Dairy Products and Processes

Abstract

The present disclosure relates to denatured whey protein compositions comprising sub-micrometer particles having a volume weighted mean diameter D(4,3) of 1 μm or less and a substantially unimodal distribution, with at least 95% by volume of the particles being 2 μm or less. The present disclosure also relates to high protein beverages, food products, and nutritional compositions comprising such denatured whey protein compositions, as well as methods for producing such denatured whey protein compositions.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 382,595, filed November 7, 2022, and U.S. Provisional Application No. 63 / 363,914, filed April 29, 2022, the entire contents of which are incorporated herein by reference.

[0002] The present invention relates to denatured whey protein compositions comprising heat stable sub-micrometer particles, methods for preparing such compositions, and the use of such compositions in food products including neutral ready-to-drink high protein beverages, high protein drinking yoghurts, and high protein snacks, protein or nutritional bars, and set or stirred yoghurts. [Background technology]

[0003] High protein food products and beverages can be produced using ingredients with high protein content. Desirable properties of ingredients with high protein content include heat stability to allow subsequent heat treatments such as retort and ultra-high temperature (UHT) treatment to ensure product safety and extended shelf life, and suspendability with minimal or substantially no sedimentation, especially in beverage applications.

[0004] Microparticulated whey protein concentrate (WPC) ingredients are known and used in the food product industry to increase the protein content of a variety of applications, such as ready-to-drink fermented beverages and food products such as bars and set or stirred yogurts.

[0005] Whey protein micronization is an advanced technology for producing whey protein particles. The size of the whey protein particles is important in achieving the desired mouthfeel. Particles between 0.1 and 3 μm give a creamy mouthfeel, whereas particles above 3 μm give a powdery or even gritty feel, and particles with a size of 0.1 μm give a watery mouthfeel. In fact, particle sizes in the range below 0.1 μm are known to give an oily taste that is unpleasant if perceived as the main tactile characteristic. Furthermore, it may be desirable to keep the amount of whey protein particles with a size below 0.5 μm to a minimum, since these smaller particles may impart an undesirable high viscosity to products containing them.

[0006] The main mechanism of action for the formation of whey protein particles involves two steps: first, whey proteins unfold during heating, and second, the unfolded protein molecules aggregate mainly through disulfide bonds and hydrophobic interactions. The processing conditions, such as temperature, heating time, pH, and shear stress, determine the reaction kinetics and the physical and chemical properties of the particles.

[0007] Conventional micronization processes incorporate shear or turbulent flow to limit the size of whey protein particles. However, when high whey protein concentrations are used to produce protein particles, micronization / aggregation reactions occur very rapidly, likely due to high molecular (protein) density and high collision efficiency. Although these techniques are effective in preventing the formation of very large micronization / aggregates, the average size of protein particles produced during conventional micronization processes ranges from 1 micrometer to 10 micrometers, with the majority of particles being larger than 1 micrometer in diameter. This particle size range still tends to cause precipitation and settling over the shelf life of the product in liquid applications, causing undesirable instability. Furthermore, it has not been technically achievable to reduce protein particle size while ensuring that all unfolded protein is formed into stable microparticles.

[0008] Singer (US 4,734,287) prepared a proteinaceous water-dispersible colloid comprising substantially non-aggregated particles of sweet whey protein coagulum having a mean diameter particle size distribution when dry ranging from greater than about 0.1 μm to less than about 2.0 μm, with less than about 2% of the total particles having a diameter greater than 3.0 μm. According to Singer, these particles are heat unstable and form fused aggregates when subjected to additional heat denaturation treatment.

[0009] McCarthy (US 5,350,590) sought to improve on the composition provided by Singer and further demonstrated the heat instability of a commercial product (designated SIMPLESSE 100D) made according to Singer. McCarthy prepared "loosely bound" whey aggregates containing whey protein and casein, and suggested that desolubilized casein substantially prevented the formation of covalent bonds. McCarthy noted that aggregates prepared without casein showed very low heat stability. As shown in Figure 7, particles prepared from whey protein alone were larger than 1 μm and even larger than 10 μm, with the peak of the particle distribution being larger than 1 μm and in some cases larger than 10 μm.

[0010] Similarly, Huss and Spiegel (US 6,767,575) prepared denatured whey protein aggregates with an average aggregate size (median) of 1-4 μm starting from a relatively dilute protein stream (<3% w / v protein). Again, the majority of particles were larger than 1 μm, with some particles having sizes larger than 2 μm and even larger than 5 μm, and the particle distribution by volume peaked at greater than 1 μm.

[0011] Villagran (US 6,605,311) utilized a similar conventional process to produce a whey protein composition having a total protein content of less than 60%, starting with a 20% by weight whey protein solution. According to Villagran, the resulting protein particles had a protein insolubility of about 80% and a mean diameter particle size distribution range of about 0.1 μm to about 3.0 μm, with less than about 5% of the total number of particles having a diameter greater than about 3.0 μm.

[0012] Arase (US20150181908) reported that the denatured protein product obtained by the Villagran method had an average particle size of 1.6 μm (defined by Arase as the particle size corresponding to 50% of the cumulative distribution of particle sizes) and lacked thermal stability. Indeed, Arase reported that the denatured protein product obtained by the Villagran method exhibited visually discernible gelation when heat treated in an autoclave at 120°C for 15 minutes. Arase sought to improve this product and utilized a raw material with a lower mass percent protein (12.5% ​​in the Arase example). The resulting product had an average particle size of 0.38-0.7 μm, a protein insolubility of 43-47%, and showed better stability than the product produced by the Villagran method. Nevertheless, the product produced by the Arase method showed thermal instability, with the average particle size increasing from 0.5 to 3.5 μm when a solution of 10 mass% protein was heated in an autoclave at 120°C for 15 minutes.

[0013] Havea (WO2010120199) provided a method for preparing a whey protein concentrate by providing an aqueous whey protein solution having a 15-50% (w / v) protein concentration at a pH of 4.7-8.5 and heat treating the solution to above 50° C. for a time sufficient to cause protein denaturation (the heat treatment comprising, for example, heating the solution under turbulent flow conditions with a Reynolds number of at least 500).

[0014] Gulla (WO2013065014) and Cakir-Fuller (WO2020104954) reported that one exemplary heat-denatured whey protein concentrate produced using the method described in Havea had a primary aggregate size D(4,3) of 1.70 μm in a 10% total solids protein solution and exhibited good heat stability as indicated by 1% aggregate growth after heating of a 10% total solids protein solution. Other exemplary heat-denatured whey protein concentrates produced using the method described in Havea had primary aggregate sizes D(4,3) ranging from 1.62 to 2.50 μm in a 10% total solids protein solution and exhibited varying degrees of heat stability.

[0015] Eriksen (WO2013117599) relates to the use of particulated whey protein in frozen confectionery products. Eriksen exemplified a particulated whey protein product in which at least about 10% of the particles are greater than 5 μm. For example, in FIG. 2, Eriksen shows a multimodal particle size distribution with one peak at 0.1-1 μm and another peak at 1-10 μm, with a d of 0.097 μm. 10 , 0.316 μm d 50 , and d of 4.579 μm 90 has been obtained.

[0016] Mikkelsen (WO2015059243) illustrates a process for producing a high protein denatured whey protein composition, in which an aqueous solution containing sweet whey protein concentrate was prepared by dissolving the whey protein concentrate in water to a dry matter content of 16% and adjusting the pH to 6.4. Denaturation and micronization were carried out in a 6+6 scraped surface heat exchanger (SSHE), an APV shear agglomerator. After passing through a retention cell (60 seconds), the product was cooled in an SSHE and subsequently cooled to 10°C by a plate heat exchanger (PHE). During the heat treatment (80°C for 10 minutes), the protein was denatured and particles with a size of 0.5-10 μm were formed. The product suspension was pumped to a storage tank, after which a portion of it was dried to a powder by spray drying. Mikkelsen reported that the resulting product suspension contained insoluble whey protein particles in the size range of 0.5-10 μm. Several additional patent applications by Aria Foods (including WO2018149869, WO2019110668, and WO2020187842) utilize the same process outlined in Mikkelsen to obtain denatured whey protein products that contain insoluble particles of denatured whey protein. Aria Foods sells micronized WPC that includes Nutrilac®-YO8075, which is discussed in detail herein.

[0017] Burling (WO2005041677) prepared whey protein gels suitable for stabilizing low-fat spreads and yogurts by low-temperature gelation using only small amounts of whey protein. Burling noted that it was particularly important to keep the calcium concentration very low, the protein concentration relatively low, and the pH value above 7 during the heat treatment. In particular, a pH > 7 during the heating step promoted the formation of soluble aggregates or filaments with a size of 20-100 nm before gel formation.

[0018] A series of patent applications by Nestec (including WO2007110421, WO2007110422, WO2007110423) refer to whey protein micelle powders made in a way that the micelles have a very sharp size distribution, with more than 80% of the micelles made having a size smaller than 1 μm, preferably between 100 nm and 900 nm, more preferably between 100 and 770 nm, and most preferably between 200 and 400 nm. Micellization occurred at pH 6.0, but not at pH 6.8, where linear aggregates (i.e., acid-gellable whey protein aggregates) were formed instead. It was found that the conversion yield of native whey protein to micelles decreased when the initial protein concentration was 12% or higher. The turbidity measured at 500 nm for a 3.4% protein solution was reported to be 21 (Example 2), and a 4% whey protein micelle dispersion was reported to have a turbidity of 80 at 500 nm (Examples 11-12).

[0019] Minor (WO2009113845) prepared pasteurized liquid compositions containing 12 g / 100 mL or 16 g / 100 mL whey protein. The average particle diameter D(4,3) of the liquid compositions obtained from static light scattering (Malvern Mastersizer 2000) is 3.7-7.7 μm before and after heat treatment. Example 4 of Minor shows that the average particle diameter D(4,3) of the whey protein composition obtained from static light scattering (Malvern Mastersizer 2000) is 0.48 μm before spray cooking and 0.29 μm after spray cooking.

[0020] Nielsen (WO2021136785) prepared whey protein nanogels from β-lactoglobulin isolate and a mixture of β-lactoglobulin isolate and whey protein isolate. Nielsen explained that whey protein nanogels have also been referred to as whey protein micelles by Nestec (e.g., WO2007 / 110421), but that their micellar nature is questionable. Nielsen emphasized that it is important to control the levels of Na, K, and Ca ions to avoid gelation of the entire sample. A secondary heating treatment at UHT temperature without shear forces caused the samples to gel. Heating at 150°C for 7 minutes and 30 seconds induced gelation of samples with 14% and 16% protein content, and heating at 150°C for 3 minutes and 10 seconds induced gelation of samples with 20% protein content (and the 14% protein content sample concentrated to 24%).

[0021] Dissanayake and Vasiljevic (J. Dairy Sci. 92, 1387-1397, 2009) prepared whey protein compositions by first heat treating a 10% (w / w) whey protein retention sample at 90°C for 20 minutes and then applying dynamic high pressure shear (microfluidization) at 140 MPa using one or five passes. The average particle size of the heat treated and microfluidized whey protein was about 10 μm. These heat treated microfluidized samples were relatively unstable, exhibiting heat set times (HCT) of 87.8 seconds for one pass and 102.5 seconds for five passes at 140°C.

[0022] The same research group investigated the functional properties of whey protein microparticulated at low pH (Dissanayake et al., J. Dairy Sci. 95, 1667-1679, 2012). The mean particle size of whey protein microparticulated at low pH was approximately 100 nm. In particular, the heat-treated microparticulated citric acid acidified (HTM-CA) and heat-treated microparticulated lactic acid acidified (HTM-LA) samples were reported to be 100 ± 6.3 nm and 101 ± 19.3 nm, respectively. Both denatured particulate whey protein samples acidified with citric acid and lactic acid showed heat setting times (HCT) of more than 3 minutes at 140 °C. However, the samples showed reduced thermal stability, with only 19.7% and 10.9% of the protein content retained in the supernatant after heating at 140 °C for 10 seconds followed by centrifugation (12,000 × g, 20 °C), respectively, which could be attributed to the effect of certain acidulants.

[0023] Although certain existing denatured whey protein compositions provide some resistance to gelation in a standard heat setting time test (e.g., HCT of 140° C. or greater for 2 minutes) at 10% protein content, these existing denatured whey protein compositions do not have a particle size distribution as defined herein, and in particular do not have a particle size distribution that remains substantially the same after a secondary heat treatment.

[0024] Thus, there remains a need for particulate whey protein compositions that are stable when exposed to a second heat treatment, particularly compositions that do not exhibit substantial change in particle size distribution when exposed to a second heat treatment. Summary of the Invention

[0025] The present disclosure relates to denatured whey protein compositions and methods for preparing and using such compositions. The denatured whey protein compositions include sub-micrometer sized particles that include denatured whey protein.

[0026] By way of example, the particle size distribution of the denatured whey protein composition has some, and preferably all, of the following characteristics: at least 55% by volume of the particles have a diameter of about 0.2 μm to about 1.0 μm; less than 45% by volume of the particles have a diameter of about 1.0 μm to about 10.0 μm; a diameter of about 0.5 to about 1.2 μm or about 0.6 to about 1.0 μm. 50 d of about 0.9 to about 1.7 μm or about 1.1 to about 1.5 μm 90a volume weighted mean diameter D(4,3) of about 0.6 to about 1.0 μm; and / or a substantially unimodal distribution in which at least 95% or at least 98% by volume of the particles have a diameter less than about 2.0 μm. Without wishing to be bound by theory, it is believed that the step of treating at least the aqueous whey protein solution and / or whey protein retentate with an oxidizing agent provides a denatured whey protein composition in which the reactivity of the denatured protein and particulates upon further heating is minimal, such that the particulates are relatively resistant to further growth upon subsequent heat treatment. Indeed, such denatured whey protein compositions are highly stable to secondary heating conditions, exhibiting minimal or essentially no primary particle size growth after heating an aqueous solution of 10% (w / w) protein content (pH 6.8) at 120° C. for 15 minutes. In certain embodiments, the denatured whey protein composition has an insolubility of at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90%. In certain embodiments, the remaining denaturable whey protein in the composition is less than 20%, less than 18%, less than 16%, less than 14%, or less than 12%. In some such embodiments, the remaining denaturable whey protein is less than 16%, less than 14%, or less than 12%. In certain embodiments, the denatured whey protein in the composition has a high degree of covalent bonds. For example, at least 60%, at least 70%, or at least 80% of the β-lactoglobulin in the denatured whey protein composition is covalently cross-linked to form multimers (e.g., dimers, trimers, etc.). As another example, the ratio of covalent interactions to non-covalent interactions in the microparticles is at least 4 to 1. Furthermore, high protein liquid applications such as drinking yogurt and liquid nutritional compositions produced using denatured whey protein compositions exhibit essentially no particle size change after heat treatment, and therefore exhibit minimal or essentially no precipitation upon long-term storage.Therefore, it is expected that high protein liquid applications enabled by the present technology will provide excellent stability over the shelf life of the product.

[0027] In one aspect, the present disclosure provides a denatured whey protein composition comprising sub-micrometer particles having a particle size distribution as described herein. The particle size distribution of such denatured whey protein compositions is stable when subjected to a secondary heat treatment. For example, after heating (e.g., in an autoclave or oil bath) a 10% (w / w) protein content aqueous solution at 120° C. for 15 minutes, the particle size distribution of the denatured whey protein composition is stable. 50 is about 0.6 to about 1.0 μm, and / or d 90 The particle size is about 0.9 to about 1.7 μm.

[0028] The present disclosure provides a heat stable denatured whey protein composition comprising microparticles comprising at least 60% total protein on a dry weight basis and denatured whey protein, the microparticles having (i) a volume weighted average diameter D(4,3) of about 1.0 μm or less, (ii) a d(4,3) of about 1.0 μm or less, and 50 , (iii) d of about 2.0 μm or less 90 and (iv) at least 92% by volume of the particles have a diameter less than about 2.0 μm, and (v) not more than 45% by volume of the fine particles have a diameter of about 1.0 to about 10.0 μm.

[0029] The present disclosure also provides liquid compositions, such as drinking yogurt, acidic beverages, neutral beverages, or liquid nutritional compositions, comprising the heat stable denatured whey protein compositions disclosed herein, and food products, such as baked food products, bars, or set or stirred yogurts, comprising the heat stable denatured whey protein compositions disclosed herein.

[0030] The present disclosure also provides a heat-treated shelf-stable high protein liquid composition comprising at least 6% (w / v) whey protein, the whey protein comprising particulates comprising denatured whey protein, (i) the liquid composition has a d particle size of about 1.0 μm or less after a secondary heat treatment applied for microbial control. 50 and d of about 2.0 μm or less90 (ii) the liquid composition has a viscosity of 100 s at 20° C. -1 (iii) the liquid composition exhibits less than 10% sedimentation after storage for 6 weeks at a temperature of about 20° C. to about 25° C.; and / or (iv) the liquid composition exhibits less than 10% sedimentation after centrifugation at 1540×g for 5 minutes.

[0031] The present disclosure also provides a liquid nutritional composition comprising at least 12% (w / v) total protein, e.g., 12%-18% (w / v) total protein, wherein at least 95% of the total protein in the liquid nutritional composition is denatured whey protein. The liquid nutritional composition may further comprise a lipid component and / or a carbohydrate component, e.g., the lipid component may be present in an amount of up to 30% (w / v) and the carbohydrate component may be present in an amount of up to 30% (w / v). In certain embodiments, substantially all of the total protein in the liquid nutritional composition is denatured whey protein. In certain embodiments, the liquid nutritional composition has a caloric density of less than 1 kcal / ml, between 1 and 2 kcal / ml, or greater than 2 kcal / ml. In certain embodiments, the liquid nutritional composition has a caloric density of between 1.5 and 2.5 kcal / ml. In certain embodiments, (i) the liquid nutritional composition has a d of about 1.0 μm or less after a secondary heat treatment applied for microbial control. 50 (ii) the liquid nutritional composition has a viscosity of 100 s at 20° C. -1 (iii) the liquid nutritional composition exhibits less than 10% settling after storage for 6 weeks at a temperature of about 20° C. to about 25° C.; and / or (iv) the liquid nutritional composition exhibits less than 10% settling after centrifugation at 1540×g for 5 minutes.

[0032] In one aspect, the disclosure provides a denatured whey protein composition comprising microparticles, the microparticles comprising denatured whey protein, at least 55% by volume of the microparticles having a diameter between about 0.2 μm and about 1.0 μm, at least 95%, at least 98%, or at least 99% by volume of the microparticles having a diameter less than about 2.0 μm, and the denatured whey protein composition exhibits substantially no change in particle size distribution when exposed to a secondary heat treatment. In a particular embodiment, an aqueous solution containing 10% (w / w) protein is heated (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes, followed by d 50 is about 0.6 to about 1.0 μm, and / or d 90 The particle size is about 0.9 to about 1.7 μm.

[0033] In another aspect, the disclosure provides a denatured whey protein composition comprising microparticles, the microparticles comprising denatured whey protein, the microparticles having a volume weighted average diameter D(4,3) of about 0.6 to about 1.0 μm, at least 98% by volume of the microparticles having a diameter less than about 2.0 μm and at least 99% by volume of the microparticles having a diameter less than about 3.0 μm, the denatured whey protein composition exhibiting substantially no particle size distribution when exposed to a secondary heat treatment. In a particular embodiment, an aqueous solution of 10% (w / w) protein content is heated (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes, followed by d 50 is about 0.6 to about 1.0 μm, and / or d 90 The particle size is about 0.9 to about 1.7 μm.

[0034] In certain embodiments of any of the aspects disclosed herein, the stability of the particle size distribution is determined by evaluating the growth of primary particles after (secondary) heat treatment of an aqueous solution of 10% (w / w) protein content at 120° C. for 15 minutes or at 140° C. for 90 seconds (e.g., in an autoclave or oil bath).

[0035] In some such embodiments, a 10% (or 12%, 14%, or 16%) (w / w) protein content solution comprising denatured whey protein exhibits minimal or essentially no change in particle size distribution when subjected to a primary particle growth test. By way of example, the primary particle growth test comprises a (secondary) heat treatment at 120° C. for 15 minutes (e.g., in an oil bath).

[0036] In some such embodiments, the d of a 10% (or 12%, 14%, or 16%) (w / w) protein content solution after a primary particle growth test that includes a heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes 50 does not increase substantially.

[0037] In some such embodiments, the d of an aqueous solution with a 10% (w / w) protein content after a primary particle growth test involving heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes 50 In some such embodiments, the d of a 10% (w / w) protein content aqueous solution after primary particle growth testing is about 1.0 μm or less. 50 is about 0.6 to about 1.0 μm.

[0038] In some such embodiments, the d of a 10% (w / w) protein content aqueous solution prior to primary particle growth testing 50 is the d after the primary particle growth test 50 differs by 20% or less, 18% or less, 16% or less, 14% or less, 12% or less, 10% or less, 9% or less, 8% or less, 7% or less, 6% or less, or 5% or less.

[0039] In some such embodiments, the d of a 10% (or 12%, 14%, or 16%) (w / w) protein content solution 90 does not increase substantially after primary particle growth testing involving heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes.

[0040] In some such embodiments, the d of an aqueous solution with a 10% (w / w) protein content after a primary particle growth test involving heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes 90 In some such embodiments, the d of a 10% (w / w) protein content aqueous solution after primary particle growth testing is less than about 2.0 μm. 90 is about 0.9 to about 1.7 μm or about 1.1 to about 1.5 μm.

[0041] In some such embodiments, the d of a 10% (w / w) protein content aqueous solution prior to primary particle growth testing 90 is the d after the primary particle growth test 90 differs from by 30% or less, 25% or less, or 20% or less.

[0042] In some such embodiments, the d of an aqueous solution with a 10% (w / w) protein content after a primary particle growth test involving heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes 50 is about 0.6 to about 1.0 μm, and the d of the 10% (w / w) protein content aqueous solution after the primary particle growth test 90 is about 0.9 to about 1.7 μm.

[0043] In certain embodiments of any aspect disclosed herein, the heat set time (HCT) of the denatured whey protein composition (the time required to observe the formation of visible aggregates in a 10% (w / w) protein content aqueous solution comprising the denatured whey protein composition during heating at 140° C. (e.g., in an oil bath)) is at least 3 minutes, at least 4 minutes, at least 5 minutes, at least 6 minutes, at least 9 minutes, at least 12 minutes, at least 15 minutes, or at least 18 minutes.

