC).

[0048] In some embodiments, the method further comprises, preferably prior to step a), contacting the whey protein source with an oxidizing agent in combination with a catalyst, such as an enzymatic or chemical catalyst, preferably a peroxidase enzyme.

[0049] In some embodiments, the method further comprises, preferably prior to step b), contacting the aqueous composition with an oxidizing agent in combination with a catalyst, such as an enzyme or chemical catalyst, preferably a peroxidase enzyme.

[0050] In some embodiments, the catalyst is an enzymatic catalyst, for example, a microbial peroxidase enzyme such as MaxiBright® (DSM Food Specialties), or a dye-decolorizing (DyP-type) peroxidase such as EfeB / YcdB from Escherichia coli O157, DyPB from Rhodococcus jostii RHA1, or DyP2 from Amycolatopsis sp. 75iv2. In some embodiments, the catalyst is a chemical catalyst such as copper, iron, zinc, or manganese.

[0051] In some embodiments, the oxidizing agent is hydrogen peroxide or benzoyl peroxide, and / or the catalyst is a peroxidase enzyme.

[0052] In some embodiments, the oxidizing agent is a food-grade oxidizing agent, ie, oxygen (O), ozone, peroxides including alkyl hydroperoxides, superoxide, peroxynitrite, peroxydisulfate, or lactoperoxidase.

[0053] In some embodiments, the oxidizing agent is a peroxide, preferably an organic peroxide. In some embodiments, the peroxide is a metal peroxide (e.g., an alkali metal peroxide such as sodium peroxide, an alkaline earth metal peroxide such as magnesium peroxide or calcium peroxide, or a transition metal peroxide such as zinc peroxide), hydrogen peroxide, or benzoyl peroxide. In some embodiments, the oxidizing agent is a perborate, such as sodium perborate.

[0054] In such embodiments, the oxidizing agent is benzoyl peroxide or hydrogen peroxide. In some embodiments, the hydrogen peroxide is food grade.

[0055] In some embodiments, the oxidizing agent is present in an amount less than 300 ppm or less than 200 ppm, or the molar ratio of oxidizing agent to β-lactoglobulin is less than 2, preferably less than 1, or from about 0.1 to about 0.85.

[0056] In some embodiments, the aqueous composition comprises a total whey protein content of at least about 14 g / 100 g, such as at least about 15 / 100 g, 16 / 100 g, 17 / 100 g, or 18 g / 100 g.

[0057] In some embodiments, the aqueous composition comprises a total β-lactoglobulin content of at least about 4g / 100g aqueous composition, such as at least about 5g / 100g aqueous composition, about 6g / 100g aqueous composition, about 7g / 100g aqueous composition, about 8g / 100g aqueous composition, or about 9g / 100g aqueous composition.

[0058] In some embodiments, the aqueous composition comprises a total protein content that is at least about 15 g / 100 g, e.g., at least about 16 g / 100 g, at least about 17 g / 100 g, at least about 18 g / 100 g, at least about 19 g / 100 g, or at least about 20 g / 100 g aqueous composition.

[0059] In some embodiments, the whey protein source and / or casein protein source are in the form of a powder. In some embodiments, the method includes reconstituting the powder(s) to provide an aqueous composition.

[0060] In some embodiments, the method further comprises homogenizing the aqueous composition, preferably at about 200 / 50 bar. In some embodiments, the method further comprises homogenizing the aqueous composition, preferably at about 200 / 50 bar, preferably at about 200 / 50 bar, prior to step b).

[0061] In some embodiments, the method further comprises adjusting the pH of the aqueous composition to a pH of 5.5 to 6.8.

[0062] In some embodiments, the method further comprises concentrating the aqueous composition, preferably by evaporation, prior to step b).

[0063] In some embodiments, the whey protein comprises a denaturable whey protein, and step b) comprises heating the aqueous composition for a sufficient time to denature at least about 65% (w / w) of the denaturable whey protein in the aqueous composition, e.g., at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% (w / w) of the denaturable whey protein in the aqueous composition. A useful range, including heat treatment, can be selected between any of these values ​​(e.g., 65% to 100%, 65% to 95%, 65% to 90%, 70% to 100%, 70% to 95%, 70% to 90%, 75% to 100%, 75% to 95%, 75% to 90%, 80% to 100%, 80% to 95%, 80% to 90%, 85% to 100%, 85% to 95%, 85% to 90%, 90% to 100%, or 90% to 95%).

[0064] In some embodiments, less than about 10% (w / w) of the total denaturable whey protein in the aqueous composition prior to step b) is denatured, preferably less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than about 1% is denatured, and useful ranges can be selected between any of these values ​​(e.g., 1%-10%, 1%-8%, 1%-5%, or 1%-4%).

[0065] In some embodiments, the thermostable protein composition comprises protein particles, wherein at least about 40% by volume of the protein particles have a particle size of less than 1 μm, e.g., at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% by volume have a particle size of less than 1 μm, with useful ranges being within this range. The total volume fraction may be selected from any of the values ​​above (e.g., about 40% by volume to about 100% by volume, about 40% by volume to about 95% by volume, about 40% by volume to about 90% by volume, about 40% by volume to about 85% by volume, about 40% by volume to about 80% by volume, about 45% by volume to about 100% by volume, about 45% by volume to about 95% by volume, about 45% by volume to about 90% by volume, about 45% by volume to about 85% by volume, about 45% by volume to about 80% by volume, about 50% by volume to about 100% by volume, about 50% by volume to about 95% by volume, about 50% by volume to about 90% by volume, about 50% by volume to about 85% by volume, or about 50% by volume to about 80% by volume).

[0066] In some embodiments, step b) comprises heat treating the aqueous solution to a temperature of about 70°C to about 150°C.

[0067] In some embodiments, the thermostable protein composition comprises at least about 70% w / w, preferably about 70% to about 90% w / w, whey protein based on the total amount of protein in the composition. In some embodiments, the thermostable protein composition comprises at least about 80% w / w, preferably about 80% to about 90% w / w, whey protein based on the total amount of protein in the composition. In some embodiments, the thermostable protein composition comprises at least about 35% w / w, preferably about 40% to about 60% w / w, β-lactoglobulin based on the total amount of protein in the composition. In some embodiments, the thermostable protein composition comprises less than about 30% w / w, preferably about 10% to about 20% w / w, casein based on the total amount of protein in the composition. In some embodiments, the thermostable protein composition comprises at least about 70% w / w whey protein based on the total amount of protein in the composition and less than about 30% w / w casein based on the total amount of protein in the composition.

[0068] In some embodiments, the method further comprises drying the thermostable protein composition.

[0069] In some embodiments, the thermostable protein composition is not subjected to mechanical shear prior to drying, other than to convert the liquid into droplets to facilitate drying.

[0070] In some embodiments, step b) comprises: i. Under turbulent flow conditions with a Reynolds number of at least about 2000; ii. at least about 1000s -1 under high wall shear conditions with a wall shear rate of iii. Heating the solution under mechanical shear conditions, preferably generated by a homogenizer, colloid mill, high pressure pump, scraped surface heat exchanger, and / or high shear mixer.

[0071] In some embodiments, the method produces a thermostable protein composition according to the first aspect.

[0072] In some embodiments, the nutritional composition comprises at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 18%, at least about 20%, at least about 21%, at least about 22%, at least about 23%, at least about 24%, at least about 25%, at least about 26%, at least about 27%, at least about 28%, at least about 29%, at least about 30%, at least about 31%, at least about 32%, at least about 33%, at least about 34%, at least about 35%, at least about 36%, at least about 37%, at least about 38%, at least about 39%, at least about 40%, at least about 41%, at least about 42%, at least about 43%, at least about 44%, at least about 45%, at least about 46%, at least about 47%, at least about 48%, at least about 49%, at least about 50%, at least about 51%, at least about 52%, at least about 53%, at least about 54%, at least about 55%, at least about 56%, at least about 57%, at least about 58%, at least about 59%, at least about 60%, at least about 61%, at least about 62%, at least about 63%, at least about 64%, at least about 65%, at least about 66%, at least about 67%, at least about 68%, at least about 69%, at least about 70%, at least about 71%, at least about 72%, at least about 73%, at least about 74%, at least about 75%, At least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 30%, at least about 40%, or at least about 50% by weight of total protein, and useful ranges can be selected between any of these values ​​(e.g., about 1% to about 40%, or about 1% to about 30%, or about 1% to about 20%, or about 1% to about 16%, about 1% to about 15%, about 1% to about 14%, or about 1% by weight to about 12% by weight, or about 1% by weight to about 10% by weight, or about 2% by weight to about 50% by weight, or about 2% by weight to about 40% by weight, or about 2% by weight to about 30% by weight, or about 2% by weight to about 20% by weight, or about 2% by weight to about 16% by weight, or about 2% by weight to about 15% by weight, or about 2% by weight to about 14% by weight, or about 2% by weight to about 12% by weight, or about 4% by weight to about 50% by weight, or about 4% by weight to about 40% by weight, or about 4% by weight to about 30% by weight, or about 4% by weight to about 20% by weight, or about 4% to about 16% by weight, about 4% to about 15% by weight, about 4% to about 14% by weight, or about 4% to about 12% by weight, or about 4% to about 10% by weight, about 5% to about 50% by weight, or about 5% to about 40% by weight, or about 5% to about 30% by weight, or about 5% to about 20% by weight, or about 5% to about 16% by weight, about 5% to about 15% by weight, about 5% to about 14% by weight, or about 5% to about 12% by weight, or about 5% to about 10% by weight In some embodiments, the nutritional compositions comprise at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 30%, at least about 40%, or at least about 50% by weight of denatured whey protein on a dry basis, and useful ranges can be selected between any of these values ​​(e.g., from about 1% to about 40%, or from about 1% to about 30%, or or about 1% to about 20%, or about 1% to about 16%, or about 1% to about 15%, or about 1% to about 14%, or about 1% to about 12%, or about 1% to about 10%, or about 2% to about 50%, or about 2% to about 40%, or about 2% to about 30%, or about 2% to about 20%, or about 2% to about 16%, or about 2% to about 15%, or about 2% to about 14%, or about 2% to about 12%, or about 4% to about 50%, or about 4% to about 40%, or about 4% to about 30%, or about 4% to about 20%, or about 4% to about 16%, about 4% to about 15%, about 4% to about 14%, or about 4% to about 12%, or about 4% to about 10%, about 5% to about 50%, or about 5% to about 40%, or about 5% to about 30%, or about 5% to about 20%, or about 5% to about 16%, about 5% to about 15%, about 5% to about 14%, or about 5% to about 12%, or about 5% to about 10%).

[0073] In some embodiments, at least about 50% (w / w) of the total protein in the nutritional composition is denatured whey protein, e.g., at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% is denatured whey protein, and useful ranges can be selected between any of these values ​​(e.g., 50%-95%, 50%-95%). %~90%, 50%~85%, 50%~80%, 55%~95%, 55%~90%, 55%~85%, 55%~80%, 60%~95%, 60%~90%, 60%~85%, 60%~80%, 65%~95%, 65%~90%, 65%~85%, 65%~80%, 70%~95%, 70%~90%, 70%~85%, 70%~80%, 75%~95%, 75%~90%, 75%~85%, 75%~80%, 80%~95%, 80%~90%, or 80%~85%).

[0074] In some embodiments, at least about 50% (w / w) of the total protein in the nutritional composition is provided by the thermostable protein composition, e.g., at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% is provided by the thermostable protein composition, and useful ranges can be selected between any of these values ​​(e.g., 50%-100%, 50%-95%, 50%-90%, 50%-85%, 50%-80%, 50%-75%, 55% Up to 100%, 55% to 95%, 55% to 90%, 55% to 85%, 55% to 80%, 55% to 75%, 60% to 100%, 60% to 95%, 60% to 90%, 60% to 85%, 60% to 80%, 60% to 75%, 65% to 100%, 65% to 95%, 65% to 90%, 65% to 85% , 65% to 80%, 65% to 75%, 70% to 100%, 70% to 95%, 70% to 90%, 70% to 85%, 70% to 80%, 70% to 75%, 75% to 100%, 75% to 95%, 75% to 90%, 75% to 85%, or 75% to 80%).

[0075] In some embodiments, at least about 40% by volume of the protein particles have a particle size of less than 1 μm, e.g., at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% by volume have a particle size of less than 1 μm, with useful ranges ranging from any of these values. may be selected (e.g., about 40% by volume to about 100% by volume, about 40% by volume to about 95% by volume, about 40% by volume to about 90% by volume, about 40% by volume to about 85% by volume, about 40% by volume to about 80% by volume, about 45% by volume to about 100% by volume, about 45% by volume to about 95% by volume, about 45% by volume to about 90% by volume, about 45% by volume to about 85% by volume, about 45% by volume to about 80% by volume, about 50% by volume to about 100% by volume, about 50% by volume to about 95% by volume, about 50% by volume to about 90% by volume, about 50% by volume to about 85% by volume, or about 50% by volume to about 80% by volume).

[0076] In some embodiments, at least about 40% by volume of the protein particles have a particle size of 0.1-1 μm, e.g., at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% by volume have a particle size of 0.1-1 μm, with useful ranges ranging from any of these values. The concentration may be selected from the following (e.g., about 40% by volume to about 100% by volume, about 40% by volume to about 95% by volume, about 40% by volume to about 90% by volume, about 40% by volume to about 85% by volume, about 40% by volume to about 80% by volume, about 45% by volume to about 100% by volume, about 45% by volume to about 95% by volume, about 45% by volume to about 90% by volume, about 45% by volume to about 85% by volume, about 45% by volume to about 80% by volume, about 50% by volume to about 100% by volume, about 50% by volume to about 95% by volume, about 50% by volume to about 90% by volume, about 50% by volume to about 85% by volume, or about 50% by volume to about 80% by volume).

[0077] In some embodiments, less than about 60% by volume of the protein particles in the nutritional composition have a particle size of 1 to 5 μm, e.g., less than about 55%, less than about 50%, less than about 45%, less than about 40%, less than about 35%, less than about 30%, or less than about 25% by volume have a particle size of 1 to 5 μm, and useful ranges can be selected between any of these values ​​(e.g., 25% to 60%, 25% to 55%, 25% to 50%, 25% to 45%, 25% to 40%, 25% to 35%, 25% to 30%, 30% to 60%, 30% to 55%, 35% to 55%, 40% to 55%, or 45% to 55% by volume).

[0078] In some embodiments, less than about 10% by volume of the protein particles in the nutritional composition have a particle size of at least 5 μm, e.g., less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, less than about 1%, or about 0% by volume have a particle size of at least 5 μm, and useful ranges can be selected between any of these values ​​(e.g., 0% to 10%, 0% to 9%, 0% to 8%, 0% to 7%, 0% to 6%, 0% to 5%, 0% to 4%, 0% to 3%, 0% to 2%, or 0% to 1% by volume).

[0079] In some embodiments, the nutritional composition has been subjected to a secondary heat treatment, preferably sterilization and / or pasteurization. In some embodiments, the nutritional composition has been sterilized and / or pasteurized. Preferably, the sterilized and / or pasteurized nutritional composition exhibits minimal bacterial growth when aseptically filled after long-term storage at a temperature of about 20°C to about 25°C for at least 2 months, at least 3 months, at least 6 months, or at least 12 months.

[0080] In some embodiments, the nutritional composition is a food product.

[0081] In some embodiments, the food product is a baked good, a bar, a set or stirred yogurt, a set gel, or a semi-solid food product.

[0082] In some embodiments, the food product is set yogurt or stirred yogurt. In some embodiments, the set yogurt or stirred yogurt comprises about 6% to about 20% (w / v) total protein.

[0083] In some embodiments, the set or stirred yogurt exhibits a reduced volume weighted mean particle size compared to a control yogurt product having the same ingredient composition and the same total protein content, except that the control yogurt product does not comprise the heat-stable protein composition of the first or third aspect.

[0084] In some embodiments, the food product is a heat-treated, high-protein, set gel. In some embodiments, the heat-treated, high-protein, set gel comprises at least about 10% (w / v) or at least about 15% (w / v) total protein.

[0085] In some embodiments, the food product is a heat-processed high-protein semi-solid food product. In some embodiments, the heat-processed high-protein semi-solid food product comprises at least about 10% (w / v) or at least about 15% (w / v) total protein.

[0086] In some embodiments, the food product is refrigerated at 20° C. for 50 seconds. -1 The viscosity measured at 1000 mPa·s is less than about 1000 mPa·s, less than about 800 mPa·s, less than about 600 mPa·s, or less than about 400 mPa·s.

[0087] In some embodiments, the nutritional composition is a liquid composition, such as a liquid nutritional composition. In some embodiments, the nutritional composition is a liquid nutritional composition.

[0088] In some embodiments, the liquid composition is refrigerated at 20° C. for 100 seconds.-1 less than about 400 mPa·s, for example, less than about 350 mPa·s, less than about 300 mPa·s, less than about 250 mPa·s, less than about 200 mPa·s, less than about 150 mPa·s, less than about 100 mPa·s, less than about 90 mPa·s, less than about 80 mPa·s, less than about 70 mPa·s, less than about 60 mPa·s, less than about 50 mPa·s, less than about 40 mPa·s, less than about 30 mPa·s, less than about 20 mPa·s, less than about 10 mPa·s, less than about 8 mPa·s, less than about 6 mPa·s, They have a viscosity of less than 5 mPa·s, less than about 4 mPa·s, less than about 3 mPa·s, or less than about 2 mPa·s, and useful ranges can be selected between any of these values ​​(e.g., 2-400 mPa·s, 2-300 mPa·s, 2-200 mPa·s, 2-100 mPa·s, 4-400 mPa·s, 4-300 mPa·s, 4-200 mPa·s, 4-100 mPa·s, 6-400 mPa·s, 6-300 mPa·s, 6-200 mPa·s, or 6-100 mPa·s).

[0089] In some embodiments, the liquid composition exhibits less than about 10% sedimentation, e.g., less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, less than about 1%, or less than about 0% sedimentation, after (a) storage at a temperature of about 20° C. to about 25° C. for at least 6 weeks, (b) storage at a temperature of about 20° C. to about 25° C. for at least 3 months, and / or (c) centrifugation at 1540×g for 5 minutes, and a useful range can be selected between any of these values ​​(e.g., 0.05%). %~10%, 0%~9%, 0%~8%, 0%~7%, 0%~6%, 0%~5%, 1%~10%, 1%~9%, 1%~8%, 1%~7%, 1%~6%, 1%~5%, 2%~10%, 2%~9%, 2%~8%, 2%~7%, 2%~6%, 2%~5%, 3%~10%, 3%~9%, 3%~8%, 3%~7%, 3%~6%, 3%~5%, 4%~10%, 4%~9%, 4%~8%, 4%~7%, 4%~6%, 4%~5%, 5%~10%, 5%~9%, 5%~8%, 5%~7%, or 5%~6% sedimentation).

[0090] In some embodiments, the liquid composition comprises: a. about 0.1 to about 30% (w / v) lipid; b. about 0.1 to about 40% (w / v), preferably about 0.1 to about 30% (w / v) of carbohydrates; c. at least one monovalent cation; d. Preferably Ca 2+ at least one divalent metal cation, optionally present in an amount of at least about 30 mg / 100 mL, at least about 50 mg / 100 mL, or at least about 100 mg / 100 mL; or e. A combination of any two or more of (a) to (d).

[0091] In some embodiments, the liquid composition comprises at least about 0.1% w / v carbohydrate, e.g., at least about 1% w / v, 2% w / v, 3% w / v, 4% w / v, 5% w / v, 6% w / v, 7% w / v, 8% w / v, 9% w / v, 10% w / v, 15% w / v, 20% w / v, 25% w / v, 30% w / v, 35% w / v, or at least about 40% w / v w / v carbohydrate, with useful ranges selected between any of these values ​​(e.g., from about 0.1% w / v to about 40% w / v, about 0.1% w / v to about 30% w / v, about 0.1% w / v to about 20% w / v, about 0.1% w / v to about 10% w / v, about 1% w / v to about 40% w / v, about 1% w / v to about 30% w / v, about 1% w / v to about 20% w / v, about 1% w / v to about 10% w / v, about 10% w / v to about 40% w / v, about 10% w / v to about 30% w / v, about 10% w / v to about 20% w / v, about 20% w / v to about 40% w / v, about 20% w / v to about 30% w / v, or about 30% w / v to about 40% w / v).

[0092] In some embodiments, the liquid composition has an energy density of at least about 0.5 kcal / mL, e.g., at least about 1.0 kcal / mL, at least about 1.5 kcal / mL, or at least about 2.0 kcal / mL, and useful ranges can be selected between any of these values ​​(e.g., 0.5-2.0 kcal / mL, 1.0-2.0 kcal / mL, 1.5-2.0 kcal / mL, 0.5-1.5 kcal / mL, or 1.0-1.5 kcal / mL).

[0093] In some embodiments, the liquid composition is a heat-treated shelf-stable liquid nutritional composition.

[0094] In some embodiments, the liquid composition is a high protein beverage or a medical food.

[0095] In some embodiments, the liquid composition is a drinking yogurt. In some embodiments, the drinking yogurt is simmered at 20° C. for 50 seconds. -1 The viscosity measured at 1000 kJ / min is less than about 400 mPa·s, less than about 300 mPa·s, less than about 200 mPa·s, or less than about 100 mPa·s, and useful ranges can be selected between any of these values ​​(e.g., 100-400 mPa·s, 100-300 mPa·s, or 100-200 mPa·s).

[0096] In some embodiments, the liquid composition is an acidic beverage. In some embodiments, the acidic beverage has a pH of about 2 to about 4.8. In some embodiments, the liquid composition is a neutral beverage. In some embodiments, the neutral beverage has a pH of about 6.5 to about 7.5. In some embodiments, the acidic or neutral beverage has a pH of about 100 s at 20°C. -1 The viscosity measured at 1000 kJ / min is less than about 400 mPa·s, less than about 300 mPa·s, less than about 200 mPa·s, or less than about 100 mPa·s, and useful ranges can be selected between any of these values ​​(e.g., 100-400 mPa·s, 100-300 mPa·s, or 100-200 mPa·s).

[0097] In some embodiments, the use of the thermostable protein composition is in the preparation of a nutritional composition according to the fourth aspect.

[0098] In some embodiments, the one or more additional ingredients include one or more lipids, one or more carbohydrates, one or more proteins, one or more vitamins, one or more minerals, one or more dairy products, water, one or more food additives, one or more polyhydric alcohols, one or more colorants, one or more fruit preparations, or a combination of any two or more of these ingredients.

[0099] In some embodiments, the one or more lipids comprise one or more plant lipids and / or one or more dairy lipids.

[0100] In some embodiments, the one or more carbohydrates comprise one or more monosaccharides, disaccharides, oligosaccharides, polysaccharides, or a combination of any two or more thereof.

[0101] In some embodiments, the one or more additional protein sources are derived from milk, whey, casein, caseinate, egg, egg white, egg yolk, vegetables, plants, alfalfa, clover, peas, beans, kidney beans, soybeans, navy beans, lupin, cocoa, hornbeams, tree nuts, peanuts, rye, grains, whole grains, rice, hemp, wheat gluten, fungal or algae protein, a protein concentrate thereof, a protein isolate thereof, a hydrolysate thereof, or a combination of any two or more thereof.

[0102] In some embodiments, the one or more vitamins include vitamin C, vitamin A, vitamin E, vitamin B12, vitamin K, riboflavin, niacin, vitamin D, vitamin B6, folic acid, pyridoxine, thiamine, pantothenic acid, biotin, or any salt, derivative, or metabolite thereof, or a combination of any two or more thereof.

[0103] In some embodiments, the one or more minerals include chloride, sodium, calcium, iron, chromium, copper, iodine, zinc, magnesium, phosphorus, or potassium, or a combination of any two or more thereof.

[0104] In some embodiments, the one or more food additives comprise one or more emulsifiers, preferably lecithin, monoglycerides, diglycerides, polyglycerol esters, milk phospholipids, citric acid esters (CITREM), polysorbate 60, glyceryl monostearate, DATEM, or a combination of any two or more thereof.

[0105] In some embodiments, the one or more food additives comprise one or more stabilizers, preferably carrageenan, gellan gum, pectin, guar gum, locust bean gum, carboxymethylcellulose, alginate, agar, oat gum, tragacanth gum, acacia gum, xanthan gum, karaya gum, tara gum, starch, modified starch and microcrystalline cellulose, gelatin, or a combination of any two or more thereof.

[0106] In some embodiments, the nutritional or liquid composition is a nutritional composition according to the fourth aspect.

[0107] The present invention may also include the parts, elements and features referred to or indicated herein individually or collectively, and any or all combinations of any two or more of the parts, elements or features, and where specific integers referred to herein have known equivalents in the art to which the present invention pertains, these known equivalents are deemed to be incorporated herein as if set forth separately.

[0108] Reference to a range of numbers disclosed herein (e.g., 1 to 10) is intended to incorporate reference to every rational number within that range (e.g., 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9, and 10), as well as any range of rational numbers within that range (e.g., 2 to 8, 1.5 to 5.5, and 3.1 to 4.7); thus, all subranges of every range explicitly disclosed herein are hereby expressly disclosed. These are merely examples of what is specifically intended, and all possible combinations of numerical values ​​between the recited lower and upper limits should be considered to be expressly stated in the same manner in this application.

[0109] Although the present invention is broadly as defined above, those skilled in the art will understand that the present invention is not limited thereto and that the embodiments exemplified in the following description are also included in the present invention. [Brief explanation of the drawings]

[0110] The invention will now be described, by way of example only, with reference to the accompanying drawings in which:

[0111] [Figure 1] The change in viscosity of an aqueous solution containing a comparative denatured whey protein composition (Sample 4) at 10% total solids and pH 7.0 is shown before heating (crosses), after heating (triangles), and after evaporation to 19% total solids (diamonds), as described in Example 1. [Figure 2] FIG. 1 shows the change in particle size distribution (by volume density) as described in Example 1 for the aqueous solution shown before heating (circles), after heating (squares), after evaporation to 19% total solids (triangles), and after spray drying to a powder and reconstitution in water at 10% total solids (diamonds). [Figure 3] 1 shows the change in particle size distribution (by volume density) of one embodiment of a thermostable protein composition of the present invention (Sample 1) at 14% protein and pH 6.8 before heating (circles) and after heating at 120°C for 10 minutes (squares) as described in Example 1. [Figure 4]1 is a flow diagram illustrating an embodiment of a process for producing a heat-stable protein composition. The whey protein source may include whey retentate, whey protein powder, or a mixture thereof. [Figure 5] FIG. 1 is a flow diagram illustrating a process embodiment for producing a liquid nutritional composition comprising a thermostable protein composition. DETAILED DESCRIPTION OF THE INVENTION

[0112] The present invention relates to a heat-stable protein composition comprising protein particles containing both whey protein and casein protein. The protein particles are sized to resist settling and have a pleasant mouthfeel (i.e., not gritty or powdery). After production, the protein particles do not require additional mechanical shearing to further reduce them to a desired size and are resistant to grinding. The protein particles exhibit minimal size change after heat treatment and / or storage. The protein composition is resistant to heat-induced coagulation or gelation. The protein composition also has a favorable flavor profile, e.g., a reduced "eggy" flavor and an increased "milk-like" flavor compared to all-whey compositions. 1.Definition

[0113] Unless otherwise specified, the singular forms "a," "an," and "the" include plural references.

[0114] As used herein, the term "about" generally refers to a range of numbers that one of ordinary skill in the art would consider equivalent to the recited value (e.g., ±5-10% of the recited value). Ranges may be expressed herein as from "about" one particular value and / or to "about" another particular value. When such a range is expressed, the range includes the recited values.

[0115] As used herein, the term "and / or" means "and," or "or," or both.

[0116] As used herein, the phrase "calcium-depleted" refers to a composition, such as a milk protein concentrate (MPC), in which the concentration of calcium bound to casein has been reduced and is lower than the concentration of calcium bound to casein in a corresponding composition that has not been subjected to calcium-depletion treatment. Such compositions may also be depleted in other divalent cations, such that the concentration of divalent cations, such as magnesium, bound to casein is lower compared to the corresponding undepleted composition. Similarly, calcium in casein proteins refers to bound calcium, which is calcium bound to casein proteins.

[0117] As used herein, the term "co-aggregates of denatured whey protein and casein" refers to aggregates that include both denatured whey protein and casein.

[0118] As used herein, the term "comprising" means "consisting at least in part of." When interpreting statements herein that include this term, all features prefaced by that term in each statement must be present, although other features may also be present. Related terms such as "comprises" and "includes" are to be interpreted similarly.

[0119] The term "casein" as used herein includes α(s1) and α(s2)-casein, β-casein, κ-casein, and mixtures thereof, and further includes caseinates such as sodium caseinate, calcium caseinate, magnesium caseinate, potassium caseinate, ammonium caseinate, etc.

[0120] The term "caseinate" refers to a compound of casein and a metal ion produced by precipitating casein with an acid and then redissolving it in an alkali containing the metal ion. Hydroxide solutions containing sodium, calcium, magnesium, potassium, and ammonium may be used to produce sodium caseinate, calcium caseinate, magnesium caseinate, potassium caseinate, and ammonium caseinate. Caseinates and methods for producing caseinates suitable for use herein are described in Fox & McSweeney, 2003, and Dairy Processing Handbook, 2003.

[0121] As used herein, the term "covalently aggregated" and related terms such as "covalent aggregate" refer to a denatured β-lactoglobulin molecule that contains at least one intermolecular covalent bond with another protein molecule. One example of such a covalent bond is a disulfide bond formed between two sulfhydryl groups, such as those in the side chains of two cysteine ​​amino acids.

[0122] As used herein, the term "denaturable whey protein" or "denaturable whey protein" refers to the total amount of denaturable whey protein. Certain whey protein sources, such as cheese whey, may contain proteins such as glycomacropeptide (GMP), a casein-related protein, and proteose peptone 5 (pp5). Heat treatment denatures bovine serum albumin (BSA), α-lactalbumin, β-lactoglobulin, lactoferrin, and immunoglobulins. In contrast, GMP and pp5 do not denature. Therefore, the term "denatured whey protein" excludes GMP and pp5. The total amount of denatured whey protein can be calculated as described in Section 1.1 of Example 1.

[0123] As used herein, the term "dry weight basis" refers to the percentage of a substance in a composition or product after the water in the product has been removed, which can be calculated by applying a correction to account for retained water in the product.

[0124] As used herein, the term "thermostable" refers to a composition that is resistant to undesirable changes due to secondary heat treatment, such as an increase in viscosity, gelation, and / or a change in particle size distribution. Thermostability is typically assessed as described in Section 7. A composition that is not thermostable may gel or exhibit an increase in viscosity when subjected to a secondary heat treatment, and may exhibit an increase in particle size distribution (e.g., an increase in the proportion of particles having a particle size of at least 5 μm, an increase in the proportion of particles having a particle size of 1-5 μm, and / or a decrease in the proportion of particles having a particle size of 0.1-1 μm). In some embodiments, a thermostable composition exhibits minimal or substantially no gelation, no substantial increase in viscosity, no substantial increase in the proportion of protein particles having a particle size of 1-5 μm and / or at least 5 μm, and / or no substantial decrease in the proportion of protein particles having a particle size of 0.1-1 μm when the thermostable composition is subjected to a secondary heat treatment.

[0125] The term "liquid nutritional composition" refers to an aqueous composition administered to a subject's digestive tract. Administration is preferably oral, but other methods, such as tube feeding, including nasogastric tube feeding and gastric tube feeding, are also possible. The term "liquid nutritional composition" includes medical foods, enteral nutritional formulas, specialized medical foods, liquid meal replacers, and supplements. The liquid nutritional compositions described herein may provide significant amounts of protein and carbohydrates, and usually also lipids. Vitamins and minerals may also be included. In various embodiments, the subject in need of nutrition may be suffering from or predisposed to a disease or condition, undergoing or having undergone treatment for a disease or condition, may be elderly, a person recovering from a disease or condition, or may be malnourished. In other embodiments, the subject may be a healthy individual, such as an athlete or an active elderly person, including, but not limited to, a person with specific nutritional requirements.

[0126] As used herein, the term "mechanical shearing process" refers to a process in which a mechanical device such as a homogenizer, colloid mill, high pressure pump, scraped surface heat exchanger, high shear mixer, ultrasonic generator, microfluidizer, or the like is used to mix a solution or break down particles within the solution.

[0127] As used herein, the term "non-dairy protein" refers to a protein that is not a dairy protein (i.e., a protein that is not derived from animal milk). Non-dairy proteins include plant-derived proteins, fungal proteins, and algae proteins.

[0128] The terms "non-denatured" and "native" refer to proteins that have not been denatured. This includes both denaturable and non-denaturable proteins.

[0129] As used herein, the term "non-whey protein" refers to a protein that is not a whey protein and includes casein and proteins from one or more non-dairy sources.

[0130] As used herein, the term "nutritional composition" refers to a composition for human or animal consumption, including foods and beverages. Consumption can be by eating or drinking. In various embodiments, the foods provided herein meet food safety standards required by the U.S. Food and Drug Administration (FDA), the U.S. Department of Agriculture, the European Food Safety Authority, and / or other state or local food regulatory agencies. The term includes compositions that can be combined with or added to other ingredients to create a composition for human or animal consumption.

[0131] As used herein, the terms "primary heating" and "primary heat treatment" refer to a heat treatment used to denature a protein during the preparation of a thermostable protein composition of the present invention. In some embodiments, the primary heat treatment may include multiple heating and / or holding steps. For example, in some embodiments, the primary heat treatment includes preheating at a first temperature followed by heating at a second temperature. The term "primary heat treatment" is intended to encompass all such heating steps used to denature a protein during the preparation of a thermostable protein composition of the present invention.

[0132] As used herein, the term "protein particles" refers to aggregates of proteins that are insoluble in aqueous solutions such as water. Such particles can be formed by denaturation, e.g., by heat, and shearing, e.g., by mechanical shearing. Protein particles that are insoluble in aqueous solutions can be separated from aqueous solutions by centrifugation, e.g., at 20,000 x g and 25°C for 1 hour. If the composition containing protein particles includes casein, centrifugation should be performed at a neutral pH to avoid precipitation of casein.

[0133] The terms "remaining undenatured" and "remaining native" refer to proteins that are not denatured after a denaturing treatment. This includes both denaturable proteins that are native, and non-denaturable proteins.

[0134] As used herein, the term "retentate" refers to the retentate fraction after ultrafiltration of whey or whey source, cow's milk, or skim milk. Such fraction has a higher percentage of protein and a lower percentage of lactose as a total solids content compared to the starting material.

[0135] As used herein, the word "(s)" following a noun refers to the plural and / or singular form of that noun.

[0136] As used herein, the terms "secondary heating" and "secondary heat treatment" refer to any additional heating step that occurs after the heat treatment used to denature proteins during the preparation of the thermostable protein compositions of the present invention. For example, in some embodiments, the thermostable protein compositions of the present invention are used to produce a nutritional composition, which is then subjected to a secondary heat treatment. Non-limiting examples of secondary heat treatments include high temperature pasteurization, ultra-high temperature (UHT) treatment, retort heating, and the like. In some embodiments, a sample of the thermostable protein composition of the present invention is subjected to a primary particle growth test as described in Section 1.4 of Example 1. The primary particle growth test comprises a secondary heat treatment. In some embodiments, the secondary heat treatment comprises heating to at least 80°C, e.g., at least 90°C, at least 110°C, at least 120°C, or at least 135°C, or at least 140°C. In some embodiments, the secondary heat treatment comprises heating to a temperature of 80-85°C for 20-30 minutes, or to a temperature of 90-95°C for 5 minutes, or to a temperature of 135-150°C for 4-10 seconds, or to a temperature of 110-130°C for 10-20 minutes. In some embodiments, the secondary heat treatment comprises heating to about 90°C for about 10 minutes, or to about 120°C for about 4 minutes, or to about 140°C for about 2 minutes.

[0137] As used herein, the term "shelf-stable" refers to a composition (e.g., a liquid composition, such as a liquid nutritional composition) that can be stored at room temperature (e.g., about 20°C to about 25°C) for an extended period of time (e.g., at least 2 months, at least 3 months, at least 6 months, or at least 12 months) without undergoing undesirable changes. For example, in some embodiments, a shelf-stable composition exhibits minimal or substantially no settling, gelling, aggregation, flocculation, coagulation, viscosity increase, increase in the volume fraction of protein particles having a particle size of 0.1 to 1 μm, increase in the volume fraction of protein particles having a particle size of 1 to 5 μm, and / or increase in the volume fraction of protein particles having a particle size of at least 5 μm after storage at a temperature of about 20°C to about 25°C for a period of at least 2 months, at least 3 months, at least 6 months, or at least 12 months. In some embodiments, a shelf-stable composition exhibits minimal or substantially no mealiness or grittiness. To inhibit bacterial growth, shelf-stable compositions are typically heat-treated, e.g., sterilized and / or pasteurized, and aseptically filled. In some embodiments, shelf-stable compositions (e.g., sterilized and / or pasteurized shelf-stable compositions) exhibit negligible bacterial growth after long-term storage at temperatures of about 20° C. to about 25° C. for at least 2 months, at least 3 months, at least 6 months, or at least 12 months.

[0138] As used herein, the term "subject" includes humans and other primates, as well as other mammals, such as livestock, game and sport animals, and pets. In certain embodiments, the subject is a human. In such embodiments, the subject is a human infant, toddler, child, or adult. In certain embodiments, the subject is in need of nutritional support.

[0139] As used herein, the term "total denaturable whey protein" refers to the sum of the amounts of denaturable serum albumin, denaturable α-lactalbumin, denaturable β-lactoglobulin, denaturable lactoferrin, and denaturable immunoglobulin (IgG). "Denaturable" in this context refers to the sum of denatured protein and denaturable non-denatured protein, and does not include non-denaturable non-denatured protein. The total amount of denaturable whey protein can be calculated as described in Example 1, Section 1.1.

[0140] As used herein, the term "denatured whey protein" refers to the sum of the amounts of denatured serum albumin, denatured α-lactalbumin, denatured β-lactoglobulin, denatured lactoferrin, and denatured immunoglobulin (IgG). The total amount of denatured whey protein can be calculated as described in Example 1, Section 1.1.

[0141] As used herein, the term "total protein" refers to the total protein content of a composition, determined by determining the total nitrogen content of the composition, without subtracting the non-protein nitrogen content, and multiplying by 6.38. The total protein content of a sample can be measured by the Kjeldahl method as described in ISO 8968-1:2014.

[0142] As used herein, the term "whey" or "whey" refers to the liquid composition remaining after the removal of casein from milk. This is due to the action of rennet enzymes, such as those used in cheese production, and the resulting whey is called "sweet whey" or "cheese whey." Alternatively, casein may be removed by acid precipitation, for example, by reducing the pH of the milk to pH 4.6 or below. Whey produced in this manner is called "acid whey" or "sour whey." Alternatively, casein may be removed by microfiltration or other suitable methods known in the art. All such wheys may be used in the present invention.

[0143] As used herein, the term "whey protein concentrate" or "WPC" refers to a fraction of whey from which lactose has been at least partially removed to increase the protein content to at least 20% by weight. In certain embodiments, WPC has at least 65%, at least 70%, at least 75%, or at least 80% by weight of its total solids (TS) as whey protein. In some instances, the proportion of whey protein is substantially unchanged relative to the proportion of whey from which the WPC is derived. In various embodiments, WPC is an evaporated whey protein retentate. As used herein, the term "WPC" includes whey protein isolate (WPI), where the context allows.

[0144] As used herein, the term "whey protein isolate" or "WPI" refers to a WPC in which at least 90% of the TS is whey protein.

[0145] As used herein, the term "yogurt" refers to an acidic or fermented food or beverage product prepared from dairy products that contains live cultures or chemical acidulants, or both. The term "yogurt" includes not only drinking yogurt, but also set or stirred yogurt, and also ambient yogurt. 2. Heat-stable protein composition

[0146] In a first aspect, the present invention provides a heat-stable protein composition comprising whey protein and casein, The composition comprises β-lactoglobulin and casein in a weight ratio of about 1.6:1 to about 5:1, the whey protein comprises at least about 65% (w / w) denaturable whey protein; at least about 40% (w / w) of the total β-lactoglobulin in the composition is covalently aggregated; the composition comprises protein particles; At least about 40% by volume of the protein particles have a particle size of less than 1 μm; and At least a portion of the protein particles comprise co-aggregates of denatured whey protein and casein.

[0147] In certain embodiments, the thermostable protein composition comprises, on a dry weight basis, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% total protein by weight, and useful ranges can be selected between any of these values ​​(e.g., 60%-95%, 65%-95%, 70%-95%, 75%-90%, 80%-85%).

[0148] In various embodiments, at least about 70% w / w of the total protein in the composition is whey protein, e.g., at least about 75%, at least about 80%, at least about 85%, or about 90% is whey protein, and useful ranges can be selected between any of these values ​​(e.g., 70%-90%, 75%-90%, or 80%-90%).

[0149] In various embodiments, less than 30% w / w of the total protein in the composition is casein, e.g., less than about 25%, less than about 20%, less than about 15%, or about 10% w / w is casein, and useful ranges can be selected between any of these values ​​(e.g., 10%-30%, 10%-25%, 10%-20%, 10%-15%, 15%-30%, 15%-25%, or 15%-20%). In one embodiment, about 80 to about 90% w / w of the total protein is whey protein, and about 10 to about 20% w / w of the total protein is casein.

[0150] The relative amounts of β-lactoglobulin and casein in the compositions of the present invention can be described using weight ratios. The ratios of β-lactoglobulin to casein described herein are generally expressed in the format "x:1," meaning that "x" weight units of β-lactoglobulin are present for every weight unit of casein. For example, a weight ratio of β-lactoglobulin to casein of 5:1 means that 5 g of β-lactoglobulin is present for every 1 g of casein, and a weight ratio of 1.6:1 means that 1.6 g of β-lactoglobulin is present for every 1 g of casein. Similarly, a weight ratio of β-lactoglobulin to casein between 1.6:1 and 5:1 means that 1.6 to 5 g of β-lactoglobulin is present for every 1 g of casein.

[0151] The weight ratio of β-lactoglobulin to casein in a composition of the "x:1" format can be calculated by dividing the weight of β-lactoglobulin present by the weight of casein present to determine the value of "x".

[0152] The ratio of β-lactoglobulin to casein can be adjusted by varying the amount of β-lactoglobulin source (such as a whey protein source) and casein source (such as caseinate). In various embodiments, the composition comprises β-lactoglobulin and casein in a weight ratio of about 1.6:1 to about 5:1, e.g., about 1.6:1 to about 4:1, about 1.6:1 to about 3.5:1, about 1.6:1 to about 3:1, about 2:1 to about 5:1, about 2:1 to about 4:1, about 2:1 to about 3.5:1, about 2:1 to about 3:1, about 2.1:1 to about 5:1, about 2.1:1 to about 4:1, about 2.1:1 to about 3.5:1, about 2.1:1 to about 3:1, about 2.2:1 to about 5:1, about 2.2:1 to about 4:1, about 2.2:1 to about 3.5:1, or about 2.2:1 to about 3:1. In various embodiments, the composition comprises β-lactoglobulin and casein in a weight ratio of at least about 1.6:1, e.g., at least about 2:1, at least about 2.1:1, at least about 2.2:1, at least about 2.4:1, at least about 2.6:1, at least about 2.8:1, or at least about 2.9:1.

[0153] In various embodiments, the composition comprises total whey protein and casein in a weight ratio of at least 3:1, at least 4:1, at least 4.1:1, at least 4.2:1, at least 4.3:1, at least 4.4:1, at least 4.5:1, at least 5:1, at least 5.5:1, at least 6:1, at least 7:1, at least 8:1, or at least 9:1. In various embodiments, the composition comprises total whey protein and casein in a weight ratio of 3:1 to 10:1, 3:1 to 9:1, 3:1 to 8:1, 3:3:1 to 7:1, 3:1 to 6:1, 3:1 to 5:1, 4:1 to 10:1, 4:1 to 9:1, 4:1 to 8:1, 4:1 to 7:1, 4:1 to 6:1, 4:1 to 5:1, 4.5:1 to 10:1, 4.5:1 to 9:1, 4.5:1 to 8:1, 4.5:1 to 7:1, 4.5:1 to 6:1, 4.5:1 to 5:1, 5:1 to 10:1, 5:1 to 9:1, 5:1 to 8:1, 5:1 to 7:1, or 5:1 to 6:1.

[0154] In some embodiments, the composition is a liquid composition, hi other embodiments, the composition is a powder, for example, a powder containing less than about 5% moisture. 3. Protein Source

[0155] In some embodiments, the protein source comprises both whey and casein (such as calcium-depleted MPC). In some embodiments, separate whey and casein protein sources are used. In some embodiments, one or more protein sources comprising both whey and casein are used with one or more whey and / or casein protein sources. This is useful, for example, to adjust the total protein content, the total amount of whey protein, the total amount of casein protein, the weight ratio of total whey protein to casein, and / or the weight ratio of β-lactoglobulin to casein. 3.1 Whey protein sources

[0156] The source of whey protein for the heat-stable protein composition can be any source providing one or more whey proteins, such as sweet whey, acid casein whey, milk whey (obtained from microfiltration of skim milk as the permeate phase), or a combination thereof. Exemplary sources of whey protein include, but are not limited to, rennet whey or cheese whey, lactic acid whey, mineral acid whey, casein whey, and microfiltered skim milk whey. In certain embodiments, the source of whey protein for the heat-stable protein composition is whey protein concentrate (WPC) or whey protein isolate (WPI). Whey protein compositions including WPC and WPI may be derived from acid casein whey or cheese whey. Alternative sources of whey protein for the heat-stable protein composition include concentrated compositions containing individual whey proteins (e.g., β-lactoglobulin-enriched compositions or α-lactalbumin-enriched compositions).

[0157] WPC is rich in whey protein, but also contains other components such as lipids, lactose, minerals / ash, and in the case of cheese whey-based WPC, glycomacropeptide (GMP), an undenaturable non-globular protein related to casein. The typical production method for whey protein concentrate utilizes membrane filtration.

[0158] WPC is often listed as "WPC" followed by the percentage (w / w) of whey protein, for example, WPC80 is WPC that contains 80% whey protein by weight.

[0159] The whey protein may be derived from any mammalian species, such as cow, sheep, goat, horse, buffalo, deer, camel, etc. Preferably, the whey protein is of bovine origin.

[0160] In certain embodiments, the whey protein source is available as a powder, preferably a WPC or WPI powder.

[0161] The heat-stable protein composition can be prepared from a mixture of cheese and / or acid WPC and / or WPI, or a mixture of proteins. In certain embodiments, the whey protein source is or comprises WPC and / or WPI. In certain embodiments, the whey protein source is or comprises a blend of WPC and / or WPI, optionally with one or more ingredients including whey protein and / or non-whey protein.

[0162] In certain embodiments, the whey protein source is cheese whey or a WPC prepared from cheese whey. In some such embodiments, coloring agents are used in the cheese-making process. For example, cheesemakers may use annatto to impart an orange-yellow color to colored cheeses such as Cheddar, Leicester, and Gloucester. Annatto color is obtained from the seeds of Bixa orellana (achiote), a shrub native to Central America. The seeds contain carotenoid pigments such as bixin, norbixin, and relin. In some such embodiments, the whey protein source includes coloring agents. 3.2 Casein protein sources

[0163] The source of casein protein for the heat-stable protein composition can be any source that provides one or more casein proteins. Preferably, the casein is non-micellar casein. Preferably, the casein protein source is a source of non-micellar casein. Exemplary sources of non-micellar casein protein include, but are not limited to, caseinates such as sodium caseinate, potassium caseinate, calcium caseinate, and / or magnesium caseinate, calcium-depleted milk protein concentrate (MPC), total milk protein (TMP), or a combination of any two or more thereof.

[0164] In some embodiments, at least about 50% of the casein in the thermostable protein composition is non-micellar casein, e.g., at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% is non-micellar casein, and useful ranges can be selected between any of these values ​​(e.g., 50%-100%, 50%-90%, 60%-100%, 60%-90%, 70%-100%, 70%-90%, 80%-100%, 80%-90%, or 90%-100%).

[0165] In some embodiments, the casein may be enriched in β-casein and / or κ-casein. For example, in some embodiments, the casein source is a β-casein and / or κ-casein enriched fraction. In various embodiments, at least about 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 90, 9%, 99, or 100% of the total casein is provided by β-casein and / or κ-casein enriched casein, and various ranges between any two of these values ​​may be selected.