[0044] In certain embodiments of any of the aspects disclosed herein, (i) the denatured whey protein composition comprises between 65% and 95%, between 75% and 90%, or between 80% and 85% total protein by weight on a dry weight basis; (ii) the remaining denaturable whey protein in the denatured whey protein composition is less than 16%, less than 14%, or less than 12%, and / or the denatured whey protein composition has an insolubility of at least 60% or at least 65%; (iii) the ratio of covalent to non-covalent interactions in the microparticles is at least 2 to 1, at least 3 to 1, or at least 4 to 1, and / or at least 60%, at least 70%, or at least 80% of the β-lactoglobulin is covalently crosslinked; and (iv) the β-lactoglobulin is a denatured whey protein. (v) the denatured whey protein composition has a lactose content of 10% or less, 8% or less, 6% or less, or 4% or less by weight; (vi) the denatured whey protein composition has a fat content of 20% or less, 18% or less, 16% or less, 14% or less, 12% or less, 10% or less, 8% or less, 6% or less, or 4% or less by weight; (vii) the denatured whey protein composition has an ash content of 10% or less, 8% or less, 6% or less, or 4% or less by weight; (viii) the denatured whey protein composition has a casein content of 5% or less, 4% or less, 3% or less, 2% or less, or 1% or less by weight; or (ix) a combination of two or more or all of (i)-(viii).

[0045] The present disclosure also provides a method for producing the denatured whey protein compositions described herein. The method comprises: (a) providing an aqueous whey protein solution or whey protein retentate; (b) contacting the aqueous whey protein solution or whey protein retentate with an oxidizing agent; and (c) subjecting the whey protein solution or whey protein retentate to heat treatment under conditions in which protein denaturation occurs, the total protein content of the aqueous whey protein solution or whey protein retentate in step (c) is preferably at least 16% (w / w). In certain embodiments, the oxidizing agent is a peroxide, such as hydrogen peroxide, and step (b) comprises contacting the aqueous whey protein solution or whey protein retentate with the peroxide in combination with a peroxidase enzyme. In certain embodiments, the oxidizing agent is optionally inactivated, removed, or consumed prior to step (c), such that the aqueous whey protein solution or whey protein retentate is substantially free of the oxidizing agent during the heat treatment of step (c). In certain embodiments, the aqueous whey protein solution or whey protein retentate is substantially free of oxidizing agents during the heat treatment of step (c) where no active steps are taken to inactivate, remove, or consume the oxidizing agents. For example, the oxidizing agents may be present in amounts where removal or consumption of the oxidizing agents is undesirable (e.g., <10 ppm). In certain embodiments, the heat treatment of step (c) comprises a temperature of at least 70° C.

[0046] The present invention also provides edible consumer products, including food products and beverages, and liquid nutritional compositions, which comprise the denatured whey protein compositions described herein.

[0047] The present disclosure also provides a method for providing nutritional supplementation to a subject in need thereof, the method comprising enterally administering to the subject a denatured whey protein composition disclosed herein or a liquid nutritional composition comprising such a denatured whey protein composition.

[0048] The present disclosure also provides a liquid composition comprising a denatured whey protein composition, wherein the liquid composition exhibits minimal or essentially no change in particle size distribution after pasteurization or sterilization. In certain embodiments, the liquid composition has a d of less than 1 μm. 50 and d less than 2 μm 90 In certain embodiments, the liquid compositions exhibit essentially no settling, retain a relatively low viscosity, exhibit a pleasant taste and mouthfeel, and are not chalky or gritty after storage for 1, 3, 6, or 12 months (e.g., at a temperature of about 20° C. to about 25° C.).

[0049] It is an object of the present invention to provide denatured whey protein compositions having improved heat stability, and / or compositions that exhibit minimal or essentially no primary particle size growth upon secondary heat treatment, and / or products comprising denatured whey protein compositions having a particle size distribution that provides improved mouthfeel, settling, and / or shelf life, and / or at least a useful choice for the public.

[0050] These and other objects of the present invention are described in the following paragraphs, which should not be construed as narrowing the scope of the present invention. [Brief description of the drawings]

[0051] [Figure 1] FIG. 1 is a process flow diagram for producing an exemplary denatured whey protein composition.

[0052] [Diagram 2] FIG. 2 is a process flow diagram for producing an exemplary liquid composition comprising a denatured whey protein composition, i.e., a high protein beverage.

[0053] [Diagram 3] FIG. 3 is a process flow diagram for producing an exemplary liquid composition including a denatured whey protein composition, namely, drinking yogurt.

[0054] [Figure 4] FIG. 4 shows the particle size distribution of high protein beverages prepared with Powder A, Powder G, or Powder H after heating at 120° C. for 15 minutes.

[0055] [Diagram 5] FIG. 5 shows the particle size distribution of protein solutions containing 10%, 12%, 14%, or 16% (w / w) Powder A after heating at 120° C. for 15 minutes.

[0056] [Figure 6] FIG. 6 shows the heat coagulation time of denatured whey protein compositions prepared at protein concentrations of 10% to 20% (w / w).

[0057] [Figure 7] FIG. 7 shows particle size distribution of high protein (15% (w / w) and 20% (w / w)) drinking yogurt with denatured whey protein concentrate at one week.

[0058] [Figure 8] FIG. 8 shows the heat set time (HCT) of denatured whey protein compositions prepared from samples treated with different peroxide dosage rates.

[0059] [Figure 9] FIG. 9 shows the protein profile of the samples after enzymatic treatment and before heat treatment by HPLC analysis.

[0060] [Figure 10] FIG. 10 shows the particle size distribution of a 12% (w / v) reduced calorie high protein beverage before and after indirect UHT heating (143° C., 6 seconds). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0061] This detailed description is intended only to acquaint others skilled in the art with the invention, its principles, and its practical applications, so that they may adapt and apply the invention in its many forms that may best suit the requirements of a particular application. This description and its specific examples are intended for illustrative purposes only. Thus, the invention is not limited to the embodiments set forth in this patent application, which may be modified in various ways.

[0062] A.Definition

[0063] As used in this specification and the accompanying claims, the following terms have the meanings indicated unless specified otherwise.

[0064] 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 to "about" another particular value. When such a range is expressed, the range includes the recited values.

[0065] The term "denatured whey protein composition" refers to a composition that contains at least some denatured whey protein, preferably a significant amount of denatured whey protein. The composition may also contain non-denatured whey protein. However, a denatured whey protein composition preferably has an insolubility of at least 50%. Alternatively or additionally, the remaining denaturable whey protein in the composition is preferably less than 20%.

[0066] The term "dry weight basis" refers to the percentage of a substance in a composition or product after the moisture in the product has been removed. This can be calculated by applying a correction for the moisture retained in the product.

[0067] The term "liquid nutritional composition" refers to an aqueous composition that is administered to the subject's stomach, preferably orally or by other means, generally by tube feeding. Such other means include nasogastric tube feeding and gastric tube feeding. The term "liquid nutritional composition" includes medical foods, enteral nutrition, food products for special medical purposes, liquid meal replacements, and supplements. The liquid nutritional compositions described herein provide significant amounts of protein and carbohydrates, and usually also lipids. They may also include vitamins and minerals. In various embodiments, the subject in need of nutrition may suffer from or be susceptible to a disease or condition, or may be undergoing or being treated for a disease or condition, is an elderly person, a person recovering from a disease or condition, or a malnourished person. In other embodiments, the subject may also be a healthy individual (including one with specific nutritional requirements), including but not limited to sportsmen or active elderly people.

[0068] The term "microparticle" refers to a population of insoluble aggregates comprising denatured whey protein. Such aggregates are formed via covalent interactions (e.g., intermolecular sulfhydryl-disulfide exchange reactions) and / or non-covalent interactions (e.g., hydrophobic interactions). Generally, individual microparticles have a particle size of about 0.1 μm to about 3.0 μm or more (e.g., up to about 20 μm), and the population of particles has a particle size distribution as further defined herein.

[0069] The term "non-dairy protein" refers to any protein that is not a dairy protein (i.e., any protein that is not derived from the milk of an animal). Non-dairy proteins include plant-derived proteins, fungal proteins, and algae proteins.

[0070] The term "non-whey protein" refers to any protein that is not a whey protein. As used herein, the term "non-whey protein" includes casein and proteins derived from one or more non-dairy sources.

[0071] The term "subject" also includes humans and other primates, as well as other mammals, such as farm animals, sports animals, and pets. In certain embodiments, the subject is a human. In some such embodiments, the subject is a human infant, a human toddler, a human child, or a human adult. In certain embodiments, the subject is in need of nutritional support.

[0072] The term "substantially non-hydrolyzed" refers to unhydrolyzed (intact) proteins and proteins having a degree of hydrolysis of less than about 10.0%, 5.0%, 4.0%, 3.5%, 3.0%, 2.5%, 2.0%, 1.5%, 1.0%, or less than about 0.5%. Unless otherwise specified, whey proteins and non-whey proteins for use herein are substantially non-hydrolyzed.

[0073] The term "total protein" relates to the total amount of protein in a composition or product, and can be determined, for example, using the Kjeldahl method and the appropriate nitrogen conversion factor for dairy protein.

[0074] The term "whey protein concentrate" or "WPC" refers to a portion 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, the WPC has a total solids (TS) of at least 35% by weight, preferably at least 60% by weight, at least 65% by weight, at least 70% by weight, at least 75% by weight, or at least 80% by weight, as whey protein. For the purposes of this specification, the term "WPC" also includes whey protein isolates (WPIs), where the context permits.

[0075] The term "whey protein isolate" used herein refers to a composition that is mainly composed of whey protein, with minimal lipid and lactose content.Therefore, the preparation of WPI typically requires more rigorous separation processes, such as a combination of microfiltration and ultrafiltration or ion exchange chromatography.It is generally recognized that WPI refers to a composition in which at least 90% by weight of solids is whey protein.

[0076] The term "yoghurt" refers to an acidic or fermented food or beverage product prepared from dairy sources and containing either viable microorganisms or chemical acidulants or both. The term "yoghurt" or "yogurt" includes set or stirred yoghurt, as well as drinking yoghurt, including ambient yoghurt.

[0077] B. Denatured Whey Protein Compositions

[0078] The denatured whey protein compositions disclosed herein may comprise sub-micrometer particles and exhibit essentially no change in particle size distribution when exposed to secondary heat treatment. Such whey protein compositions are highly suitable for applications involving secondary heat conditions, such as high temperature pasteurization, ultra-high temperature (UHT) treatment, or retort heating applied for sterilization and microbial control. The denatured whey protein compositions disclosed herein may provide a thinner, less chalky mouthfeel than previously known compositions, and exhibit minimal or essentially no grittiness or sandiness.

[0079] 1. Characteristics of particle size distribution

[0080] As discussed herein, the particle size distribution characteristics are predicted based on the particle size distribution for a reconstituted liquid comprising the denatured whey protein compositions disclosed herein.

[0081] In certain embodiments, the composition comprises microparticles comprising denatured whey protein, the microparticles having a volume weighted average diameter D(4,3) of about 1.0 μm or less. In some such embodiments, the microparticles have a volume weighted average diameter D(4,3) of about 0.6 to about 1.0 μm or about 0.7 to about 1.0 μm. In some such embodiments, at least 91% by volume of the microparticles have a diameter less than about 2.0 μm. Alternatively, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by volume of the microparticles have a diameter less than about 2.0 μm. In some such embodiments, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by volume of the microparticles have a diameter less than about 1.5 μm. In some such embodiments, at least 98% by volume of the microparticles have a diameter less than about 3.0 μm. Alternatively, at least 99% by volume of the microparticles have a diameter less than about 3.0 μm.

[0082] In certain embodiments, the composition comprises microparticles comprising denatured whey protein, wherein at least 40% by volume of the microparticles have a diameter of about 0.1 μm to about 1.0 μm. In some such embodiments, at least 45% by volume of the microparticles have a diameter of about 0.1 μm to about 1.0 μm. In some such embodiments, at least 50% by volume of the microparticles have a diameter of about 0.1 μm to about 1.0 μm. In some such embodiments, at least 55% by volume of the microparticles have a diameter of about 0.1 μm to about 1.0 μm. In some such embodiments, at least 60% by volume of the microparticles have a diameter of about 0.1 μm to about 1.0 μm. In some such embodiments, at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% by volume of the microparticles have a diameter of less than about 1.5 μm. In some such embodiments, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by volume of the microparticles have a diameter less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% by volume of the microparticles have a diameter less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% by volume of the microparticles have a diameter less than about 3.0 μm.

[0083] In certain embodiments, the composition comprises microparticles comprising denatured whey protein, wherein at least 40% by volume of the microparticles have a diameter of about 0.2 μm to about 1.0 μm. In some such embodiments, at least 45% by volume of the microparticles have a diameter of about 0.2 μm to about 1.0 μm. In some such embodiments, at least 50% by volume of the microparticles have a diameter of about 0.2 μm to about 1.0 μm. In some such embodiments, at least 55% by volume of the microparticles have a diameter of about 0.2 μm to about 1.0 μm. In some such embodiments, at least 60% by volume of the microparticles have a diameter of about 0.2 μm to about 1.0 μm. In some such embodiments, at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% by volume of the microparticles have a diameter of less than about 1.5 μm. In some such embodiments, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by volume of the microparticles have a diameter less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% by volume of the microparticles have a diameter less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% by volume of the microparticles have a diameter less than about 3.0 μm.

[0084] In certain embodiments, the composition comprises microparticles comprising denatured whey protein, wherein at least 40% by volume of the microparticles have a diameter of about 0.3 μm to about 1.0 μm. In some such embodiments, at least 45% by volume of the microparticles have a diameter of about 0.3 μm to about 1.0 μm. In some such embodiments, at least 50% by volume of the microparticles have a diameter of about 0.3 μm to about 1.0 μm. In some such embodiments, at least 55% by volume of the microparticles have a diameter of about 0.3 μm to about 1.0 μm. In some such embodiments, at least 60% by volume of the microparticles have a diameter of about 0.3 μm to about 1.0 μm. In some such embodiments, at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% by volume of the microparticles have a diameter of less than about 1.5 μm. In some such embodiments, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by volume of the microparticles have a diameter less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% by volume of the microparticles have a diameter less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% by volume of the microparticles have a diameter less than about 3.0 μm.

[0085] In certain embodiments, the composition comprises microparticles comprising denatured whey protein, wherein at least 40% by volume of the microparticles have a diameter of about 0.4 μm to about 1.0 μm. In some such embodiments, at least 45% by volume of the microparticles have a diameter of about 0.4 μm to about 1.0 μm. In some such embodiments, at least 50% by volume of the microparticles have a diameter of about 0.4 μm to about 1.0 μm. In some such embodiments, at least 55% by volume of the microparticles have a diameter of about 0.4 μm to about 1.0 μm. In some such embodiments, at least 60% by volume of the microparticles have a diameter of about 0.4 μm to about 1.0 μm. In some such embodiments, at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% by volume of the microparticles have a diameter of less than about 1.5 μm. In some such embodiments, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by volume of the microparticles have a diameter less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% by volume of the microparticles have a diameter less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% by volume of the microparticles have a diameter less than about 3.0 μm.

[0086] In certain embodiments, the composition comprises microparticles comprising denatured whey protein, wherein at least 40% by volume of the microparticles have a diameter of about 0.5 μm to about 1.0 μm. In some such embodiments, at least 45% by volume of the microparticles have a diameter of about 0.5 μm to about 1.0 μm. In some such embodiments, at least 50% by volume of the microparticles have a diameter of about 0.5 μm to about 1.0 μm. In some such embodiments, at least 55% by volume of the microparticles have a diameter of about 0.5 μm to about 1.0 μm. In some such embodiments, at least 60% by volume of the microparticles have a diameter of about 0.5 μm to about 1.0 μm. In some such embodiments, at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% by volume of the microparticles have a diameter of less than about 1.5 μm. In some such embodiments, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by volume of the microparticles have a diameter less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% by volume of the microparticles have a diameter less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% by volume of the microparticles have a diameter less than about 3.0 μm.

[0087] In certain embodiments, the composition comprises microparticles comprising denatured whey protein, the microparticles having a d of about 1.2 μm or less, about 1.1 μm or less, about 1.0 μm or less. 50 In some such embodiments, the microparticles have a d of about 0.5 to about 1.2 μm, about 0.6 to about 1.1 μm, about 0.6 to about 1.0 μm, or about 0.7 to about 1.0 μm. 50In some such embodiments, at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% by volume of the microparticles have a diameter less than about 1.5 μm. In some such embodiments, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by volume of the microparticles have a diameter less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% by volume of the microparticles have a diameter less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% by volume of the microparticles have a diameter less than about 3.0 μm.

[0088] In certain embodiments, a composition comprises microparticles comprising denatured whey protein, the microparticles having a d of about 2.0 μm or less, about 1.9 μm or less, or about 1.8 μm or less. 90 In some such embodiments, the microparticles have a d of about 0.6 to about 2.0 μm, about 0.7 to about 1.9 μm, about 0.8 to about 1.8 μm, about 0.9 to about 1.7 μm, about 1.0 to about 1.6 μm, or about 1.1 to about 1.5 μm. 90 In some such embodiments, at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% by volume of the microparticles have a diameter less than about 1.5 μm. In some such embodiments, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by volume of the microparticles have a diameter less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% by volume of the microparticles have a diameter less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% by volume of the microparticles have a diameter less than about 3.0 μm.

[0089] In certain embodiments, the composition comprises microparticles comprising denatured whey protein, the microparticles having a d of about 0.5 to about 1.2 μm. 50 and d of about 0.9 to about 1.7 μm 90 In some such embodiments, the microparticles have a d of about 0.6 to about 1.0 μm. 50 and d of about 0.9 to about 1.7 μm 90 In some such embodiments, the microparticles have a d of about 1.1 to about 1.5 μm. 90 Thus, in some such embodiments, the microparticles have a d of about 0.6 to about 1.0 μm. 50 and d of about 1.1 to about 1.5 μm 90 In some such embodiments, at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% by volume of the microparticles have a diameter less than about 1.5 μm. In some such embodiments, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by volume of the microparticles have a diameter less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% by volume of the microparticles have a diameter less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% by volume of the microparticles have a diameter less than about 3.0 μm.

[0090] In certain embodiments, the composition comprises microparticles comprising denatured whey protein, the microparticles having a d of about 0.2 to about 0.8 μm. 10 , d of about 0.5 to about 1.2 μm 50 , and d of about 0.9 to about 1.7 μm 90 In some such embodiments, the microparticles have a d of about 0.3 to about 0.7 μm or about 0.4 to about 0.6 μm. 10 In some such embodiments, the microparticles have a d of about 0.6 to about 1.0 μm. 50 In some such embodiments, the microparticles have a d of about 1.1 to about 1.5 μm.90 Thus, in some such embodiments, the microparticles have a d of about 0.4 to about 0.6 μm. 10 , d of about 0.6 to about 1.0 μm 50 and d of about 1.1 to about 1.5 μm 90 In some such embodiments, at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% by volume of the microparticles have a diameter less than about 1.5 μm. In some such embodiments, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by volume of the microparticles have a diameter less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% by volume of the microparticles have a diameter less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% by volume of the microparticles have a diameter less than about 3.0 μm.

[0091] In certain embodiments, the composition comprises microparticles comprising denatured whey protein, wherein 45% or less by volume of the microparticles have a diameter of about 1.0 to about 10.0 μm. In some such embodiments, at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% by volume of the microparticles have a diameter of less than about 1.5 μm. In some such embodiments, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by volume of the microparticles have a diameter of less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% by volume of the microparticles have a diameter of less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% by volume of the microparticles have a diameter of less than about 3.0 μm.

[0092] As used herein, a particle size of about 0.4 μm includes 0.35-0.44 μm, a particle size of about 0.5 μm includes 0.45-0.54 μm, a particle size of about 0.6 μm includes 0.55-0.64 μm, a particle size of about 0.7 μm includes 0.65-0.74 μm, a particle size of about 0.8 μm includes 0.75-0.84 μm, a particle size of about 0.9 μm includes 0.85-0.94 μm, and a particle size of about 1.0 μm includes 0.95-1.04 μm.

[0093] Methods for evaluating particle size are known in the art. For example, particle size can be measured using a Malvern Mastersizer2000 (Malvern Instruments Ltd, Worcs, UK) with a particle refractive index of 1.46 and a solvent refractive index of 1.33. For example, in an exemplary embodiment, the primary aggregate size of the reconstituted powder is determined by homogenizing a suspension of 10% total solids (TS) at neutral pH (e.g., 150 / 50 bar) and determining the average particle size (characterized by D[4,3]) using a Malvern Mastersizer2000 (Malvern Instruments Ltd, Worcs, UK) with a particle refractive index of 1.46 and a solvent refractive index of 1.33 as described above.

[0094] 2.Stable particle size distribution

[0095] In certain embodiments, the particle size distribution of the denatured whey protein composition does not substantially change after a primary particle growth test involving heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes.

[0096] In certain embodiments, the denatured whey protein composition exhibits minimal or essentially no change in particle size distribution when subjected to a primary particle growth test. In certain embodiments, the primary particle growth test comprises a (secondary) heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes. For example, an aqueous solution containing a 10% (w / w) protein content of the denatured whey protein composition exhibits minimal or essentially no change in particle size distribution after the primary particle growth test.

[0097] In some such embodiments, the d of a 10% (or 12%, 14%, or 16%) (w / w) protein content solution 50 does not increase substantially after primary particle growth testing involving heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes.

[0098] In some such embodiments, the d of an aqueous solution with a 10% (w / w) protein content after a primary particle growth test involving heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes 50 In some such embodiments, the d of a 10% (w / w) protein content aqueous solution after primary particle growth testing is about 1.0 μm or less. 50 is about 0.6 to about 1.0 μm.

[0099] In some such embodiments, the d of a 10% (w / w) protein content aqueous solution prior to primary particle growth testing 50 is the d after the primary particle growth test 50 differs by 20% or less, 18% or less, 16% or less, 14% or less, 12% or less, 10% or less, 9% or less, 8% or less, 7% or less, 6% or less, or 5% or less.

[0100] In some such embodiments, the denatured whey protein compositions disclosed herein comprise a 50 is stable when subjected to primary particle growth tests at protein contents up to 16% (w / w).

[0101] In some such embodiments, the d of a 10% (or 12%, 14%, or 16%) (w / w) protein content solution 90 does not increase substantially after primary particle growth testing involving heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes.

[0102] In some such embodiments, the d of an aqueous solution with a 10% (w / w) protein content after a primary particle growth test involving heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes 90 In some such embodiments, the d of an aqueous solution containing 10% (w / w) protein after primary particle growth testing is about 2.0 μm or less, about 1.9 μm or less, or about 1.8 μm or less. 90 is about 0.6 to about 2.0 μm, about 0.7 to about 1.9 μm, about 0.8 to about 1.8 μm, about 0.9 to about 1.7 μm, about 1.0 to about 1.6 μm, or about 1.1 to about 1.5 μm.

[0103] In some such embodiments, the d of a 10% (w / w) protein content aqueous solution prior to primary particle growth testing 90 is the d after the primary particle growth test 90 differs by 30% or less, 28% or less, 26% or less, 24% or less, 22% or less, 20% or less, 18% or less, or 16% or less.