[0166] As used herein, the term "β-casein-enriched fraction" or the like refers to a casein fraction having a higher ratio of β-casein to α-casein than that of skim milk. As used herein, the term "κ-casein-enriched fraction" or the like refers to a casein fraction having a higher ratio of κ-casein to α-casein than that of skim milk. The casein ratio can be measured by polyacrylamide gel electrophoresis followed by staining with Coomassie blue and densitometry. Other suitable analytical methods are known to those skilled in the art. Preferably, the ratio of β-casein to α-casein is greater than 1:1, more preferably greater than 1.2:1, more preferably greater than 1.4:1, more preferably greater than 1.5:1, more preferably greater than 1.6:1, and most preferably greater than 1.7:1. Preferably, the ratio of κ-casein to α-casein is greater than 0.2:1, more preferably greater than 0.25:1, more preferably greater than 0.3:1, more preferably greater than 0.35:1, and most preferably greater than 0.4:1. - The casein-enriched fraction and the κ-casein-enriched fraction are enriched in β-casein and κ-casein, respectively, relative to the casein in the casein source (generally cow's milk) from which they were prepared. 4. Protein Particles

[0167] The present invention provides a thermostable protein composition comprising protein particles, at least a portion of the protein particles comprising co-aggregates of denatured whey protein and casein.

[0168] The protein particles present in the compositions of the present invention have a large proportion of particles with a small particle size (e.g., less than 1 μm in some embodiments) and are heat stable. Larger insoluble protein particles, especially particles greater than 3 μm, are often undesirable in some applications, as they can cause an unpleasant gritty or sandy mouthfeel. Larger particles are also more likely to be present in liquid compositions, especially low viscosity liquid compositions (e.g., 100 s at 20° C.). -1The heat-stable particles tend to settle in a wide range of applications (e.g., compositions having a viscosity of less than about 400 mPa·s as measured by HPLC). Particles that are not heat-stable may gel upon secondary heat treatment (e.g., pasteurization, ultra-high temperature (UHT) processing, or retort heating), resulting in a high-viscosity product. The inventors have found that co-aggregates of denatured whey protein and casein have small particle sizes (e.g., less than 1 μm in some embodiments) with improved heat stability compared to aggregates of denatured whey protein alone. The heat-stable protein compositions of the present invention can be used to produce high-protein, heat-stable nutritional compositions, including food products, and beverages with low viscosity and low sedimentation.

[0169] The particle size distribution characteristics of protein particles of the thermostable protein compositions of the present invention are based on the particle size distribution of an aqueous composition comprising the thermostable protein composition. In some embodiments, the thermostable protein composition is an aqueous composition, such as a composition produced by the methods of the present invention, before drying. In these embodiments, the particle size distribution characteristics can be measured directly. In other embodiments, if the thermostable protein composition is, for example, a powder, it must be reconstituted in a liquid before measuring the particle size. Particle size may be measured using a Malvern Mastersizer 2000 or 3000, as described in Section 1.3 of Example 1. Other suitable methods for measuring particle size will be apparent to those skilled in the art.

[0170] In various embodiments, the particle size distribution of the protein particles of the thermostable protein composition is substantially unimodal. Monomodality can be assessed by various means known in the art, such as using a dip test (Hartigan & Hartigan, 1985, Ann. Stat. 13(1), 70-84). In some embodiments, the particle size distribution has a p-value of less than 0.10, preferably less than 0.05, as determined by the dip test.

[0171] In some embodiments, at least about 40% by volume of the protein particles have a particle size of less than 1 μm, e.g., at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% have a particle size of less than 1 μm, and are useful. A suitable range can be selected from between any of these values ​​(e.g., about 40% to about 100%, about 40% to about 95%, about 40% to about 90%, about 40% to about 85%, about 40% to about 80%, about 45% to about 100%, about 45% to about 95%, about 45% to about 90%, about 45% to about 85%, about 45% to about 80%, about 50% to about 100%, about 50% to about 95%, about 50% to about 90%, about 50% to about 85%, or about 50% to about 80%).

[0172] In various embodiments, at least 40% by volume of the protein particles in the composition have a particle size of about 0.1 to about 1 μm, or at least 45% by volume, or at least 50% by volume, or at least 55% by volume, or at least 60% by volume, or at least 65% by volume, or at least 70% by volume, or at least 75% by volume, or at least 80% by volume, or at least 85% by volume, or at least 90% by volume, or at least 95% by volume, or 100% by volume of the protein particles have a particle size of about 0.1 to about 1 μm. It has a particle size of 1 to about 1 μm, and a useful range can be selected from between any of these values ​​(e.g., 40% to 100% by volume, 40% to 95% by volume, 40% to 90% by volume, 40% to 85% by volume, 40% to 80% by volume, 45% to 100% by volume, 45% to 95% by volume, 45% to 90% by volume, 45% to 85% by volume, 45% to 80% by volume, 50% to 100% by volume, 50% to 95% by volume, 50% to 90% by volume, 50% to 85% by volume, or 50% to 80% by volume).

[0173] In various embodiments, less than 5% by volume of the protein particles in the composition have a particle size of at least about 5 μm, or less than 4%, or less than 3%, or less than 2%, or less than 1%, or about 0% by volume of the protein particles in the composition have a particle size of at least about 5 μm, and useful ranges can be selected between any of these values ​​(e.g., 0% to 5%, 0% to 4%, 0% to 3%, 0% to 2%, or 0% to 1% by volume).

[0174] In various embodiments, less than 5% by volume of the protein particles in the composition have a particle size of at least about 5 μm, or less than 4% by volume, or less than 3% by volume, or less than 2% by volume, or less than 1% by volume, or about 0% by volume have a particle size of at least about 5 μm, and useful ranges can be selected between any of these values ​​(e.g., 0% to 5% by volume, 0% to 4% by volume, 0% to 3% by volume, 0% to 2% by volume, or 0% to 1% by volume).

[0175] Particle size may be expressed as the volume-weighted mean diameter (D[4,3]). Means for determining D[4,3] are known in the art. Briefly, D[4,3] is calculated based on the volume of the particle, assuming the particle is spherical. It is based on the principle that particles of different sizes scatter light at different angles, with larger particles scattering light at smaller angles. Angular scattering intensity data is measured with an appropriate device and used to calculate particle size using Mie theory. In various embodiments, the [4,3] particle size distribution of the particles in the composition is less than about 5 μm, e.g., less than about 4 μm, less than about 3 μm, less than about 2 μm, less than about 1.5 μm, less than about 1.2 μm, less than about 1.1 μm, less than about 1.0 μm, or less than about 0.9 μm, and useful values ​​can be selected between any of these values ​​(e.g., 0.9 μm to 5 μm, 0.9 μm to 4 μm, 0.9 μm to 3 μm, 0.9 μm to 2 μm, 0.9 μm to 1.5 μm, 0.9 μm to 1.2 μm, 1.0 μm to 5 μm, 1.0 μm to 4 μm, 1.0 μm to 3 μm, 1.0 μm to 2 μm, 1.0 μm to 1.5 μm, or 1.0 μm to 1.2 μm).

[0176] The composition also includes soluble casein that is not bound to particles, and may include whey protein that is not bound to particles (eg, non-denatured whey protein). 5. Denaturation

[0177] Methods for measuring the degree of denaturation of whey proteins are well known in the art. One exemplary method is shown in Section 1.1 of Example 1. Other methods include those relying on an Agilent 2100 Bioanalyzer (Agilent Technologies, Inc. 2000, 2001-2007, Waldbronn, Germany) and a microfluidic chip, utilizing Agilent 2100 Expert software (e.g., Anema, (2009) International Dairy J, 19(4), 198-204), and polyacrylamide gel electrophoresis (e.g., Patel et al, (2007) Le Lait, 87, 251-268).

[0178] If a pre-denatured sample (ie, a sample of the protein composition before the heat treatment step) is available, the method set forth in Example 1, Section 1.1 is preferred.

[0179] If pre-denatured samples are not available, the degree of denaturation of whey proteins can be estimated by the following method: If necessary, the sample is reconstituted to 3% protein with 0.1 M NaCl, after which the sample is divided into three fractions.

[0180] For Fraction 1, total crude protein is measured by Kjeldahl as total nitrogen x 6.38, which includes casein, denatured whey protein aggregates, and soluble undenatured whey protein.

[0181] Fraction 2 is centrifuged at 7,000 x g for 20 minutes without adjusting the pH. This step removes whey protein aggregates, leaving the soluble native whey proteins and casein in solution. The supernatant is collected and the protein content is measured, again using the Kjeldahl method, as total nitrogen x 6.38. This measurement includes casein and soluble native whey proteins.

[0182] Fraction 3 is acidified to pH 4.6 with 15% acetic acid, and the amount added to correct for the resulting dilution is recorded. This fraction is centrifuged again at 7,000 x g for 20 minutes. This step removes casein and whey protein aggregates, leaving the soluble native whey protein in solution. The supernatant is collected and the protein content is measured again using the Kjeldahl method as total nitrogen x 6.38, adjusted to account for the dilution caused by the addition of acetic acid. This measurement includes casein and soluble native whey protein.

[0183] Casein content can be calculated as Fraction 2 - Fraction 3, denatured whey protein aggregate content can be calculated as Fraction 1 - Fraction 2, and total whey protein content can be calculated as (Fraction 1 - Fraction 2) + Fraction 3.

[0184] The soluble protein fractions can be characterized by examining fractions 1 and 3 by HPLC to determine what protein sources may be blended.

[0185] The denaturation rate of whey protein can be calculated using the following formula:

number

[0186] In certain embodiments, less than about 20% w / w of the total protein in the composition is residual denatured whey protein, e.g., less than about 18%, less than about 16%, or less than about 14%, or less than about 12%, and useful ranges can be selected between any of these values ​​(e.g., 12%-20%, 12%-18%, 12%-16%, or 12%-14%). One exemplary method for determining the amount of residual denatured whey protein is provided in Example 1, section 1.1. 6. Covalent aggregation

[0187] Bonds formed during heating of compositions containing whey proteins include covalent and non-covalent bonds, such as hydrogen bonds, ionic bonds, hydrophobic bonds and van der Waals interactions. Without wishing to be bound by theory, it is believed that covalent interactions lead to irreversible aggregation of whey proteins.

[0188] Covalent bonds in heated whey compositions include inter- and intramolecular disulfide bonds formed via sulfhydryl-disulfide exchange or sulfhydryl oxidation (Havea, 2006, Int. Dairy J., 16(5), 415-422). α-Lactalbumin, β-Lactoglobulin, κ-Casein, and αs2-Casein may participate in sulfhydryl-disulfide interactions (Anema & McKenna, 1996, J. Agric. Food Chem., 44, 422-428). β-Lactoglobulin has two intramolecular disulfide (SS) groups and one free sulphide group, making it highly susceptible to covalent aggregation. BSA has 17 SS bonds and one sulphide group, allowing it to undergo covalent aggregation. α-Lactalbumin contains four SS bonds but no free sulphide groups to initiate covalent aggregation.

[0189] The covalent aggregates in the protein particles of the present invention can include covalent bonds formed between any proteins containing cysteine ​​amino acids. Covalent bonds have greater binding strength than non-covalent interactions. Covalent bonds can be cleaved using strong reducing agents such as 2-mercaptoethanol, dithiothreitol (DTT), and tris(2-carboxyethyl)phosphine (TCEP).

[0190] Non-covalently aggregated proteins can form, for example, through hydrophobic interactions (Glani & Apenten, 1999, Int. J. Food Sci., 34(5-6), 467-476) or calcium bridge-mediated protein-protein interactions (Anema & McKenna, 1996, J. Agric. Food Chem., 44(2), 422-428).

[0191] In the protein particles of the present invention, one or more covalent bonds can be formed between two different molecules of the same protein (e.g., between two β-lactoglobulin molecules) and / or between two different proteins (e.g., between β-lactoglobulin and α-lactalbumin), BSA in whey proteins, and either αS2- or κ-casein in caseins. Of the caseins, only αS2- and κ-casein can form covalent bonds with whey proteins. αS2- and κ-casein account for approximately 20% of the casein in the compositions described herein.

[0192] Without wishing to be bound by theory, it is believed that the higher the proportion of β-lactoglobulin that is covalently aggregated in the presence of casein molecules, the more heat stable and small the particles (e.g., particles less than 1 μm).

[0193] Because the compositions of the invention comprise whey proteins, and β-lactoglobulin is the most abundant whey protein, it is convenient to determine the level of covalent aggregation in terms of the weight amount of covalently aggregated denatured β-lactoglobulin. Without wishing to be bound by theory, this is expected to indicate the overall degree of covalent aggregation for all proteins in the composition that are capable of covalent aggregation.

[0194] In various embodiments, the denatured whey protein comprises denatured β-lactoglobulin. In some embodiments, at least about 40% (w / w) of the total β-lactoglobulin in the composition is covalently aggregated, or at least about 50%, or at least about 55%, or at least about 60%, or at least about 65%, or at least about 70%, or at least about 75%, or at least about 80% is covalently aggregated, and useful ranges can be selected between any of these values ​​(e.g., 40%-80%, 50%-80%, 60%-80%, 70%-80%, 40%-70%, 50%-70%, 60%-70%, or 55%-65%).

[0195] Methods for measuring the percentage of covalently aggregated β-lactoglobulin by weight are well known in the art. One exemplary method used herein relies on the use of a Bioanalyzer using the method of Anema 2009, (International Dairy J, 19(4), 198-204), modified as described in Section 1.2 of Example 1. 7.Thermal stability

[0196] Methods for assessing thermal stability are known in the art. In certain embodiments, thermal stability is assessed by subjecting a sample to a secondary heat treatment, such as the primary particle growth test described in Section 1.4 of Example 1. The particle size distribution can be assessed after the primary particle growth test, as described in Section 1.3. The particle size distribution can be assessed directly for the primary particle growth test sample or compared to the particle size distribution of a control sample. The control sample is a sample of the composition used in the primary particle growth test that has not been subjected to the secondary heat treatment.

[0197] The primary particle growth test is typically performed on an aqueous composition containing the protein composition to be tested. Preferably, an aqueous composition containing a sufficient amount of the protein composition to be tested to provide a total protein content of 15% (w / w) is used. The aqueous composition is then subjected to a secondary heat treatment.

[0198] In some embodiments, the primary particle growth test comprises subjecting an aqueous composition comprising a sufficient amount of the thermostable protein composition to a total protein content of 15% (w / w) to a secondary heat treatment of (a) 90°C for 10 minutes, (b) 120°C for 4 minutes, or (c) 140°C for 2 minutes.

[0199] In some embodiments, the particle size distribution characteristics of the protein particles of the thermostable protein composition are substantially unchanged when subjected to a primary particle growth test.

[0200] In some embodiments, the proportion of protein particles having a particle size of 0.1 to 1 μm does not decrease substantially when subjected to a primary particle growth test. In some embodiments, after the primary particle growth test, the volumetric proportion of protein particles having a particle size of 0.1 to 1 μm is at least about 70% of that of the control sample, e.g., at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98%, or at least about 99%, and a useful range can be selected between any of these values ​​(e.g., 70% to 99%, 75% to 99%, 80% to 99%, 85% to 99%, 90% to 99%, 70% to 95%, 75% to 95%, 80% to 95%, 85% to 95%, 90% to 95%, 70% to 90%, 75% to 90%, 80% to 90%, or 85% to 90%).

[0201] In some embodiments, after a primary particle growth test, at least about 40% by volume of the protein particles in the aqueous composition have a particle size of 0.1 to 1 μm, e.g., at least about 42%, at least about 44%, at least about 45%, at least about 46%, at least about 48%, at least about 50%, at least about 52%, at least about 54%, or about 55% by volume have a particle size of 0.1 to 1 μm, and a useful range can be selected from between any of these values ​​(e.g., 40% to 55%, 42% to 55%, 44% to 55%, 46% to 55%, 48% to 55%, 50% to 55%, 52% to 55%, 40% to 50%, 42% to 50%, 44% to 50%, 46% to 50%, or 48% to 50% by volume).

[0202] In one specifically contemplated embodiment, an aqueous composition comprising a sufficient amount of the thermostable protein composition to provide a 15% w / w total protein content, which is subjected to a secondary heat treatment at 120°C for 4 minutes, comprises at least about 40% by volume, preferably at least about 45% by volume, and more preferably at least about 50% by volume, of the protein particles having a particle size of 0.1 to 1 μm.

[0203] In some embodiments, the volume fraction of protein particles having a particle size of 1 to 5 μm does not increase substantially when subjected to a primary particle growth test. In some embodiments, after a primary particle growth test, the volume fraction of protein particles having a particle size of 1 to 5 μm is less than about 130% of the volume fraction of the control sample, e.g., less than about 125%, less than about 120%, less than about 115%, less than about 110%, less than about 105%, less than about 103%, less than about 102%, or less than about 101% of the volume fraction of the control sample.

[0204] In some embodiments, less than about 60% by volume of the protein particles in the aqueous composition have a particle size of 1-5 μm, e.g., less than about 58%, less than about 56%, less than about 55%, less than about 54%, less than about 53%, less than about 52%, less than about 51%, less than about 50%, less than about 49%, less than about 48%, less than about 47%, less than about 46%, or less than about 45% have a particle size of 1-5 μm, and useful ranges can be selected between any of these values ​​(e.g., 45%-60%, 45%-56%, 45%-55%, 45%-52%, or 45%-50%).

[0205] In one specifically contemplated embodiment, an aqueous composition comprising a sufficient amount of a thermostable protein composition to provide a 15% w / w total protein content, which is subjected to a secondary heat treatment at 120°C for 4 minutes, comprises less than about 60% by volume, preferably less than about 55% by volume, and more preferably less than about 50% by volume, of the protein particles having a particle size of 1-5 μm.

[0206] In some embodiments, the proportion of protein particles having a particle size of at least 5 μm does not increase substantially when subjected to a primary particle growth test. In some embodiments, after a primary particle growth test, the volume fraction of protein particles having a particle size of at least 5 μm is less than about 130% of the volume fraction of the control sample, e.g., less than about 125%, less than about 120%, less than about 115%, less than about 110%, less than about 105%, less than about 103%, less than about 102%, or less than about 101% of the volume fraction of the control sample.

[0207] In some embodiments, the volume fraction of protein particles having a particle size of at least 5 μm is less than about 10%, e.g., less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, less than about 1%, or about 1%, and useful ranges can be selected between any of these values ​​(e.g., 0%-10%, 0%-8%, 0%-6%, 0%-4%, 0%-2%, 1%-10%, 1%-8%, 1%-6%, 1%-4%, or 1%-2%).

[0208] In one specifically contemplated embodiment, an aqueous composition comprising a sufficient amount of a thermostable protein composition to provide a 15% w / w total protein content, which is subjected to a secondary heat treatment at 120°C for 4 minutes, comprises protein particles in which less than about 10% by volume, preferably less than about 5% by volume, more preferably less than about 2% by volume, and most preferably less than about 1% by volume of the protein particles have a particle size of at least 5 μm.

[0209] In some embodiments, the D[4,3] of the protein particles does not increase substantially upon subjection to the primary particle growth test. In some embodiments, after the primary particle growth test, the D[4,3] is less than about 140%, e.g., less than about 135%, less than about 130%, less than about 120%, less than about 110%, less than about 105%, less than about 103%, less than about 102%, or less than about 101% of that of the control sample.

[0210] In some embodiments, the D[4,3] of the protein particle is less than about 5 μm, e.g., less than about 4 μm, less than about 3 μm, less than about 2.5 μm, less than about 2 μm, less than about 1.5 μm, less than about 1 μm, or about 0.9 μm, and a useful range can be selected between any of these values ​​(e.g., 0.9 μm to 5 μm, 0.9 μm to 4 μm, 0.9 μm to 3 μm, 0.9 μm to 2 μm, 0.9 μm to 1.5 μm, or 0.9 μm to 1 μm).

[0211] In some embodiments, after the primary particle growth test, the volume ratio of protein particles having a particle size of 0.1-1 μm is at least about 80% of the volume ratio of the unheated sample, and the volume ratio of protein particles having a particle size of 1-5 μm is less than about 105% of the volume ratio of the unheated sample. In some embodiments, after the primary particle growth test, at least about 40% of the protein particles have a particle size of 0.1-1 μm, less than about 55% of the protein particles have a particle size of 1-5 μm, and less than about 3% of the protein particles have a particle size of at least 5 μm.

[0212] Thermal stability can also be evaluated by the state of the aqueous solution after the primary particle growth test. In some embodiments, after the primary particle growth test, the aqueous solution containing the thermostable protein composition exhibits minimal or substantially no gelation, sedimentation, or aggregation. The gelation of a liquid composition is considered to be a change in state from a liquid to a soft to a solid. When the solution no longer flows after heating, it is considered to have gelled.

[0213] In some embodiments, an aqueous composition comprising a thermostable protein composition in an amount sufficient to provide a 15% w / w total protein content, and subjected to a secondary heat treatment at 90°C for 10 minutes or 120°C for 4 minutes, exhibits minimal or substantially no gelling or solidification.

[0214] In certain embodiments, the particle size distribution of the thermostable protein composition is stable even during a secondary heat treatment (e.g., high-temperature pasteurization, UHT treatment, or retort heating). High-temperature pasteurization requires temperatures of 80-85°C for 20-30 minutes or 90-95°C for 5 minutes. UHT treatment typically involves sterilizing the composition at temperatures above 135°C, e.g., 135-150°C. Typical UHT hold times are 4-10 seconds (or longer). Retort treatment typically involves sterilizing the composition in a sealed can at temperatures of 110-130°C for 10-20 minutes.

[0215] The thermal stability can also be evaluated by the viscosity of the aqueous solution after the primary particle growth test. The viscosity of the sample can be measured by methods known in the art, such as the method shown in Section 1.6 of Example 1.

[0216] In some embodiments, after the primary particle growth test, the aqueous solution comprising the thermostable protein composition exhibits minimal or substantially no increase in viscosity. In some embodiments, after the primary particle growth test, the aqueous solution comprising the thermostable protein composition with a protein content of 15% (w / w) exhibits a viscosity increase of 100 s. -1 Viscosity of less than about 100 mPa·s at 20°C at a shear rate of, e.g., 100 s -1 and a viscosity of less than about 90 mPa·s, less than about 80 mPa·s, less than about 70 mPa·s, less than about 60 mPa·s, less than about 50 mPa·s, less than about 40 mPa·s, less than about 30 mPa·s, less than about 20 mPa·s, less than about 15 mPa·s, less than about 10 mPa·s, less than about 5 mPa·s, or about 4 mPa·s at a shear rate of 100 mPa·s, and a useful range can be selected between any of these values ​​(e.g., 4-100, 4-80, 4-60, 4-50, 4-20, 4-15, 4-10, 5-100, 5-80, 5-60, 5-50, 5-20, 5-15, or 5-10 mPa·s).

[0217] In some embodiments, thermal stability can be assessed by measuring the thermal clot time (HCT). For example, high thermal stability can be indicated by a long thermal clot time.

[0218] Methods for determining heat clotting time (HCT) are known in the art. One exemplary method for determining HCT is provided in Example 1, section 1.5.

[0219] While not wishing to be bound by any particular theory, Applicants believe that liquid compositions with a heat setting time of less than 60 seconds at 140°C pose a high risk of extensive fouling and blocking of UHT heating equipment, while liquid compositions with an HCT of 65-80 seconds at 140°C pose a potential risk of fouling. As described herein, liquid compositions with a heat setting time of 80 seconds or more are considered stable to UHT heating at 140°C for 5 seconds. Alternatively, or additionally, samples with an HCT of less than 3 minutes after heating in an oil bath at 120°C pose a high risk of gelling and clumping in retort cans.

[0220] In some embodiments, an aqueous composition comprising a sufficient amount of a thermostable protein composition to provide a 10% w / w total protein content has an HCT of at least about 60 seconds, e.g., at least about 70 seconds, about 80 seconds, about 90 seconds, about 100 seconds, about 110 seconds, about 120 seconds, about 130 seconds, about 140 seconds, about 150 seconds, about 160 seconds, about 170 seconds, about 180 seconds, about 190 seconds, about 200 seconds, about 210 seconds, about 220 seconds, about 230 seconds, about 240 seconds, about 250 seconds, or about 260 seconds, with useful ranges selected therebetween (e.g., 60-260 seconds, 80-260 seconds, 80-220 seconds, 100-260 seconds, 100-220 seconds, 120-260 seconds, or 100-220 seconds).

[0221] In some embodiments, an aqueous composition comprising a sufficient amount of a thermostable protein composition to provide a total protein content of 15% (w / w) and subjected to a secondary heat treatment at 90°C for 10 minutes, 120°C for 4 minutes, or 140°C for 2 minutes shows no visible signs of gelation. 8. Methods for Preparing Thermostable Protein Compositions

[0222] Applicants have surprisingly found that heat treatment of aqueous solutions containing high concentrations of whey protein and casein protein under conditions of high shear stress can produce protein particles comprising denatured whey protein and casein with a high proportion of covalently aggregated denatured β-lactoglobulin, which particles are small in size (e.g., less than 1 μm in some embodiments), heat stable, and do not require a mechanical shearing step to further break down the particles.

[0223] Without wishing to be bound by theory, a total whey protein concentration of at least about 18 g / 100 g aqueous composition, a β-lactoglobulin concentration of at least about 9 g / 100 g aqueous composition, and a total protein concentration of at least about 20 g / 100 g aqueous composition, combined with a weight ratio of β-lactoglobulin to casein of about 1.6:1 to about 5:1, favors the formation of small particles (e.g., at least 40% by volume of the protein particles are less than 1 μm in size) with a high proportion of total β-lactoglobulin that is covalently aggregated (e.g., at least 40% by weight of the total β-lactoglobulin that is covalently aggregated).

[0224] Advantageously, such protein compositions are heat stable, i.e., when subjected to a secondary heat treatment, there is no or minimal increase in particle size in the composition.

[0225] In various embodiments, when the protein composition is subjected to a second heat treatment: a) there is minimal or no reduction in the volume percent of particles in the protein composition having a particle size of 0.1 to 1 μm (e.g., the volume percent of protein particles having a particle size of 0.1 to 1 μm in the protein composition may be at least 70% of the volume percent of a control sample that is not subjected to the secondary heat treatment); b) there is minimal or no increase in the volume percentage of particles having a particle size of 1-5 μm in the protein composition (e.g., the volume percentage of protein particles having a particle size of 1-5 μm may be less than 130% of the volume percentage of a control sample not subjected to the secondary heat treatment); and / or c) there is minimal or no increase in the volume percentage of particles in the protein composition having a particle size of at least 5 μm (e.g., the volume percentage of protein particles having a particle size of at least 5 μm may be less than 130% of the volume percentage of a control sample not subjected to the secondary heat treatment).

[0226] Such compositions may contain, after a secondary heat treatment, at least about 40% by volume of protein particles having a particle size of 0.1-1 μm, less than about 60% by volume having a particle size of 1-5 μm, and / or less than 10% by volume having a particle size of at least 5 μm. Such compositions may also experience minimal or no increase in viscosity upon secondary heat treatment. Such compositions may also experience minimal or substantially no gelation or solidification upon secondary heat treatment.

[0227] Thus, in one embodiment, the present invention provides a method for preparing a thermostable protein composition, the method comprising: a. An aqueous composition having a pH of 5.5 to 6.8, i. a total whey protein content of at least about 18 g / 100 g aqueous composition; ii. a beta-lactoglobulin content that is at least about 9 g / 100 g aqueous composition; iii. a total protein content that is at least about 20 g / 100 g aqueous composition; and iv. providing an aqueous composition comprising β-lactoglobulin and casein in a weight ratio of about 1.6:1 to about 5:1; and b. heat-treating the aqueous composition to at least about 70°C for a time sufficient to cause protein denaturation, wherein the heat-treating comprises heating the aqueous composition under high shear stress to provide a thermostable protein composition.

[0228] In another aspect, the present invention provides a method for preparing a thermostable protein composition, the method comprising: a. contacting a whey protein source with an oxidizing agent in combination with a catalyst, such as an enzymatic or chemical catalyst, preferably a peroxidase enzyme; b. An aqueous composition having a pH of 5.5 to 6.8 obtained by contacting a whey protein source with a casein protein source, i. a total whey protein content of at least about 14 g / 100 g aqueous composition; ii. a β-lactoglobulin content in a ratio that is at least about 4 g / 100 g aqueous composition; iii. a total protein content that is at least about 15 g / 100 g aqueous composition; and iv. providing an aqueous composition comprising β-lactoglobulin and casein in a weight ratio of about 1.6:1 to about 5:1; and c. heat-treating the aqueous composition to at least about 70°C for a time sufficient to cause protein denaturation, wherein the heat-treating comprises heating the aqueous composition under high shear stress to provide a thermostable protein composition.

[0229] In another aspect, the present invention provides a method for preparing a thermostable protein composition, the method comprising: a. An aqueous composition having a pH of 5.5 to 6.8, i. a total whey protein content of at least about 14 g / 100 g aqueous composition; ii. a β-lactoglobulin content in a ratio that is at least about 4 g / 100 g aqueous composition; iii. a total protein content that is at least about 15 g / 100 g aqueous composition; and iv. providing an aqueous composition comprising β-lactoglobulin and casein in a weight ratio of about 1.6:1 to about 5:1; b. contacting the aqueous composition with an oxidizing agent in combination with a catalyst, such as an enzymatic or chemical catalyst, preferably a peroxidase enzyme; and c. heat-treating the aqueous composition to at least about 70°C for a time sufficient to cause protein denaturation, wherein the heat-treating comprises heating the aqueous composition under high shear stress to provide a thermostable protein composition.

[0230] In some embodiments, the thermostable protein composition produced by the method is a composition according to the first aspect.

[0231] An exemplary method for preparing a thermostable protein composition is shown in Figure 4. Suitable modifications of the methods described herein to achieve the thermostable protein compositions will be apparent to those skilled in the art.

[0232] As shown in Figure 4, in some embodiments, the method includes contacting, e.g., mixing, a whey protein source and a casein protein source to provide the aqueous composition of step a). The whey protein source and / or casein protein source can be any suitable source as described herein. In some embodiments, the whey protein source comprises or consists of whey protein concentrate (WPC), whey protein isolate (WPI), or a combination thereof. In some embodiments, the casein protein source comprises or consists of caseinate, calcium-depleted milk protein concentrate (MPC), total milk protein (TMP), or a combination of any two or more thereof. Examples of caseinate include sodium caseinate, potassium caseinate, calcium caseinate, and / or magnesium caseinate. Alternatively, or additionally, other non-micellar casein sources may be used. Alternatively, or additionally, a protein source containing both whey and casein (e.g., calcium-depleted MPC or total milk protein (TMP)) may be used.

[0233] In some embodiments, the whey protein source and / or casein protein source are in the form of a powder. When the whey protein source and / or casein protein source are in the form of a powder, they generally need to be reconstituted to provide the aqueous composition of step a). Thus, in some such embodiments, the method comprises reconstituting the powder(s) to provide the aqueous composition, for example, reconstituting the powder(s) in water.

[0234] In some embodiments, reconstitution involves stirring for a time sufficient to fully hydrate the powder, for example, 60 minutes. In some embodiments, stirring may be performed at an elevated temperature, such as 50°C. Alternatively, or additionally, the powder may be reconstituted by homogenization. In some embodiments, the method further comprises homogenizing the aqueous composition, preferably at about 200 / 50 bar, preferably prior to step b).

[0235] In various embodiments, the powder is added to water in an amount sufficient to provide the required concentration. It will be appreciated that by reformulating different amounts of whey protein source and / or casein protein source, different concentrations of total whey protein, β-lactoglobulin, and total protein can be achieved. It will also be appreciated that by varying the relative amounts of whey protein source and casein protein source used, different weight ratios of β-lactoglobulin to casein can be achieved.

[0236] In some embodiments, the whey protein source and / or the casein protein source are in the form of a solution. For example, the whey protein source may include whey retentate.

[0237] In one exemplary embodiment, the method comprises providing a whey protein source, such as cheese whey, clarifying, separating, and / or thermally (pasteurizing) the cheese whey to produce a whey protein solution, subjecting the whey protein solution to ultrafiltration and / or diafiltration to produce a whey protein retentate, and contacting the whey protein retentate with a casein protein source to produce the aqueous solution of step a).

[0238] Cheese whey recovered after cheese making is clarified and then heated (pasteurized) for microbial control and to inactivate rennet enzymes and starter cultures left over from cheese production.

[0239] Typical conditions for thermalization (pasteurization) are known in the art and include, but are not limited to, about 63°C for 30 minutes (also known as the batch holding method), about 72°C for 15 seconds (also known as the high temperature, short time (HTST) method), about 89°C for 1 second, or any alternative thermal and non-thermal treatments that have equivalent sterilizing effects to the above-mentioned treatments.

[0240] The whey protein solution can be subjected to microfiltration and / or ultrafiltration, and optionally diafiltration, to obtain a retentate. In certain embodiments, the whey protein retentate has a total solids (TS) content of about 10% to about 40%. In some such embodiments, the whey protein retentate has a TS content of about 13% to about 38%, about 18% to about 33%, or about 23% to about 30%. In some such embodiments, the whey protein retentate has a TS content of at least 18%, at least 20%, at least 22%, or at least 24%.

[0241] In certain embodiments, the filtration step is carried out to provide a retentate having about 65% to about 95% total protein by weight. In some such embodiments, the total protein content of the whey protein retentate is about 75% to about 90% by weight, or about 80% to about 85% by weight. In some such embodiments, the total protein content of the whey protein retentate is at least 65%, at least 70%, at least 75%, or at least 80% by weight.

[0242] At this stage, it may be desirable to ensure that the whey protein solution and / or whey protein retentate remains substantially undenatured (e.g., less than 10% denatured), particularly during thermal treatment. Without wishing to be bound by theory, the uncontrolled formation of denatured aggregates may lead to the formation of larger than desired aggregates during the thermal denaturation step.

[0243] In some such embodiments, the whey protein source and / or whey protein retentate have a low level of denaturation prior to step b). For example, the level of denaturation of the whey protein source and / or whey protein retentate prior to step b) may be less than about 10%, such as about 3% to about 8%. Alternatively, the level of denaturation of the whey protein source and / or whey protein retentate prior to the denaturing heat treatment step may be less than 9%, less than 8%, less than 7%, less than 6%, or less than 5%. In some such embodiments, the level of denaturation of the whey protein source and / or whey protein retentate prior to step b) is about 5%, about 4%, about 3%, about 2%, or about 1%.

[0244] In various embodiments, the aqueous composition has a low level of denaturation prior to step b). In various embodiments, less than about 10% (w / w) of the total denaturable whey protein in the aqueous composition prior to step b) is denatured, e.g., less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than about 1% is denatured, and a useful range can be selected between any of these values ​​(e.g., 1%-10%, 1%-9%, 1%-8%, 1%-7%, 1%-6%, 1%-5%, 1%-4%, 1%-3%, or 1%-2%). In some embodiments, about 5% of the total denaturable whey protein in the aqueous composition prior to step b) is denatured, e.g., about 4%, about 3%, about 2%, or about 1% is denatured.

[0245] In various embodiments, the aqueous composition of step a) comprises a total whey protein content that is at least about 18 g / 100 g aqueous composition, or at least about 20 g, or at least about 22 g, or at least about 24 g, or at least about 26 g / 100 g aqueous composition, and a useful range can be selected between any of these values ​​(e.g., about 18 g / 100 g to about 26 g / 100 g, about 18 g / 100 g to about 24 g / 100 g, about 18 g / 100 g to about 22 g / 100 g, about 20 g / 100 g to about 26 g / 100 g, about 20 g / 100 g to about 24 g / 100 g, or about 20 g / 100 g to about 22 g / 100 g).

[0246] In various embodiments, the aqueous composition of step a) comprises a total whey protein content that is at least about 9 g / 100 g aqueous composition, or at least about 10 g, or at least about 11 g, or at least about 12 g, or at least about 13 g / 100 g aqueous composition, and a useful range can be selected between any of these values ​​(e.g., about 9 g / 100 g to about 13 g / 100 g, about 9 g / 100 g to about 12 g / 100 g, about 10 g / 100 g to about 13 g / 100 g, about 10 g / 100 g to about 12 g / 100 g).

[0247] In various embodiments, the aqueous composition of step a) comprises a total whey protein content that is at least about 20 g / 100 g aqueous composition, or at least about 22 g, or at least about 24 g aqueous composition, or at least about 26 g, or at least about 28 g, at least about 30 g, at least about 32 g / 100 g aqueous composition, and useful ranges can be selected between any of these values ​​(e.g., from about 20 g / 100 g to about 32 g / 100 g, from about 20 g / 100 g to about 32 g / 100 g, from about 20 g / 100 g to about 32 g / 100 g). 0g / 100g, about 20g / 100g to about 28g / 100g, about 22g / 100g to about 32g / 100g, about 22g / 100g to about 30g / 100g, about 22g / 100g to about 28g / 100g, about 24g / 100g to about 32g / 100g, about 24g / 100g to about 30g / 100g, about 24g / 100g to about 28g / 100g, about 26g / 100g to about 32g / 100g, about 26g / 100g to about 30g / 100g, or about 26g / 100g to about 28g / 100g).

[0248] In various embodiments, the aqueous composition of step a) comprises a weight ratio of β-lactoglobulin to casein of about 1.6:1 to about 5:1, e.g., about 1.6:1 to about 4:1, about 1.6:1 to about 3.5:1, about 1.6:1 to about 3:1, about 2:1 to about 5:1, about 2:1 to about 4:1, about 2:1 to about 3.5:1, about 2:1 to about 3:1, about 2.1:1 to about 5:1, about 2.1:1 to about 4:1, about 2.1:1 to about 3.5:1, about 2.1:1 to about 3:1, about 2.2:1 to about 5:1, about 2.2:1 to about 4:1, about 2.2:1 to about 3.5:1, or about 2.2:1 to about 3:1. In various embodiments, the aqueous solution of step a) comprises β-lactoglobulin and casein in a weight ratio of at least about 1.6:1, such as at least about 2:1, at least about 2.1:1, at least about 2.2:1, at least about 2.4:1, at least about 2.6:1, at least about 2.8:1, or at least about 2.9:1, or at least about 3:1, or at least about 3.5:1, or at least about 4:1, or at least about 4.5:1, or at least about 5:1.

[0249] In some embodiments, the method further comprises, preferably prior to step a), contacting the whey protein source with an oxidizing agent in combination with a catalyst, such as an enzymatic or chemical catalyst, preferably a peroxidase enzyme.

[0250] In some embodiments, the method further comprises, preferably prior to step b), contacting the aqueous composition with an oxidizing agent in combination with a catalyst, such as an enzyme or chemical catalyst, preferably a peroxidase enzyme.

[0251] In some embodiments, the oxidizing agent is a peroxide, such as hydrogen peroxide, and the method comprises contacting the aqueous composition, or the whey protein source, with the peroxide in combination with a peroxidase enzyme.

[0252] In certain embodiments, the amount of oxidizing agent is less than 300 ppm, for example, from about 5 to about 250 ppm, for example, from about 5 to about 200 ppm, or from about 10 to about 150 ppm, or from about 20 to about 120 ppm, from about 10 to about 200 ppm, for example, from about 40 to about 190 ppm, or from about 140 ppm to about 190 ppm, or from about 20 to about 220 ppm, for example, from about 20 to about 120 ppm, or from about 30 to about 100 ppm.

[0253] In a particular embodiment, the amount of oxidizing agent is 1200×10 whey protein -6 kg / kg, e.g., about 20 x 10 whey protein -6 kg / kg ~ Whey protein approx. 900 x 10 -6 kg / kg, whey protein approx. 40 x 10 -6 kg / kg ~ whey protein approx. 800 x 10 -6 kg / kg, or approximately 60 x 10 whey protein -6 kg / kg ~ Whey protein approx. 700 x 10 -6 kg / kg, e.g., whey protein approximately 80 x 10 -6 kg / kg ~ Whey protein approx. 600 x 10 -6 kg / kg, e.g., about 100 × 10 -6kg / kg ~ Whey protein approx. 400 x 10 -6 kg / kg.

[0254] In certain embodiments, the amount of oxidizing agent is 2000×10 of the denaturable whey protein. -6 kg / kg, e.g., less than about 30×10 -6 kg / kg ~ approx. 1400 x 10 of denaturable whey protein -6 kg / kg, approximately 60 × 10 of denaturable whey protein -6 kg / kg ~ approx. 1200 x 10 of denaturable whey protein -6 kg / kg, or approximately 90 x 10 of denaturable whey protein -6 kg / kg ~ approx. 1000 x 10 of denaturable whey protein -6 kg / kg, e.g., about 120 × 10 of denaturable whey protein -6 kg / kg ~ approx. 800 x 10 of denaturable whey protein - 6 kg / kg, e.g., about 150 x 10 of denaturable whey protein -6 kg / kg ~ approx. 600 x 10 of denaturable whey protein -6 kg / kg.

[0255] In certain embodiments, the amount of oxidizing agent is 2400×10 of β-lactoglobulin protein. -6 kg / kg, e.g., about 40 x 10 of β-lactoglobulin protein -6 kg / kg ~ approx. 1800 x 10 -6 kg / kg, approximately 80 × 10 of β-lactoglobulin protein -6 kg / kg ~ approx. 1600 x 10 -6 kg / kg, or approximately 120 × 10 of β-lactoglobulin protein -6 kg / kg ~ approx. 1400 x 10 -6 kg / kg, for example, about 160 × 10 -6 kg / kg ~ approx. 1200 x 10 -6 kg / kg, e.g., about 200 × 10 -6 kg / kg ~ approx. 800 x 10 -6kg / kg.

[0256] In certain embodiments, the molar ratio of oxidizing agent to β-lactoglobulin protein is less than 2, for example, from about 0.04 to about 1.8, from about 0.08 to about 1.5, from about 0.12 to about 1.2, from about 0.16 to about 0.9, from about 0.2 to about 0.7, from about 0.24 to about 0.6, or from about 0.28 to about 0.5.

[0257] In certain embodiments, the amount of oxidizing agent is added in one step. More preferably, the total amount of peroxide is added in one or more steps, e.g., 2, 3, 4, 5 or more steps, to avoid inhibition or deactivation of the catalyst. More preferably, the amount of peroxide is added continuously. Those skilled in the art can determine the optimal rate of oxidizing agent addition as a function of catalyst / enzyme activity and protein concentration in the aqueous solution or whey protein source.

[0258] In some embodiments, the oxidizing agent is a food-grade oxidizing agent, hi some such embodiments, the oxidizing agent is oxygen (O), ozone, peroxides including alkyl hydroperoxides, superoxide, peroxynitrite, peroxydisulfate, or lactoperoxidase.

[0259] In some embodiments, the oxidizing agent is a peroxide, preferably an organic peroxide. Exemplary peroxides include, but are not limited to, metal peroxides (e.g., alkali metal peroxides such as sodium peroxide, alkaline earth metal peroxides such as magnesium peroxide or calcium peroxide, or transition metal peroxides such as zinc peroxide), hydrogen peroxide, and benzoyl peroxide. In some embodiments, the oxidizing agent is a perborate, such as sodium perborate.

[0260] In some embodiments, the oxidizing agent is benzoyl peroxide or hydrogen peroxide. Hydrogen peroxide is available in a variety of concentrations and purities. In one embodiment, the hydrogen peroxide is food grade.

[0261] In some embodiments, the oxidizing agent is used in conjunction with a catalyst, which may be an enzymatic or chemical catalyst. In some embodiments, the oxidizing agent is used in conjunction with a catalytic enzyme, such as a microbial peroxidase enzyme. An exemplary fungal peroxidase enzyme is MaxiBright® (DSM Food Specialties). Other exemplary microbial peroxidase enzymes include dye-decolorizing (DyP-type) peroxidases, such as EfeB / YcdB from Escherichia coli O157, DyPB from Rhodococcus jostii RHA1, and DyP2 from Amycolatopsis sp. 75iv2. Exemplary chemical catalysts include, but are not limited to, copper, iron, zinc, manganese, and the like.

[0262] In some embodiments, the oxidizing agent is added over a period of time. For example, an initial amount of oxidizing agent may be added, and subsequent amounts of oxidizing agent may be added as the initial amount is consumed. Thus, the total amount of oxidizing agent may be added in one or more steps, e.g., two, three, four, five or more steps, or may be added continuously over a period of time.

[0263] Exemplary oxidizing agents, including hydrogen peroxide, have previously been employed in the dairy industry to bleach whey.

[0264] Hydrogen peroxide (H2O2) is a clear, colorless liquid with a slightly pungent odor. Hydrogen peroxide is one of two bleaching agents currently approved for bleaching whey in the United States, for decolorizing whey compositions prepared from annatto-containing cheeses.

[0265] In some embodiments, the oxidizing agent may be optionally inactivated, removed, or consumed prior to step (b), such that the aqueous composition is substantially free of oxidizing agent during the heat treatment of step (b). In certain embodiments, no active steps are taken to inactivate, remove, or consume the oxidizing agent during the heat treatment of step (b), and the aqueous composition is substantially free of oxidizing agent. For example, the oxidizing agent may be present in an amount such that removal or consumption of the oxidizing agent is undesirable (e.g., <10 ppm).

[0266] In certain embodiments, the oxidizing agent is used in conjunction with a catalytic enzyme, such as a microbial peroxidase enzyme, and the amounts of oxidizing agent and peroxidase enzyme are such that the oxidizing agent is completely consumed by the enzyme. Alternatively, when the oxidizing agent is hydrogen peroxide, a catalase enzyme may be used to catalyze the decomposition of hydrogen peroxide into water and oxygen. In certain embodiments, no catalase enzyme is used.

[0267] The aqueous composition may be sampled to confirm that it is substantially free of oxidizing agents. By way of example, the aqueous composition may be sampled and tested for detectable peroxides using peroxide test strips available from Merck / MilliporeSigma.

[0268] As shown in Figure 4, in some embodiments, the method further comprises adjusting the pH of the aqueous composition to a pH of 5.5 to 6.8. The pH adjustment can be performed by adding a food-safe acid(s) or base(s) to achieve the required pH. In various embodiments, the pH is adjusted using NaOH, KOH, and / or HCl.

[0269] In various embodiments, the method further comprises adjusting the pH of the aqueous composition to a pH of 5.5 to 6.8, or 5.6 to 6.7, or 5.7 to 6.6, or 5.8 to 6.5, or 5.9 to 6.4, or 6.0 to 6.3, or 6.1 to 6.3. Preferably, the method comprises adjusting the pH of the aqueous composition to a pH of 6.1 to 6.3.

[0270] In some embodiments, the method includes concentrating the aqueous composition, preferably by evaporation, prior to step b).

[0271] The aqueous composition is subjected to a heat treatment, as shown in Figure 4. The heat treatment is carried out to impart the required modifications.

[0272] The heat treatment of step b) is carried out at a temperature and for a time sufficient to denature a portion of the whey protein into insoluble aggregates. In some embodiments, step b) comprises heat treating the aqueous composition for a time sufficient to denature at least about 65% (w / w) of the denaturable whey protein in the aqueous composition, e.g., at least about 70%, or at least about 75%, or at least about 80%, such as at least about 85%, or at least about 90%, or at least about 95%, or about 100% (w / w), and a useful range can be selected between any of these values ​​(e.g., about 65%). to about 100%, about 70% to about 100%, about 75% to about 100%, about 80% to about 100%, about 85% to about 100%, about 90% to about 100%, about 65% to about 95%, about 70% to about 95%, about 75% to about 95%, about 80% to about 95%, about 85% to about 95%, about 90% to about 95%, about 65% to about 90%, about 70% to about 90%, about 75% to about 90%, about 80% to about 90%, about 85% to about 90%, about 65% to about 85%, about 70% to about 85%, about 75% to about 85%, or about 80% to about 85%).

[0273] Temperature and time can be varied depending on the level of protein denaturation required: if a higher temperature is used, a shorter time may be sufficient to provide the same level of denaturation as a lower temperature and longer time.

[0274] In various embodiments, step b) comprises heating the aqueous composition to a temperature of at least about 70° C., or at least about 75° C., or at least about 80° C., or at least about 85° C., or at least about 90° C. In various embodiments, step b) comprises heating the solution to a temperature of from about 70° C. to about 150° C., or from about 70° C. to about 140° C., or from about 70° C. to about 130° C., or from about 70° C. to about 120° C., or from about 70° C. to about 110° C., or from about 70° C. to about 100° C., or from about 70° C. to about 90° C., or from about 70° C. to about 85° C., or from about 70° C. to about 80° C., or from about 75° C. to about 150° C., or from about 75° C. to about 140° C., or from about and heating to a temperature of 75°C to about 130°C, or about 75°C to about 120°C, or about 75°C to about 110°C, or about 75°C to about 100°C, or about 75°C to about 90°C, or about 80°C to about 150°C, or about 80°C to about 140°C, or about 80°C to about 130°C, or about 80°C to about 120°C, or about 80°C to about 110°C, or about 80°C to about 100°C, or about 80°C to about 90°C. Preferably, step b) comprises heating the solution to a temperature of about 80°C to about 90°C.