[0104] In some such embodiments, the denatured whey protein compositions disclosed herein comprise a 90 is stable when subjected to primary particle growth tests at protein contents up to 16% (w / w)

[0105] In some such embodiments, the d of an aqueous solution with a 10% (w / w) protein content after a primary particle growth test involving heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes 50 is about 0.6 to about 1.0 μm, and the d of the 10% (w / w) protein content aqueous solution after the primary particle growth test 90 is about 1.1 to about 1.5 μm.

[0106] Methods for evaluating the growth of primary particles are known in the art. In certain embodiments, the growth of primary particles is determined as described herein. Briefly, 10% (w / w) protein solution (pH 6.8) is heated at 120°C for 15 minutes (e.g., in an autoclave or oil bath). After heat treatment, particle size distribution can be evaluated.

[0107] The stability of the particle size distribution of the denatured whey protein composition or liquid composition comprising the denatured whey protein composition includes having minimal or essentially no gelation, sedimentation, or aggregation after the second heat treatment. The gelation of a liquid composition is considered to be a change in state from liquid to soft solid to hard solid. A solution is considered to have gelled when it no longer flows after heating.

[0108] In certain embodiments, the particle size distribution of the denatured whey protein composition is stable upon sterilization and / or pasteurization (e.g., high temperature pasteurization, UHT processing, or retort heating). High temperature pasteurization requires a temperature of 80-85°C for 20-30 minutes or a temperature of 90-95°C for 5 minutes. UHT processing typically involves subjecting the composition to a temperature above 135°C (such as 135°C to 150°C) to sterilize. Typical holding times in UHT are 4-10 seconds (or more). Two variations of UHT processing are typically used: direct processing and indirect processing. In direct UHT heating systems, steam is mixed directly with the liquid composition, while in indirect UHT heating systems, the liquid composition is heated by contacting it with steam or superheated water over the entire metal surface of a heat exchanger (Zadow, 1986). Retort processing typically involves subjecting the composition to a temperature of 110°C to 130°C for 10-20 minutes in a sealed can to sterilize.

[0109] 3. Heat solidification time

[0110] In certain embodiments, the heat set time (HCT) of the aqueous solution containing 10% (w / w) protein content comprising the denatured whey protein composition is at least 3 minutes, at least 4 minutes, at least 5 minutes, at least 6 minutes, at least 7 minutes, at least 8 minutes, at least 9 minutes, at least 10 minutes, at least 11 minutes, or at least 12 minutes. In some such embodiments, the HCT of the aqueous solution containing 10% (w / w) protein content comprising the denatured whey protein composition is at least 14 minutes. The HCT is defined as the time required to observe the formation of visible (i.e., visible to the naked eye) aggregates during heating in a 140° C. oil bath.

[0111] Methods for determining heat set time (HCT) are known in the art. The HCT method involves sealing 1 mL of sample in a glass tube, clipping it to a platform, and placing it in a thermostatically controlled silicone oil bath at 140° C. with a defined rocking rate. The time elapsed from placing the container in the oil bath until visible aggregates begin to form is defined as HCT (Singh H & Creamer LK (1992), Determination of heat stability, In: Advanced Dairy Chemistry ed Fox PF Elsevier)).

[0112] Applicant does not wish to be bound by any theory, but believes based on its own experience, including that described herein, that any liquid composition with a thermal setting time less than 60 seconds has a risk of extensive fouling and clogging of UHT heating equipment, while any liquid composition with an HCT of 65-80 seconds at 140°C has a potential risk of fouling. As described herein, liquid compositions with a thermal setting time greater than 80 seconds are considered stable to UHT heat treatment 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 121°C have a high risk of gelling and clumping in retort cans.

[0113] In some such embodiments, the denatured whey protein compositions disclosed herein exhibit minimal or essentially no gelling or coagulation after treatment at 140° C. for 2 minutes at protein contents up to 16% (w / w).

[0114] In certain embodiments, the HCT of a 12%, 14%, or 16% (w / w) protein content solution comprising the denatured whey protein composition is at least 2 minutes. In some such embodiments, the HCT of a 12% (w / w) protein content solution comprising the denatured whey protein composition is at least 8 minutes. In some such embodiments, the HCT of a 14% (w / w) protein content solution comprising the denatured whey protein composition is at least 4 minutes. In some such embodiments, the HCT of a 16% (w / w) protein content solution comprising the denatured whey protein composition is at least 2 minutes.

[0115] 4.High protein content

[0116] In certain embodiments, the denatured whey protein composition comprises between 60% and 95% total protein on a dry weight basis. In some such embodiments, the denatured whey protein composition comprises between 65% and 95% total protein on a dry weight basis. In some such embodiments, the denatured whey protein composition comprises between 70% and 95% total protein on a dry weight basis. In some such embodiments, the denatured whey protein composition comprises between 75% and 90% total protein on a dry weight basis. In some such embodiments, the denatured whey protein composition comprises between 80% and 85% total protein by weight of the total protein on a dry weight basis.

[0117] In certain embodiments, the denatured whey protein composition comprises at least 60%, at least 65%, at least 70%, at least 75%, or at least 80% total protein on a dry weight basis. In some such embodiments, the denatured whey protein composition comprises at least 70%, at least 75%, or at least 80% total protein on a dry weight basis.

[0118] Methods for assessing protein concentration are known in the art. For example, protein concentration can be measured using the Kjeldahl method. This method is based on nitrogen determination, and protein concentration is calculated by multiplying the total nitrogen result by the nitrogen conversion factor for dairy protein, 6.38.

[0119] Optionally, non-protein nitrogen (NPN) may also be determined according to ISO 8968-4 / IDF020-4 - Milk - Determination of nitrogen content - Part 4 (Determination of the non-protein nitrogen content and true protein content of a sample is (m 総窒素 -m NPN ) × 6.38). Unless otherwise specified herein, total protein content is reported without consideration of NPN.

[0120] 5. Severe denaturation

[0121] Whey proteins include bovine serum albumin (BSA), α-lactalbumin, β-lactoglobulin, lactoferrin, immunoglobulins, and other minor proteins. Cheese whey-based ingredients may contain additional proteins such as the casein-related proteins glycomacropeptide (GMP) and proteose peptone 5 (pp5). Heat treatment of whey or WPC results in denaturation of BSA, α-lactalbumin, β-lactoglobulin, lactoferrin, and immunoglobulins. In contrast, GMP and pp5 are undenatured.

[0122] As noted herein, GMP-free acid casein whey can be used as a source of whey protein. For example, exemplary materials can be made using 60% cheese WPC and 40% acid WPC, which provide higher levels of heat denaturable protein than corresponding materials made from cheese WPC alone.

[0123] Methods for determining the degree of protein denaturation are known in the art. An exemplary method used herein relies on HPLC (Elgar et al (2000) J Chromatography A, 878, 183-196), and other methods suitable for use include those that rely on the Agilent 2100 Bioanalyzer (Agilent Technologies, Inc. 2000, 2001-2007, Waldbronn, Germany) and microfluidic chips, and utilize Agilent 2100 Expert software (e.g., Anema, (2009) International Dairy J, 19, 198-204) and polyacrylamide gel electrophoresis (e.g., Patel et al, (2007) Le Lait, 87, 251-268).

[0124] Denatured whey protein compositions may be characterized by measuring the remaining denaturable protein as a percentage of total protein (TN x 6.38) according to the following formula:

number

[0125] Denaturable whey protein is measured as Σ(bovine serum albumin+α-lactalbumin+β-lactoglobulin+lactoferrin+immunoglobulins).

[0126] In certain embodiments, the remaining denaturable whey protein in the composition is less than 20%, less than 18%, less than 16%, or less than 14%, or less than 12%. In some such embodiments, the remaining denaturable whey protein is less than 16%, or less than 14%, or less than 12%.

[0127] For an exemplary denatured whey protein composition, the percentage of denaturable protein that is denatured can be estimated according to the following formula:

number

[0128] The denatured whey protein composition may also be characterized in terms of insolubility. For example, a suspension containing the denatured whey protein composition in NaCl is treated with acetic acid and centrifuged. The total protein content of the supernatant and the total protein content of the suspension without centrifugation are determined, and the percent insolubility of the sample can be determined as follows:

number

[0129] In certain embodiments, the denatured whey protein composition has an insolubility of at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%.

[0130] In certain embodiments, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% of the heat-denaturable whey protein in the composition is present in a denatured state. In some such embodiments, at least 60% of the heat-denaturable whey protein is present in a denatured state, at least 70% of the heat-denaturable whey protein is present in a denatured state, or at least 80% of the heat-denaturable whey protein is present in a denatured state.

[0131] Denatured whey protein compositions may also be characterized in terms of the ratio of non-denaturable protein (e.g., GMP) to remaining denaturable protein. For example, in certain embodiments, the ratio of GMP to the sum of soluble α-lactalbumin and soluble β-lactoglobulin is at least 1, at least 2, or at least 3.

[0132] 6.Covalent bond / Non-covalent bond

[0133] Both covalent and non-covalent interactions can be involved in the formation of whey protein microparticles. Covalent interactions involve the sharing of electrons and are relatively strong bonds. Non-covalent interactions involved in the formation of whey protein microparticles include van der Waals forces, hydrophobic interactions, and hydrogen bonds. Without wishing to be bound by theory, it is believed that covalent interactions cause irreversible aggregation of whey protein.

[0134] The presence of free sulfhydryl (-SH) or intramolecular disulfide (SS) groups in the secondary structure of whey proteins promotes the formation of covalent interactions. β-Lactoglobulin contains two intramolecular disulfide (SS) groups and one free SH group, which makes it highly reactive for covalent interactions. BSA contains 17 SS bonds and one SH group, which also makes it capable of initiating covalent aggregation. α-Lactalbumin contains four SS bonds but no free SH group that can serve as the starting point for the covalent aggregation reaction.

[0135] In certain embodiments, the denatured whey protein in the composition has a high degree of covalent bonds.

[0136] For example, at least 60%, at least 70%, or at least 80% of the β-lactoglobulin in the denatured whey protein composition is covalently cross-linked to another protein (e.g., another β-lactoglobulin molecule and / or BSA and / or α-lactalbumin) to form multimers (e.g., dimers, trimers, etc.). In some such embodiments, at least 85% of the β-lactoglobulin in the denatured whey protein composition is covalently cross-linked to another protein.

[0137] As another example, the ratio of covalent to non-covalent interactions in the microparticles is at least 2:1, at least 3:1, or at least 4:1.

[0138] 7. Further Characterization of Denatured Whey Protein Composition

[0139] In certain embodiments, alpha-lactalbumin represents no more than 60%, no more than 50%, or no more than 40% of the total protein in the denatured whey protein composition.

[0140] In certain embodiments, the denatured whey protein composition comprises from about 5% to about 30% alpha-lactalbumin, from about 10% to about 25% alpha-lactalbumin, or from about 15% to about 20% alpha-lactalbumin relative to the total protein in the denatured whey protein composition.

[0141] In certain embodiments, β-lactoglobulin represents no more than 80%, no more than 75%, or no more than 70% of the total protein in the denatured whey protein composition.

[0142] In certain embodiments, the denatured whey protein composition comprises from about 30% to about 80% β-lactoglobulin, from about 35% to about 75%, from about 40% to about 65%, or from about 45% to about 60% β-lactoglobulin based on the total protein in the denatured whey protein composition.

[0143] In certain embodiments, the denatured whey protein composition has a ratio of α-lactalbumin to β-lactoglobulin of from about 1:15 to about 10:1, from about 1:10 to about 5:1, or from about 1:5 to about 1:1.

[0144] In certain embodiments, the denatured whey protein composition exhibits a high degree of turbidity when in aqueous solution. For example, an aqueous solution containing a 3.4% (w / w) protein content comprising the denatured whey protein composition exhibits a turbidity value measured at 500 nm and 20° C. of at least 50 absorbance units. In some such embodiments, an aqueous solution containing a 3.4% (w / w) protein content comprising the denatured whey protein composition exhibits a turbidity value measured at 500 nm of at least 75 or at least 100 absorbance units. As another example, an aqueous solution containing a 4% (w / w) protein content comprising the denatured whey protein composition exhibits a turbidity value measured at 500 nm of at least 90 absorbance units. In some such embodiments, an aqueous solution containing a 4% (w / w) protein content comprising the denatured whey protein composition exhibits a turbidity value measured at 500 nm of at least 100, at least 110, or at least 120 absorbance units.

[0145] To assess turbidity, it may be necessary to further dilute the aqueous solution. For example, a 3.4% (w / w) protein content aqueous solution can be diluted to a protein content of less than 1%, less than 0.5%, less than 0.1%, or less than 0.05%, such as 0.025%. In such cases, the dilution factor can be used to obtain the absorbance units of the original solution.

[0146] In certain embodiments, the denatured whey protein composition has a total concentration of monovalent metal cations of at least 25 mM or at least 30 mM. In some such embodiments, the concentration of monovalent metal cations in the denatured whey protein composition is from about 25 mM to about 200 mM, e.g., from about 30 mM to about 150 mM. The monovalent metal cations include Na + and K + Includes:

[0147] In certain embodiments, the denatured whey protein compositions comprise less than about 10%, 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 0.1% lactose by weight. In some such embodiments, the denatured whey protein compositions have a lactose content of up to 10%, up to 8%, up to 6%, or up to 4% by weight.

[0148] In certain embodiments, the denatured whey protein composition has a whey protein to lactose ratio of at least 10 to 1, at least 15 to 1, at least 20 to 1, at least 25 to 1, at least 30 to 1, at least 35 to 1, at least 40 to 1, at least 45 to 1, or at least 50 to 1.

[0149] In certain embodiments, the denatured whey protein composition has been processed to reduce the lactose concentration, such as by lactose hydrolysis, or is derived from a source (e.g., WPC) that has been so processed.

[0150] In certain embodiments, the denatured whey protein composition has a whey protein to carbohydrate ratio of at least 10 to 1, at least 15 to 1, at least 20 to 1, at least 25 to 1, at least 30 to 1, at least 35 to 1, at least 40 to 1, at least 45 to 1, or at least 50 to 1.

[0151] In certain embodiments, the denatured whey protein compositions comprise less than about 20%, less than about 15%, less than about 10%, 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 0.1% fat by weight. In some such embodiments, the denatured whey protein compositions have a fat content of up to 10%, up to 8%, up to 6%, or up to 4%.

[0152] In certain embodiments, the denatured whey protein compositions have a whey protein to fat ratio of at least 4 to 1, at least 6 to 1, at least 8 to 1, at least 10 to 1, at least 12 to 1, at least 14 to 1, at least 16 to 1, at least 18 to 1, at least 20 to 1, at least 22 to 1, at least 24 to 1, at least 26 to 1, at least 28 to 1, or at least 30 to 1.

[0153] In certain embodiments, the denatured whey protein compositions comprise less than about 12%, less than about 11%, less than about 10%, less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, or less than about 4% by weight ash. In some such embodiments, the denatured whey protein compositions have an ash content of up to 10%, up to 8%, up to 6%, or up to 4% by weight.

[0154] In certain embodiments, the denatured whey protein composition has a whey protein to ash ratio of at least 10:1, at least 12:1, at least 14:1, at least 16:1, at least 18:1, or at least 20:1.

[0155] In certain embodiments, the denatured whey protein composition comprises less than about 10% by weight, less than about 9% by weight, less than about 8% by weight, less than about 7% by weight, less than about 6% by weight, less than about 5% by weight, less than about 4% by weight, less than about 3% by weight, less than about 2% by weight, less than about 1% by weight, or less than about 0.1% by weight of casein. In some such embodiments, the denatured whey protein composition has a casein content of up to 5% by weight, up to 4% by weight, up to 3% by weight, up to 2% by weight, or up to 1% by weight. In some such embodiments, the denatured whey protein composition is substantially free of casein.

[0156] In certain embodiments, the denatured whey protein composition has a whey protein to casein ratio of at least 10:1, at least 15:1, at least 20:1, at least 25:1, at least 30:1, or at least 35:1.

[0157] Each feature of the denatured whey protein compositions described herein may be combined with any other feature unless expressly stated otherwise. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature indicated as being preferred or advantageous.

[0158] By way of example and not limitation, in certain embodiments, the denatured whey protein composition comprises microparticles comprising denatured whey protein and has a substantially stable particle size distribution after secondary heat treatment. The particle size distribution of the denatured whey protein composition has some, and preferably all, of the following characteristics: at least 55% by volume of the particles have a diameter of about 0.2 μm to about 1.0 μm; less than 45% by volume of the particles have a diameter of about 1.0 μm to about 10.0 μm; a diameter of about 0.5 to about 1.2 μm or about 0.6 to about 1.0 μm; 50 d of about 0.9 to about 1.7 μm or about 1.1 to about 1.5 μm 90 a volume weighted mean diameter D(4,3) of about 0.6 to about 1.0 μm; and / or a substantially unimodal distribution in which at least 95% or at least 98% by volume of the particles have a diameter less than about 2.0 μm.

[0159] After a primary particle growth test involving heat treatment (e.g., in an autoclave or oil bath) at 120° C. for 15 minutes, some, preferably all, of the aforementioned particle size distribution characteristics remain substantially the same. For example, the d 50 is approximately 1.0 μm or less, and the d of the 10% (w / w) protein content aqueous solution after the primary particle growth test 90 In some such embodiments, the d of an aqueous solution containing 10% (w / w) protein after primary particle growth testing is about 2.0 μm or less, about 1.9 μm or less, or about 1.8 μm or less. 50 is about 0.6 to about 1.0 μm, and the d of the 10% (w / w) protein content aqueous solution after the primary particle growth test 90 is about 1.1 to about 1.5 μm.

[0160] In some such embodiments, the denatured whey protein compositions have a particle size distribution that is substantially stable and exhibits minimal or essentially no primary particle size growth after heating an aqueous solution of 10% (w / w) protein content (pH 6.8) at 120° C. for 15 minutes, and further have the following characteristic or combination of characteristics:

[0161] (i) the denatured whey protein composition comprises, on a dry weight basis, 65%-95%, 75%-90%, or 80%-85% total protein;

[0162] (ii) the denatured whey protein composition has less than 16%, less than 14%, or less than 12% residual denatured whey protein, and / or the denatured whey protein composition has an insolubility of at least 60% or at least 65%;

[0163] (iii) the ratio of covalent to non-covalent interactions in the microparticle is at least 2:1, at least 3:1, or at least 4:1, and / or at least 60%, at least 70%, or at least 80% of the β-lactoglobulin is covalently cross-linked;

[0164] (iv) beta-lactoglobulin comprises no more than 80% or no more than 75% of the total protein in the denatured whey protein composition;

[0165] (v) the denatured whey protein composition has a lactose content of up to 10% by weight, up to 8% by weight, up to 6% by weight, or up to 4% by weight;

[0166] (vi) a fat content of not more than 20% by weight, not more than 18% by weight, not more than 16% by weight, not more than 14% by weight, not more than 12% by weight, not more than 10% by weight, not more than 8% by weight, not more than 6% by weight, or not more than 4% by weight;

[0167] (vii) the denatured whey protein composition has an ash content of up to 10% by weight, up to 8% by weight, up to 6% by weight, or up to 4% by weight;

[0168] (viii) the denatured whey protein composition has a casein content of up to 5%, up to 4%, up to 3%, up to 2%, or up to 1% by weight; or

[0169] (ix) A combination of two or more or all of (i) to (viii).

[0170] In certain embodiments, the denatured whey protein composition is heat stable (e.g., an aqueous solution of 10% (w / w) protein content exhibits an HCT of at least 3 minutes, at least 6 minutes, at least 9 minutes, or at least 12 minutes) and comprises microparticles comprising denatured whey protein, wherein at least 50%, at least 55%, or at least 60% by volume of the microparticles have a diameter of about 0.1 μm to about 1.0 μm.

[0171] C. Whey Protein Source

[0172] The whey protein source for the denatured whey protein composition may be any source that provides one or more whey proteins, such as sweet whey, or acid casein whey, or milk whey (obtained as a permeate phase by microfiltration of skim milk), 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 whey protein source for the denatured whey protein composition is a whey protein concentrate (WPC) or a whey protein isolate (WPI). Whey protein compositions including WPC and WPI may be derived from acid casein whey or cheese whey. Alternative whey protein sources for the denatured whey protein composition include enriched compositions that include individual whey proteins (e.g., β-lactoglobulin-enriched compositions or α-lactalbumin-enriched compositions).

[0173] WPCs are rich in whey proteins but also contain other components such as lipids, lactose, minerals / ash, and, in the case of cheese whey-based WPCs, glycomacropeptide (GMP), a non-denaturable casein-related non-globular protein.Typical production methods for whey protein concentrates utilize membrane filtration.

[0174] WPC is often listed with the percentage (w / w) of whey protein appended to "WPC." For example, WPC80 is a WPC with 80% whey protein by weight.

[0175] The whey protein may be derived from any mammalian species, such as, for example, cows, sheep, goats, horses, buffalo, deer, and camels. Preferably, the whey protein is of bovine origin.

[0176] In certain embodiments, the denatured whey protein can be obtained according to the processes specified by US6,767,575 (Huss & Spiegel), US2006 / 0204643 (Merrill et al), US4,734,827 (Singer et al), US5,494,696 (Holst et al), US2012 / 0114795 (Havea et al; also published as WO 2010 / 120199), EP0412590 (Hakkaart et al), and US5,188,842 (Visser et al; also published as EP0347237). Each method of producing denatured whey protein components imparts different characteristics, and users of the present invention will need to select the protein material most suitable for their process.

[0177] In certain embodiments, the whey protein source is available as a powder, preferably a WPC or WPI powder.

[0178] The denatured whey protein composition may be prepared from a mixture of cheese and / or acidic WPC and / or WPI, or from a mixture of proteins. In certain embodiments, the whey protein source is or comprises a WPC and / or WPI. In certain embodiments, the whey protein source is or comprises a mixture with a WPC and / or WPI, and optionally one or more ingredients that contain whey protein and / or non-whey protein.

[0179] In certain embodiments, the whey protein source is cheese whey or a WPC prepared from cheese whey. In some such embodiments, colorants are used during the cheese making process. For example, cheese makers may use annatto to impart an orange-yellow color to colored cheeses such as Cheddar, Leicester, and Gloucester. Annatto colorant is derived from the seeds of Bixa orellana (achiote), a shrub native to Central America. The seeds contain carotenoid pigments, including bixin, norbixin, and orellin. In some such embodiments, the whey protein source contains colorants.

[0180] D. Manufacturing method

[0181] In at least one embodiment, the present disclosure provides a method for producing a denatured whey protein composition. An exemplary method for producing a denatured whey protein composition is described below and illustrated in Figure 1. Suitable alternative methods for achieving the denatured whey protein composition described herein will be apparent to those skilled in the art.

[0182] In certain embodiments, the method comprises (a) providing an aqueous whey protein solution or whey protein retentate, (b) contacting the aqueous whey protein solution or whey protein retentate with an oxidizing agent, and (c) subjecting the whey protein solution or whey protein retentate to a heat treatment under conditions capable of producing the desired protein denaturation, wherein the aqueous whey protein solution or whey protein retentate has a total protein content of preferably at least 16% (w / w). In some such embodiments, the denaturing heat treatment is carried out at a temperature above 70° C. under conditions of mechanical shear and / or turbulence, as known in the art.