[0275] In various embodiments, step b) comprises heating the aqueous composition for about 1 second to about 30 minutes, or about 1 second to about 20 minutes, or about 1 second to about 15 minutes, or about 1 second to about 10 minutes, or about 1 second to about 5 minutes, or about 1 second to about 1 minute, or about 1 second to about 45 seconds, or about 1 second to about 30 seconds, or about 1 second to about 15 seconds, or about 1 second to about 5 seconds, or about 5 seconds to about 30 minutes, or about 5 seconds to about 20 minutes, or about 5 seconds to about 15 minutes, or about 5 seconds to about 10 minutes, or about The method may include heat-treating the aqueous composition for 5 seconds to about 5 minutes, or for about 5 seconds to about 1 minute, or for about 5 seconds to about 45 seconds, or for about 5 seconds to about 30 seconds, or for about 5 seconds to about 15 seconds, or for about 5 seconds to about 10 seconds, or for about 10 seconds to about 30 minutes, or for about 10 seconds to about 20 minutes, or for about 10 seconds to about 15 minutes, or for about 10 seconds to about 10 minutes, or for about 10 seconds to about 5 minutes, or for about 10 seconds to about 1 minute, or for about 10 seconds to about 45 seconds, or for about 10 seconds to about 30 seconds, or for about 10 seconds to about 15 seconds. Preferably, step b) includes heat-treating the aqueous composition for about 10 seconds to about 30 seconds.

[0276] In various embodiments, step b) comprises preheating to 55°C followed by heating to 85°C for 10-15 seconds.

[0277] Step b) comprises heating the aqueous composition under high shear stress conditions.Shear stress can be generated by various means known in the art.For example, in certain embodiments, step b) is carried out under high shear stress conditions generated by increasing serum viscosity, or wall shear rate, or changing flow pattern to turbulent flow, or applying mechanical shearing process, or any combination thereof.

[0278] In certain embodiments, step b) is carried out while maintaining a high wall shear rate, optionally under laminar flow, for example for at least about 1000 s -1 In some such embodiments, the wall shear rate is about 1000 s. -1 ~about 10000s-1 , or about 1500s -1 ~about 5000s -1 , or about 1500s -1 ~approximately 4000s -1 , or about 2000s -1 ~about 3000s -1 is.

[0279] In certain embodiments, step b) comprises heating the aqueous composition to at least 70° C. under turbulent flow conditions, e.g., conditions having a Reynolds number of at least about 2000. In some such embodiments, the Reynolds number is from about 2000 to about 20,000, from about 2000 to about 10,000, or from about 2000 to about 5,000. In some such embodiments, the Reynolds number is from about 2000 to about 2500, or from about 2100 to about 2300.

[0280] Turbulent flow is defined as flow with sufficient mass flow rate through the heated pipe to result in a Reynolds number, called Re, of at least 2000. Such Reynolds numbers are characteristic of turbulent flow and are known in the field of fluid mechanics. Determining Re depends on the mass velocity of the fluid and its maximum viscosity at the target heating temperature. This viscosity is defined as the nominal viscosity, determined using the Hagen-Poiseuille equation from measurements of pressure drop along a known length of a horizontal pipe of known uniform circular cross section at a known flow rate of the fluid heated at a uniform temperature before drying. To calculate Re for a given process, Newtonian fluid formulas can be used, provided the maximum viscosity of the fluid heated at the target temperature is used. This means that other viscosities (i.e., lower viscosities) will result in a higher Re and fall into the turbulent zone.

[0281] In certain embodiments, step b) comprises heat treating the aqueous composition under mechanical shear conditions to at least about 70° C. In some such embodiments, the mechanical shear is generated by a homogenizer, a colloid mill, a high-pressure pump, a scraped surface heat exchanger, a high-shear mixer, or the like.

[0282] In certain embodiments, step b) comprises heat-treating the aqueous composition to a temperature of about 70° C. to about 90° C. under conditions of high shear stress resulting from increased serum viscosity, high wall shear rate, turbulent flow, mechanical shear, or a combination of two or more thereof. In some such embodiments, step b) comprises heating the solution at pH 6.1 to pH 6.3 to a temperature of about 80° C. to about 90° C. under conditions of sufficiently high shear stress resulting from increased serum viscosity, high wall shear rate, turbulent flow, mechanical shear, or a combination of two or more thereof.

[0283] Heating can be accomplished by a variety of means, as will be appreciated by those skilled in the art. Some exemplary heating means are described in US20120114795A1, which is incorporated herein by reference. In a preferred embodiment, a long tubular thermal reactor is used. The thermal reactor typically has a nominal retention time of between 1 second and 1000 seconds.

[0284] The temperature of the aqueous composition at the end of the reactor may be between about 70°C and about 150°C, preferably between about 75°C and about 120°C, and more preferably between about 80°C and about 90°C.

[0285] Following step b), the thermostable protein composition may be dried. Methods for drying aqueous compositions are well known in the art and include spray drying, freeze drying, drum drying, and fluidized bed drying.

[0286] Spray drying is currently preferred. Preferably, the heat treatment zone is directly connected to a spray dryer equipped with a nozzle or nozzles, rotary atomizer or ultrasonic atomizer to produce a stream of droplets.

[0287] In various embodiments, the protein composition is not subjected to mechanical shear prior to drying, other than to convert the liquid into droplets to facilitate drying.

[0288] In some embodiments, the protein composition may be used directly in the preparation of a high protein product without drying, hi some embodiments, the product exiting the flow channel is used as an ingredient in preparing a food product.

[0289] In certain embodiments, the protein composition is not subjected to further particle size reduction or particle size selection procedures, such as microfiltration, to achieve the particle size distribution(s) described herein.

[0290] In various embodiments, the heat-treated material is not subjected to a particle size reduction treatment before drying. In various embodiments, the heat-treated material is subjected to a particle size reduction treatment before drying.

[0291] In some embodiments, the method further comprises cooling the heat-treated aqueous composition, which can be accomplished by any suitable method, such as refrigeration, a heat exchanger, a chilled water bath, or the like. 9. Nutritional Composition

[0292] The thermostable protein compositions of the present invention are useful in the production of a variety of nutritional compositions.

[0293] Thus, in one aspect, the present invention provides a nutritional composition comprising the thermostable protein composition of the first or third aspect.

[0294] In some embodiments, the nutritional compositions have an improved flavor compared to all-whey compositions, for example, the compositions may exhibit an increased "milky" flavor and / or a reduced "eggy" flavor compared to all-whey compositions.

[0295] In various embodiments, the nutritional composition may include one or more lipids. In various embodiments, the nutritional composition may include one or more carbohydrates. In various embodiments, the nutritional composition may include one or more lipids and one or more carbohydrates.

[0296] In various embodiments, the nutritional compositions may comprise, on a dry basis, at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 30%, about 40%, or at least about 50% by weight of total protein, and useful ranges can be selected between any of these values ​​(e.g., from about 1% to about 40%, or from about 1% to about 30%, or from about 1% to about 20%, or from about 1% to about 16% by weight, about 1% to about 15% by weight, about 1% to about 14% by weight, or about 1% to about 12% by weight, or about 1% to about 10% by weight, or about 2% to about 50% by weight, or about 2% to about 40% by weight, or about 2% to about 30% by weight, about 4% to about 14% by weight, or about 4% to about 12% by weight, or about 4% to about 10% by weight, about 5% to about 50% by weight, or about 5% to about 40% by weight, or about 5% to about 30% by weight, or about 5% to about 20% by weight, or about 5% to about 16% by weight, about 5% to about 15% by weight, about 5% to about 14% by weight, or about 5% to about 12% by weight, or about 5% to about 10% by weight.

[0297] In various embodiments, the nutritional compositions may comprise, on a dry basis, at least about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 30%, about 40%, or at least about 50% by weight of denatured whey protein, and useful ranges can be selected between any of these values ​​(e.g., from about 1% to about 40% by weight, or from about 1% to about 30% by weight, or from about 1% to about 20% by weight, or from about 1% to about 50% by weight). about 16% by weight, about 1% to about 15% by weight, about 1% to about 14% by weight, or about 1% to about 12% by weight, or about 1% to about 10% by weight, or about 2% to about 50% by weight, or about 2% to about 40% by weight, or about 2% to about 30% by weight, about 4% to about 14% by weight, or about 4% to about 12% by weight, or about 4% to about 10% by weight, about 5% to about 50% by weight, or about 5% to about 40% by weight, or about 5% to about 30% by weight, or about 5% to about 20% by weight, or about 5% to about 16% by weight, about 5% to about 15% by weight, about 5% to about 14% by weight, or about 5% to about 12% by weight, or about 5% to about 10% by weight.

[0298] In various embodiments, at least about 50% (w / w) of the total protein in the nutritional composition is denatured whey protein, e.g., at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, or at least about 80% is denatured whey protein, and useful ranges can be selected between any of these values ​​(e.g., about 50% to about 80%, about 50% to about 70%, about 50% to about 60%, about 60% to about 80%, about 60% to about 70%, or about 70% to about 80%).

[0299] In various embodiments, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% (w / w) of the total protein in the nutritional composition is provided by the thermostable protein composition, with useful ranges ranging from these values. It can be selected from any of the following (e.g., about 50% to about 100%, about 50% to about 90%, about 50% to about 80%, about 50% to about 70%, about 50% to about 60%, about 60% to about 100%, about 60% to about 90%, about 60% to about 80%, about 60% to about 70%, about 70% to about 100%, about 70% to about 90%, about 70% to about 80%, about 80% to about 100%, about 80% to about 90%, or about 90% to about 100%).

[0300] In various embodiments, at least about 40% by volume of the protein particles in the nutritional composition have a particle size less than about 1 μm, or at least about 45%, or at least about 50%, or at least about 55%, or at least about 60%, or at least about 65%, or at least about 70%, or at least about 75%, or at least about 80%, or at least about 85%, or at least about 90%, or at least about 95%, or about 100% have a particle size less than about 1 μm, and useful ranges can be selected therebetween (e.g., 40%-100%, 45%-100%, 45%-95%, 45%-90%, 45%-85%, 45%-80%, 50%-100%, 50%-95%, 50%-90%, 50%-85%, or 50%-80%).

[0301] In various embodiments, at least about 40% by volume of the protein particles in the nutritional composition have a particle size of about 0.1 to about 1 μm, or at least about 45%, or at least about 50%, or at least about 55%, or at least about 60%, or at least about 65%, or at least about 70%, or at least about 75%, or at least about 80%, or at least about 85%, or at least about 90%, or at least about 100%. At least about 95%, or about 100%, have a particle size of about 0.1 to about 1 μm, and useful ranges can be selected between any of these values ​​(e.g., 40% to 100%, 40% to 95%, 40% to 90%, 40% to 85%, 40% to 80%, 45% to 100%, 45% to 95%, 45% to 90%, 45% to 85%, 45% to 80%, 50% to 100%, 50% to 95%, 50% to 90%, 50% to 85%, or 50% to 80%).

[0302] In various embodiments, less than about 55% by volume of the protein particles in the nutritional composition have a particle size of about 1 to about 5 μm, or less than about 50%, or less than about 45%, or less than about 40%, or less than about 35%, or less than about 30%, or less than about 25% have a particle size of about 1 to about 5 μm, and useful ranges can be selected between any of these values ​​(e.g., 25%-55%, 25%-50%, 25%-45%, 25%-40%, 25%-35%, 25%-30%, 30%-55%, 35%-55%, 40%-55%, or 45%-55%).

[0303] In various embodiments, less than about 5% by volume of the protein particles in the nutritional composition have a particle size of at least about 5 μm, or less than about 4% by volume, or less than about 3% by volume, or less than about 2% by volume, or less than about 1% by volume, or about 0% by volume have a particle size of at least about 5 μm, and useful ranges can be selected between any of these values ​​(e.g., 0% to 5% by volume, 0% to 4% by volume, 0% to 3% by volume, 0% to 2% by volume, or 0% to 1% by volume).

[0304] In various embodiments, the nutritional composition may comprise at least about 0.1% lipid by weight on a dry basis, for example, about 0.1%, about 0.2%, or about 0.5%, or about 1%, or about 3%, or about 5%, or about 10% lipid by weight on a dry basis. In various embodiments, the nutritional composition may comprise between about 0.1% and 40% lipid by weight on a dry basis. , a useful range can be selected between any of these values ​​(e.g., about 0.1 to about 40% by weight, or about 0.5 to about 40% by weight, or about 1 to about 40% by weight, or about 3 to about 40% by weight, or about 5 to about 40% by weight, or about 10 to about 40% by weight, or about 15 to about 40% by weight, or about 20 to about 40% by weight, or about 0.1 to about 35% by weight, or about 0.5 to about 35% by weight, or about 1 to about 35% by weight, or about 3 to about 35% by weight, or about 5 to about 35% by weight, or about 10 to about 35% by weight, or about 15 to about 40% by weight). About 35% by weight, or about 20 to about 35% by weight, or about 0.1 to about 30% by weight, or about 0.5 to about 30% by weight, or about 1 to about 30% by weight, or about 3 to about 30% by weight, or about 5 to about 30% by weight, or about 10 to about 30% by weight, or about 15 to about 30% by weight, or about 20 to about 30% by weight, or about 0.1 to about 20% by weight, or about 0.5 to about 20% by weight, or about 1 to about 20% by weight, or about 3 to about 20% by weight, or about 5 to about 20% by weight, or about 10 to about 20% by weight, or about 15 to about 20% by weight.

[0305] In various embodiments, the nutritional compositions may comprise at least about 0.1% carbohydrate by weight on a dry basis, e.g., about 0.1%, or about 0.5%, or about 1%, or about 3%, or about 5%, or about 10% carbohydrate by weight on a dry basis. In various embodiments, the nutritional compositions may comprise from about 0.1% to about 40%, or from about 0.1% to about 80% carbohydrate by weight on a dry basis, and useful ranges may be selected between any of these values ​​(e.g., from about 0.1% to about 80%, from about 0.1% to about 70%, from about 0.1% to about 65%, from about 0.1% to about 60%, from about 0.1% to about 50%, from about 0.1% to about 40%, or about 0.5%. % to about 40% by weight, or about 1% to about 40% by weight, or about 3% to about 40% by weight, or about 5% to about 40% by weight, about 10% to about 80% by weight, about 10% to about 70% by weight, about 10% to about 65% by weight, about 10% to about 60% by weight, about 10% to about 50% by weight, or about 10% to about 40% by weight, or about 15% to about 40% by weight, about 20% to about 80% by weight, about 20% to about 70% by weight, about 20% to about 65% by weight %, about 20% by weight to about 60% by weight, about 20% by weight to about 50% by weight, or about 20% by weight to about 40% by weight, about 30% by weight to about 80% by weight, about 30% by weight to about 70% by weight, about 30% by weight to about 65% by weight, about 30% by weight to about 60% by weight, about 30% by weight to about 50% by weight, about 40% by weight to about 80% by weight, about 40% by weight to about 70% by weight, about 40% by weight to about 65% by weight, about 40% by weight to about 60% by weight, about 40% by weight to about 50% by weight, about 50% by weight to about 80% by weight , about 50% by weight to about 70% by weight, about 50% by weight to about 65% by weight, about 50% by weight to about 60% by weight, or about 0.1% by weight to about 35% by weight, or about 0.5% by weight to about 35% by weight, or about 1% by weight to about 35% by weight, or about 3% by weight to about 35% by weight, or about 5% by weight to about 35% by weight, or about 10% by weight to about 35% by weight, or about 15% by weight to about 35% by weight, or about 20% by weight to about 35% by weight, or about 0.1% by weight to about 30% by weight, or about 0.5% to about 30% by weight, or about 1% to about 30% by weight, or about 3% to about 30% by weight, or about 5% to about 30% by weight, or about 10% to about 30% by weight, or about 15% to about 30% by weight, or about 20% to about 30% by weight, or about 0.1% to about 20% by weight, or about 0.5% to about 20% by weight, or about 1% to about 20% by weight, or about 3% to about 20% by weight, or about 5% to about 20% by weight, or about 10% to about 20% by weight, or about 15% to about 20% by weight.

[0306] In various embodiments, the nutritional compositions may comprise at least about 20% lactose by weight on a dry basis, e.g., at least about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, or about 80% lactose by weight, and useful ranges can be selected between any of these values ​​(e.g., about 20% to about 80%, about 20% to about 70%, about 20% to about 60%, About 20% by weight to about 55% by weight, about 20% by weight to about 50% by weight, about 30% by weight to about 80% by weight, about 30% by weight to about 70% by weight, about 30% by weight to about 60% by weight, about 30% by weight to about 55% by weight, about 30% by weight to about 50% by weight, about 40% by weight to about 80% by weight, about 40% by weight to about 70% by weight, about 40% by weight to about 60% by weight, about 40% by weight to about 55% by weight, about 40% by weight to about 50% by weight, about 45% by weight to about 80% by weight, about 45% by weight to about 70% by weight, about 45% by weight to about 60% by weight, or about 45% by weight to about 55% by weight.

[0307] In various embodiments, the thermostable protein composition is formulated into a nutritional composition in powder form, such as a protein powder, shake mix, or supplement, suitable for combination with a liquid and reconstitution into a beverage.

[0308] In various embodiments, the thermostable protein composition is formulated into a nutritional composition by wet blending. In various embodiments, the nutritional composition is a wet blend nutritional composition, such as a wet blend meal replacement composition, a wet blend infant formula, or a wet blend medical food.

[0309] In various embodiments, the nutritional composition contains at least about 30 mg / 100 g of Ca. 2+ In various embodiments, the nutritional composition may contain at least about 30 mg / 100 ml of Ca, for example, at least about 40 mg / 100 g, at least about 50 mg / 100 g, at least about 60 mg / 100 g, at least about 75 mg / 100 g, or at least about 100 mg / 100 g of divalent cations such as Ca. 2+ and the like, for example, at least about 40 mg / 100 ml, at least about 50 mg / 100 ml, at least about 60 mg / 100 ml, at least about 75 mg / 100 ml, or at least about 100 mg / 100 ml.

[0310] In certain embodiments, the nutritional composition is a medical food. By way of example only, U.S. federal law and Food and Drug Administration (FDA) regulations define a medical food as "a food formulated to be taken or administered enterally under the supervision of a physician and intended for the specific dietary management of a disease or condition for which distinctive nutritional requirements have been established based on accepted scientific principles through medical evaluation" (§ 5(b) of the 1988 Orphan Drug Act (21 U.S.C. 360ee(b)(3)) and FDA regulation 21 CFR 101.9(j)(8)).

[0311] Described herein are methods for providing nutritional support to a subject, comprising enterally administering to the subject a nutritional composition comprising a thermostable protein composition of the present invention. In certain embodiments, the subject is a subject in need of nutritional support. Accordingly, the nutritional compositions described herein are for use in methods for providing nutritional support to a subject in need thereof. In some embodiments, the subject is a human. In some embodiments, the subject is a human infant or toddler. In other embodiments, the subject is a human adult. In some embodiments, the subject is a pregnant female. Enteral administration can be oral administration or tube administration (e.g., nasogastric feeding, or gastric feeding).

[0312] In certain embodiments, the nutritional composition is enterally administered to a subject to maintain or increase muscle protein synthesis, maintain or increase muscle mass, prevent or reduce muscle mass loss, maintain or enhance growth, prevent or reduce muscle catabolism, prevent or treat cachexia, prevent or treat sarcopenia, increase the rate of glycogen resynthesis, regulate blood glucose levels, increase insulin response to elevated blood glucose levels, reduce satiety, reduce satiety, increase food intake, increase calorie intake, improve glucose metabolism, speed up post-operative recovery, increase the effect of pre-rehabilitation before surgery or chemotherapy, speed up recovery after injury, speed up recovery after exercise, enhance sports performance, and / or provide nutrition. Enteral administration can be oral administration or tube administration (e.g., nasogastric feeding or gastric feeding).

[0313] In certain embodiments, nutritional compositions comprising the thermostable protein compositions of the present invention are used to maintain or increase muscle protein synthesis, maintain or increase muscle mass, prevent or reduce muscle mass loss, maintain or enhance growth, prevent or reduce muscle catabolism, prevent or treat cachexia, prevent or treat sarcopenia, increase the rate of glycogen resynthesis, regulate blood glucose levels, increase insulin response to elevated blood glucose levels, reduce satiety, reduce satiation, increase food intake, increase caloric intake, improve glucose metabolism, speed post-operative recovery, increase the effectiveness of pre-rehabilitation before surgery or chemotherapy, speed recovery after injury, speed recovery after exercise, enhance sports performance, and / or provide nutrition.

[0314] In certain embodiments, the nutritional compositions are sterilized and / or pasteurized by a method suitable for commercial use. In some embodiments, the sterilization and / or pasteurization comprises a secondary heat treatment. In other embodiments, the sterilization and / or pasteurization comprises a non-heat treatment. Exemplary techniques for sterilization and / or pasteurization include, but are not limited to, secondary heat treatments such as high-temperature sterilization, ultra-high temperature (UHT) treatment, and retort heating.

[0315] In a further aspect, the present invention provides a method for preparing a nutritional composition, the method comprising: a) a thermostable protein composition of the present invention; b) one or more additional components.

[0316] Other procedures for preparing and packaging the nutritional compositions will depend on the nutritional composition being produced and will be known to those skilled in the art.

[0317] In various embodiments, the one or more additional components include one or more lipids, one or more carbohydrates, one or more monovalent cations, and / or one or more divalent metal cations.

[0318] In various embodiments, the method can include providing an aqueous composition comprising the thermostable protein composition. In some embodiments, the method can include reconstituting the powdered thermostable protein composition, optionally with one or more additional dry ingredients, to produce the aqueous composition. In other embodiments, the powdered thermostable protein composition can be used without reconstitution.

[0319] The thermostable protein compositions of the present invention are useful for producing powder or ready-to-mix compositions, particularly wet blend compositions, wet blend meal replacement compositions, wet blend infant formulas, and / or wet blend medical foods. A wet blend composition is a composition prepared using a wet mixing step in which one or more ingredients are combined as an aqueous solution, optionally subsequently dried and powdered. Wet mixing may be required, for example, for ready-to-mix nutritional compositions that require a wet mixing step to incorporate a fat source into the composition.

[0320] The thermostable protein compositions of the present invention are particularly useful in wet blend applications because the low viscosity of the compositions allows high total solids contents to be achieved during evaporation. If the evaporation step can achieve high total solids contents, less water must be removed during spray drying, resulting in a corresponding reduction in energy usage. Furthermore, the thermostability of the compositions of the present invention may reduce line fouling during evaporation and drying, resulting in a corresponding reduction in production downtime and an increase in plant throughput and product yield.

[0321] Methods for preparing a wet blend composition will be known to those skilled in the art. Briefly, methods for preparing a wet blend composition may include combining two or more components to form an aqueous solution. The aqueous solution may optionally be subjected to one or more additional steps, such as homogenization, standardization, heat treatment (e.g., pasteurization and / or UHT treatment), homogenization, evaporation, and / or spray drying. The aqueous solution may comprise the thermostable protein composition of the present invention and, optionally, one or more additional components selected from one or more lipids, one or more carbohydrates, one or more additional components described herein, or any combination thereof. 9.1 Food

[0322] In some embodiments, the nutritional composition is a food product.

[0323] In various embodiments, the food product may be ice cream, a fermented product, buttermilk, cheese, processed cheese, a cheese analog, quark, a pudding, a frozen dessert, a coffee whitener, a gel, a bar, or a baked good.

[0324] Foods prepared using the thermostable protein compositions of the present invention exhibit only a slight increase in undesirable flavors (e.g., bitter, savory, or eggy flavors) compared to a control food having the same ingredient composition and protein content as the food of the present invention, except that the control food does not contain the thermostable protein composition of the present invention. 9.2 Liquid composition

[0325] The thermostable protein compositions of the present invention are useful for producing liquid compositions, particularly liquid nutritional compositions such as high-protein beverages. In such compositions, the protein compositions of the present invention can provide high protein content while maintaining acceptable viscosity, resisting particle size change after secondary heat treatment, exhibiting reduced settling, and a good mouthfeel (e.g., little grittiness or chalkiness). In some embodiments, the liquid compositions also have improved flavor compared to all-whey compositions, such as increased "milky" flavor and / or decreased "eggy" flavor.

[0326] Thus, in one aspect, the present invention provides a liquid composition comprising the thermostable protein composition of the present invention. In some embodiments, the liquid composition is a liquid nutritional composition.

[0327] Examples of suitable liquid compositions include, but are not limited to, high-protein beverages, such as aqueous liquids (e.g., concentrates) that can be diluted, ready-to-drink (RTD) beverages, etc. In certain embodiments, the liquid composition is a shake, smoothie, dairy drink, UHT milk, milk powder, sports drinks, including dairy and non-dairy sports drinks, protein shots, fruit juice, medical foods, or milk powders, such as infant formula, follow-on formula, and growth formula. In certain embodiments, the liquid composition is a neutral high-protein beverage. In certain embodiments, the liquid composition is an acidic high-protein beverage.

[0328] Exemplary liquid compositions include sports drinks, which are typically high in protein, low in fat, and contain added vitamins and minerals, flavorings, sweeteners, stabilizers, and / or salt.

[0329] In some embodiments, the liquid composition comprises at least about 6% (w / w) total protein by weight of the liquid composition, e.g., at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 21%, at least about 22%, at least about 24%, at least about 26%, at least about 28%, or at least about 30% (w / w) total protein, with useful ranges being selected between any of these values ​​(e.g., 6%-30%, 7%-28%, 8%-26%, or 10%-24%). The total protein in a liquid composition is the sum of all protein contributed by all protein-containing components in the composition. For example, in embodiments where a liquid composition comprises a heat-stable protein composition and other non-dairy and / or dairy proteins, e.g., from skim milk powder (SMP) or a milk protein concentrate (MPC), the total protein in the liquid composition is the sum of the total protein present in the heat-stable protein composition and the SMP and / or MPC.

[0330] In some embodiments, the liquid composition comprises at least about 6% (w / w) denatured whey protein by weight of the liquid composition, e.g., at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 21%, at least about 22%, at least about 24%, at least about 26%, at least about 28%, or at least about 30% (w / w) denatured whey protein, with useful ranges being selected between any of these values ​​(e.g., 6%-30%, 7%-28%, 8%-26%, or 10%-24%).

[0331] In some embodiments, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% of the total protein in the liquid composition is denatured whey protein, and useful ranges can be selected between any of these values ​​(e.g., 50%-95%, 60%-90%, or 70%-85%). In some embodiments, the liquid composition includes other dairy proteins and / or non-dairy proteins. In some embodiments, the liquid composition includes whey protein from two or more sources, such as a heat-stable protein composition of the present invention, a denatured whey protein composition, a non-denatured whey protein composition, a whey protein hydrolysate, or a component that includes both whey and casein (e.g., MPC). In such embodiments, the total whey protein in the composition is the sum of the total whey protein present in the heat-stable protein composition of the present invention, the denatured whey protein composition, the non-denatured whey protein composition, the whey protein hydrolysate, and / or the MPC.

[0332] In certain embodiments, the liquid composition exhibits minimal or substantially no sedimentation under conditions favorable for sedimentation. Sedimentation can be assessed by storing the liquid composition at a temperature of about 20°C to about 25°C and conducting shelf-life tests for 1 month, 6 weeks, 3 months, 6 months, or 12 months. To test for sedimentation, the product is inverted three times and carefully poured out. The container is then left upside down for 30 minutes, and the weight of the container and sediment is recorded. The amount of sediment is calculated by subtracting the weight of the empty container. The percentage of sediment is calculated as the ratio of the weight of the sediment to the total weight of the product in the container.

[0333] In some embodiments, the liquid composition (e.g., 100 s at 20°C) -1 In some embodiments, liquid compositions (e.g., compositions having a viscosity of less than about 400 mPa·s measured at 20° C.) exhibit less than about 10% settling after storage at a temperature of about 20° C. to about 25° C. for at least 6 weeks, e.g., at least 3 months, at least 6 months, or at least 12 months. -1The composition (having a viscosity of less than about 400 mPa·s as measured by HPLC) exhibits less than about 10% settling, e.g., less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than about 1%, or substantially no settling, after storage at a temperature of about 20°C to about 25°C for 3 months.

[0334] In various embodiments, the liquid composition is heat stable, e.g., the composition is in a liquid state in which substantially no gelation, flocculation, aggregation, coagulation, sedimentation, increase in viscosity, decrease in the volume fraction of protein particles having a particle size of 0.1 to 1 μm, increase in the volume fraction of protein particles having a particle size of 1 to 5 μm, or increase in the volume fraction of protein particles having a particle size of at least 5 μm is observed in the composition after the secondary heat treatment.

[0335] In various embodiments, the liquid composition is storage stable, e.g., the composition is in a liquid state in which, after storage at a temperature of about 20°C for an extended period of time, e.g., at least 3 months, preferably at least 6 months or 12 months, substantially no gelation, flocculation, aggregation, coagulation, sedimentation, increase in viscosity, increase in the volume fraction of protein particles having a particle size of 0.1 to 1 μm, increase in the volume fraction of protein particles having a particle size of 1 to 5 μm, or increase in the volume fraction of protein particles having a particle size of at least 5 μm, is observed in the composition.

[0336] Gelation of a liquid composition is considered to be a change in state from a liquid, soft to a solid. A solution is considered to have gelled when it no longer flows.

[0337] In certain embodiments, the liquid composition is sterilized and / or pasteurized by a method suitable for commercial use. In some embodiments, the sterilization and / or pasteurization comprises a secondary heat treatment. In other embodiments, the sterilization and / or pasteurization comprises a non-heat treatment. Exemplary techniques for sterilization and / or pasteurization include, but are not limited to, secondary heat treatments such as high-temperature pasteurization, ultra-high temperature (UHT) treatment, and retort heating.

[0338] A secondary heat treatment (eg, sterilization and / or pasteurization) may be useful to provide a liquid composition that is shelf stable and can be stored at room temperature for extended periods of time.

[0339] In some embodiments, the liquid composition is shelf-stable after a secondary heat treatment (e.g., sterilization and / or pasteurization). In some embodiments, the liquid composition exhibits minimal or substantially no gelation, flocculation, aggregation, coagulation, sedimentation, viscosity increase, decrease in the volume fraction of protein particles having a particle size of 0.1 to 1 μm, increase in the volume fraction of protein particles having a particle size of 1 to 5 μm, or increase in the volume fraction of protein particles having a particle size of at least 5 μm after a secondary heat treatment (e.g., sterilization and / or pasteurization). In some embodiments, the sterilized and / or pasteurized liquid composition exhibits minimal bacterial growth when aseptically filled after long-term storage at a temperature of about 20° C. to about 25° C. for at least 2 months, at least 3 months, at least 6 months, or at least 12 months. In some such embodiments, the liquid composition exhibits minimal or substantially no grittiness or mealiness. Thus, for example, the liquid composition can be a heat-treated shelf-stable liquid composition.

[0340] In some embodiments, the volume fraction of protein particles in the liquid composition having a particle size of 0.1-1 μm does not decrease after a secondary heat treatment (e.g., sterilization and / or pasteurization). In some embodiments, after the secondary heat treatment, the volume fraction of protein particles having a particle size of 0.1-1 μm is at least 70% of the composition before the secondary heat treatment, e.g., at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99% of the composition before the secondary heat treatment.

[0341] In some embodiments, at least about 40% by volume of the protein particles in the heat-treated liquid composition have a particle size of 0.1 to 1 μm, e.g., at least about 42%, at least about 44%, at least about 45%, at least about 46%, at least about 48%, at least about 50%, or at least about 52%, or at least about 54%, or about 55% by volume have a particle size of 0.1 to 1 μm, and useful ranges can be selected from any of these values ​​(e.g., 40% to 55%, 42% to 55%, 44% to 55%, 46% to 55%, 48% to 55%, or 50% to 55% by volume).

[0342] In some embodiments, the volume fraction of protein particles in the liquid composition having a particle size of 1-5 μm does not substantially increase after secondary heat treatment (e.g., sterilization and / or pasteurization). For example, in some embodiments, after secondary heat treatment, the volume fraction of protein particles in the liquid composition having a particle size of 1-5 μm is less than 130% of the composition before secondary heat treatment, e.g., less than 125%, less than 120%, less than 115%, less than 110%, less than 105%, less than 103%, less than 102%, or less than 101% of the composition before secondary heat treatment.

[0343] In some embodiments, less than 60% by volume of the protein particles in the heat-treated liquid composition have a particle size of 1-5 μm, e.g., less than about 58%, less than about 56%, less than about 55%, less than about 54%, less than about 53%, less than about 52%, less than about 51%, less than about 50%, less than about 49%, less than about 48%, less than about 47%, less than about 46%, or less than about 45% have a particle size of 1-5 μm, and useful ranges can be selected between any of these values ​​(e.g., 45%-60%, 45%-58%, 45%-56%, 45%-54%, or 45%-52%).

[0344] In some embodiments, the volume fraction of protein particles in the liquid composition having a particle size of at least 5 μm does not substantially increase after secondary heat treatment (e.g., sterilization and / or pasteurization). For example, in some embodiments, after secondary heat treatment, the volume fraction of protein particles in the liquid composition having a particle size of 1-5 μm is less than 130% of the composition before secondary heat treatment, e.g., less than 125%, less than 120%, less than 115%, less than 110%, less than 105%, less than 103%, less than 102%, or less than 101% of the composition before secondary heat treatment.

[0345] In some embodiments, less than about 10% by volume of the protein particles in the heat-treated liquid composition have a particle size of at least 5 μm, e.g., less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, less than about 1%, or about 0% by volume have a particle size of at least 5 μm, and useful ranges can be selected between any of these values ​​(e.g., 0% to 10%, 0% to 8%, 0% to 6%, 0% to 4%, 0% to 2%, 1% to 10%, 1% to 8%, 1% to 6%, 1% to 4%, or 1% to 2% by volume).

[0346] In some embodiments, the D[4,3] of the protein particles in the liquid composition does not increase substantially after a secondary heat treatment (e.g., sterilization and / or pasteurization). In some embodiments, after the secondary heat treatment, the D[4,3] of the composition is less than 170% of the D[4,3] of the composition before the secondary heat treatment, e.g., less than 160%, less than 150%, less than 140%, less than 130%, less than 120%, less than 110%, less than 105%, less than 103%, less than 102%, or less than 101% of the D[4,3] of the composition before the secondary heat treatment.

[0347] In some embodiments, the D[4,3] of the protein particles in the heat-treated liquid composition is less than about 20 μm, e.g., less than about 15 μm, less than about 10 μm, less than about 5 μm, less than about 4 μm, less than about 3 μm, less than about 2.5 μm, less than about 2 μm, less than about 1.5 μm, less than about 1 μm, or less than about 0.9 μm.

[0348] In some embodiments, the viscosity of the heat-treated liquid composition is 100 s -1 At a shear rate of less than about 400 mPa·s at 20°C, e.g., 100 s -1 at a shear rate of less than about 350 Pa·s, less than about 300 Pa·s, less than about 250 Pa·s, less than about 200 Pa·s, less than about 180 Pa·s, less than about 160 Pa·s, less than about 140 Pa·s, less than about 120 Pa·s, less than about 100 Pa·s, less than about 80 Pa·s, less than about 60 Pa·s, less than about 50 Pa·s, less than about 40 Pa·s, less than about 30 Pa·s, less than about 20 Pa·s, less than about 15 Pa·s, less than about 10 Pa·s, or less than about 5 Pa·s at 20°C.

[0349] In some embodiments, when the liquid composition is a neutral high-protein beverage, the viscosity is 100 s -1 In some embodiments, when the liquid composition is an acidic high-protein beverage (e.g., drinking yogurt), the viscosity is less than about 200 mPa·s at a shear rate of 100 s or less at 20° C. -1 At a shear rate of 20°C, the shear strength is less than approximately 400 mPa·s.

[0350] In some embodiments, after a secondary heat treatment (e.g., sterilization and / or pasteurization), the proportion of protein particles in the liquid composition having a particle size of 0.1-1 μm is at least about 80% of the proportion in the composition before the secondary heat treatment, and the proportion of protein particles in the liquid composition having a particle size of 1-5 μm is less than about 105% of the proportion in the composition before the secondary heat treatment. In some embodiments, at least about 40% of the protein particles in the heat-treated liquid composition have a particle size of 0.1-1 μm, less than about 55% of the protein particles in the heat-treated liquid composition have a particle size of 1-5 μm, and less than about 3% of the protein particles in the heat-treated liquid composition have a particle size of at least 5 μm.

[0351] It is well known that the lethal effect of high temperature on microorganisms depends on both temperature and holding time, with increasing temperature shortening the time required to kill the same number of microorganisms. The time required to reduce the initial microbial population by a specific amount at a specific temperature is commonly referred to as the "F value." The F value is the equivalent time (in minutes) at a specified temperature that provides the same thermal lethality as 121°C. As is known in the relevant art, the F value required to achieve commercial sterility in acidic products is generally lower than in products processed at neutral pH.

[0352] In certain embodiments, the liquid composition comprises, in addition to the heat-stable protein composition of the present invention, a non-whey protein and / or a non-dairy protein. Exemplary non-whey protein sources include, but are not limited to, skim milk powder (SMP), whole milk powder, milk protein concentrate (MPC), milk protein isolate (MPI), micellar casein concentrate (MCC), etc. As another example, casein may be added to the liquid composition in the form of sodium caseinate, potassium caseinate, calcium caseinate, or magnesium caseinate.

[0353] In certain embodiments, the liquid composition comprises one or more, two or more, or three or more non-dairy proteins. Non-dairy proteins suitable for inclusion in the liquid composition include algal proteins, fungal proteins (e.g., mycoproteins), plant proteins, and animal proteins, and hydrolysates thereof. In some such embodiments, the liquid composition comprises soy protein, rice protein, and / or pea protein.

[0354] Furthermore, the liquid composition may contain whey protein from more than one source. For example, the liquid composition may contain a mixture of WPC and WPI, one or both of which may be heat-denatured. As another example, the liquid composition may contain a mixture of WPCs produced by different methods or with different properties.

[0355] In addition to the methods disclosed herein, exemplary methods for preparing thermally denatured whey protein compositions suitable for use in liquid compositions are provided in PCT International Application Nos. PCT / NZ2007 / 000059 (published as WO2007 / 108709) and PCT / NZ2010 / 000072 (published as WO2010 / 120199) and PCT International Application No. PCT / IB2012 / 056103 (published as WO2013 / 065014), each of which is incorporated herein by reference in its entirety.

[0356] In certain embodiments, one or more of the protein components, such as the heat-stable protein composition or liquid composition, can be treated to reduce the lactose content. In some such embodiments, the heat-stable protein composition or liquid composition is treated with an enzyme such as β-galactosidase or subjected to filtration to remove lactose. Suitable enzyme treatment and filtration protocols for reducing lactose content will be apparent to those skilled in the art.

[0357] In certain embodiments, the liquid composition comprises less than about 10% (w / w) lactose by weight of the liquid composition, e.g., less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, less than about 1%, or about 0.1% (w / w) lactose, and useful ranges can be selected between any of these values ​​(e.g., 0.1%-10%, 0.1%-8%, 0.1%-6%, 0.1%-4%, 0.1%-2%, 1%-10%, 1%-8%, 1%-6%, 1%-4%, or 1%-2%). In some such embodiments, the liquid composition has a lactose content of up to 10% (w / w), up to 8% (w / w), up to 6% (w / w), or up to 4% (w / w) by weight of the liquid composition.

[0358] In a further aspect, the present invention provides a method for preparing a liquid composition, the method comprising: a) a thermostable protein composition of the present invention; b) one or more additional components.

[0359] In various embodiments, the one or more additional ingredients comprise one or more additional protein sources, one or more lipids, one or more carbohydrates, one or more vitamins, one or more minerals, one or more food additives including one or more emulsifiers, one or more food additives including one or more stabilizers, or a combination of any two or more thereof. In some embodiments, the mineral component comprises a monovalent cation and / or a divalent metal cation.

[0360] Obviously, the exact method will vary depending on the high protein beverage being produced. Such variations will be apparent to those skilled in the art. An example of a method for producing a high protein beverage is shown in Figure 5.

[0361] As shown in Figure 5, in one embodiment (e.g., when the liquid composition is an acidic beverage), the one or more additional ingredients may include pectin. In such an embodiment, the method may include hydrating the pectin in an aqueous solution at an elevated temperature for a time sufficient to hydrate the pectin. In one embodiment, the method includes hydrating the pectin at about 80°C.

[0362] In some embodiments, the one or more additional ingredients comprise dry ingredients (e.g., powders), including, for example, a protein ingredient, a carbohydrate ingredient, a mineral ingredient, an acidity regulator, a stabilizer, and / or an emulsifier. In some embodiments, the thermostable protein composition is a dry ingredient (e.g., powder). In some embodiments, the method comprises contacting and / or mixing two or more dry ingredients, such as a thermostable protein composition and one or more additional ingredients.

[0363] In some embodiments, the thermostable protein composition and / or one or more dry ingredients are hydrated in an aqueous solution, e.g., water. In some embodiments, the method includes a rehydration step, which includes hydrating the thermostable protein composition and / or one or more dry ingredients in an aqueous solution (e.g., water) for a time sufficient to hydrate the thermostable protein composition and / or one or more dry ingredients.

[0364] The conditions required to rehydrate the dry ingredients will vary depending on the dry ingredients used. Such variations will be apparent to those skilled in the art. In some embodiments, the thermostable protein composition and / or one or more dry ingredients are hydrated for about 60 minutes. The rehydration step may also include stirring the aqueous solution, which aids in rehydration. In some embodiments, an antifoaming agent is used to reduce or prevent foaming during the rehydration step. For example, in some embodiments, the rehydration step includes adding an antifoaming agent to the aqueous solution before or during rehydration.

[0365] In some embodiments, the aqueous solution used to hydrate the thermostable protein composition and / or one or more dry ingredients is at an elevated temperature, e.g., about 55° C. In some embodiments, the rehydration step comprises heating the aqueous solution (e.g., water) to an elevated temperature (e.g., about 55° C.) before or during rehydration.

[0366] The lipid component(s) may be added before, during, or after the rehydration step, or alternatively, if no rehydration step is used (e.g., if the thermostable protein composition is in the form of an aqueous solution), the lipid component(s) may be added to the thermostable protein composition.

[0367] In some embodiments, the one or more additional ingredients comprise one or more vitamins and / or one or more minerals. In some embodiments, the method comprises contacting the thermostable protein composition with the vitamins and / or minerals. The amount of vitamins and / or minerals used in the liquid composition may be typical of meal replacement products known to those skilled in the art. Micronutrient requirements for various subgroups of the population are also known. Recommended daily amounts of vitamins and minerals can be specified for various subgroups. See, for example, Dietary Reference Intakes (DRIs): Recommended Dietary Allowances and Adequate Intakes, United States National Academy of Sciences, Institute of Medicine, Food and Nutrition Board (2010).

[0368] In some embodiments, the method includes a pH adjustment step. In other embodiments, pH adjustment may not be necessary. pH adjustment may involve adding a food-safe acid or base to achieve the desired pH. An acidity adjuster may also be used to adjust the pH. The desired pH will vary depending on the liquid composition being produced. In one embodiment, the pH is neutral (e.g., about 6.5 to about 7.5). In another embodiment, the pH is acidic (e.g., about 2 to about 4.8). In one embodiment, the method includes adjusting the pH to about 6.8, for example, by adding KOH.

[0369] In some embodiments, the method includes a shearing step. For example, the liquid composition may be sheared in an ultrasonicator at 7,000 rpm for 3 minutes.

[0370] In some embodiments, the method includes a homogenization step. For example, the liquid composition may be passed once through a two-stage homogenizer at 200 / 50 bar.

[0371] In some embodiments, the method includes a shearing step. For example, the liquid composition may be cooled using an ice bath.

[0372] In some embodiments, the method includes a secondary heat treatment step, such as a UHT step, a pasteurization step, a sterilization step, or a retort step. For example, in some embodiments, the method includes UHT treatment, followed by optional post-homogenization, followed by aseptic packaging. In one embodiment (such as a neutral composition), the method includes UHT treatment at about 145°C for about 4 seconds. In one embodiment (such as an acidic composition), the method includes UHT treatment at about 110°C for about 4 seconds. In other embodiments, the method includes filling the liquid composition into a container (such as a can) followed by sterilization. In one embodiment, sterilization is at about 120°C for about 15 minutes.

[0373] Other steps for preparing and packaging a liquid composition (such as a high protein beverage) will vary depending on the composition being produced and will be known to those skilled in the art.

[0374] Some exemplary methods for producing liquid compositions in the form of neutral and acidic high-protein beverages are described below. 1. Neutral high-protein beverage

[0375] The required proteins, carbohydrates, minerals, and stabilizers are dry blended and hydrated in water heated to 55°C for 60 minutes. Oil is added to the mixture. The pH is adjusted to approximately pH 6.8 ± 0.1 using 5% KOH. The mixture is homogenized at 55°C and 150 / 50 bar. The beverage may be sterilized in one of two ways:

[0376] (1) The homogenized mixture is filled into a retort can and heated at 120°C for 15 minutes.

[0377] (2) The homogenized mixture is UHT sterilized at 145°C for 4 seconds and then homogenized at 150 / 50 bar. The product is packaged aseptically. 2. Acidic, high-protein drinks

[0378] The same process as for neutral drinks is followed, except that a pectin solution is prepared at 80°C and added to the water before adding the proteins, carbohydrates, minerals and stabilizers. The pH is adjusted to pH 4.0 ± 0.1 with 15% hydrochloric acid. UHT sterilisation is carried out at 110°C for 4 seconds, followed by homogenisation at 150 / 50 bar. The product is packaged aseptically. 9.3 Additional Ingredients

[0379] The nutritional compositions, food products, and / or liquid compositions of the present invention may comprise one or more additional ingredients described herein. The methods for preparing the nutritional compositions and / or methods for preparing the liquid compositions may comprise contacting the thermostable protein compositions of the present invention with one or more additional ingredients described herein.

[0380] In various embodiments, the one or more additional ingredients may include lipids, carbohydrates, proteins, flavorings, vitamins, minerals, dairy products, water, food additives, polyalcohols, colors, fruit preparations, or combinations of any two or more of these ingredients.

[0381] In various embodiments, lipids can be plant lipids or animal lipids, including dairy lipids.Plant oils are often cited as exemplary lipids because they are easy to prepare and have low saturated fatty acid content.Examples of plant oils include canola (rapeseed) oil, corn oil, sunflower oil, olive oil, soybean oil, hydrogenated vegetable oil, or sn-2 palmitoyl triacylglycerol.

[0382] In various embodiments, the dairy lipids include cream, butter, shortening, anhydrous milk fat (AMF), buttermilk, its hydrolyzed products, combinations of hydrolyzed and / or non-hydrolyzed compositions, one or more stages of hard milk fat extract from milk fat fraction (including hard (H), soft-hard (SH), and soft-soft-hard (SSH) extract), one or more stages of soft milk fat extract from milk fat fraction (including soft (S), soft-soft (SS), and soft-soft-soft (SSS) extract), combinations of hard milk fat extracts, combinations of soft milk fat extracts, combinations of soft milk fat extracts, hard milk types, or any combination of multiple. These compositions may be derived from whole milk or colostrum, and any derivative of whole milk or colostrum, including cream, cultured cream, and whey cream (milk lipids derived from whey, including acid whey or cheese whey, preferably cheese whey). Cultured cream is cream from whole milk or colostrum fermented with acid-producing microorganisms, preferably lactic acid bacteria.

[0383] In various embodiments, the vegetable oil may be coconut oil, corn oil, cottonseed oil, canola oil, rapeseed oil, olive oil, palm oil, peanut oil, safflower oil, sesame oil, soybean oil, sunflower oil, hazelnut oil, almond oil, cashew oil, macadamia nut oil, pecan nut oil, pistachio oil, walnut oil, oil derived from melon and gourd seeds, pumpkin seed oil, apricot oil, glycerin, avocado oil, flaxseed oil, linseed oil, grapeseed oil, hemp oil, linseed oil, rice bran oil, wheat germ oil, or any combination of two or more thereof. In some embodiments, the vegetable oil may be hydrogenated coconut oil.

[0384] In various embodiments, carbohydrates may include monosaccharides, disaccharides, oligosaccharides, and polysaccharides, as well as mixtures thereof, including sugars, sucrose, and sucralose. Many of these are commercially available starches, modified starches, maltodextrins (3-20 glucose equivalents (DE)), or long-chain carbohydrates such as corn syrup (>20 DE). Non-digestible carbohydrates may also be included, such as fructooligosaccharides, inulin, and galactooligosaccharides. In various embodiments, carbohydrates may include polyhydric alcohols selected from the group including, for example, glycerol (glycerin), maltitol, erythritol, sorbitol, and any combination of any two or more thereof.