[0183] In certain embodiments, the oxidizing agent is a peroxide, such as hydrogen peroxide, and step (b) comprises contacting the aqueous whey protein solution or whey protein retentate with the peroxide in combination with a peroxidase enzyme.

[0184] 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.

[0185] In a particular embodiment, the amount of oxidizing agent is 1200×10 per kg of whey protein. -6 kg, e.g., about 20 x 10 per kg of whey protein -6 kg ~ Approx. 900 x 10 per kg of whey protein -6 kg, approximately 40 x 10 per kg of whey protein -6 kg ~ Approx. 800 x 10 per kg of whey protein -6 kg, or approximately 60 x 10 per kg of whey protein -6 kg ~ Approx. 700 x 10 per kg of whey protein -6 kg, e.g., about 80 x 10 per kg of whey protein-6 kg ~ Approx. 600 x 10 per kg of whey protein -6 kg, e.g., about 100 x 10 per kg of whey protein -6 kg ~ Approx. 400 x 10 per kg of whey protein -6 kg.

[0186] In a particular embodiment, the amount of oxidizing agent is 2000×10 per kg of denaturable whey protein. -6 kg, e.g., about 30×10 per kg of denaturable whey protein -6 kg ~ Approximately 1400 × × 10 per kg of denaturable whey protein -6 kg, approximately 60 x 10 per kg of denaturable whey protein -6 kg ~ Approx. 1200 × × 10 per kg of denaturable whey protein -6 kg, or approximately 90 x 10 per kg of denaturable whey protein -6 kg ~ approx. 1000 x 10 -6 kg, e.g., about 120 x 10 per kg of denaturable whey protein -6 kg ~ Approximately 800 x 10 per kg of denaturable whey protein -6 kg, e.g., about 150 x 10 per kg of denaturable whey protein -6 kg ~ Approximately 600 x 10 per kg of denaturable whey protein -6 kg.

[0187] In a particular embodiment, the amount of oxidizing agent is 2400×10 per kg of β-lactoglobulin protein. -6 kg, e.g., about 40 × 10 per kg of β-lactoglobulin protein -6 kg ~ 1800 x 10 per kg of β-lactoglobulin protein -6 kg, approximately 80 × 10 per kg of β-lactoglobulin protein -6 kg ~ 1600 x 10 per kg of β-lactoglobulin protein -6 kg, or approximately 120 x 10 per kg of β-lactoglobulin protein -6 kg ~ 1400 x 10 per kg of β-lactoglobulin protein-6 kg, e.g., about 160 × 10 per kg of β-lactoglobulin protein -6 kg ~ 1200 x 10 per kg of β-lactoglobulin protein -6 kg / kg, e.g., about 200×10 per kg of β-lactoglobulin protein -6 kg / kg ~ 800 × 10 per kg of β-lactoglobulin protein -6 kg.

[0188] 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, from about 0.28 to about 0.5.

[0189] In some such embodiments, the aqueous whey protein solution or whey protein retentate comprises a whey protein source, such as a WPC and / or a WPI, hi some such embodiments, the whey protein source comprises a colorant.

[0190] In certain embodiments, the oxidizing agent is a peroxide, such as hydrogen peroxide, and step (b) comprises contacting the aqueous whey protein solution or whey protein retentate with the peroxide in combination with a peroxidase enzyme.

[0191] In a particular embodiment, an amount of oxidizing agent is added in one step. More preferably, and to avoid inhibition and inactivation of the catalyst, the total amount of peroxide is added in two or more steps, for example, 2, 3, 4, 5 or more steps. More preferably, an amount of peroxide is added continuously. Those skilled in the art can determine the optimal oxidizing agent addition rate as a function of catalyst / enzyme activity and protein concentration in the whey solution.

[0192] In certain embodiments, the oxidizing agent is optionally inactivated, removed, or consumed prior to step (c), such that the aqueous whey protein solution or whey protein retentate is substantially free of oxidizing agent during the heat treatment of step (c). In certain embodiments, the aqueous whey protein solution or whey protein retentate is substantially free of oxidizing agent during the heat treatment of step (c) where no active steps are taken to inactivate, remove, or consume the oxidizing agent. For example, the oxidizing agent may be present in an amount (e.g., less than 10 ppm) where removal or consumption of the oxidizing agent is undesirable.

[0193] In certain embodiments, the denaturing heat treatment step is carried out at temperatures above 70° C. under conditions of high shear stress / force, which can be produced by increasing serum viscosity or wall shear rate, changing the flow pattern to turbulent, or applying mechanical shear, or a combination thereof.

[0194] In one exemplary embodiment, as shown in FIG. 1, a method includes providing a whey protein source, such as cheese whey, clarifying, separating, and / or heat treating (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, contacting the retentate with an oxidizing agent (e.g., hydrogen peroxide, optionally in combination with a peroxidase enzyme), and subjecting the whey protein retentate to a heat treatment step (preferably when no detectable oxidizing agent remains) to cause protein denaturation.

[0195] As shown in Figure 1, cheese whey collected after cheese making is clarified, subjected to microbial control, and heat treated (pasteurized) to inactivate any residual rennet enzymes and starter cultures from the cheese making.

[0196] Common conditions for heat treatment (pasteurization) are known in the art and include, but are not limited to, about 63° C. (about 145° F.) for 30 minutes (also known as the batch holding method), about 72° C. (about 161° F.) for 15 seconds (also known as the high temperature, short time (HTST) method), about 89° C. (about 192° F.) for 1 second, or any alternative heat and non-heat treatments that have an equivalent killer effect to these treatments.

[0197] As shown in Figure 1, the whey protein solution is 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 28%. 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%.

[0198] In certain embodiments, the filtration step provides a retentate comprising about 65% to about 95% total protein by weight. In some such embodiments, the whey protein retentate has a total protein content of about 75% to about 90% or about 80% to about 85% by weight. In some such embodiments, the whey protein retentate has a total protein content of at least 65%, at least 70%, at least 75%, or at least 80% by weight.

[0199] 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 heat treatment. Without wishing to be bound by theory, uncontrolled formation of denatured aggregates may cause the formation of larger than desired aggregates during the heat denaturation step.

[0200] In some such embodiments, the whey protein solution and / or whey protein retentate has a low level of denaturation before the denaturing heat treatment step. For example, the level of denaturation of the whey protein solution and / or whey protein retentate before the denaturing heat treatment step can be less than 10%, for example, about 3% to about 8%. Alternatively, the level of denaturation of the whey protein solution and / or whey protein retentate before the denaturing heat treatment step can 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 solution and / or whey protein retentate before the denaturing heat treatment step is about 5%, about 4%, about 3%, about 2%, or about 1%.

[0201] As shown in Figure 1, the retentate (and / or whey protein solution) is contacted with the oxidizing agent. The timing of treating the whey protein stream with the oxidizing agent may vary. For example, a diluted whey protein composition (i.e., pre-UF / DF) is contacted with the oxidizing agent.

[0202] In some such embodiments, the oxidizing agent is any food product grade oxidizing agent, hi some such embodiments, the oxidizing agent is oxygen (O2), ozone, peroxides including alkyl hydroperoxides, superoxide, peroxynitrite, peroxodisulfate, or lactoperoxidase.

[0203] In some such 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, e.g., sodium peroxide; alkaline earth metal peroxides, e.g., magnesium peroxide or calcium peroxide; or transition metal peroxides, e.g., zinc peroxide), hydrogen peroxide, and benzoyl peroxide. In some such embodiments, the oxidizing agent is a perborate, e.g., sodium perborate.

[0204] In some such embodiments, the oxidizing agent is benzoyl peroxide or hydrogen peroxide. Hydrogen peroxide can be obtained in a variety of concentrations and purities. In one embodiment, the hydrogen peroxide is food product grade.

[0205] In some such embodiments, the oxidizing agent is used with a catalyst, which may be either an enzyme catalyst or a chemical catalyst. In some such embodiments, the oxidizing agent is used with a catalytic enzyme, such as, for example, 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, or manganese.

[0206] In some such embodiments, the oxidizing agent is added over a period of time. For example, an initial amount of oxidizing agent may be added, and then subsequent amounts of oxidizing agent may be added as the initial amount is consumed. Thus, the total amount of peroxide may be added in two or more steps, for example, 2, 3, 4, 5, or more steps, or may be added continuously over a period of time.

[0207] Exemplary oxidizing agents include hydrogen peroxide, which are conventionally used in the dairy industry to decolorize whey.

[0208] Hydrogen peroxide (H2O2) is a clear, colorless liquid with a slightly pungent odor. Hydrogen peroxide is one of two decolorizing agents currently approved in the United States for the decolorization of whey, for example for the decolorization of whey compositions prepared from cheeses containing annatto.

[0209] Previous reports have provided conflicting information regarding the effect of hydrogen peroxide on whey protein denaturation. For example, Kramer et al., J. Agric. Food Chem, 65, 10258-10269 (2017) reported that enhanced aggregation of β-lactoglobulin (β-Lg) alone and a mixture of β-Lg and α-lactalbumin (α-La) was observed when low levels of hydrogen peroxide (e.g., 500 pM, i.e., a 5:1 molar ratio compared to protein) were present during the heating step. Similarly, Kang et al., J. Dairy Sci., 93, 3891-3901 (2010) reported that concentrations of hydrogen peroxide above the legal limit (500 ppm) increased the amount of whey protein denaturation. On the other hand, there are some reports that hydrogen peroxide reduced whey protein denaturation (see, for example, Fish and Mickelsen, Journal of Dairy Science, 50(7), 1045-1048 (1967) (stating that treating skim milk with hydrogen peroxide at 89°C for 30 minutes prior to denaturation reduced the denaturation of acidic whey proteins); Sutariya and Patel, Food Chemistry, 223, 114-120 (2017) (suggesting that adding hydrogen peroxide to whey protein isolates prevented whey protein denaturation and aggregation); see also Figure 8 in US2016 / 0235082). Furthermore, Bechtle (US3,818,109) reported that by combining moderate heat (e.g., temperatures of 60°C to 70°C) with hydrogen peroxide treatment, significant denaturation of whey proteins was avoided while achieving sterilization of the medium. Notably, these reports appear to describe the application of heat to samples containing hydrogen peroxide.

[0210] On the other hand, in certain embodiments, the methods for producing denatured whey protein compositions disclosed herein provide for removing or consuming the oxidizing agent prior to the denaturing heat treatment step. Thus, in some such embodiments, the oxidizing agent is consumed prior to subjecting the aqueous whey protein solution or whey protein retentate to a denaturing heat treatment step under conditions capable of causing protein denaturation. Thus, the aqueous whey protein solution or whey protein retentate may be substantially free of oxidizing agent during the denaturing heat treatment step.

[0211] In certain embodiments, the oxidizing agent is used with a catalytic enzyme, such as, for example, a microbial peroxidase enzyme, and the amount of oxidizing agent and peroxidase enzyme is such that the oxidizing agent is completely consumed by the enzyme. Alternatively, if 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, a catalase enzyme is not used.

[0212] The aqueous whey protein solution or whey protein retentate may be sampled to confirm that it is substantially free of oxidizing agents. By way of example, the aqueous whey protein solution or whey protein retentate may be sampled and tested for detectable peroxides using colorimetric test strips available from Merck / MilliporeSigma.

[0213] In some such embodiments, the method further comprises adjusting the pH of the aqueous whey protein solution or whey protein retentate prior to the denaturing heat treatment step. In some such embodiments, the pH of the aqueous whey protein solution or whey protein retentate prior to the denaturing heat treatment step is at least 5.6, at least 5.8, at least 6.0, or at least 6.2. In some such embodiments, the pH of the aqueous whey protein solution or whey protein retentate prior to the denaturing heat treatment step is from about 5.8 to about 7.0, from about 6.0 to about 6.8, or from about 6.2 to about 6.6.

[0214] The whey protein solution or whey protein retentate is subjected to a heat treatment as shown in Figure 1. Heat treatment is applied to obtain the required denaturation.

[0215] In certain embodiments, the denaturing heat treatment step is carried out under conditions of high wall shear rate, even under laminar flow, for example at least 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 ~About 4000s -1 , or about 2000s -1 ~about 3000s -1 It is.

[0216] In certain embodiments, the denaturing heat treatment step comprises heating the aqueous whey protein solution or whey protein retentate to greater than about 70° C. under turbulent flow conditions, e.g., under turbulent flow conditions with a Reynolds number of at least 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 5000. In some such embodiments, the Reynolds number is from about 2000 to about 2500 or from about 2100 to about 2300.

[0217] Turbulent flow is defined as having sufficient mass flow rate in the heated tube to provide a Reynolds number (referred to as Re) of at least 2000. Such Reynolds numbers are characteristic of turbulent flow and are known in the field of fluid mechanics. The determination of Re depends on the mass velocity of the fluid and the maximum viscosity at the target heating temperature, which is defined as the nominal viscosity determined 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 a fluid heat treated at a uniform temperature before drying, using the Hagen-Poiseuille equation. To calculate Re for a given process, the equation for a Newtonian fluid can be used, provided that the maximum viscosity of the heat treated fluid at the target temperature is used. This means that Re at all other viscosities (i.e. lower viscosities) is higher and falls within the turbulent region.

[0218] In certain embodiments, the denaturing heat treatment step comprises heating the aqueous whey protein solution or whey protein retentate in combination with mechanical shear to above about 50° C. In some such embodiments, homogenizers, colloid mills, high pressure pumps, scraped surface heat exchangers, high shear mixers, and the like are used to mix the solution or to break down aggregates as they form or after they have formed.

[0219] In certain embodiments, the denaturing heat treatment step comprises heating the aqueous whey protein solution or whey protein retentate to a temperature of about 70° C. to about 90° C. at a pH between 6 and 7 under conditions of sufficiently high shear stress / force resulting from increasing serum viscosity, high wall shear rate, turbulence, mechanical shear, or a combination of two or more thereof. In some such embodiments, the denaturing heat treatment step comprises heating the aqueous whey protein solution or whey protein retentate to a temperature of about 80° C. to about 85° C. at a pH between 6.3 and 6.7 under conditions of sufficiently high shear stress / force resulting from increasing serum viscosity, high wall shear rate, turbulence, mechanical shear, or a combination of two or more thereof.

[0220] Known factors such as heating temperature and pH can affect protein aggregation, however, as reported herein, variation in temperature and pH within the defined ranges does not dramatically affect particle size, indicating that the methods disclosed herein are robust and reproducible.

[0221] In certain embodiments, the aqueous whey protein solution or whey protein retentate is substantially free of oxidizing agents during the heat treatment step that may cause protein denaturation, e.g., the oxidizing agents may be inactivated, consumed, and / or removed prior to the heat treatment step.

[0222] One of the main mechanisms of protein aggregation is considered to be the formation of intermolecular disulfides accompanied by sulfhydryl-disulfide exchange. Without wishing to be bound by theory, it is believed that pretreatment with oxidizing agents affects aggregation pathways and kinetics by promoting intermolecular sulfhydryl-disulfide exchange. Without wishing to be bound by theory, it is believed that the microparticles disclosed herein are primarily disulfide-bonded with low levels of thiols. Additionally or independently, limiting the pre-denaturation of whey protein (denaturation level before the denaturation heat treatment step) may also enable the formation of small particles.

[0223] After the denaturing heat treatment step, the denatured whey protein composition may be dried or, alternatively, the denatured whey protein composition may be used directly in the preparation of a high protein product without being dried.

[0224] In certain embodiments, the denatured whey protein compositions are not subjected to particle size selection procedures, such as particle size reduction or further microfiltration, to achieve the particle size distribution described herein.

[0225] E. Uses and Methods of Use

[0226] In at least one aspect, the present disclosure provides ingredients for use in the preparation of liquid nutritional compositions, such as edible consumer products, beverages, or medical foods. In some such embodiments, the ingredients are provided in liquid or dry (powder) form, or a mixture thereof.

[0227] Examples of suitable edible consumer products include, but are not limited to, baked products (e.g., muffins, cookies, brownies), puffed and / or extruded snack products, protein-enriched snacks, confectionery products (including chocolates, gels, ice cream, caramel, snack bars, spreads, sauces, dips), dairy products (including drinking yogurt (including ambient drinking yogurt), stirred yogurt, and cheese), food product additives such as protein sprinkles, and nutritional supplement products (including daily tablets).

[0228] An exemplary edible consumer product is a solid set gel, such as a set yogurt. A set yogurt prepared using the denatured whey protein compositions provided herein may exhibit reduced firmness compared to a control yogurt having the same ingredient composition and protein content, provided that the control yogurt does not include the denatured whey protein compositions provided herein.

[0229] Another exemplary edible consumer product is a semi-solid food product, such as stirred yogurt. Stirred yogurt prepared using the denatured whey protein compositions provided herein may exhibit reduced viscosity compared to a control yogurt having the same ingredient composition and protein content.

[0230] In certain embodiments, the set or stirred yogurt exhibits a reduced gritty texture in the mouth (or reduced grittiness) and / or a minimal increase in undesirable flavors or no increase in undesirable flavors compared to a suitable control yogurt.

[0231] Examples of suitable beverages include, but are not limited to, ready-to-drink fermented (RTD) beverages, as well as aqueous liquids (e.g., concentrates) that may be diluted, and powders that may be reconstituted to provide such beverages. In certain embodiments, the beverage is drinking yogurt (including ambient drinking yogurt), milk, milk powder, sports supplements, including dairy and non-dairy based sports supplements, fruit juice, or formulas, such as infant formulas, follow-on formulas, or growing-up formulas, in powder or liquid form. In certain embodiments, the beverage is a neutral high-protein beverage. In certain embodiments, the beverage is an acidic high-protein beverage.

[0232] An exemplary drink is drinking yoghurt. Drinking yoghurt is a low viscosity fermented milk drink. The drinking yoghurt may be an ambient drinking yoghurt.

[0233] Another exemplary beverage is a sports drink, which typically contains a high protein content, has a low fat content, and has added vitamins, minerals, flavors, sweeteners, stabilizers, and / or salt.

[0234] In certain embodiments, the denatured whey protein compositions disclosed herein are used as ingredients in the preparation of nutritional compositions, preferably liquid nutritional compositions (e.g., medical foods). By way of example, U.S. federal law and Food and Drug Administration regulations define a medical food as "a food product formulated to be ingested 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 based on recognized scientific principles have been established by medical evaluation" (§5(b) of the Orphan Drug Act of 1988 (21 U.S.C. 360ee(b)(3)) and FDA regulations 21 CFR 101.9(j)(8)). In some such embodiments, the liquid nutritional composition further comprises a fat and / or carbohydrate component.

[0235] In certain embodiments, the denatured whey protein compositions disclosed herein are incorporated into a nutritional composition by wet mixing. In some such embodiments, the nutritional composition is a wet mixed nutritional composition, such as a wet mixed food replacement composition or a wet mixed infant formula.

[0236] The denatured whey protein compositions disclosed herein can be useful for the manufacture of powder compositions or ready-to-mix compositions, particularly wet mix compositions, wet mix meal replacement compositions, and / or wet mix infant formulas.Wet mix compositions are compositions prepared using a wet mix process, in which two or more ingredients are combined as an aqueous solution, and optionally subsequently dried into a powder.Wet mix may be necessary, for example, for ready-to-mix nutritional compositions, which require a wet mix process to incorporate a fat source into the composition.

[0237] The modified whey protein composition disclosed herein is particularly useful for wet blend applications, because it can achieve high total solids content during evaporation due to low composition viscosity.When the solids content is high during evaporation, less water must be removed during spray drying, and energy usage is accordingly reduced.In addition, the thermal stability of the modified whey protein composition disclosed herein can reduce line fouling during evaporation and drying, thereby reducing production downtime and increasing plant throughput and product yield.

[0238] Methods for preparing wet mix compositions are known to those skilled in the art. Briefly, the method for preparing wet mix compositions may include combining two or more materials 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 include the thermostable protein composition of the present invention and one or more additional materials, optionally selected from one or more lipids, one or more carbohydrates, one or more additional materials described herein, or any combination thereof.

[0239] In at least one aspect, the present disclosure provides a method for providing nutritional support to a subject, comprising enterally administering to the subject a nutritional composition comprising the denatured whey protein composition disclosed herein. In certain embodiments, the subject is a subject in need of nutritional support. Thus, the nutritional compositions described herein are for use in a method for providing nutritional support to a subject in need of nutritional support. In some such embodiments, the subject is a human. In some such embodiments, the subject is a human infant or toddler. In other such embodiments, the subject is an adult. In some such embodiments, the subject is a pregnant woman. Enteral administration may be orally or via a tube (e.g., nasogastric tube feeding or gavage).

[0240] 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 increase muscle mass loss, maintain or increase growth, prevent or reduce muscle catabolism, prevent or treat cachexia, prevent or treat sarcopenia, increase glycogen resynthesis rate, 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, increase recovery rate after surgery, increase prehabilitation effect before surgery or chemotherapy, increase recovery rate after injury, increase recovery rate after exercise, increase sports performance, and / or provide nutrition. Enteral administration may be oral or via a tube (e.g., nasogastric tube or gastric tube).

[0241] In certain embodiments, nutritional compositions comprising the denatured whey protein compositions disclosed herein are for use to maintain or increase muscle protein synthesis, maintain or increase muscle mass, prevent or increase muscle mass loss, maintain or increase 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 concentrations, reduce satiety, increase food intake, increase caloric intake, improve glucose metabolism, increase the rate of recovery after surgery, increase the effect of prehabilitation before surgery or chemotherapy, increase the rate of recovery after injury, increase the rate of recovery after exercise, increase sports performance, and / or provide nutrition.

[0242] F. High Protein Liquid Compositions

[0243] In at least one aspect, the present disclosure provides a high-protein liquid composition comprising denatured whey protein.The high-protein liquid composition can be an acidic or neutral pH composition, including a neutral pH high-protein beverage or an acidic high-protein beverage, a drinking yogurt, or an acidic or neutral pH liquid nutritional composition, each of which is discussed in more detail herein.In certain embodiments, the high-protein liquid composition comprises at least 6% (w / v) denatured whey protein. In some such embodiments, the high protein liquid composition comprises at least 7% (w / v), at least 8% (w / v), at least 9% (w / v), at least 10% (w / v), at least 11% (w / v), at least 12% (w / v), at least 13% (w / v), at least 14% (w / v), at least 15% (w / v), at least 16% (w / v), at least 17% (w / v), at least 18% (w / v), at least 19% (w / v), at least 20% (w / v), at least 21% (w / v), at least 22% (w / v) denatured whey protein. In some such embodiments, the liquid composition comprises between 6% and 30% (w / v) denatured whey protein, e.g., between 7% and 28% (w / v) denatured whey protein, between 8% and 26% (w / v) denatured whey protein, or between 10% and 24% (w / v) denatured whey protein.