[0385] In various embodiments, the additional protein may be a dairy protein or a dairy protein. In various embodiments, the additional protein may be milk, whey, casein, caseinate, egg, egg white, egg yolk, vegetable, plant, alfalfa, clover, pea, legume, kidney bean, soybean, kidney bean, lupin, cocoa, hornbeam, tree nut, peanut, rye, grain, whole grain, rice, hemp, wheat gluten, fungal or algae protein, protein concentrate thereof, protein isolate thereof, hydrolysate thereof, or a combination of any two or more thereof.

[0386] In various embodiments, the additional protein may be a protein powder. The protein powder may be obtained from any of the protein sources described above. The protein powder may be non-agglomerated, agglomerated, roll-pressed, freeze-dried, drum-dried, spray-dried, or foam-spray-dried. In various embodiments, the protein powder comprises whey protein concentrate (WPC) or whey protein isolate (WPI). In various embodiments, the protein powder comprises whole milk powder, skim milk powder, or milk protein concentrate (MPC).

[0387] In various embodiments, the one or more additional ingredients may be flavorings, including but not limited to sweeteners, natural flavors, artificial flavors, herbs, and flavorings.

[0388] In various embodiments, the one or more additional ingredients may include nuts and / or seeds.

[0389] In various embodiments, the one or more additional ingredients may be vitamins. The vitamins may include fat-soluble or water-soluble vitamins. Suitable vitamins include, but are not limited to, vitamin C, vitamin A, vitamin E, vitamin B12, vitamin K, riboflavin, niacin, vitamin D, vitamin B6, folic acid, pyridoxine, thiamine, pantothenic acid, and biotin. Forms of vitamins may include salts of vitamins, derivatives of vitamins, compounds having the same or similar activity as vitamins, and metabolites of vitamins.

[0390] In various embodiments, the one or more additional components may be minerals, including but not limited to chloride, sodium, calcium, iron, chromium, copper, iodine, zinc, magnesium, phosphorus, potassium, chromium, etc. Suitable forms of minerals include soluble mineral salts, sparingly soluble mineral salts, insoluble mineral salts, chelated minerals, mineral complexes, carbonyl minerals, non-reactive minerals such as reduced minerals, and combinations thereof.

[0391] In various embodiments, the nutritional compositions, food products, and / or liquid compositions may comprise, for example, at least about 10, at least about 20, at least about 25, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 75, at least about 80, at least about 90, or 100% of the Recommended Daily Intakes (RDIs) of vitamins and minerals set by European (FSMP) or USDRA regulations, in a 100 mL, 250 mL, 500 mL, or 1 liter serving size.

[0392] In various embodiments, the one or more additional ingredients may include a dairy product. In some embodiments, the dairy product may be selected from the group including milk powder protein concentrate, skim milk powder, whole milk powder, whey protein concentrate, whey protein isolate, caseinate, milk fat, cream, curd casein, cheese, or cream cheese. In various embodiments, the one or more additional ingredients may include other dairy products such as milk powder protein concentrate, skim milk powder, whole milk powder, whey protein concentrate, whey protein isolate, caseinate, milk fat, or cream.

[0393] In various embodiments, the one or more additional ingredients may comprise food or beverage additives, including, but not limited to, rennet, antifoaming agents, stabilizers, emulsifiers, preservatives, fiber, probiotics, antioxidants, flavor enhancers, colorants, acidity regulators, or emulsifying salts. In various embodiments, the one or more additional ingredients may be food additives, including, but not limited to, rennet, antifoaming agents, stabilizers, emulsifiers, preservatives, fiber, probiotics, antioxidants, flavor enhancers, colorants, acidity regulators. A useful preservative is potassium sorbate in acidified products.

[0394] In various embodiments, the one or more additional ingredients may be a polyhydric alcohol, which may be selected from arabitol, erythritol, glycerol, isomalt, isomaltulose, lactitol, maltitol, mannitol, sorbitol, xylitol, and hydrogenated starch hydrolysates.

[0395] In certain embodiments, one or more additional components may comprise a stabilizer or emulsifier. Suitable emulsifiers include lecithin, monoglycerides, diglycerides, polyglycerol esters, milk phospholipids, citric acid esters (CITREM), polysorbate 60, glyceryl monostearate, and DATEM. Suitable stabilizers include, but are not limited to, carrageenan, gellan gum, pectin, guar gum, locust bean gum, carboxymethylcellulose, alginate, agar, oat gum, tragacanth gum, acacia gum, xanthan gum, karaya gum, tara gum, starch, modified starch, and microcrystalline cellulose, gelatin, or combinations thereof. Those skilled in the art will recognize that many different gel forms are suitable for use in the liquid compositions disclosed herein, in addition to those listed above.

[0396] In various embodiments, the one or more additional components may include a salt or acidity adjuster such as sodium chloride, potassium chloride, ethylenediaminetetraacetic acid (EDTA) salt, lactic acid, acetic acid, citric acid, potassium hydroxide, phosphate salts such as dipotassium phosphate and disodium phosphate, citrate salts such as disodium citrate, dipotassium citrate, and tripotassium citrate. In some embodiments, the citrate salt may be selected from the group including disodium citrate, dipotassium citrate, tripotassium citrate, and trisodium citrate. In some embodiments, the phosphate salt may be selected from the group including dipotassium phosphate, disodium phosphate, n-phosphate, diphosphate, and polyphosphate.

[0397] In various embodiments, the one or more additional components may include a source of amino acids, amino acid precursors, or amino acid metabolites, or any combination of any two or more thereof, preferably free amino acids, amino acid precursors, or amino acid metabolites.

[0398] The method for contacting the thermostable protein composition with one or more additional ingredients to produce a nutritional, food, or liquid composition will vary depending on the nutritional, food, or liquid composition being formed, and will be well known to those skilled in the art.

[0399] The following non-limiting examples are offered to illustrate the present invention and in no way limit its scope. [Example]

[0400] 1. Example 1: General Procedure 1.1 Measurement of denaturation

[0401] The method used herein relies on HPLC (Elgar et al (2000) J Chromatography A, 878, 183-196). Two samples are used: a sample of the modified composition to be tested (modified sample), and a sample of the composition before heat treatment (native sample).

[0402] Step 1: Remove casein and insoluble whey protein aggregates. The pH of both samples was lowered to 4.6 to precipitate the casein. The samples were then centrifuged to remove the precipitated casein and insoluble whey protein aggregates.

[0403] Step 2: Reverse-phase HPLC. The remaining soluble denatured whey protein in the denatured sample can be measured using reverse-phase HPLC (Elgar et al., 2000) as Σ(bovine serum albumin + α-lactalbumin + β-lactoglobulin + lactoferrin + immunoglobulins) and expressed as grams protein / 100 grams total solids.

[0404] The total soluble and denaturable whey protein in the native sample is determined in the same manner as in step 1.

[0405] Step 3: Calculate denatured whey protein. First, normalize the remaining soluble and denaturable whey protein and total soluble and denaturable whey protein values ​​using the relative levels of glycomacropeptide (GMP) in each sample, taking into account possible concentration changes during processing. If a sample does not contain GMP (e.g., if the whey protein source is acid whey), normalize the values ​​using the total solids content of the two samples from step 1 instead.

[0406] The total denatured whey protein as a percentage of the total denaturable whey protein can then be calculated using the following formula:

number

[0407] The weight percentage of covalently aggregated β-lactoglobulin was measured using the 2100 Agilent Bioanalyzer System (Agilent, USA) with microfluidic SDS electrophoresis technology, following the method of Anema (2009, International Dairy J, 19(4), 198-204) with the following modifications.

[0408] The percentage of remaining native, covalently aggregated, and non-covalently aggregated β-lactoglobulin was determined by dissolving the denatured whey protein composition in different bond-dissociating buffers and measuring the change in soluble protein after a centrifugation step.

[0409] To measure the remaining undenatured β-lactoglobulin content, the samples before and after heating were dissolved or diluted in water to 10% (w / w) protein, and a 300 mg aliquot of this sample was dissolved in 1.2 mL of acetate buffer (0.2 M, pH 4.35).

[0410] To measure the content of noncovalently aggregated β-lactoglobulin, 300 mg of sample was dissolved in 1.2 mL of phosphate / urea / SDS buffer (PSU buffer) at a final concentration of 2% (w / v) in 0.1 M phosphate buffer (pH 6.7) prepared from 0.0281 mol NaH2PO4.2H2O and 0.0218 mol Na2HPO4. Urea was added to a final concentration of 8 M.

[0411] The prepared sample was shaken in a test tube shaker for 1 hour.

[0412] The samples were centrifuged at 20,000 xg and 25°C for 1 hour.

[0413] A 100 μL aliquot was carefully removed from the supernatant of each sample, prepared according to Anema, and run using the Bioanalyzer System. The area under the curve of the β-lactoglobulin peak was automatically integrated by the software (2100 Bioanalyzer Expert Software package). The peak area was used to calculate the type of conjugation based on the sample preparation described above.

[0414] The acetate buffer used in this experiment precipitates denatured whey proteins or residual casein in solution. Unheated samples are expected to contain less denatured protein. Heated samples are expected to contain more denatured protein. Residual undenatured β-lactoglobulin in the heated samples was calculated as a percentage of the soluble β-lactoglobulin in the unheated (raw) solution.

[0415]

number

[0416]

number

[0417]

number

[0418] If the composition to be measured was a powder, it was reconstituted with water before particle size measurement. A 10% (w / w) protein concentration solution was prepared by combining the protein composition with deionized water at room temperature (~20°C) using a magnetic stirrer. The protein powder(s) was gradually added to the water under vortex stirring, taking care to avoid foaming. Once the powder was completely dispersed, hydration was continued for 30 minutes with continuous stirring. The solution was homogenized at 150 / 50 bar, and particle size was measured as described below.

[0419] Particle size was measured using a Malvern Mastersizer 2000 or 3000 (Malvern Instruments Ltd, Worcs, UK). Samples were dispersed in deionized water (refractive index (RI) = 1.33) with a refractive index of 1.46 as the dispersed phase. Samples were added dropwise until an obscuration value of 10-15% was achieved. Particle size is reported as the percentage (volume) of particles within the particle size range under test. The volume percentage of particles within a defined size range was determined by summing the relevant size classes in the raw data reported by the Mastersizer software. 1.4 Primary particle growth test

[0420] The protein composition, or a liquid composition comprising the protein composition, can be tested for thermal stability by the primary particle growth test, as described herein.

[0421] The primary particle growth test involves subjecting an aqueous solution containing a protein composition to a secondary heat treatment at elevated temperature for a holding time, followed by immediate cooling. This can be accomplished, for example, by preparing an aqueous solution containing a protein composition at pH 6.8, placing a 5 mL aliquot in an 8 mL glass vial, placing the vial in a rocking device immersed in a high-temperature silicone oil bath for a holding time, and then immediately cooling in ice water. Particle size analysis is performed as described in Section 1.3. Alternatively, or additionally, other parameters, such as viscosity, can be measured.

[0422] In some embodiments, the aqueous solution comprising the protein composition has a protein content of 10%, 14%, 15%, 18%, or 20% (w / w). In one embodiment, the elevated temperature is 90°C and the hold time is 10 minutes. In another embodiment, the elevated temperature is 120°C and the hold time is 4 minutes. In another embodiment, the elevated temperature is 140°C and the hold time is 2 minutes.

[0423] For example, in some embodiments, the primary particle growth test comprises heating an aqueous solution comprising the protein composition having a protein content of 14% (w / w) to 120°C for 10 minutes. In some embodiments, the primary particle growth test comprises heating an aqueous solution comprising the protein composition having a protein content of 10%, 15%, or 18% (w / w) to 90°C for 10 minutes. In some embodiments, the primary particle growth test comprises heating an aqueous solution comprising the protein composition having a protein content of 10%, 15%, or 18% (w / w) to 120°C for 4 minutes. In some embodiments, the primary particle growth test comprises heating an aqueous solution comprising the protein composition having a protein content of 10% or 15% (w / w) to 140°C for 2 minutes. 1.5 Measurement of thermal clotting time (HCT)

[0424] The thermal coagulation time (HCT) can be determined by the following method.

[0425] A 10% protein (w / w) solution is prepared from the powdered protein composition using deionized water at room temperature (approximately 20°C) on a magnetic stirrer. The powder is gradually added to the water under vortex stirring conditions, taking care to avoid foaming. Once the powder is completely dispersed, hydration is continued for 30 minutes with continuous stirring. The solution is homogenized at 150 / 50 bar. 1 ml of the protein solution (pH 6.8) is placed in a glass vial, clipped to a platform, and placed in a thermostatically controlled silicone oil bath at 140°C for a secondary heating treatment at a gentle rocking speed.

[0426] The thermal coagulation time is defined as the time (minutes) elapsed from the time the container is placed in the oil bath until the onset of visible aggregate formation (Singh H & Creamer LK (1992), Determination of heat stability, In: Advanced Dairy Chemistry ed. Fox PF Elsevier). 1.6 Viscosity measurement

[0427] Unless otherwise specified, the viscosity of the composition was measured using a rheometer such as that manufactured by Anton Paar, using a cup and bob assembly at a shear rate of 100 s -1 Measured at 20° C. It will be appreciated that other methods of measuring or estimating viscosity are well known in the art and may be employed where appropriate. 2. Example 2

[0428] This example describes the preparation of a thermostable protein composition of the present invention. 2.1 Preparation of Protein Compositions

[0429] Cheese whey protein concentrate powder was used as the whey protein source, and sodium caseinate powder was used as the casein source. The composition of these ingredients is shown in Table 1. [Table 1]

[0430] The cheese whey protein concentrate and sodium caseinate component powders were reconstituted as detailed in Table 2 by adding the powders to water at 50°C and stirring slowly with an overhead stirrer for 60 minutes to produce an aqueous composition containing 30% total solids by weight.

[0431] The compositions were homogenized at 200 / 50 bar to fully hydrate the powder. Samples 3, 4, 5, and 6 were diluted to 10% or 20.15% total solids after homogenization, as shown in Table 2.

[0432] The pH was adjusted with NaOH, KOH or HCl to the final pH shown in Table 2.

[0433] The composition was then heated in a two-stage heater. The feed composition was preheated to 55°C in the first stage (tube length 28.84 m, tube inner diameter 4.5 mm), then heated to 85°C in the second stage (tube length 15.84 m, tube inner diameter 4.5 mm), and then held in a holding tube (capillary tubes were placed on both sides of the holding tube). The feed flow rate was 0.5 L / min, and the product residence time in the second stage at the target temperature of 85°C was 10-15 seconds. The maximum heater pressure was 42 bar.

[0434] Sample 1 is a composition of the present invention. Samples 2-6 are comparative compositions. Sample 2 lacks casein. [Table 2]

[0435] The properties of the protein composition are shown in Table 3 below. [Table 3]

[0436] 1 Measurement according to Example 1.1.

[0437] 2 Measurement according to Example 1.3.

[0438] 3 Measurement according to Example 1.2. 2.2 Preparation of dry powder

[0439] The protein composition of Sample 4 was further concentrated by evaporation in preparation for subsequent spray drying (a dispersion with 10% total solids is too inefficient during drying and must be concentrated before spray drying). Sample 4 formed mostly submicron particles, but upon evaporation and concentration, the viscosity of the evaporated product increased significantly, and fouling within the evaporator was observed. The product obtained after evaporation only reached 19% total solids. The viscosity of the evaporated product is shown in Figure 1. The product with 10% total solids had a low viscosity (Newtonian flow) after initial heating, but increased significantly after evaporation.

[0440] The evaporated product was spray-dried to a powder and reconstituted with 10% water as the phase solids. As shown in Figure 2, a significant change in particle size distribution was observed, indicating that particle dispersions prepared with casein at low solids (and therefore low whey protein concentrations) are not stable to further processing, such as evaporation or secondary heat treatment.

[0441] Because evaporation caused undesirable changes to the particle properties of Sample 4, all heated solutions were not concentrated but instead freeze-dried to powder. 2.3 Heat Stability Test of Liquid Compositions Containing Protein Compositions

[0442] The lyophilized powder was reconstituted in water and the pH adjusted to 6.8 to obtain a 14% w / w liquid composition. A 5 ml sample was subjected to the primary particle growth test described in Section 1.4 of Example 1. Briefly, the solution was placed in a glass vial and immersed in an oil bath set at 120°C for 10 minutes for secondary heating. The vial was then removed and immediately cooled to room temperature in ice water. The particle size and viscosity of the heated product were measured. The results are shown in Table 4. The particle size distribution of Sample 1 before and after heating is shown in Figure 3. The particle size distribution of Sample 1 was essentially unimodal and showed no substantial change after secondary heating. [Table 4]

[0443] Various β-lactoglobulin:casein ratios were also tested. A 1.2:1 ratio containing 9 g / 100 g of β-lactoglobulin resulted in a composition that was too concentrated for further processing. A 1.6:1 ratio containing 9 g / 100 g of β-lactoglobulin resulted in a composition with properties similar to Sample 1. 3. Example 3

[0444] This example illustrates the use of a thermostable protein composition of the present invention in a heat-treated high-protein liquid composition. 3.1 Preparation of thermostable protein compositions.

[0445] A thermostable protein composition was prepared as described for Sample 1 in Section 2.1, except that 32% w / w total solids was used. The composition at the time of the primary heat treatment is shown in Table 5. [Table 5]

[0446] A description of the properties of the thermostable protein compositions is provided in Table 6 below. [Table 6]

[0447] 1 Measurement according to Example 1.1.

[0448] 2 Measurement according to Example 1.3.

[0449] 3 Measurement according to Example 1.2. 3.2 Preparation of high protein liquid compositions.

[0450] The thermostable protein composition (Sample 7) was diluted with water to obtain liquid compositions with final protein contents of 10, 15, and 18% (w / w). The final pH was adjusted to 6.82 ± 0.02 using a mixture of NaOH / KOH and subjected to primary particle growth testing as described in Section 1.4 of Example 1. Briefly, 5 mL samples of the liquid compositions were placed in glass vials and subjected to secondary heat treatment by immersion in an oil bath at 90°C for 10 minutes, 120°C for 4 minutes, or 140°C for 2 minutes. The solutions were removed from the oil bath immediately after the designated time and cooled in ice water. Viscosity and particle size distribution were measured.

[0451] The protein content, heat treatment conditions, and properties of the high-protein liquid composition are shown in Table 7. [Table 7]

[0452] 1 Measurement according to Example 1.3. 4. Example 4

[0453] This example describes the preparation of a heat-stable protein composition of the present invention using a mixture of cheese whey retentate and mineral acid whey protein concentrate, and the use of the heat-stable protein composition in a heat-treated high-protein liquid nutritional composition. 4.1 Preparation of Thermostable Protein Compositions

[0454] The thermostable protein composition was prepared according to the steps shown in Figure 4, as summarized below.

[0455] The heat-stable protein composition was prepared by first combining cheese whey retentate (60% w / w of the protein in the final composition), mineral acid whey retentate (24% w / w of the protein in the final composition), and recombined sodium caseinate solution (16% w / w of the protein in the final composition) to a total solids concentration of approximately 17% (w / w). Table 8 details the protein composition of the mineral acid whey protein concentrate. [Table 8]

[0456] *Percentage of total protein.

[0457] The pH was adjusted to 6.30. The mixture was evaporated to a total solids content of 28% and subjected to the following primary heating process: The protein composition was preheated to 55°C using a heat exchanger heated with hot water. The protein composition was then denatured using the method described in WO2010120199. The protein composition was fed in series into two identical single-tube, high-pressure steam-heated shell-and-tube heat exchangers using a high-pressure pump with a delivery pressure of 250-350 bar at a flow rate high enough to achieve a Reynolds number of ≥ 2100. The concentrate was discharged from the first high-pressure heater at 71-73°C and from the second high-pressure heater at 80-85°C.

[0458] A summary of the protein composition during the primary heat treatment is shown in Table 9. [Table 9]

[0459] The thermostable protein composition exiting the second heater passed through a holding tube and piping before reaching the nozzle bank at the top of the spray dryer, which had the same tubing configuration as the high-pressure heater. This ensured turbulent flow. The length of the holding tube, when used in conjunction with the piping leading to the nozzle bank, was selected to provide an additional 20 seconds of residence time for the heated stream before spray drying. As a result, the temperature loss throughout the piping from the outlet of the second high-pressure heater to the nozzle bank was less than 1°C. Therefore, no additional mechanical shearing steps were used during heating, after the heater-reactor system, and before spray drying.

[0460] In the spray dryer, the thermostable protein composition was fed into a bank containing three nozzles and atomized into a droplet spray at pressures above 160 bar. An inlet hot air temperature of ~200°C and an outlet chamber temperature of 70-80°C were used. This powder was further dried and cooled in a vibrating fluidized bed, after which the material was sieved and packed to produce a powder with less than 5% moisture. The particle types and particle sizes in the powder are shown in Table 10. [Table 10]

[0461] 1 Measurement according to Example 1.1.

[0462] 2 Measurement according to Example 1.3.

[0463] 3 Measurement according to Example 1.2. 4.2 Heat coagulation time (HCT)

[0464] The HCT of a 10% (w / w) protein solution prepared from powdered heat-stable protein was measured according to section 1.5 of Example 1 using a secondary heat treatment temperature of 140° C. The HCT was determined to be 3 minutes 22 seconds. 4.3 Preparation of high-protein drinks

[0465] This heat-stable protein composition was used to prepare a liquid nutritional composition in the form of a heat-treated high-protein beverage formula with a final protein content of 14% (w / w). The beverage was prepared according to the formula in Table 11. The beverage was prepared according to the process in Figure 5, described below. The final composition is shown in Table 12. [Table 11] [Table 12]

[0466] The steps are as follows: 1. All dry ingredients, i.e., sodium chloride, tripotassium citrate monohydrate, gellan gum, cellulose gum, lecithin, were pre-weighed and pre-blended.

[0467] 2. The reverse osmosis water required for each batch was weighed into a stainless steel container and heated to 55°C in a water bath using an overhead stirrer.

[0468] 3. 80% of the antifoam was added to the water.

[0469] 4. The heat stable protein composition was added to the water.

[0470] 5. The dry blended powders were slowly added to the stainless steel container over a period of 15 minutes, stirring to create a vortex.

[0471] 6. Oil was added to each container.

[0472] 7. The stirring speed was adjusted to give good mixing but not to create a vortex.

[0473] 8. The mixture was stirred at 55°C ± 2°C for 60 minutes.

[0474] 9. The mixture was sheared in an ultraturrex at 7,000 rpm for 3 minutes.

[0475] 10. If necessary, water was added to the drink to bring the final weight to 20 kg.

[0476] 11. The remaining 20% ​​of the antifoam was added.

[0477] 12. Check product weight and adjust with water if necessary.

[0478] 13. The mixture was heated to 55°C ± 1°C and passed once through a two-stage homogenizer at 200 / 50 bar (total 250 bar).

[0479] 14. The product was cooled to approximately 20°C ± 5°C by placing the container in ice water for pH measurement.

[0480] 15. The pH was adjusted to 6.8 ± 0.1 with 5% KOH as needed.

[0481] 16. The mixture was UHT treated at approximately 146°C for 6.5 seconds and then aseptically packaged. The preheat temperature was 85°C, achieved using a plate heat exchanger. The product was held at that temperature for 30 seconds. The final heat treatment temperature was then increased to 146°C using direct steam injection (direct heat treatment). The first stage was cooled to 80°C using a flash vessel, and the final stage was cooled to approximately 20-25°C using a plate heat exchanger.

[0482] 17. The product was immediately filled into 200 mL PET bottles at a temperature of about 24-25°C and capped.

[0483] 18. The product was then stored at approximately 25°C.

[0484] Secondary heat treatment (UHT treatment) of the high-protein beverage was also successful, and no fouling was observed, indicating that the protein composition in the heat-treated high-protein beverage was stable. After UHT treatment, the viscosity of the beverage was 100 s -1 The viscosity was 28 mPa·s. The beverage had a "milky" flavor and a smooth texture without any grittiness or powderiness, and the organoleptic properties were also satisfactory. 5. Example 5

[0485] This example describes how to prepare an exemplary high protein drinking yogurt using Sample 8 powder according to the method of the present invention.

[0486] The thermostable protein composition (Sample 8) and skim milk powder (SMP) were reconstituted with water at 10°C to produce an aqueous composition containing 15% protein by weight, 12% protein from the thermostable protein composition. 9.2% (w / w) SMP was reconstituted and 15% (w / w) of the thermostable protein composition was added. The balance to 100% was achieved by adding water. The mixture was stirred for 60 minutes. The reconstituted mixture was preheated to 60°C and homogenized at 150 / 50 bar. The mixture was heat-treated at 95°C for 6 minutes and 40 seconds. The mixture was cooled to 43°C and fermented with 0.02% (w / w) bacterial culture. The inoculated mixture was fermented overnight at 43°C to achieve a pH below 4.6. The yogurt was cooled to 20°C and smoothed. The yogurt was filled, cooled to below 4°C, and stored at 4°C for further testing.

[0487] High-protein drinking yogurt was successfully produced. The viscosity of the high-protein drinking yogurt reached 50 s after 7 days of production. -1 The viscosity was 181 mPa·s. The volume fraction of protein particles with a particle size of 0.1 to 1 μm was 41.5%, the volume fraction of protein particles with a particle size of 1 to 5 μm was 49.5%, and the volume fraction of protein particles with a particle size of >5 μm was 9%. Drinking yogurt produced using the composition of the present invention showed significantly less sedimentation than high-protein drinking yogurt produced using conventional microparticulated whey by the same method and protein content. 6. Example 6

[0488] This example describes the preparation of a thermostable protein composition using a peroxidase enzyme and an oxidizing agent. Two methods (Method A and Method B) for producing a thermostable protein composition and a comparative method (Method C) for producing a casein-free whey protein composition are described. 6.1 Stock solution

[0489] A whey protein concentrate stock solution of 31.8% (w / w) total solids is prepared by reconstituting uncolored dairy cheese whey protein concentrate (total solids: 96.00%, total protein: 79.5%, β-lactoglobulin: 41.7%) with water at 50°C and holding for 0.5 hours.

[0490] A sodium caseinate stock solution of 11.4% (w / w) total solids is prepared by dissolving sodium caseinate powder (containing 93% w / w protein) in water at 55°C and holding for 1 hour. 6.2 Method A

[0491] Method A describes the preparation of a heat stable protein composition by combining whey protein concentrate and sodium caseinate, treating with peroxidase and peroxide, and heat treating: 1. Combine whey protein concentrate and sodium caseinate stock solutions in appropriate ratios to form a composite solution with a β-lactoglobulin to casein weight ratio of 2.7:1.

[0492] 2. Dilute the solution with water to a total protein content of 21.5g / 100g, whey protein content of 18.0g / 100g, and β-lactoglobulin of 9.46g / 100g.

[0493] 3. The solution is homogenized at 200 / 50 bar.

[0494] 4. Adjust the pH to 6.2 using a 50:50 blend of 1M KOH and NaOH.

[0495] 5. Divide the protein solution into two samples, Sample 1 and Sample 2. Sample 1 is treated with peroxidase enzyme and an oxidizing agent as in steps a-c below, and Sample 2 is not treated with peroxidase enzyme and an oxidizing agent.

[0496] a. Add peroxidase enzyme from Aspergillus niger (MaxiBright®) to Sample 1 according to the manufacturer's instructions and mix at 150 rpm at 4°C.

[0497] b. Hydrogen peroxide is added slowly to Sample 1 to achieve a concentration of 75 ppm (equivalent to 0.375 mol H2O2 / mol β-lactoglobulin) to avoid enzyme inhibition.

[0498] c. Sample 1 is tested using peroxide indicator test strips (Merck MQuant® Peroxide Test Strips, responds to 0.5-25 mg / L H2O2) to ensure there is no residual peroxide.

[0499] 6. Both samples are preheated to 55°C in a heat exchanger with a scraped surface.

[0500] 7. Both samples are then heated to 85°C in a steam-heated tubular heat exchanger at a flow rate of 0.5 L / min.

[0501] 8. Both samples are held in holding tubes at 85°C for 10-15 seconds.

[0502] 9. Cool and retrieve the sample. 6.3 Method B

[0503] Method B describes the preparation of a heat-stable protein composition by treating whey protein concentrate with peroxidase and peroxide, combining with sodium caseinate, and heat treating: 1. Use whey protein concentrate concentrate directly with a total solid content of 31.8% (w / w) and a protein content of 26.3 / 100g.

[0504] 2. The solution is homogenized at 200 / 50 bar.

[0505] 3. Adjust the pH to 6.2 using a 50:50 blend of 1M KOH and NaOH.

[0506] 4. Add peroxidase enzyme from Aspergillus niger (MaxiBright®) to the solution according to the manufacturer's instructions and mix at 150 rpm at 4°C.

[0507] 5. Hydrogen peroxide is added slowly to a concentration of 75 ppm (equivalent to 0.375 mol H2O2 / mol β-lactoglobulin protein) to avoid enzyme inhibition.

[0508] 6. Test the solution using peroxide indicator test strips (Merck MQuant® Peroxide Test Strips, responds to 0.5-25 mg / L H2O2) to ensure there is no residual peroxide.

[0509] 7. Combine the whey solution with the sodium caseinate stock solution in an appropriate ratio to form a composite solution with a total protein content of 21.5g / 100g, a whey protein content of 18.0g / 100g, a β-lactoglobulin content of 9.46g / 100g, and a weight ratio of β-lactoglobulin and casein of 2.7:1.

[0510] 8. Heat treat the composite solution as in steps 6-9 of Method A to produce a thermostable protein composition (Sample 3). 6.4 Method C

[0511] Method C describes the preparation of a comparative sample containing no casein: 1. A whey protein concentrate stock solution with a total solids content of 31.8% (w / w) is diluted with water to a total protein content of 21.5g / 100g.

[0512] 2. Treat the whey protein solution as described in steps 3-9 of Method A to produce a composition with peroxidase enzyme and oxidizing agent treatment (Sample 4) and a composition without peroxidase enzyme and oxidizing agent treatment (Sample 5).

[0513] Methods A to C are repeated, with the weight ratio of β-lactoglobulin and casein set to 5:1, to produce samples 6 to 10. 6.5 Sample characteristics

[0514] Samples 1-5 are evaluated for whey protein denaturation, covalent aggregation of β-lactoglobulin, particle size distribution, HCT, and viscosity as described in Example 1.

[0515] Samples 1-3 and Samples 6-8 comprise denaturable whey protein, wherein at least about 65% (w / w) of the denaturable whey protein is denatured and at least about 40% (w / w) of the total β-lactoglobulin in Samples 1-3 is covalently aggregated. Additionally, Samples 1-3 and Samples 6-8 comprise protein particles, wherein at least about 40% by volume of the protein particles have a particle size of less than 1 μm, and at least a portion of the protein particles comprise co-aggregates of denatured whey protein and casein.

[0516] An aqueous composition containing a sufficient amount of sample to give a total protein content of 15% w / w is prepared and subjected to a second heat treatment at 120°C for 4 minutes.

[0517] After the second heat treatment of the aqueous composition containing Samples 1-3 and 6-8, at least about 40% by volume of the protein particles in the aqueous composition have a particle size of 0.1-1 μm, less than about 60% by volume of the protein particles in the aqueous composition have a particle size of 1-5 μm, less than about 10% by volume of the protein particles in the aqueous composition have a particle size of at least 5 μm, the D[4,3] of the protein particles in the aqueous composition is less than about 5 μm, and / or the viscosity of the aqueous composition is less than 100 s -1 The HCT at 140°C of aqueous compositions containing Samples 1-3 and Samples 6-8 in amounts sufficient to provide a total protein content of 10% w / w is less than about 100 mPa·s. The aqueous compositions containing Samples 1-3 and Samples 6-8 show no visible signs of gelation. The HCT at 140°C of aqueous compositions containing Samples 1-3 and Samples 6-8 in amounts sufficient to provide a total protein content of 10% w / w is at least about 60 seconds.

[0518] Samples 5 and 10 do not have the high thermal stability seen in Samples 1-3 and 6-8, ie, resistance to increases in particle size and viscosity due to secondary heat treatment.

[0519] All documents cited herein, including but not limited to patents, patent applications, journals, books, and the like, are incorporated herein by reference in their entirety. The section headings used herein are for organizational purposes only and should not be construed as limiting the subject matter described.

[0520] Although the present invention has been described by way of example and with reference to specific embodiments, it will be understood that modifications and / or improvements may be made without departing from the scope or spirit of the invention. 7. Example 7

[0521] Thermostable protein compositions and comparative samples were prepared as described in Example 6. Samples 1 and 2 were prepared by Method A described in Section 6.2, Sample 3 was prepared by Method B described in Section 6.3, and Comparative Samples 4 and 5 were prepared by Method C described in Section 6.4.

[0522] The sample overview is shown in Table 12, and its properties are shown in Table 13. [Table 13] [Table 14]

[0523] a Measurement according to Example 1.1.

[0524] b Measurement according to Example 1.2.

[0525] c Measurement according to Example 1.3.

[0526] d Thermal coagulation time (seconds) of a 10% protein solution at 140°C. [Industrial Applicability]

[0527] The thermostable protein compositions described herein are useful for producing nutritional compositions with small particle size (e.g., less than 1 μm) and good thermostability, which are resistant to gelation, sedimentation, aggregation, and increases in particle size and viscosity when subjected to a secondary heat treatment such as UHT.

Claims

1. 1. A heat-stable protein composition comprising whey protein and casein, the composition comprises β-lactoglobulin and casein in a weight ratio of about 1.6:1 to about 5:1; the whey protein comprises at least about 65% (w / w) denatured denaturable whey protein; at least about 40% (w / w) of the total β-lactoglobulin in said composition is covalently aggregated; the composition comprises protein particles; At least about 40% by volume of the protein particles have a particle size of less than 1 μm; and The heat-stable protein composition, wherein at least a portion of the protein particles comprise co-aggregates of denatured whey protein and casein.

2. 10. The thermostable protein composition of claim 1, wherein the composition comprises non-micellar casein.

3. a. at least about 40% by volume of the protein particles have a particle size between 0.1 and 1 μm; b. less than about 55% by volume of the protein particles have a particle size between 1 and 5 μm; c. less than about 5% by volume of the protein particles have a particle size of at least 5 μm; or d. The thermostable protein composition of claim 1 or 2, comprising a combination of two or more of (a) to (c).

4. 4. A thermostable protein composition according to any one of claims 1 to 3, wherein at least about 50% by weight of the total β-lactoglobulin is covalently aggregated, preferably at least about 60% by weight of the total β-lactoglobulin is covalently aggregated.

5. a. β-lactoglobulin and casein in a weight ratio of about 1.8:1 to about 5:1, preferably about 1.8:1 to about 4:1, more preferably about 2:1 to about 4:1, and most preferably about 2:1 to about 3.5:1; and / or b. The thermostable protein composition of any one of claims 1 to 4, comprising a weight ratio of total whey protein to casein of at least about 3:1, preferably from about 3:1 to about 10:1, more preferably from about 4:1 to about 8:1, and most preferably from about 4.5:1 to about 6:

1.

6. a. at least about 70% w / w, preferably about 80% to about 90% w / w, of the total protein in the composition is whey protein; b. at least about 35% w / w, preferably about 40% to about 60% w / w, of the total protein in the composition is β-lactoglobulin; c. about 10% to about 20% w / w of the total protein in the composition is casein, or d. The thermostable protein composition of any one of claims 1 to 5, comprising a combination of two or more of (a) to (c).

7. an aqueous composition comprising a thermostable protein composition in an amount sufficient to provide a total protein content of 15% w / w, the aqueous composition having been subjected to a second heat treatment at 120°C for 4 minutes; a. at least about 40% by volume, preferably at least about 45% by volume, and more preferably at least about 50% by volume, of the protein particles in the aqueous composition have a particle size between 0.1 and 1 μm; b. less than about 60% by volume, preferably less than about 55% by volume, and more preferably less than about 50% by volume, of the protein particles in the aqueous composition have a particle size between 1 and 5 μm; c. at least about 10% by volume of the protein particles in the aqueous composition have a particle size of 0.1 to 1 μm, preferably at least about 5% by volume, and more preferably at least about 2% by volume; d. the D[4,3] of the protein particles in the aqueous composition is less than about 5 μm, preferably less than about 4, more preferably less than about 3, even more preferably less than about 2, and most preferably less than about 1 μm; e. The viscosity of the aqueous composition is 100 s －１ less than about 100 mPa·s, preferably less than 100 s －１ less than about 40 mPa·s, more preferably less than 100 s －１ less than about 30 mPa·s, and even more preferably less than 100 s －１ is less than about 20 mPa s; or f. Any combination of any two or more of (a) to (e).

8. 1. A method for preparing a thermostable protein composition, comprising: a. an aqueous composition having a pH of 5.5 to 6.8, i. a total whey protein content of at least about 18 g / 100 g aqueous composition; ii. a β-lactoglobulin content of at least about 9 g / 100 g aqueous composition; iii. a total protein content of at least about 20 g / 100 g aqueous composition; iv. providing the aqueous composition comprising β-lactoglobulin and casein in a weight ratio of about 1.6:1 to about 5:1; and b. heat-treating the aqueous composition to at least about 70°C for a time sufficient to cause protein denaturation, wherein the heat-treating comprises heating the aqueous composition under high shear stress to provide the thermostable protein composition.

9. 10. The method of claim 8, comprising contacting a whey protein source with a casein protein source to provide the aqueous composition of step a).

10. 10. The method of claim 9, wherein the whey protein source comprises or consists of whey protein concentrate (WPC), whey protein isolate (WPI), or a combination thereof.

11. 11. The method of claim 9 or 10, wherein the casein protein source comprises or consists of caseinate, calcium-depleted milk protein concentrate (MPC), total milk protein (TMP), or a combination of any two or more thereof.

12. 12. The method of any one of claims 9 to 11, further comprising, preferably prior to step a), contacting the whey protein source with an oxidizing agent in combination with a catalyst, such as an enzymatic or chemical catalyst, preferably a peroxidase enzyme.

13. 12. The method of any one of claims 8 to 11, further comprising, preferably prior to step b), contacting the aqueous composition with an oxidizing agent in combination with a catalyst, such as an enzymatic or chemical catalyst, preferably a peroxidase enzyme.

14. 14. The method of claim 12 or 13, wherein the oxidizing agent is hydrogen peroxide or benzoyl peroxide and / or the catalyst is a peroxidase enzyme.

15. 15. A method according to any one of claims 12 to 14, wherein the oxidizing agent is present in an amount of less than 300 ppm or less than 200 ppm, or the molar ratio of oxidizing agent to β-lactoglobulin is less than 2, preferably less than 1, or from about 0.1 to about 0.

85.

16. 15. The method of any one of claims 12 to 14, wherein the whey protein source and / or the casein protein source are in powder form, and the method comprises reformulating the powder(s) to provide the aqueous composition.

17. 17. The method according to any one of claims 8 to 16, further comprising homogenising the aqueous composition, preferably at about 200 / 50 bar and preferably before step b).

18. The method of any one of claims 8 to 17, further comprising adjusting the pH of the aqueous composition to a pH of 5.5 to 6.

8.

19. The method of any one of claims 8 to 18, further comprising concentrating the aqueous composition, preferably by evaporation, before step b).

20. 20. The method of any one of claims 8 to 19, wherein the whey protein comprises a denaturable whey protein, and step b) comprises heat-treating the aqueous composition for a time sufficient to denature at least about 65% (w / w) of the denaturable whey protein in the aqueous composition.

21. 21. The method according to any one of claims 8 to 20, wherein less than about 10% (w / w), preferably less than about 5% (w / w) of the total denaturable whey protein in the aqueous composition prior to step b) is denatured.

22. 22. The method of any one of claims 8 to 21, wherein the thermostable protein composition comprises protein particles, and at least about 40% by volume of the protein particles have a particle size of less than 1 μm.

23. 23. The method of any one of claims 8 to 22, wherein step b) comprises heat treating the solution to a temperature of from about 70°C to about 150°C.

24. 24. The method of any one of claims 8 to 23, wherein the thermostable protein composition comprises at least about 70% w / w whey protein based on the total protein in the composition, and less than about 30% w / w casein based on the total protein in the composition.

25. 25. The method of any one of claims 8 to 24, further comprising drying the thermostable protein composition.

26. 26. The method of any one of claims 25, wherein the thermostable protein composition is not subjected to a mechanical shearing step prior to drying other than to convert the liquid into droplets to facilitate drying.

27. Step b) is i. under turbulent flow conditions having a Reynolds number of at least about 2000; ii. at least about 1000 s －１ under high wall shear conditions with a wall shear rate of iii. The method of any one of claims 8 to 26, comprising heating the solution under mechanical shear conditions, preferably under mechanical shear conditions generated by a homogenizer, a colloid mill, a high pressure pump, a scraped surface heat exchanger, an ultrasonic generator, a microfluidizer, and / or a high shear mixer.

28. A method according to any one of claims 8 to 27, producing a thermostable protein composition according to any one of claims 1 to 7.

29. A thermostable protein composition produced by the method of any one of claims 8 to 28.

30. 30. A nutritional composition comprising the thermostable protein composition of any one of claims 1-7 or 29.

31. 31. The nutritional composition of claim 30, comprising at least about 6%, at least about 8%, at least about 10%, at least about 12%, at least about 14%, at least about 16%, at least about 18%, or at least about 20% (w / w) total protein on a dry basis.

32. 32. The nutritional composition of claim 30 or 31, comprising at least about 6%, at least about 8%, at least about 10%, at least about 12%, at least about 14%, at least about 16%, at least about 18%, or at least about 20% (w / w) denatured whey protein on a dry basis.

33. 33. The nutritional composition of any one of claims 30 to 32, wherein at least about 50% (w / w), or at least about 60% (w / w) of the total protein in the nutritional composition is denatured whey protein.

34. 34. The nutritional composition of any one of claims 30 to 33, wherein at least about 50% (w / w) of the total protein in the nutritional composition is provided by the thermostable protein composition.

35. 35. The nutritional composition of any one of claims 30 to 34, wherein at least about 40% by volume, preferably at least about 45% by volume, or at least about 50% by volume, of the protein particles in the nutritional composition have a particle size of 0.1 to 1 μm.

36. 36. A nutritional composition according to any one of claims 30 to 35, wherein less than about 60%, preferably less than about 55%, or less than about 50% by volume of the protein particles in the nutritional composition have a particle size of 1 to 5 μm.

37. 37. A nutritional composition according to any one of claims 30 to 36, wherein less than about 10% by volume of the protein particles in the nutritional composition have a particle size of at least 5 μm, preferably less than about 5% or less than about 1%.

38. A nutritional composition according to any one of claims 30 to 37, which has been subjected to a secondary heat treatment, preferably sterilisation and / or pasteurisation.

39. The nutritional composition according to any one of claims 30 to 38, which is a food product.

40. 40. The nutritional composition of claim 39, wherein the food product is a baked good, a bar, a set or stirred yogurt, a set gel, or a semi-solid food product.

41. 41. The nutritional composition of claim 39 or 40, wherein the food product is set or stirred yogurt, optionally comprising from about 6% to about 20% (w / v) total protein.

42. 42. The nutritional composition of claim 41, wherein the set yogurt or the stirred yogurt exhibits a reduced volume weighted mean particle size compared to a control yogurt product having the same ingredient composition and the same total protein content, except that the control yogurt product does not comprise the heat-stable protein composition of any one of claims 1 to 7 or 29.

43. 41. The nutritional composition of claim 39 or 40, wherein the food product is a heat-treated, high-protein set gel, optionally comprising at least about 10% (w / v) or at least about 15% (w / v) total protein.

44. 41. The nutritional composition of claim 39 or 40, wherein the food product is a heat-processed, high-protein, semi-solid food product, optionally comprising at least about 10% (w / v) or at least about 15% (w / v) total protein.

45. The food was stored at 20°C for 50 seconds. －１ 45. The nutritional composition of claim 43 or 44, having a viscosity measured at 1000 mPa.s, less than about 800 mPa.s, less than about 600 mPa.s, or less than about 400 mPa.s.

46. A nutritional composition according to any one of claims 30 to 38, which is a liquid composition, preferably a liquid nutritional composition.

47. 100 s at 20°C －１ 47. The nutritional composition of claim 46, having a viscosity measured at 1000 kJ / min, of less than about 400 mPa.s, preferably less than about 200 mPa.s.

48. 48. The nutritional composition of claim 46 or 47, wherein the liquid composition exhibits less than about 10% sedimentation after storage at a temperature of about 20°C to about 25°C for at least 6 weeks, and preferably less than about 10% sedimentation after storage at a temperature of about 20°C to about 25°C for at least 3 months.

49. The liquid composition a. about 0.1 to about 30% (w / v) lipid; b. about 0.1 to about 40% (w / v) carbohydrate, preferably about 0.1 to about 30% (w / v) carbohydrate; c. at least one monovalent cation; d. optionally present in an amount of at least about 30 mg / 100 mL, at least about 50 mg / 100 mL, or at least about 100 mg / 100 mL, preferably Ca ２＋ at least one divalent metal cation, e. Any combination of any two or more of (a) to (d).

50. 50. The nutritional composition of any one of claims 46 to 49, wherein the liquid composition has an energy density of at least about 0.5 kcal / mL, preferably at least about 1.0, at least about 1.5, or at least about 2.0 kcal / mL.

51. 51. The nutritional composition of any one of claims 46 to 50, wherein the liquid composition is a heat-treated, shelf-stable liquid nutritional composition.

52. 52. The nutritional composition of any one of claims 46 to 51, wherein the liquid composition is a high-protein drink or a medical food.

53. The liquid composition is drinking yogurt, and is preferably simmered at 20°C for 50 seconds. －１ 52. The nutritional composition of any one of claims 46 to 51, having a viscosity measured by a sieve of less than about 400 mPa.s, less than about 300 mPa.s, less than about 200 mPa.s, or less than about 100 mPa.s.

54. A nutritional composition according to any one of claims 46 to 51, wherein the liquid composition is an acidic beverage, preferably having a pH of from about 2 to about 4.

8.

55. 52. The composition of any one of claims 46 to 51, wherein the liquid composition is a neutral beverage, preferably having a pH of about 6.5 to about 7.

5.

56. The liquid composition is －１ 56. The nutritional composition of claim 54 or 55, wherein the viscosity measured at 1000 kJ / min is less than about 400 mPa.s, less than about 300 mPa.s, less than about 200 mPa.s, or less than about 100 mPa.s.

57. 30. Use of a thermostable protein composition according to any one of claims 1 to 7 or 29 in the preparation of a nutritional composition.

58. 58. The use according to claim 57, wherein the nutritional composition is a nutritional composition according to any one of claims 30 to 56.

59. 57. A method for providing nutrition to a subject in need thereof, comprising administering to the subject a nutritional composition according to any one of claims 30 to 56.

60. 1. A method for preparing a nutritional composition, comprising: a. the thermostable protein composition of any one of claims 1-7 or 29, and b. one or more additional ingredients.

61. 1. A method for preparing a liquid composition, comprising: a. the thermostable protein composition of any one of claims 1-7 or 29, and b. one or more additional components.

62. said one or more additional ingredients being: a. one or more lipids, preferably one or more vegetable lipids and / or one or more dairy lipids; b. one or more carbohydrates, preferably one or more monosaccharides, disaccharides, oligosaccharides, polysaccharides, or a combination of any two or more thereof; c. one or more additional protein sources, preferably derived from milk, whey, casein, caseinate, egg, egg white, egg yolk, vegetables, plants, alfalfa, clover, peas, beans, kidney beans, soybeans, navy beans, lupin, cocoa, hornbeams, tree nuts, peanuts, rye, grains, whole grains, rice, hemp, wheat gluten, fungal or algae protein, a protein concentrate thereof, a protein isolate thereof, a hydrolysate thereof, or a combination of any two or more thereof; d. one or more vitamins, preferably vitamin C, vitamin A, vitamin E, vitamin B12, vitamin K, riboflavin, niacin, vitamin D, vitamin B6, folic acid, pyridoxine, thiamine, pantothenic acid, biotin, or any salt, derivative, or metabolite thereof, or a combination of any two or more thereof; e. one or more minerals, preferably chloride, sodium, calcium, iron, chromium, copper, iodine, zinc, magnesium, phosphorus, or potassium, or a combination of any two or more thereof; f. one or more food additives, preferably comprising one or more emulsifiers, which are lecithin, monoglycerides, diglycerides, polyglycerol esters, milk phospholipids, citrate esters (CITREM), polysorbate 60, glyceryl monostearate, DATEM, or a combination of any two or more thereof; g. one or more food additives, preferably comprising one or more stabilizers selected from carrageenan, gellan gum, pectin, guar gum, locust bean gum, carboxymethylcellulose, alginate, agar, oat gum, tragacanth gum, acacia gum, xanthan gum, karaya gum, tara gum, starch, modified starch and microcrystalline cellulose, gelatin, or combinations thereof; (f) any combination of two or more of (a) to (g).

63. A method according to any one of claims 60 to 62, wherein the nutritional composition or the liquid composition is a nutritional composition according to any one of claims 30 to 56.

Images