[0244] In certain embodiments, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, or at least 95% of the total protein in the high protein liquid composition is denatured whey protein. In some such embodiments, substantially all of the total protein in the high protein liquid composition is denatured whey protein. In other such embodiments, the high protein liquid composition comprises other dairy proteins and / or non-dairy proteins. In other such embodiments, the high protein liquid composition comprises whey protein from two or more sources, such as a heat denatured whey protein composition, a non-denatured whey protein composition, a whey protein hydrolysate, or a material that contains both whey and casein (such as an MPC), and the total whey protein in the composition is the sum of the total whey protein present in the heat denatured whey protein composition, the non-denatured whey protein composition, the whey protein hydrolysate, and / or the MPC.

[0245] In certain embodiments, the high protein liquid composition comprises at least 5%, at least 10%, at least 15%, or at least 20% (w / v) total protein. The total protein in a liquid composition is the sum of all proteins contributed by all protein-containing materials in the composition. For example, in an embodiment where the liquid composition comprises a heat-denatured whey protein composition and other non-dairy and / or dairy proteins, e.g., from skim milk powder (SMP) or milk protein concentrate (MPC), the total protein in the liquid composition is the sum of the total proteins present in the heat-denatured whey protein composition and the SMP and / or MPC. In some such embodiments, the liquid composition comprises at least 7% (w / v), at least 8% (w / v), at least 9% (w / v), at least 10% (w / v), at least 11% (w / v), at least 12% (w / v), at least 13% (w / v), at least 14% (w / v), at least 15% (w / v), at least 16% (w / v), at least 17% (w / v), at least 18% (w / v), at least 19% (w / v), at least 20% (w / v), at least 21% (w / v), at least 22% (w / v) total protein. In some such embodiments, the liquid composition comprises between 6% and 30% (w / v) total protein, e.g., between 7% and 28% (w / v) total protein, between 8% and 26% (w / v) total protein, or between 10% and 24% (w / v) total protein. In some such embodiments, the high protein liquid composition comprises 10% (w / v), 11% (w / v), 12% (w / v), 13% (w / v), 14% (w / v), 15% (w / v), 16% (w / v), 17% (w / v), 18% (w / v), 19% (w / v), 20% (w / v), 21% (w / v), or 22% (w / v) total protein.

[0246] In certain embodiments, less than about 5%, less than about 4%, less than about 3%, less than about 2%, less than about 1%, or less than about 0.1% of the total protein in the high protein liquid composition is casein. In some such embodiments, up to 5%, up to 4%, up to 3%, up to 2%, up to 1%, or up to 0.1% of the total protein in the high protein liquid composition is casein. In some such embodiments, the high protein liquid composition is substantially free of casein. In certain embodiments, the high protein liquid composition comprises 12% (w / v) or less total protein, e.g., 5% (w / v), 6% (w / v), 7% (w / v), 8% (w / v), 9% (w / v), 10% (w / v), or 11% (w / v) total protein. In some such embodiments, at least 90% or at least 95% of the total protein in the high protein liquid composition is denatured whey protein. In some such embodiments, substantially all of the total protein in the high protein liquid composition is denatured whey protein.

[0247] In certain embodiments, the high protein liquid composition comprises at least 12% (w / v) total protein, e.g., 12% (w / v), 13% (w / v), 14% (w / v), 15% (w / v), 16% (w / v), 17% (w / v), 18% (w / v), 19% (w / v), or 20% (w / v) total protein. In some such embodiments, the high protein liquid composition comprises between 12% and 18% (w / v) total protein. In some such embodiments, at least 90% or at least 95% of the total protein in the high protein liquid composition is denatured whey protein. In some such embodiments, substantially all of the total protein in the high protein liquid composition is denatured whey protein.

[0248] In certain embodiments, the average particle size (characterized by D(4,3)) of the high protein liquid composition after sterilization or pasteurization is less than 5 μm, less than 4 μm, less than 3 μm, less than 2 μm, or less than 1 μm. In some such embodiments, the average particle size (characterized by D(4,3)) of the high protein liquid composition comprising 7%-25% (w / v) total protein is about 0.1 μm to about 5 μm or about 0.5 μm to about 3 μm, preferably about 1 μm. In some such embodiments, the average particle size (characterized by D(4,3)) of the high protein liquid composition comprising 9%-20% (w / v) total protein is less than 1 μm.

[0249] In certain embodiments, the liquid composition is sterilized and / or pasteurized in a manner suitable for commercial use. In some such embodiments, the sterilization and / or pasteurization comprises a thermal (i.e., heating) treatment. In other such embodiments, the sterilization and / or pasteurization comprises a non-thermal treatment. Exemplary techniques of sterilization and / or pasteurization include, but are not limited to, high temperature pasteurization, ultra-high temperature (UHT) treatment, and retort heat treatment. In some such embodiments, the particle size distribution of the liquid composition after such sterilization or pasteurization is D(4,3) less than 1 μm, d ... 50 , and d less than 5 μm 90 In some such embodiments, the particle size distribution of the liquid composition after such sterilization or pasteurization includes a D(4,3) of less than 1 μm, a d(4,5) of less than 1 μm, 50 , and d less than 3 μm 90 Includes.

[0250] It is well known that the lethal effect of high temperature on microorganisms depends on both temperature and holding time, with the time required to kill the same number of microorganisms decreasing as the temperature increases. The time it takes to reduce an initial number of microorganisms by a particular amount at a specified temperature is commonly referred to as the "F value." The F value is the equivalent time (in minutes) for a specified temperature that gives the same thermal lethality as 121°C. As is known in the relevant art, acidic products generally require a lower F value than products processed at neutral pH to achieve commercial sterilization.

[0251] In certain embodiments, the liquid composition is shelf-stable after sterilization and / or pasteurization. In some such embodiments, the liquid composition exhibits minimal precipitation, gelling, or clumping, or essentially no precipitation, gelling, or clumping, and negligible bacterial growth, when aseptically packaged after extended 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 essentially no chalkiness or grittiness. Thus, for example, the liquid composition can be a heat-treated shelf-stable liquid composition.

[0252] In certain embodiments, the liquid composition exhibits minimal or essentially no settling under conditions favoring settling.

[0253] In certain embodiments, the high-protein liquid composition comprises non-whey and / or non-dairy proteins in addition to the denatured whey protein. Exemplary sources of non-whey proteins include, but are not limited to, skim milk powder (SMP), whole milk powder, milk protein concentrate (MPC) or milk protein isolate (MPI), or micellar casein concentrate (MCC). As another example, casein may be included in the high-protein liquid composition in the form of sodium caseinate, potassium caseinate, calcium caseinate, or magnesium caseinate.

[0254] In certain embodiments, the high protein 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 algae proteins, fungal proteins (e.g., mycoproteins), plant proteins, and animal proteins, and hydrolyzed forms thereof. In some such embodiments, the liquid composition comprises soy protein, rice protein, and / or pea protein.

[0255] Furthermore, the high protein liquid composition may contain whey protein from two or more sources.For example, the liquid composition may contain a mixture of WPC and WPI, one or both of which are heat denatured.As another example, the liquid composition may contain a mixture of WPCs that are produced in different ways or have different properties.

[0256] In addition to the methods disclosed herein, exemplary methods for preparing thermally denatured whey protein compositions suitable for use in liquid compositions are described in PCT International Application Nos. PCT / NZ2007 / 000059 (published as WO2007 / 108709) and PCT / NZ2010 / 000072 (published as WO2010 / 120199), as well as PCT International Application No. PCT / IB2012 / 056103 (published as WO2013 / 065014), each of which is incorporated herein by reference in its entirety.

[0257] In certain embodiments, one or more of the protein materials, such as the heat denatured whey protein composition or the high protein liquid composition, may be treated to reduce lactose content. In some such embodiments, the protein materials or the high protein liquid composition are treated with an enzyme such as beta-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.

[0258] In certain embodiments, the high protein liquid composition comprises less than about 10% (w / v), less than about 9% (w / v), less than about 8% (w / v), less than about 7% (w / v), less than about 6% (w / v), less than about 5% (w / v), less than about 4% (w / v), less than about 3% (w / v), less than about 2% (w / v), less than about 1% (w / v), or less than about 0.1% (w / v) lactose. In some such embodiments, the high protein liquid composition has a lactose content of up to 10% (w / v), up to 8% (w / v), up to 6% (w / v), or up to 4% (w / v).

[0259] Exemplary high-protein liquid compositions may also include fat and / or carbohydrate in addition to protein. In certain embodiments, the high-protein liquid composition also includes a lipid component. In some such embodiments, the lipid component is present in an amount of up to 30% (w / v). In certain embodiments, the high-protein liquid composition also includes a carbohydrate component. In some such embodiments, the carbohydrate component is present in an amount of up to 30% (w / v), and in certain embodiments, the high-protein liquid composition has a caloric density of 1 kcal / ml or less, 1-2 kcal / ml, or more than 2 kcal / ml.

[0260] In certain embodiments, the high protein liquid composition comprises at least one monovalent cation and / or at least one divalent metal cation, hi some such embodiments, the high protein liquid composition comprises at least 30 mg / 100 mL, at least 50 mg / 100 mL, or at least 100 mg / 100 mL of a divalent metal cation.

[0261] Unless expressly stated to the contrary, each feature of the high protein liquid composition described herein may be combined with other features. In particular, any feature indicated as being preferred or advantageous may be combined with other features indicated as being preferred or advantageous. One example of a liquid composition is a liquid nutritional composition. A liquid nutritional composition can provide enteral nutrition by oral administration or by administration through a tube (e.g., nasogastric or gastric tube feeding). In addition to protein, an exemplary liquid nutritional composition also typically contains fat and / or carbohydrate at a level and combination to achieve a caloric value of at least 0.5 kcal / g or kcal / ml. For example, a liquid nutritional composition may have a caloric density of up to 3 kcal / g or more. Until now, it has been difficult to achieve such a high caloric density with low viscosity and sufficient protein.

[0262] In certain embodiments, the liquid nutritional composition also includes a lipid component. The lipid component may include vegetable lipids or animal lipids (e.g., bovine, porcine, etc.), including dairy lipids and fish oils. In some such embodiments, vegetable oils are preferred due to their ease of formulation and low saturated fatty acid content. Exemplary vegetable oils include canola (rapeseed) oil, corn oil, sunflower oil, olive oil, or soybean oil.

[0263] In various embodiments, the liquid nutritional composition comprises at least about 0.01, 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or at least about 30 grams of lipid per 100 mL of composition, and various ranges can be selected between these values, for example, from about 0.01 to about 30, from about 0.1 to about 30, from about 1 to about 30, or from about 1 to about 30, per 100 mL of composition. , about 0.01 to about 25, about 0.1 to about 25, about 1 to about 25, about 0.01 to about 20, about 0.1 to about 20, about 0.5 to about 20, about 1 to about 20, about 2 to about 20, about 3 to about 20, about 5 to about 20 g, about 0.01 to about 15, about 0.1 to about 15, about 1 to about 15, about 2 to about 15, about 5 to about 15, about 0.01 to about 10, about 0.1 to about 10, about 1 to about 10, or about 2 to about 10 g of lipid.

[0264] In certain embodiments, the liquid nutritional composition also includes a carbohydrate component. The carbohydrate component may include easily digestible carbohydrates, indigestible carbohydrates, or a combination thereof. The carbohydrate component may include monosaccharides, disaccharides, oligosaccharides, polysaccharides, and mixtures thereof. Suitable oligosaccharides include, but are not limited to, glucose oligosaccharides. Suitable non-digestible carbohydrates include, but are not limited to, fructooligosaccharides, inulin, galactooligosaccharides, and the like.

[0265] In certain embodiments, the liquid nutritional composition comprises at least about 0.01, 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40 g, or at least about 45 g carbohydrate per 100 mL of the composition, and various ranges can be selected between these values, for example, from about 0.01 to about 4, from about 0.01 to about 45, from about 0.1 to about 45, from about 1 to about 45, about 4 to about 45, about 5 to about 45, about 0.01 to about 40, about 0.1 to about 40, about 1 to about 40, about 4 to about 40, about 5 to about 40, about 0.01 to about 30, about 1 to about 30, about 1 to about 30, about 4 to about 30, about 5 to about 30, 0.01 to about 20, about 0.1 to about 20, about 0.5 to about 20, about 0.1 to about 15, about 1 to about 20, about 2 to about 20, about 3 to about 20, about 4 to about 20, or about 5 to about 20 g of carbohydrates.

[0266] In certain embodiments, the liquid nutritional composition also includes a stabilizer or emulsifier. Suitable emulsifiers include lecithin, mono- and diglycerides, polyglycerol esters, milk phospholipids, citric acid esters (CITREM), and diacetyl tartaric acid esters of mono- and diglycerides (DATEM). These emulsifiers may be added in amounts of about 0.003 g to about 0.06 g per gram of lipid. Suitable stabilizers include, but are not limited to, carrageenan, gellan gum, pectin, guar gum, locust bean gum, xanthan gum, carboxymethylcellulose, and microcrystalline cellulose, and combinations thereof. One skilled in the art will recognize that many different gum forms, in addition to those listed above, are suitable for use in the liquid compositions disclosed herein.

[0267] In certain embodiments, the liquid nutritional composition also comprises a source of amino acids, amino acid precursors, amino acid metabolites, or any combination of any two or more thereof, preferably a source of free amino acids, amino acid precursors, or amino acid metabolites.

[0268] In certain embodiments, the liquid nutritional composition also contains vitamins and / or minerals necessary to maintain the nutrition of the patient for a long period of time, as well as trace ingredients such as antioxidants, flavorings, and / or colorings. The amount of vitamins and / or minerals used in the liquid nutritional composition is a typical amount for meal replacement products known to those skilled in the art. The micronutrient requirements of various subgroups of the population are also known. Recommended daily requirements of vitamins and minerals can be identified for various subgroups of the population. 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).

[0269] In certain embodiments, the liquid nutritional composition is a neutral pH liquid nutritional composition comprising at least 12% (w / v) total protein, for example, 12%-18% (w / v) total protein. In some such embodiments, at least 90% or at least 95% of the total protein in the neutral pH liquid nutritional composition is denatured whey protein. In some such embodiments, substantially all of the total protein in the neutral pH liquid nutritional composition is denatured whey protein. Thus, in certain embodiments, the neutral pH liquid nutritional composition may comprise 12%-18% (w / v) total protein, with substantially all of the total protein coming from denatured whey protein. In some such embodiments, the neutral pH liquid nutritional composition comprises fat and / or carbohydrate at a level and combination to achieve a caloric density of 1-3 kcal / ml, for example, 1.5-2.5 kcal / ml. An exemplary neutral pH liquid nutritional composition comprises 12% (w / v) total protein, with substantially all of the total protein coming from denatured whey protein, and has a caloric density of 1.60 kcal / ml. Another exemplary neutral pH liquid nutritional composition comprises 15% (w / v) total protein, substantially all of which is from denatured whey protein, and has a caloric density of 2.25 kcal / ml.

[0270] Another exemplary liquid composition is a high protein drinking yogurt.Methods for producing liquid compositions such as drinking yogurt are generally known in the art.

[0271] 3 shows an exemplary process for producing drinking yogurt. In an exemplary process, the ingredients (e.g., skim milk powder and whey protein composition) are mixed and added to water at 60°C. The mixture is then homogenized and heated, for example, to 95°C for 6 minutes, to 85°C for 15 minutes, or to 80°C for 20 minutes. The mixture is then cooled to, for example, 38°C or 42°C. The mixture is then inoculated with a bacterial starter culture. The mixture is then fermented until at least the pH is less than 4.6, after which it is cooled to 20°C. The gel thus formed is broken, sheared, and then packed and cooled to 4°C.

[0272] In a particular embodiment, the high protein drinking yogurt comprises 15% total protein and the drinking yogurt comprises at least 12% denatured whey protein (i.e., at least 80% (w / w) of the total protein is derived from denatured whey protein). In a particular embodiment, the high protein drinking yogurt comprises 20% total protein and the drinking yogurt comprises at least 17% denatured whey protein (i.e., at least 85% (w / w) of the total protein is derived from denatured whey protein).

[0273] In certain embodiments, the high protein drinking yoghurt is pasteurised and / or pasteurised, for example the drinking yoghurt may be pasteurised.

[0274] In certain embodiments, the average particle size (characterized by D(4,3)) of the drinking yogurt is less than 3.0 μm, less than 2.5 μm, less than 2.0 μm, or less than 1.5 μm. In some such embodiments, the average particle size (characterized by D(4,3)) of the drinking yogurt is from about 0.6 μm to about 1.5 μm, or from about 0.8 μm to about 1.3 μm, or from about 0.9 μm to about 1.2 μm. In some such embodiments, the average particle size (characterized by D(4,3)) of the drinking yogurt (wherein at least 75%, at least 80%, or at least 85% of the total protein of the drinking yogurt is from the denatured whey protein composition) is from about 0.6 μm to about 1.5 μm, or from about 0.8 μm to about 1.3 μm, or from about 0.9 μm to about 1.2 μm.

[0275] In a particular embodiment, the drinking yogurt 50 In some such embodiments, the d of the drinking yogurt is less than 2.0 μm, less than 1.5 μm, or less than 1.0 μm. 50 In some such embodiments, the d of the drinking yogurt (wherein at least 75%, at least 80%, or at least 85% of the total protein of the drinking yogurt is from the denatured whey protein composition) is from about 0.5 μm to about 1.3 μm, or from about 0.6 μm to about 1.2 μm, or from about 0.7 μm to about 1.1 μm. 50 is about 0.5 μm to about 1.3 μm, or about 0.6 μm to about 1.2 μm, or about 0.7 μm to about 1.1 μm.

[0276] In a particular embodiment, the drinking yogurt 90 In some such embodiments, the d of the drinking yogurt is less than 3.0 g, less than 2.5 g, or less than 2.0 g. 90 In some such embodiments, the d of the drinking yogurt (wherein at least 75%, at least 80%, or at least 85% of the total protein of the drinking yogurt is from the denatured whey protein composition) is from about 1.3 gm to about 2.5 gm, or from about 1.4 gm to about 2.4 gm, or from about 1.5 gm to about 2.3 gm. 90is about 1.3 μm to about 2.5 μm, or about 1.4 μm to about 2.4 μm, or about 1.5 μm to about 2.3 μm.

[0277] In certain embodiments, the high protein drinking yogurt exhibits minimal or essentially no sedimentation over the shelf life of the product, e.g., up to 6 weeks for refrigerated drinking yogurt and up to 6-9 months for ambient drinking yogurt. For example, the high protein drinking yogurt exhibits minimal or essentially no sedimentation over the shelf life of the product (e.g., up to 6 weeks). In some such embodiments, the high protein drinking yogurt exhibits less than 20% sedimentation over the shelf life of the product. In some such embodiments, the high protein drinking yogurt exhibits less than 15%, less than 12%, less than 10%, less than 8%, or less than 6% sedimentation over the shelf life of the product. In some such embodiments, the high protein drinking yogurt exhibits about 2% to about 6% or about 3% to about 5% sedimentation over the shelf life of the product.

[0278] As another example, centrifugation tests can be used to assess sedimentation. Thus, in some such embodiments, the high protein drinking yogurt exhibits less than 20% sedimentation after centrifugation at 1540×g for 5 minutes. In some such embodiments, the liquid composition exhibits less than 15%, less than 12%, less than 10%, less than 8%, or less than 6% sedimentation after centrifugation. In some such embodiments, the liquid composition exhibits about 2% to about 6% or about 3% to about 5% sedimentation after centrifugation.

[0279] In certain embodiments, the viscosity of the high protein drinking yogurt 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. In certain embodiments, the viscosity of the high protein drinking yogurt is less than about 300 mPa·s, less than about 200 mPa·s, or less than about 100 mPa·s. In some such embodiments, the drinking yogurt has a viscosity of about 20 to about 400 mPa·s. In some such embodiments, the high protein drinking yogurt comprising 10% to 20% (w / v) total protein has a viscosity of about 20 to about 400 mPa·s, about 20 to about 300 mPa·s, about 20 to about 200 mPa·s, about 20 to about 100 mPa·s, about 30 to about 90 mPa·s, or about 40 to about 80 mPa·s.

[0280] The viscosity of a sample may be measured by methods known in the art. For example, one method uses an MCR302 rheometer (Anton-Paar) and subjects the sample to a pre-shear (e.g., 300 s -1 The sample was subjected to a shear rate sweep (e.g., from 0.001 to 398 s for 1 min) followed by a rest period. -1 Up to (50s -1 and / or 100s -1 Alternatively, the apparent viscosity can be measured at 10° C. using a Haake viscometer (Haake Mess-Technik, GmbH & Co., Karlsruhe, Germany). Viscosity measurements are performed by varying the shear rate from 0 to 120 s over a period of 180 s. -1 The shear rate was then increased to 0 s for 30 s. -1 The apparent viscosity is reduced to 50s. -1 The shear rate is measured at 100 .mu.m.

[0281] The compositions, methods, and uses described herein will be better understood by reference to the following illustrative embodiments and examples, which are included as illustrations of, but not intended to limit, the scope of the invention.

[0282] G. High Protein Set Yogurt and Stirred Yogurt

[0283] In at least one embodiment, the present disclosure provides a high-protein food product comprising denatured whey protein.The high-protein food product can be yoghurt, such as set yoghurt and stirred yoghurt, each of which is discussed in more detail herein.The methods for producing set yoghurt and stirred yoghurt are generally known in the art.

[0284] In certain embodiments, the high protein food product comprises at least 6% (w / w) denatured whey protein. In some such embodiments, the high protein food product comprises at least 7% (w / w), at least 8% (w / w), at least 9% (w / w), at least 10% (w / w), at least 11% (w / w), at least 12% (w / w), at least 13% (w / w), at least 14% (w / w), or at least 15% (w / w) denatured whey protein. In some such embodiments, the food product comprises between 6% and 30% (w / w) denatured whey protein, e.g., between 7% and 25% (w / w) denatured whey protein, between 8% and 20% (w / w) denatured whey protein, or between 10% and 15% (w / w) denatured whey protein.

[0285] In certain embodiments, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, or at least 95% of the total protein in the high protein food product is denatured whey protein. In some such embodiments, substantially all of the total protein in the high protein food product is denatured whey protein. In other such embodiments, the high protein food product comprises other dairy proteins and / or non-dairy proteins. In other such embodiments, the high protein food product comprises whey protein from two or more sources, such as a thermally denatured whey protein composition, a non-denatured whey protein composition, a whey protein hydrolysate, or a material that contains both whey and casein (such as an MPC), and the total whey protein in the composition is the sum of the total whey protein present in the thermally denatured whey protein composition, the non-denatured whey protein composition, the whey protein hydrolysate, and / or the MPC.

[0286] In certain embodiments, the high protein food product comprises at least 5%, at least 10%, at least 15%, or at least 20% (w / w) total protein. The total protein in a food product is the sum of all protein contributed by all protein-containing materials in the product. For example, in embodiments where the food product comprises a thermally denatured whey protein composition and other non-dairy and / or dairy proteins, e.g., from skim milk powder (SMP) or milk protein concentrate (MPC), the total protein in the food product is the sum of the total protein present in the thermally denatured whey protein composition and the SMP and / or MPC. In some such embodiments, the food product comprises at least 7% (w / w), at least 8% (w / w), at least 9% (w / w), at least 10% (w / w), at least 11% (w / w), at least 12% (w / w), at least 13% (w / w), at least 14% (w / w), or at least 15% (w / w) total protein. In some such embodiments, the food product comprises between 6% and 30% (w / w) total protein, for example between 8% and 25% (w / w) total protein, or between 10% and 15% (w / w) total protein. In some such embodiments, the high protein food product comprises 10% (w / w), 11% (w / w), 12% (w / w), 13% (w / w), 14% (w / w), 15% (w / w), 16% (w / w), 17% (w / w), 18% (w / w), 19% (w / w), or 20% (w / w) total protein.

[0287] In certain embodiments, the food product is sterilized and / or pasteurized in a manner suitable for commercial use. In some such embodiments, the sterilization and / or pasteurization comprises a thermal (i.e., heat) treatment. In other such embodiments, the sterilization and / or pasteurization comprises a non-thermal treatment.

[0288] In certain embodiments, the high protein food product comprises a set gel, such as a set yogurt. In some such embodiments, the set gel comprises at least 6% (w / w), at least 7% (w / w), at least 8% (w / w), at least 9% (w / w), at least 10% (w / w), at least 11% (w / w), at least 12% (w / w), at least 13% (w / w), at least 14% (w / w), or at least 15% (w / w) whey protein. The whey protein comprises microparticles comprising denatured whey protein.

[0289] In some such embodiments, the set gel exhibits a hardness of less than 1000 g·sec, or less than 900 g·sec, or less than 800 g·sec, or less than 700 g·sec, hi some such embodiments, the set gel exhibits a breaking strength of less than 50 g, less than 45 g, or less than 40 g.

[0290] In certain embodiments, the high protein food product is a semi-solid food product, such as stirred yogurt. In some such embodiments, the particle size distribution of the semi-solid food product after sterilization or pasteurization is less than 1 μm. 50 and d less than 5 μm 90 In some such embodiments, the semi-solid food product comprises at least 6% (w / w), at least 7% (w / w), at least 8% (w / w), at least 9% (w / w), at least 10% (w / w), at least 11% (w / w), at least 12% (w / w), at least 13% (w / w), at least 14% (w / w), or at least 15% (w / w) whey protein. The whey protein comprises microparticles comprising denatured whey protein.

[0291] In some such embodiments, the semi-solid food product is storable at 20° C. for 50 s. -1 The viscosity of the composition is less than 500 mPa.s or less than 400 mPa.s, as measured at 100.degree.

[0292] H. Exemplary Embodiments

[0293] In one aspect, the present disclosure provides a heat stable denatured whey protein composition comprising microparticles comprising denatured whey protein, wherein at least 50% or at least 55% by volume of the particles have a diameter between 0.2 μm and 1.0 μm.

[0294] In another aspect, the disclosure provides a heat stable denatured whey protein composition comprising microparticles comprising denatured whey protein, wherein the microparticles have a volume weighted mean diameter D(4,3) of about 1.0 μm or less, with at least 95% of the particles by volume having a diameter less than about 2.0 μm.

[0295] In certain embodiments of any aspect disclosed herein, the disclosure provides a heat stable denatured whey protein composition comprising microparticles comprising denatured whey protein, wherein the microparticles exhibit minimal or essentially no primary particle size growth after heating an aqueous solution of 10% (w / w) protein content (pH 6.8) at 120° C. for 15 minutes. In some such embodiments, the d of an aqueous solution of 10% (w / w) protein content after a primary particle growth test comprising heat treatment at 120° C. for 15 minutes is 50 In some such embodiments, the d of a 10% (w / w) protein content aqueous solution after a primary particle growth test including a heat treatment at 120° C. for 15 minutes is about 1.0 μm or less, preferably about 0.6 to about 1.0 μm. 90 is less than about 2.0 μm, preferably about 0.9 to about 1.7 μm or about 1.1 to about 1.5 μm.

[0296] In certain embodiments of any aspect disclosed herein, the whey protein has a degree of hydrolysis of less than about 10%. In some such embodiments, the whey protein has a degree of hydrolysis of less than about 5%. In some such embodiments, the whey protein is substantially unhydrolyzed.

[0297] In certain embodiments of any of the aspects disclosed herein, the heat stable denatured whey protein composition comprises at least 65%, at least 70%, at least 75%, or at least 80% total protein on a dry weight basis.

[0298] In certain embodiments of any of the aspects disclosed herein, the heat stable denatured whey protein composition comprises at least 70%, at least 80%, or at least 90% (w / w) whey protein relative to the total protein.

[0299] In certain embodiments of any aspect disclosed herein, the remaining denatured whey protein in the denatured whey protein composition is less than 16%, less than 14%, or less than 12%.

[0300] In certain embodiments of any of the aspects disclosed herein, the denatured whey protein composition has an insolubility of at least 60% or at least 65%.

[0301] In certain embodiments of any aspect disclosed herein, the heat stable denatured whey protein composition has a lactose content of up to 10% by weight, up to 8% by weight, up to 6% by weight, or up to 4% by weight.

[0302] In certain embodiments of any of the aspects disclosed herein, the heat stable denatured whey protein composition has a fat content of up to 20%, up to 18%, up to 16%, up to 14%, up to 12%, up to 10%, up to 8%, or up to 6%.

[0303] In certain embodiments of any aspect disclosed herein, the heat stable denatured whey protein composition has an ash content of up to 10% by weight, up to 8% by weight, up to 6% by weight, or up to 4% by weight.

[0304] In certain embodiments of any aspect disclosed herein, the heat stable denatured whey protein composition comprises less than about 10% by weight casein.

[0305] In certain embodiments of any aspect disclosed herein, the heat stable denatured whey protein composition is substantially free of casein.

[0306] Exemplary embodiment 1. A microparticle comprising at least 60% total protein on a dry weight basis and denatured whey protein, the microparticle having (i) a volume weighted average diameter D(4,3) of about 1.0 μm or less, (ii) a d(4,3) of about 1.0 μm or less. 50 , (iii) d of about 2.0 μm or less 90 and (iv) at least 92% by volume of the particles having a diameter less than about 2.0 μm, and (v) not more than 45% by volume of the particulates having a diameter of about 1.0 to about 10.0 μm.

[0307] Exemplary embodiment 2. A particle size distribution having (i) a volume weighted average diameter D(4,3) of about 1.0 μm or less, e.g., about 0.6 to about 1.0 μm, and (iii) a volume weighted average diameter D(4,3) of about 2.0 μm or less, e.g., about 0.9 to about 1.7 μm. 90 The heat-stable denatured whey protein composition of exemplary embodiment 1, comprising:

[0308] Exemplary embodiment 3. The particle size distribution is (ii) about 1.0 μm or less, e.g., about 0.6 to about 1.0 μm d 50 (iii) about 2.0 μm or less, for example, about 0.9 to about 1.7 μm d 90 The heat-stable denatured whey protein composition of exemplary embodiment 1, comprising:

[0309] Exemplary embodiment 4. The heat stable denatured whey protein composition of exemplary embodiment 3, wherein the particle size distribution comprises: (i) a volume weighted average diameter D(4,3) of about 1.0 μm or less, e.g., about 0.6 to about 1.0 μm.

[0310] Exemplary Embodiment 5. The heat stable denatured whey protein composition of any one of Exemplary Embodiments 1-4, wherein the particle size distribution comprises: (iv) at least 92%, at least 95%, or at least 98% by volume of the particles have a diameter less than about 2.0 μm.

[0311] Exemplary embodiment 6. The heat stable denatured whey protein composition of any one of exemplary embodiments 1-5, wherein the particle size distribution comprises: (v) no more than 45% by volume of the particulates have a diameter of about 1.0 to about 10.0 μm.

[0312] Exemplary embodiment 7. The heat stable denatured whey protein composition of any one of exemplary embodiments 1-6, wherein the particle size distribution does not change substantially after heating an aqueous solution of 10% (w / w) protein content comprising the denatured whey protein composition at 120° C. for 15 minutes.

[0313] Exemplary embodiment 8.d 50 The heat stable denatured whey protein composition of any one of exemplary embodiments 1-7, wherein the average particle size is about 0.6 to about 1.0 μm after a 10% (w / w) protein content aqueous solution comprising the denatured whey protein composition is heated at 120° C. for 15 minutes.

[0314] Exemplary embodiment 9.d 90 The heat-stable denatured whey protein composition of any one of exemplary embodiments 1-8, wherein the average particle size is about 2.0 μm or less, e.g., about 0.9 to about 1.7 μm, after a 10% (w / w) protein content aqueous solution comprising the denatured whey protein composition is heated at 120° C. for 15 minutes.

[0315] Exemplary embodiment 10. The heat stable denatured whey protein composition of any one of exemplary embodiments 1 to 9, having a volume weighted mean diameter D(4,3) of about 1.0 μm or less, e.g., about 0.6 to about 1.0 μm, after a 10% (w / w) protein content aqueous solution comprising the denatured whey protein composition is heated at 120° C. for 15 minutes.

[0316] Exemplary embodiment 11. The heat stable denatured whey protein composition of any one of exemplary embodiments 1-10, wherein an aqueous solution of 10% (w / w) protein content comprising the denatured whey protein composition has a heat set time (HCT) at 140°C of at least 3 minutes, at least 4 minutes, at least 5 minutes, at least 6 minutes, at least 9 minutes, at least 12 minutes, at least 15 minutes, or at least 18 minutes.

[0317] Exemplary Embodiment 12. The heat stable denatured whey protein composition of any one of Exemplary Embodiments 1-11, wherein the composition comprises less than about 10% by weight casein.

[0318] Exemplary Embodiment 13. The heat stable denatured whey protein composition of any one of Exemplary Embodiments 1-12, wherein the composition is substantially free of casein.

[0319] Exemplary embodiment 14. The heat-stable denatured whey protein composition of any one of exemplary embodiments 1-13, wherein the composition comprises from about 30% to about 80% beta-lactoglobulin, from about 35% to about 75% beta-lactoglobulin, or from about 40% to about 65% beta-lactoglobulin relative to the total protein in the composition.

[0320] Exemplary Embodiment 15. The heat-stable denatured whey protein composition of any one of Exemplary Embodiments 1-14, wherein the composition comprises at least 70% (w / w) whey protein based on total protein, at least 80% (w / w) whey protein based on total protein, or at least 90% (w / w) whey protein based on total protein.

[0321] Exemplary embodiment 16. The heat-stable denatured whey protein composition of any one of exemplary embodiments 1-15, wherein the residual denatured whey protein in the denatured whey protein composition is less than 16%.

[0322] Exemplary embodiment 17. The heat-stable denatured whey protein composition of any one of exemplary embodiments 1-15, wherein the denatured whey protein composition has an insolubility of at least 60%.

[0323] Exemplary embodiment 18. A liquid composition comprising the heat stable denatured whey protein composition of any one of exemplary embodiments 1-17.

[0324] Exemplary embodiment 19. The liquid composition of exemplary embodiment 18, wherein the liquid composition is a drinking yogurt.

[0325] Exemplary embodiment 20. A food product comprising the heat stable denatured whey protein composition of any one of exemplary embodiments 1-17.

[0326] Exemplary embodiment 21. The food product of exemplary embodiment 20, wherein the food product is a baked food product, a bar, or a set or stirred yogurt.

[0327] Exemplary embodiment 22. The food product of exemplary embodiment 20, wherein the food product is a set yogurt, the set yogurt optionally comprising about 6% to about 20% (w / v) total protein, and optionally wherein at least 50% (w / w) of the total protein is derived from the denatured whey protein composition, and wherein the set yogurt exhibits reduced firmness and / or reduced volume weighted mean particle size compared to a control set yogurt product having the same ingredient composition and the same protein content (with the proviso that the control set yogurt product does not comprise the denatured whey protein composition of any of exemplary embodiments 1-17).

[0328] Exemplary embodiment 23. The food product of exemplary embodiment 20, wherein the food product is a stirred yogurt, the stirred yogurt optionally comprising about 6% to about 20% (w / v) total protein, and optionally wherein at least 50% (w / w) of the total protein is derived from the denatured whey protein composition, and wherein the stirred yogurt exhibits a reduced viscosity and / or a reduced volume weighted mean particle size compared to a control stirred yogurt product having the same ingredient composition and the same protein content (with the proviso that the control stirred yogurt product does not comprise the denatured whey protein composition of any of exemplary embodiments 1-17).

[0329] Exemplary embodiment 24. A nutritional composition comprising the heat stable denatured whey protein composition of any one of exemplary embodiments 1-17.

[0330] Exemplary embodiment 25. A liquid nutritional composition comprising the heat stable denatured whey protein composition of any one of exemplary embodiments 1-17.

[0331] Exemplary embodiment 26. A method for providing nutrition to a subject in need thereof, comprising administering to the subject the nutritional composition of exemplary embodiment 24 or the liquid nutritional composition of exemplary embodiment 25.

[0332] Exemplary embodiment 27. The nutritional composition of exemplary embodiment 24 or the liquid nutritional composition of exemplary embodiment 25 for use in providing nutrition to a subject in need thereof.

[0333] Exemplary embodiment 28. A method for preparing a heat stable denatured whey protein composition, comprising: (a) providing an aqueous whey protein solution or whey protein retentate; (b) contacting the aqueous whey protein solution or whey protein retentate with an oxidizing agent; and (c) subjecting the whey protein solution or whey protein retentate to a heat treatment under conditions of high shear stress / force, wherein the total protein content of the aqueous whey protein solution or whey protein retentate in step (c) is preferably at least 16% (w / w), and the high shear stress / force optionally results in an increase in serum phase viscosity, 1000 s -1 The method of claim 1, wherein the wall shear rate is generated by a turbulent flow pattern of at least 2000 Re, by application of mechanical shear, or a combination thereof.

[0334] Exemplary Embodiment 29 The method of Exemplary Embodiment 28, wherein the oxidizing agent is hydrogen peroxide or benzoyl peroxide.

[0335] Exemplary embodiment 30. The method of exemplary embodiment 29, wherein step (b) further comprises contacting the aqueous whey protein solution or whey protein retentate with an oxidizing agent in combination with an enzyme or chemical catalyst, preferably a catalyst such as a peroxidase enzyme.

[0336] Exemplary Embodiment 31. The method of any one of Exemplary Embodiments 28-30, wherein the oxidizing agent is present in step (b) in an amount of less than 300 ppm or less than 200 ppm.

[0337] Exemplary embodiment 32. The oxidizing agent in step (b) is 1200×10 per kg of whey protein. -6 kg, 2000 x 10 per kg of denatured whey protein -6 kg or 2400 x 10 per kg of β-lactoglobulin protein -6 The method of any one of Exemplary Embodiments 28-30, wherein the compound is present in an amount of less than kg.

[0338] Exemplary Embodiment 33. The method of any one of Exemplary Embodiments 28-32, wherein the oxidizing agent is inactivated, removed, or consumed prior to step (c), such that the whey protein solution or whey protein retentate is substantially free of oxidizing agent during the heat treatment of step (c).

[0339] Exemplary Embodiment 34. The method of any one of Exemplary Embodiments 28-33, wherein the aqueous whey protein solution or whey protein retentate has a low level of denaturation prior to step (c).

[0340] Exemplary embodiment 35. The method of exemplary embodiment 34, wherein the level of denaturation of the whey protein solution or whey protein retentate prior to step (c) is less than 10%, for example, from about 3% to about 8%.

[0341] Exemplary embodiment 36. The method of any one of exemplary embodiments 28-35, wherein the heat-stable denatured whey protein composition is the heat-stable denatured whey protein composition of any one of exemplary embodiments 1-17.

[0342] Exemplary embodiment 37. A heat stable denatured whey protein composition prepared by the method of any one of exemplary embodiments 28-35.

[0343] Exemplary embodiment 38. A method for preparing a liquid composition, comprising combining a thermostable denatured whey protein composition of any one of exemplary embodiments 1-17 and exemplary embodiment 37 with at least one additional component, such as a lipid component or a carbohydrate component, and heating the liquid composition to a temperature above 80°C to inhibit microbiological activity.

[0344] Exemplary embodiment 39. A heat-treated shelf-stable high protein liquid composition comprising at least 6% (w / v) whey protein, the whey protein comprising microparticles comprising denatured whey protein, (i) the liquid composition has a d of about 1.0 μm or less after a secondary heat treatment for microbial control. 50 and / or d of about 2.0 μm or less 90 (ii) the liquid composition has a viscosity of 100 s at 20° C. -1 (iii) the liquid composition exhibits less than 10% precipitation after storage at a temperature of about 20° C. to about 25° C. over the shelf-life of the liquid composition; and / or (iv) the liquid composition exhibits less than 10% precipitation after centrifugation at 1540×g for 5 minutes.

[0345] Exemplary embodiment 40. A liquid composition or liquid nutritional composition or a heat-treated shelf-stable liquid composition, comprising: d less than approx. 1.0μm 50 and d of about 2.0 μm or less 90 having comprises at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) of total protein; and / or The liquid composition of exemplary embodiment 18, the liquid nutritional composition of exemplary embodiment 25, or the heat-treated shelf-stable liquid composition of exemplary embodiment 39, comprising at least 6%, at least 8%, at least 10%, at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) denatured whey protein.

[0346] Exemplary embodiment 41. The heat-treated, shelf-stable liquid composition of exemplary embodiment 39, wherein the liquid composition comprises at least 10%, at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) denatured whey protein.

[0347] Exemplary embodiment 42. The heat-treated, shelf-stable liquid composition of exemplary embodiment 39, wherein the liquid composition is a drinking yogurt, and the drinking yogurt comprises at least 6%, at least 8%, at least 10%, at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) total protein and at least 6%, at least 8%, at least 10%, at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) denatured whey protein.

[0348] Exemplary embodiment 43. Drinking yogurt has a viscosity of 50° C. at 20° C. s-1 The heat-treated shelf-stable liquid composition of exemplary embodiment 42, having a viscosity 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, as measured by HPLC.

[0349] Exemplary embodiment 44. The heat-treated shelf-stable liquid composition of exemplary embodiment 42 or exemplary embodiment 43, wherein the drinking yogurt comprises at least 9%, at least 12%, at least 15%, at least 18%, or at least 20% (w / v) total protein.

[0350] Exemplary Embodiment 45. The heat-processed shelf-stable liquid composition of exemplary embodiment 39, wherein the liquid composition is an acidic beverage.

[0351] Exemplary Embodiment 46. The heat-treated shelf-stable liquid composition of exemplary embodiment 45, wherein the acidic beverage has a pH of about 2 to about 4.8.

[0352] Exemplary embodiment 47. The heat-treated shelf-stable liquid composition of exemplary embodiment 45, wherein the acidic beverage comprises at least 6%, at least 8%, at least 10%, at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) total protein and at least 6%, at least 8%, at least 10%, at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) denatured whey protein.

[0353] Exemplary embodiment 48. An acidic beverage is aged for 100 seconds at 20°C. -1 The heat-treated shelf-stable liquid composition of exemplary embodiment 47, having a viscosity 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, as measured by HPLC.

[0354] Exemplary embodiment 49. The heat-treated shelf-stable liquid composition of exemplary embodiment 47 or exemplary embodiment 48, wherein the acidic beverage comprises at least 9%, at least 12%, at least 15%, at least 18%, or at least 20% (w / v) total protein.

[0355] Exemplary Embodiment 50. The heat-processed shelf-stable liquid composition of exemplary embodiment 39, wherein the liquid composition is a neutral beverage.

[0356] Exemplary embodiment 51. The heat-treated shelf-stable liquid composition of exemplary embodiment 50, wherein the neutral beverage has a pH of about 6.5 to 7.5.

[0357] Exemplary embodiment 52. The heat-treated shelf-stable liquid composition of exemplary embodiment 50 or exemplary embodiment 51, wherein the neutral beverage comprises at least 6%, at least 8%, at least 10%, at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) total protein and at least 6%, at least 8%, at least 10%, at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) denatured whey protein.

[0358] Exemplary embodiment 53. The heat-treated shelf-stable liquid composition of exemplary embodiment 39, wherein the liquid composition is a liquid nutritional composition for use in providing nutrition to a subject in need thereof.

[0359] Exemplary embodiment 54. The heat-treated shelf-stable liquid composition of exemplary embodiment 53, wherein the liquid nutritional composition comprises at least 30 mg / 100 mL, at least 50 mg / 100 mL, or at least 100 mg / 100 mL of a divalent cation, e.g., Ca++.

[0360] Exemplary embodiment 55. The heat-treated shelf-stable liquid composition of exemplary embodiment 53 or exemplary embodiment 54, wherein the neutral beverage comprises at least 6%, at least 8%, at least 10%, at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) total protein and at least 6%, at least 8%, at least 10%, at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) denatured whey protein.

[0361] Exemplary Embodiment 56. The heat-treated, shelf-stable liquid composition of any one of Exemplary Embodiments 53-55, wherein the liquid nutritional composition comprises a lipid component and a carbohydrate component.

[0362] Exemplary embodiment 57. The heat-processed shelf-stable liquid composition of any one of exemplary embodiments 53-56, wherein the liquid composition has an energy density of at least 0.5, at least 1.0, at least 1.5, or at least 2.0 kcal / ml.

[0363] Exemplary embodiment 58. The heat-treated, shelf-stable liquid composition of any one of exemplary embodiments 39 to 57, further comprising a non-whey protein, preferably selected from the group consisting of casein, caseinate, micellar casein isolate, and mixtures thereof.

[0364] Exemplary embodiment 59. A liquid nutritional composition comprising at least 6% (w / v) whey protein, the whey protein comprising microparticles comprising denatured whey protein, the liquid nutritional composition comprising a lipid component, optionally present in an amount of up to 30% (w / v), a carbohydrate component, optionally present in an amount of up to 30% (w / v), at least one monovalent cation, and at least one divalent metal cation, the liquid nutritional composition comprising at least 30 mg / 100 mL, at least 50 mg / 100 mL, or at least 100 mg / 100 mL of divalent metal cations.

[0365] Exemplary embodiment 60. The liquid nutritional composition of exemplary embodiment 59, wherein the composition comprises at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / w) whey protein.

[0366] Exemplary embodiment 61. A method for providing nutrition to a subject in need thereof, comprising administering to the subject a liquid nutritional composition of any one of exemplary embodiments 25, 53-57, or 59-60.

[0367] Exemplary embodiment 62. A heat-processed high protein set gel, the set gel comprising at least 10% or at least 15% (w / v) total protein, wherein at least 50% or at least 60% (w / w) of the total protein is denatured whey protein, the set gel exhibiting (i) a firmness of less than 1000 g·sec and / or (ii) a breaking strength of less than 50 g.

[0368] Exemplary embodiment 63. The heat-processed high protein set gel of exemplary embodiment 62, wherein the set gel comprises yogurt.

[0369] Exemplary embodiment 64. The heat-treated high protein set gel of exemplary embodiment 62 or exemplary embodiment 63, wherein the set gel comprises at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / w) whey protein.

[0370] Exemplary embodiment 65. A heat-treated high protein semi-solid food product, said semi-solid food product comprising at least 10% or at least 15% (w / v) total protein, wherein at least 50% or at least 60% (w / v) of the total protein is denatured whey protein, said semi-solid food product being heat-treated at 20° C. for 50 s. -1 4. A heat-treated high protein semi-solid food product having a viscosity measured at 1000 mPa·s, 800 mPa·s, 600 mPa·s, or 400 mPa·s.

[0371] Exemplary embodiment 66. The heat-processed high protein semi-solid food product of exemplary embodiment 65, wherein the semi-solid food product comprises stirred yogurt.

[0372] Exemplary embodiment 67. The heat-treated high protein semi-solid food product of exemplary embodiment 65 or exemplary embodiment 66, wherein the semi-solid food product comprises at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / w) whey protein.

[0373] Exemplary embodiment 68. A liquid nutritional composition comprising microparticles comprising denatured whey protein, wherein the liquid nutritional composition comprises 12% (w / v) or less total protein, wherein at least 95% of the total protein in the liquid nutritional composition is denatured whey protein, and wherein the liquid nutritional composition optionally further comprises a lipid component and / or a carbohydrate component.

[0374] Exemplary embodiment 69. The liquid nutritional composition of exemplary embodiment 68, wherein substantially all of the total protein in the high protein liquid composition is denatured whey protein.

[0375] Exemplary embodiment 70. The liquid nutritional composition of exemplary embodiment 68 or 69, wherein the lipid component is optionally present in an amount of up to 30% (w / v) and the carbohydrate component is optionally present in an amount of up to 30% (w / v).

[0376] Exemplary embodiment 71. The liquid nutritional composition of any one of exemplary embodiments 68 to 70, further comprising at least one divalent metal cation, wherein the liquid nutritional composition comprises at least 30 mg / 100 mL, at least 50 mg / 100 mL, or at least 100 mg / 100 mL of a divalent metal cation.

[0377] Exemplary embodiment 72. The liquid nutritional composition of any one of exemplary embodiments 68-71, wherein the liquid nutritional composition has a caloric density of less than 1 kcal / ml.

[0378] Exemplary embodiment 73. The liquid nutritional composition of any one of exemplary embodiments 68-71, wherein the liquid nutritional composition has a caloric density of 1-2 kcal / ml.

[0379] Exemplary embodiment 74. The liquid nutritional composition of any one of exemplary embodiments 68-71, wherein the liquid nutritional composition has a caloric density of greater than 2 kcal / ml.

[0380] Exemplary embodiment 75. The liquid nutritional composition of any one of exemplary embodiments 68-71, wherein the liquid nutritional composition has 1.5-2.5 kcal / ml.

[0381] Exemplary embodiment 76. (i) the liquid composition has a d of about 1.0 μm or less after a secondary heat treatment is applied for microbial control. 50 (ii) the liquid composition has a viscosity of 100 s at 20° C. -1 The liquid nutritional composition of any one of exemplary embodiments 68-75, wherein the liquid composition has a viscosity of less than 400 mPa·s or less than 200 mPa·s measured at 20° C.; (iii) the liquid composition exhibits less than 10% precipitation after storage for at least 3 months at a temperature of about 20° C. to about 25° C.; and / or (iv) the liquid composition exhibits less than 10% precipitation after centrifugation at 1540×g for 5 minutes.

[0382] Exemplary embodiment 77. A liquid nutritional composition comprising microparticles comprising denatured whey protein, wherein the liquid nutritional composition comprises at least 12% (w / v) total protein, and wherein at least 95% of the total protein in the liquid nutritional composition is denatured whey protein, and wherein the liquid nutritional composition further comprises a lipid component and / or a carbohydrate component.

[0383] Exemplary embodiment 78. The liquid nutritional composition of exemplary embodiment 77, wherein substantially all of the total protein in the liquid nutritional composition is denatured whey protein.

[0384] Exemplary embodiment 79. The liquid nutritional composition of exemplary embodiment 77 or 78, wherein the liquid nutritional composition has a neutral pH.

[0385] Exemplary embodiment 80. The liquid nutritional composition of any one of exemplary embodiments 77-79, wherein the liquid nutritional composition comprises 12%-18% (w / v) total protein, and substantially all of the total protein is derived from denatured whey protein.

[0386] Exemplary embodiment 81. The liquid nutritional composition of any one of exemplary embodiments 77-80, wherein the lipid component is optionally present in an amount of up to 30% (w / v) and the carbohydrate component is optionally present in an amount of up to 30% (w / v).

[0387] Exemplary embodiment 82. The liquid nutritional composition of any one of exemplary embodiments 77 to 81, further comprising at least one divalent metal cation, wherein the liquid nutritional composition comprises at least 30 mg / 100 mL, at least 50 mg / 100 mL, or at least 100 mg / 100 mL of a divalent metal cation.

[0388] Exemplary embodiment 83. The liquid nutritional composition of any one of exemplary embodiments 77-82, wherein the liquid nutritional composition has a caloric density of less than 1 kcal / ml.

[0389] Exemplary embodiment 84. The liquid nutritional composition of any one of exemplary embodiments 77-82, wherein the liquid nutritional composition has a caloric density of 1-2 kcal / ml.

[0390] Exemplary embodiment 85. The liquid nutritional composition of any one of exemplary embodiments 77-82, wherein the liquid nutritional composition has a caloric density of greater than 2 kcal / ml.

[0391] Exemplary embodiment 86. The liquid nutritional composition of any one of exemplary embodiments 77-82, wherein the liquid nutritional composition has a caloric density of 1.5-2.5 kcal / ml.

[0392] Exemplary embodiment 87. (i) the liquid composition has a d of about 1.0 μm or less after a secondary heat treatment for microbial control. 50 (ii) the liquid composition has a viscosity of 100 s at 20° C. -1 The liquid nutritional composition of any one of exemplary embodiments 77-86, wherein the liquid composition has a viscosity of less than 400 mPa·s or less than 200 mPa·s measured at 20° C.; (iii) the liquid composition exhibits less than 10% precipitation after storage for at least 3 months at a temperature of about 20° C. to about 25° C.; and / or (iv) the liquid composition exhibits less than 10% precipitation after centrifugation at 1540×g for 5 minutes. EXAMPLES

[0393] I. Working Example

[0394] In order to demonstrate the practice of the subject matter disclosed herein, the following examples have been prepared and tested, but should not be construed as limiting the scope of the invention.

[0395] methodology

[0396] Preparation and evaluation of denatured whey protein powders

[0397] All powders are recombined at a protein concentration of 10% (w / w) using deionized water at ambient temperature (-20°C) and a magnetic stirrer. The protein powder is gradually added to the water under vortex mixing conditions, taking care to avoid foaming. Once all powders are dispersed, hydration is continued for 30 minutes with continuous stirring. The solution is homogenized at 150 / 50 bar and tested for primary particle size, primary particle size growth, heat setting time, insolubility, and denaturation as described below.

[0398] The primary particle size of the reconstituted powder is measured using a Malvern Mastersizer 2000 or Malvern Mastersizer 3000 (Malvern Instruments Ltd, Worcs, UK) with a refractive index of the particles of 1.46 and a refractive index of the solvent of 1.33. The volume-weighted mean particle size d[4,3], D 50 , and D 90 Percentiles of values ​​were calculated using Mastersizer software.

[0399] The volume percent of particles in a defined size range is determined by taking the sum of the relevant size class of the raw data reported by the Mastersizer software.

[0400] Primary particle size growth is determined after heating a 10% protein solution (pH 6.8) in a silicon oil bath at 120° C. for 15 minutes. A 5 mL aliquot of the 10% protein solution is placed in a closed 8 mL glass vial and placed in a rocking unit, which is then placed in a 120° C. oil bath and the sample is gently rocked during the 15 minutes of heating. The sample is immediately cooled by placing in ice water and particle size analysis is performed as described above.

[0401] Heat coagulation time: A 1 ml aliquot of 10% protein (w / w) solution (pH 6.8) is placed in a glass vial, clipped to a platform, and placed in a thermostatically controlled silicon oil bath at 140° C. with a gentle rocking speed. The time (in minutes) elapsed from placing the vessel in the oil bath until visible aggregates begin to form is defined as the heat coagulation time (HCT).

[0402] Insolubility (%) = 7.5 g of denatured whey protein powder is dissolved in 192.5 g of 0.1 M NaCl solution by mixing for 30 min using a magnetic stirrer. A 50 g portion of this suspension is adjusted to pH 4.6 using 15% acetic acid (the amount of acetic acid added is recorded for the dilution correction factor). 40 g of this suspension is transferred to a 50 ml centrifuge tube and centrifuged at 7250 g for 20 min (Beckman). The supernatant of this solution is analyzed for total protein by testing the total nitrogen content by the Kjeldahl method (ISO8968-1 / 2|IDF020-1 / 2-Milk-Determination of nitrogen content-Part 1 / 2: Determination of nitrogen content using the Kjeldahl method). The total protein content is calculated by multiplying the total nitrogen result by the conversion factor 6.38. Another (not centrifuged) portion of the suspension is also analyzed for total protein.

number

[0403] Denatured and soluble proteins are analyzed based on the method described by Elgar et al (2000) J Chromatography A, 878, 183-196.

[0404] The powders are characterized by measuring the remaining denaturable protein as a percentage of the total protein according to the following formula:

number

[0405] Soluble whey protein was measured using reverse phase HPLC and is expressed as grams of protein / 100 grams of powder.

[0406] Denaturable whey protein is measured as Σ bovine serum albumin + α-lactalbumin + β-lactoglobulin + lactoferrin + immunoglobulins.

[0407] The denaturation percentage of a denaturable protein can be calculated based on the following formula:

number

[0408] The percentage of covalently cross-linked β-lactoglobulin was determined using a 2100 Agilent Bioanalyzer System (Agilent, USA), which utilizes microfluidic SDS electrophoresis technology using a modified method of Anema (2009) (SG Anema, 2009, The use of “lab-on-a-chip” microfluidic SDS electrophoresis technology for the separation and quantification of milk proteins. International Dairy Journal, Volume 19, Issue 4, Pages 198-204).

[0409] The percentage of remaining native protein, covalently aggregated protein, and non-covalently aggregated protein is determined by dissolving the denatured whey protein composition in a buffer, thereby dissociating the different bonds, and determining the change in soluble protein after a centrifugation step.

[0410] To determine the remaining native protein content, dissolve or dilute the pre- and post-heated samples in water to a 10% (w / w) protein concentration in water. Dissolve a 300 mg aliquot of the sample in 1.2 mL of acetate buffer (0.2 M, pH 4.35).

[0411] Prepare 0.1 M phosphate buffer (pH = 6.7) from 0.0281 mol NaH2PO4 2H2O and 0.0218 mol Na2HPO4, add urea to a final concentration of 8 M, and add sodium dodecyl sulfate (SDS) to a final concentration of 2% (w / v). Dissolve a 300 mg aliquot of the sample in 1.2 mL of phosphate / urea / SDS buffer (PSU buffer).

[0412] The prepared sample is shaken on a test tube shaker for 1 hour.

[0413] The samples are centrifuged at 20,000 x g for 1 hour at 25°C.

[0414] Carefully take a 100 μL aliquot of the supernatant and run it using the bioanalyzer system, prepared according to the method of Anema. The areas under the curve of the protein peaks are automatically integrated by the software (2100 Bioanalyzer Expert software package). The peak areas are used to calculate the binding type based on the sample preparation as described above.

[0415] basis

[0416] The acetate buffer used in this experiment precipitates any denatured whey protein or remaining casein in solution. Unheated samples are expected to have low levels of denatured protein. Heated samples are expected to have high levels of denaturation. The remaining native protein in the heated samples is calculated as a percentage of the unheated starting native protein.

number

[0417] Urea can break hydrophobic bonds and SDS can break non-covalent bonds such as hydrogen bonds and hydrophobic bonds. Heated samples added to phosphate buffer containing urea and SDS cause solubilization of non-covalent bonds, thus allowing calculation of non-covalent aggregated protein compared to total soluble protein in non-heated solution, taking into account remaining native-like protein.

number

[0418] Therefore, the covalently aggregated protein can be calculated as the remaining protein that was not solubilized by the PSU buffer.

number

[0419] Preparation and Analysis of High Protein Liquid Nutritional Compositions and / or Beverages

[0420] A flow chart of the entire production process of a high-protein beverage is shown in Figure 2.

[0421] Neutral high protein beverage

[0422] The required proteins, carbohydrates, minerals and stabilizers are dry mixed and hydrated in water heated to 55°C for 60 minutes, with antifoam added if required. 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 can be pasteurized in two ways:

[0423] (1) The homogenized mixture is filled into a retort can and heat-treated at 120°C for 15 minutes.

[0424] (2) The homogenized mixture is UHT sterilized at 145°C for 4 seconds, followed by homogenization at 150 / 50 bar. The product is aseptically packed.

[0425] Acidic high protein beverage

[0426] The same process was carried out according to the process for neutral beverages, but the pectin solution was prepared at 80°C and added to the water, followed by the addition of proteins, carbohydrates, minerals and stabilizers. The pH is adjusted to 4.0±0.1 with 15% hydrochloric acid. UHT sterilization is carried out at 110°C for 4 seconds, followed by homogenization at 150 / 50 bar. The product is aseptically packed.

[0427] Unless otherwise stated, measurements were performed using a rheometer such as an Anton Paar instrument using a cup and bob assembly for 100 s -1 The viscosity of the formulation is measured at a shear rate of 1000 μm and 20° C. It will be appreciated that other methods of measuring or estimating viscosity are well known in the art and may be used where appropriate.

[0428] The particle size distribution of the beverage is measured in the same manner as described for the primary particle size, and the volume percent analysis of the particles is performed in the same manner as above.

[0429] Sedimentation Test: After suitable heat treatment of the beverage, store the product at 25°C for shelf life testing at 1 month, 3 months, 6 months, or 12 months shelf life. To test for sediment, invert the product 3 times and carefully pour out. Place the container upside down for 30 minutes, record the weight of the container and sediment, and calculate the amount of sediment by subtracting the weight of the empty container. The % sediment is then calculated by the ratio of the sediment weight to the total weight of the product in the container.

[0430] Preparation and analysis of high protein drinking yoghurt, set yoghurt and stirred yoghurt

[0431] A flow chart of the yogurt production process is shown in Figure 3. The denatured whey protein particle concentrate, skim milk powder (SMP) and optionally MPC are recombined in water at 10°C for 60 minutes using an overhead stirrer. The composition is held at 4°C overnight. The mixture is preheated to 60°C followed by homogenization at 150 / 50 bar. The composition is then heated to 95°C and held for 6 minutes or 80°C for 20 minutes or equivalent and then cooled to 42°C. The mixture is inoculated with a starter culture (Chr Hansen YF-L702). The mixture is incubated at 42°C for about 9-16 hours to a final pH of about 4.6 to form the yogurt. The drinking yogurt and / or stirred yogurt is cooled to 20°C and treated to break the gel. The yogurt is packed into containers and cooled to about 4°C. Immediately after inoculation the set yogurt is filled into pottles and fermented to produce the set yogurt.

[0432] The pH is measured at approximately 10° C. using a pH meter (model pHM210, Radiometer Analytical SAS, France) calibrated against pH 7.0 and pH 4.0 buffers before use. On the seventh day, the pH of each sample is measured at least twice from separate bottles.

[0433] The apparent viscosity is measured at 10° C. using a Haake viscometer (Haake Mess-Technik, GmbH & Co., Karlsruhe, Germany). Viscosity measurements are performed by varying the shear rate from 0 to 120 s over a period of 180 s. -1 and then the shear rate was increased to 0 s for 30 s. -1 This is done by lowering the -1 The apparent viscosity at shear rate is reported. On the seventh day, tests are performed in duplicate from separate sample bottles.

[0434] Sedimentation by centrifugation is determined by centrifuging the drinking yogurt samples at 3000 rpm (1540 x g) for 5 min at a temperature of 10-15 °C using a Sorvall Evolution RC centrifuge (Rotor F13S-14 × 50cY, rotor code is C = SLA600TC Angle Rotor). The supernatant is carefully removed and the sides of the tube wall are dried. The sediment as a percentage of the total weight of the original sample in the tube is calculated as Sediment %.

[0435] Sediment is determined by carefully pouring the drinking yoghurt out of the container, thereby allowing the liquid to pour off. Sediment is determined by measuring the sediment in the pot and the height of the yoghurt in the pot and calculating the sediment as a percentage of the total sample. Tests are performed in duplicate from separate sample pots after 1 week and 6 weeks of storage.

[0436] Phase separation is determined by measuring the height of the clear layer formed on top of the yogurt in the pot and the height of the yogurt and calculating the phase separation as a percentage of the total sample. Tests are performed in duplicate from separate sample pots after 1 week and 6 weeks of storage.

[0437] Breaking strength and firmness are assessed using a TAHD Plus Texture Analyser from Stable Micro Systems (Godalming, England). 1.27 cm Perspex cylinders are used in a single compression test. The initial force to break the surface of the set yogurt is recorded as breaking strength (g) and the area under the curve is recorded as firmness (gs). The sensory attributes of the food products are evaluated using the following sensory procedure: The food products are presented in transparent sample cups that are labeled with randomly selected three-digit blind codes. The samples are presented to the panel at a temperature appropriate for the particular food product. The sensory evaluation is carried out by eight expert panelists who are experienced in tasting specific food products. Participants rate the texture and flavor characteristics and intensity of the food product samples. A consensus approach is used to collate the characteristics that best describe each sample.

[0438] FIG. 2 is an exemplary process flow diagram for producing an exemplary liquid composition, i.e., a high protein beverage, comprising a denatured whey protein concentrate.

[0439] FIG. 3 is an exemplary process flow diagram for the production of an exemplary liquid composition comprising a denatured whey protein composition, namely, drinking yogurt.

[0440] Example 1A: Preparation of a denatured whey protein particle composition of the present invention

[0441] Annatto colored dairy cheese whey was concentrated to a maximum protein concentration of about 22-24% (w / w) using standard commercial ultrafiltration / diafiltration technology. Table 1 shows the specific conditions used to produce powders A, B, C, and D from the colored whey. The colored whey retentate was decolorized by pre-charging the retentate silo with the fungal peroxidase enzyme Maxi Bright and adding 50-80 ppm total hydrogen peroxide. The ratio of hydrogen peroxide to retentate, rate of hydrogen peroxide addition, and mixed hold time of the retentate in the silo after hydrogen peroxide addition were selected to provide optimal improvement in denatured whey protein composition while providing the desired whiteness and preventing off-flavor formation. This step was completed at a temperature below 10°C. Prior to further processing, the retentate was sampled and tested for detectable peroxide using Merck peroxide indicator test strips (MQuant Peroxide test strips, 0.5-25 mg / L H2O2), and no peroxide was detected.

[0442] The pH of the whey protein concentrate was adjusted using 2% (w / v) NaOH solution as shown in Table 1 and preheated to 54°C using a hot water heated heat exchanger. The denatured whey protein composition was then prepared according to the method described in WO2010120199. The whey protein concentrate was fed into two identical single-tube high-pressure steam heated shell-and-tube heat exchangers in series at a flow rate high enough to achieve a Reynolds number of 2100 or higher using a high-pressure pump with a delivery pressure of 250-350 bar. As shown in Table 1, 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.

[0443] After being discharged from the second heater, the heat-treated whey protein concentrate passes through a holding tube and pipe to a nozzle bank at the top of the spray dryer, which has the same tube specifications as the high pressure heater, indicating that turbulence is maintained. The length of the holding tube is selected in such a way that, in combination with the pipe to the nozzle bank, it provides an additional residence time of 20 seconds for the heated stream before spray drying with less than 1°C temperature loss over the entire pipe from the outlet of the second high pressure heater to the nozzle bank. This means that no additional mechanical shear inducer is used after the heater reactor system and before spray drying during heating.

[0444] In the spray dryer, the heat-treated concentrate was fed to a bank of three nozzles and atomized at pressures above 200 bar into a droplet spray. An inlet hot air temperature of 210-220°C and an outlet chamber temperature of 60-70°C were used. The powder was further dried and then cooled in a vibrating fluidized bed before the material was sieved and packed to produce a powder with less than 5% moisture. Several colorless whey protein concentrates (powders E, F, G) were also processed without the addition of hydrogen peroxide and peroxidase enzymes under turbulent flow with Reynolds numbers above 2100 according to the method described in WO2010120199. A commercially available micronized WPC (NutrilacYO-8075) (powder H) from Arla Foods was obtained and tested in comparison using the method described in the methodology section.

[0445] [Table 1]

[0446] [Table 2]

[0447] [Table 3]

[0448] Example 1B: Turbidity of Denatured Whey Protein Concentrate Powder After Reformulation

[0449] The turbidity of the denatured whey protein concentrate was measured by reconstituting the powder prepared according to Powder C (above) in water. The absorbance of the 0.1% protein solution measured at 500 nm was out of range (>1). The solution was further diluted to 0.025% protein (w / w) to obtain a reading within the measurement range of the spectrophotometer. The absorbance of the 0.025% protein solution at 500 nm was 0.857 at approximately 20° C. Taking into account the dilution factor, the turbidity of the 0.1% protein solution was 3.4 absorbance units.

[0450] Example 2: Preparation of a shelf-stable beverage with 9% protein (w / v)

[0451] This example describes the preparation of an exemplary high protein liquid beverage comprising a powder (Powder A) using the methods of the present invention compared to the comparative powders (Powder F, G, H) described in Example 1. The beverages were prepared under neutral conditions following the procedures in the methodology section. The formulas used to prepare the beverages are shown in Table 4, and the nutritional composition of the beverages is summarized in Table 5.

[0452] [Table 4]

[0453] [Table 5]

[0454] Table 6 shows the properties of the high protein beverages prepared using different denatured whey protein concentrates. The high protein beverages containing Powder A showed minimal changes in particle size characteristics after heating at 120°C for 15 minutes compared to the characteristics of Powder A summarized in Table 3 of Example 1. The high protein beverages prepared with Powder A had a uniform consistency, were smooth in appearance, and were very thin and pourable. Powder F caused gelation of the high protein beverage during heat treatment, and large aggregates with a grainy appearance were formed in the solution, thus showing significant instability to sterilization conditions. Powders G and H showed significant particle growth upon heat treatment, which resulted in an undesirable very gritty / chalky mouthfeel. This difference in particle size distribution after sterilization can be clearly seen in Figure 4. Figure 4 shows the particle size distribution of the high protein beverages prepared with Powder A, Powder G, or Powder H after heating at 120°C for 15 minutes.

[0455] [Table 6]

[0456] Example 3: Primary particle size growth

[0457] This example shows the particle size of a denatured whey protein concentrate of the present invention and a comparative denatured whey protein concentrate after heating at 120° C. for 15 minutes.

[0458] All powders were prepared according to the reformulation method described in the methodology, but protein solutions of 12, 14, 16, and 18% (w / w) protein were also prepared. Particle size was measured according to the primary particle size growth method described in the methodology.

[0459] Table 7 shows the D of the protein solution after heating. 50 and D. 90 These values ​​can be compared to Table 3 of Example 1, which shows the particle size of each powder before heating at 120° C. for 15 minutes. The appearance of the denatured whey protein compositions after heating is also shown in the table, along with whether they are in a liquid, thick liquid, or gelled state.

[0460] [Table 7]

[0461] The powders of the present invention did not show significant change in particle size after 15 minutes of heat treatment at 120°C for solutions containing up to 16% (w / w) protein, and were still liquid at 18% (w / w) protein. Comparative compositions gelled after the same heat treatment at 10% (w / w) or 12% (w / w) protein content. The denatured whey protein compositions of the present invention exhibited excellent heat stability even at high protein concentrations, and showed minimal or no change in particle size distribution compared to the comparative denatured whey protein compositions. Figure 5 shows the particle size distribution of Powder A-containing protein solutions of 10%-16% (w / w) protein after heating at 120°C for 15 minutes.

[0462] D less than 1μm after heat treatment 50 and D less than 2 μm 90 is an important feature to enable good shelf life stability, minimal settling, low viscosity, and good mouthfeel properties such as lack of chalkiness and grittiness. This was typical for all powders of the present invention.

[0463] FIG. 5 shows particle size distribution of Powder A-containing protein solutions at 10%, 12%, 14% or 16% (w / w) protein after heating at 120° C. for 15 minutes.

[0464] Example 4: Heat setting times at high protein concentrations and use of the denatured whey protein concentrate of the present invention in high protein liquid nutritional compositions

[0465] This example shows the heat setting time (140° C.) of protein solutions containing a denatured whey protein concentrate of the present invention and a comparative whey protein concentrate at high protein concentrations.

[0466] All compositions were prepared according to the reformulation methodology described in Methodology, but also at protein concentrations of 12, 14, 16, 18% and 20% (w / w) protein. Heating was performed according to the heat set time methodology described in Methodology.

[0467] FIG. 6 shows that the denatured whey protein compositions of the present invention are significantly more heat stable than the comparative denatured whey protein compositions. Based on our experience, it is known that it takes at least 1 to 1.5 minutes for a sample to reach 140° C., and therefore samples that do not show a heat setting time of more than 1 minute are unlikely to be stable during the UHT heat treatment process. All of the denatured whey protein compositions of the present invention had a heat setting time of more than 1 minute at a protein content of up to 20% (w / w). Powders G and H showed a heat setting time of less than 1 minute when prepared at a concentration of 14% (w / w) or more. Powder F showed a heat setting time of less than 1 minute when prepared at a concentration of 10% (w / w) or more.

[0468] Figure 6: Thermal coagulation times of denatured whey protein compositions prepared at protein concentrations between 10% and 20% (w / w).

[0469] As a next step, the particle size distribution of protein solutions containing Powder C of the invention at 10%-18% (w / w) protein was measured after heating at 140° C. for 90 seconds and rapid cooling. The values ​​in Table 8 can be compared with the respective powders in Table 3 of Example 1, which show the particle size before heating. Powder C not only remains liquid after such heating conditions, but also shows minimal particle size change at protein concentrations up to 18% (w / w) when heated at 140° C. for 90 seconds.

[0470] [Table 8]

[0471] Based on these results, several high protein liquid nutritional compositions were prepared using the powder of the present invention (Powder A), as described in Tables 9, 10, and 11. Because the comparable denatured whey protein concentrates at concentrations of 14% (w / w) and above were not stable to heating at 140° C., further testing of these samples was not performed in the high protein nutritional compositions. The beverages were prepared according to the procedure given in the methodology section.

[0472] [Table 9]

[0473] [Table 10]

[0474] [Table 11]

[0475] Additional high protein liquid nutritional compositions were prepared using Powder A, as described in Tables 12, 13, and 14.

[0476] [Table 12]

[0477] [Table 13]

[0478] [Table 14]

[0479] Powder A was used to prepare pH neutral and acidic beverages with various compositions, which showed minimal particle size growth after UHT heat treatment. The particle size of the beverages after heating can be compared to Table 3 in Example 1. Powder A showed very low viscosity in the high protein beverages after UHT heat treatment, all exhibiting a homogenous consistency with no evidence of graininess or separation in the pack.

[0480] Example 5: Use of the improved denatured whey protein particle composition of the present invention in high protein drinking yogurt

[0481] This example describes the preparation of several exemplary high protein drinking yogurts containing Powders B and C using the method according to the invention, compared to other denatured whey protein concentrates. Table 15 shows the nutritional composition and characteristics of the yogurts containing denatured whey protein concentrates.

[0482] The flow chart of the yogurt production process is shown in Figure 3. The denatured whey protein concentrate and skim milk powder (SMP) were reformulated in water to produce an aqueous composition with 15% protein by weight (12% is protein from the denatured whey protein particle composition) or 20% protein by weight (17% is from the denatured whey protein particle composition). For each batch, 9.2% (w / w) SMP was reformulated and topped off with 15% or 20% (w / w) denatured whey protein concentrate depending on the final target protein. 0.02% (w / w) bacterial culture was added for fermentation. Additional water was used to achieve 100% balance.

[0483] [Table 15]

[0484] When powders B and C were used to prepare 15% (w / w) protein drinking yogurt, the yogurt showed minimal changes in particle size characteristics after 1 week of production compared to the powder properties in Table 3 of Example 1. Comparative powders E, F, and G showed an increase in particle size after 1 week of yogurt production. Yogurts prepared with powders B and C also showed less sediment (%) than the comparative yogurt. Similarly, for powders B and C, the sediment (%) remained low at the end of the shelf life after 6 weeks of production and no phase separation was observed. The overall appearance of the yogurt was smooth, uniform, very thin and pourable, without a chalky or gritty texture. In comparison, powders E and F showed a thicker texture and a chalky / gritty mouthfeel. Drinking yogurts prepared with powders E, F, and G showed significant sediment formation at the end of the 6-week shelf life. The sediment was a very hard gel layer that could not be redispersed in the drinking yogurt by shaking.

[0485] Powder C was used to prepare a 20% (w / w) protein drinking yogurt, which showed minimal changes in particle size characteristics after one week of yogurt production, compared to the powder properties in Table 3 of Example 1. The yogurt was smooth, homogenous, very thin and pourable. Powders F and G were also used to prepare a 20% (w / w) protein drinking yogurt, but they were found to be unsuitable for use during heat treatment at 85° C. for 15 minutes, as the samples caused the formation of large gelled particles and lumps, and the viscosity increased significantly. The samples had a granular texture, which resulted in clogging of the processing equipment.

[0486] FIG. 7 shows the particle size distribution of 15% (w / w) and 20% (w / w) protein yogurt one week after production, clearly showing the smaller particle size distribution of the denatured whey protein concentrate of the present invention.

[0487] Figure 7: Particle size distribution of high protein (15% (w / w) and 20% (w / w)) drinking yogurt with denatured whey protein concentrate at 1 week.

[0488] Example 6: Use of Improved Denatured Whey Protein Concentrate in High Protein Set and Stirred Yogurts

[0489] This example describes the preparation of exemplary high protein set and stirred yogurts containing the denatured whey protein composition of Powder C of the present invention compared to Powder F. The formulas, compositions, and yogurt properties of the set and stirred yogurts are summarized in Table 16. The yogurts were prepared according to Figure 3 with some modifications as mentioned in the methodology.

[0490] [Table 16]

[0491] Stirred yogurt using Powder C showed lower viscosity compared to Powder F and produced a soft, spoonable, glossy and smooth looking yogurt whereas yogurt using Powder F had a coarse texture. Set yogurt prepared with Powder C showed lower breaking strength and firmness compared to Powder F.

[0492] Example 7: Use of Improved Denatured Whey Protein Concentrate in Protein Bars

[0493] An exemplary method is described illustrating the use of denatured whey protein concentrate in a protein bar.

[0494] Protein bars were made by combining maltodextrin and Powder C. Glucose syrup, glycerin, and water were combined and heated to 50-55°C. The glucose syrup and glycerin mixture was added to the maltodextrin and protein mixture, followed by the oil and lecithin mixture. The confectionery fat and lecithin were heated until the fat was melted. The mixture was mixed using a Hobart mixer (Model N-50) at speed 1 for 90 seconds, then the bowl was scraped down. Mixing (speed 2) was continued until a homogenous mass was obtained.

[0495] The mixture was poured into a bar frame (approximately 16mm deep), spread evenly, and extended to the same height as the frame, and the excess was trimmed off. The mixture was left to harden overnight. The mixture was removed from the frame and cut into 30mm x 100mm bars. The bars were packed in foil bags for storage until use. The composition and breaking strength (g) properties of the protein bars are listed in Table 17.

[0496] The breaking strength (g) of the bars was evaluated using a TAHD Plus texture analyzer from Stable Micro Systems (Godalming, England). Texture measurements were made by penetration. Force was measured over a set penetration depth of 12 mm. A 5 mm stainless steel cylindrical probe was pressed into the bar to a depth of 12 mm at a constant rate of 1 mm / s and then withdrawn at a rate of 10 mm / s. Force (g) versus time (s) was measured for the movement of the probe. Three compressions were performed on the surface of each bar sample. Two bars were evaluated for each sample. Samples were removed from storage at 20°C and texture measurements were performed at 20°C in a temperature controlled room.

[0497] [Table 17]

[0498] Powder C produced a protein bar that was acceptable in terms of texture. The organoleptic characteristics of the bar were acceptable in terms of textural characteristics including hardness, firmness, cohesiveness, and graininess. No off-flavor characteristics were detected in the protein bar. The hardness of the protein bar was 2310 g after one month storage, with a hardness of less than about 4000 g being preferred after 12 months storage. The water activity value of the bar was 0.51 after one month storage, which is below the microbial stability limit of 0.65. The results indicate that the denatured whey protein concentrate is suitable for use in protein bars.

[0499] Example 8: Effect of Addition Rate on Improving the Thermal Stability of Denatured Whey Protein Particle Compositions

[0500] Colorless dairy cheese whey solutions were prepared by reconstituting dairy cheese whey protein (WPC80) powder with protein concentrations up to 20-22% (w / w). The composition of the WPC80 powder is summarized in Table 18. The solutions were homogenized at 200 / 50 bar and then divided into four batches. Three batches were pH adjusted to 6.2 using a 1 M KOH:NaOH (50:50) mixture and one sample was pH adjusted to 5.3 using 6 M HC1. pH 5.3 is close to the optimal pH for maximum activity of MaxiBright, which allows for faster dosing of peroxide without inhibiting enzyme activity. Sufficient equal amounts of commercial MaxiBright solution were added to all four samples and the samples were mixed using an overhead stirrer at 150 rpm in a 4 °C refrigerator. The sample designations are as follows: 1) control-no peroxide added, 2) pH 6.2-75 ppm peroxide added all in one step, 3) pH 6.2-75 ppm peroxide added stepwise at a rate of 10 ppm / hr, 4) pH 5.3-75 ppm peroxide added stepwise at a rate of 30 ppm / hr. 75 ppm peroxide corresponds to 0.375 moles of H2O2 per mole of β-lactoglobulin protein. The enzymatic dairy decolorization activity according to DSM's COA was 5000 U / g. After dosing of the peroxide was completed and no peroxide was found to remain in the solution, sample 4 was adjusted to pH 6.2 using the aforementioned KOH:NaOH mixture before heating. A combination of a scraped surface heat exchanger and a steam heated pilot scale tubular heat exchanger was then used to produce a heat stable denatured whey protein particle composition. The samples were preheated to 55°C in a scraped surface heat exchanger and then heated to 85°C in a steam-heated tubular heat exchanger. -1The flow rate was adjusted to achieve a wall shear rate of 85°C. A holding tube with tubing specifications similar to the steam heated tubular heat exchanger was used to hold the sample temperature at 85°C for an additional 20 seconds to provide the necessary residence time to achieve the desired HCT. Samples were then cooled, collected, and tested for HCT as described in Section 3 - Thermal Coagulation Time. All samples were tested for protein profile before and after heat treatment as described in Section 5 - Extensive Denaturation.

[0501] FIG. 8: Thermal coagulation times of denatured whey protein compositions prepared from samples treated with different peroxide dosage rates.

[0502] Figure 9: Protein profile of the samples after enzymatic treatment and before heat treatment analyzed by HPLC, including peaks for glycomacropeptide (gmp), proteose peptone 5 (pp5), α-lactalbumin (alac), lactoferrin (Lf), BSA, β-lactoglobulin (blac), and immunoglobulin (igg).

[0503] [Table 18]

[0504] Example 9: Use of the improved denatured whey protein particle composition of the present invention in a neutral pH, low calorie, high protein liquid nutritional composition.

[0505] This example describes the preparation of a neutral pH, low calorie, high protein beverage containing the Powder A denatured whey protein composition of the present invention. The formula, composition, and properties of the neutral pH, low calorie, high protein beverage are summarized in Table 19. A beverage with a protein level of 12% (w / v) was prepared according to Figure 2 with some modifications to the UHT conditions. Indirect UHT conditions of 143°C for 6 seconds were used.

[0506] [Table 19]

[0507] [Table 20]

[0508] [Table 21]

[0509] [Table 22]

[0510] Figure 10: Particle size distribution of a 12% (w / v) reduced calorie high protein beverage before and after indirect UHT heating (143°C for 6 seconds).

[0511] From the above results, powder A is -1 It can be seen that the Formula 6 beverage, having a viscosity of less than 100 mPa.s at 25°C, exhibited minimal particle size growth during the shelf life after indirect UHT heat treatment. The Formula 6 beverage exhibited a homogenous consistency with no separated and visible protein particles or aggregates in the pack.

[0512] Example 10: Use of Improved Denatured Whey Protein Concentrate in High Protein Cookies

[0513] This example describes the preparation of high protein cookies containing the Powder C modified whey protein composition of the present invention. Based on the recipe and method of Cooper et al., J. Food Sci., 49(2), 376-379 (1984), high protein (about 20%) cookies were made to compare the performance of the modified whey protein composition of the present invention with another insoluble whey protein material, lactalbumin, on an equal protein basis. The recipe of the high protein cookies is summarized in Table 23.

[0514] [Table 23]

[0515] Butter at 20°C was mixed for 30 seconds in a Hobart mixer (Model N50, Hobart Corporation, US) with a flat beater at speed 2. Sugar was added over 30 seconds, the bowl scraped down and mixed for an additional 60 seconds. Protein ingredients were added over the next 60 seconds at speed 1, scraped down and mixed at speed 2 for an additional 60 seconds. Sifted flour and baking powder were added over 60 seconds at speed 1. Water (55g) was added over 60 seconds at speed 1. If the dough is very dry and does not form a coherent mass, add more water while mixing. The dough was rolled out to a thickness of 5mm and cut into disks with a diameter of 50mm. The disks were placed on a metal tray lined with baking paper and baked at 180°C for 12.5 minutes until the bottom was brown. The baked cookies were cooled on a wire rack for 30 minutes before evaluation. The experiment was performed in duplicate.

[0516] The cookies were subjectively evaluated for color, surface appearance (smooth to rough), initial texture (appearance when broken in half: close to open texture), crispness (ease of breaking at first bite), and residual mouthfeel (fine to powdery, powdery, residual). Spreadability (width / thickness) was determined using six cookies baked according to the AACC method (American Association of Cereal Chemists (1984) Approved Methods of the American Association of Cereal Chemists. Vol I. (8th edn). Method 10-50D, revised 1986. Baking quality of cookie flour. St Paul, Minnesota, US). Evaluation of the baked cookies can be found in Table 24.

[0517] [Table 24]

[0518] Cookies containing Powder C (Example 1) as the protein source were browner, crispier and had a better mouthfeel than the comparative ingredient.

[0519] Example 11: Use of Improved Denatured Whey Protein Concentrate in High Protein Ambient Drinking Yogurt

[0520] This example describes the preparation of an exemplary high protein ambient drinking yogurt comprising a denatured whey protein composition of the present invention compared to Powder F. The formulation and composition of the ambient drinking yogurt is summarized in Table X. The yogurt is prepared according to FIG. 3 with modifications. Amidated low methoxyl (LMA) pectin and gellan gum are added during reformulation of the dry ingredients, and cream is added during the last 10 minutes of reformulation. After stirring to break the gel, the ambient drinking yogurt is heat treated at 75° C. for 30 seconds, packed, and then stored at ambient temperature.

[0521] [Table 25]

[0522] The ambient drinking yogurt produced is evaluated for pH, flavor, viscosity, particle size distribution, % visual sediment at end of shelf life, % phase separation at end of shelf life, and sediment after centrifugation using the methods described herein.

[0523] The pH is expected to be acceptable for a yogurt product. The viscosity and sedimentation of ambient drinking yogurt prepared with Powder C is expected to remain low over the shelf life and have acceptable organoleptic properties.

[0524] It should be understood that the foregoing detailed description and the accompanying examples are merely illustrative and do not limit the scope of the present invention, which is defined only by the appended claims and their equivalents. Various changes and modifications to the embodiments of the present disclosure will be apparent to those skilled in the art. Such changes and modifications (including, but not limited to, changes and modifications related to chemical structure, substituents, derivatives, intermediates, synthesis, formulation, or method, or any combination of such changes and modifications of the use of the present invention) may be made without departing from the spirit and scope of the present invention.

[0525] All references (patent and non-patent) cited above are incorporated by reference into this patent application. The discussion of these references is intended only to summarize what their authors assert. It is not an admission that any reference (or any portion of a reference) is relevant prior art (or prior art). Applicant reserves the right to challenge the accuracy and pertinence of the cited references.

Claims

1. A heat-stable denatured whey protein composition comprising fine particles containing at least 60% total protein and denatured whey protein on a dry weight basis, wherein the fine particles have (i) a volume-weighted average diameter D(4,3) of about 1.0 μm or less, and (ii) a diameter of about 1.0 μm or less. ５０ (iii) d of about 2.0 μm or less ９０ A heat-stable denatured whey protein composition having a particle size distribution comprising at least two features selected from the group consisting of (iv) at least 92 volume percent of the particles having a diameter of less than about 2.0 μm, and (v) 45 volume percent or less of the fine particles having a diameter of about 1.0 to about 10.0 μm.

2. The aforementioned particle size distribution is (i) a volume-weighted average diameter D(4,3) of approximately 1.0 μm or less, for example, approximately 0.6 to approximately 1.0 μm, and (iii) a volume-weighted average diameter D(4,3) of approximately 2.0 μm or less, for example, approximately 0.9 to approximately 1.7 μm. ９０ , or (ii) Approximately 1.0 μm or less, for example, d of approximately 0.6 to approximately 1.0 μm ５０ , and (iii) d of about 2.0 μm or less, for example, about 0.9 to about 1.7 μm ９０ A heat-stable denatured whey protein composition according to claim 1, comprising:

3. The heat-stable denatured whey protein composition according to claim 1, wherein the particle size distribution comprises (iv) at least 92 volume%, at least 95 volume%, or at least 98 volume% of the particles having a diameter of less than about 2.0 μm, and / or (v) 45 volume% or less of the fine particles having a diameter of about 1.0 to about 10.0 μm.

4. A heat-stable denatured whey protein composition comprising fine particles containing at least 60% total protein and denatured whey protein on a dry weight basis, - After heating the 10% (w / w) protein-containing aqueous solution containing the denatured whey protein composition at 120°C for 15 minutes, d ５０ However, it is approximately 0.6 to approximately 1.0 μm. - After heating the 10% (w / w) protein-containing aqueous solution containing the denatured whey protein composition at 120°C for 15 minutes, d ９０ However, it is approximately 2.0 μm or less, for example, approximately 0.9 to approximately 1.7 μm. - After heating a 10% (w / w) protein-containing aqueous solution containing the denatured whey protein composition at 120°C for 15 minutes, the volume-weighted average diameter D(4,3) is approximately 1.0 μm or less, for example, approximately 0.6 to approximately 1.0 μm, and / or - An aqueous solution containing 10% (w / w) protein, comprising the denatured whey protein composition, is subjected to a heat coagulation time (HCT) of at least 3 minutes, at least 4 minutes, at least 5 minutes, at least 6 minutes, at least 9 minutes, at least 12 minutes, at least 15 minutes, or at least 18 minutes at 140°C. A heat-stable denatured whey protein composition.

5. The microparticles have at least two characteristics selected from the group consisting of: (i) a volume-weighted average diameter D(4,3) of about 1.0 μm or less, (ii) a d of about 1.0 μm or less, ５０ , (iii) a d of about 2.0 μm or less, ９０ , (iv) at least 92 volume % of the particles having a diameter of less than about 2.0 μm, and (v) 45 volume % or less of the microparticles having a diameter of from about 1.0 to about 10.0 μm, and having a particle size distribution, a heat-stable modified whey protein composition according to claim 4.

6. The heat-stable denatured whey protein composition according to any one of claims 1 to 5, wherein the composition comprises less than about 10% by weight of casein, and preferably the composition is substantially free of casein.

7. The heat-stable denatured whey protein composition according to any one of claims 1 to 5, wherein the composition comprises about 30% to about 80% β-lactoglobulin, about 35% to about 75% β-lactoglobulin, or about 40% to about 65% β-lactoglobulin, relative to the total protein in the composition.

8. The heat-stable denatured whey protein composition according to any one of claims 1 to 5, wherein the composition comprises at least 70% (w / w) of whey protein relative to total protein, at least 80% (w / w) of whey protein relative to total protein, or at least 90% (w / w) of whey protein relative to total protein.

9. The heat-stable denatured whey protein composition according to any one of claims 1 to 5, wherein the residual denaturable whey protein in the denatured whey protein composition is less than 16%.

10. A 3.4% (w / w) protein-containing aqueous solution comprising the heat-stable denatured whey protein composition exhibits a turbidity value of at least 50 absorbance units, preferably at least 100 absorbance units, measured at 500 nm at approximately 20°C, and / or a 4% (w / w) protein-containing aqueous solution comprising the denatured whey protein composition exhibits a turbidity value of at least 90 absorbance units, preferably at least 100 absorbance units, measured at 500 nm at approximately 20°C, according to any one of claims 1 to 5.

11. The heat-stable denatured whey protein composition according to any one of claims 1 to 5, wherein the denatured whey protein composition has an insolubility of at least 60%.

12. A composition comprising the heat-stable denatured whey protein composition according to any one of claims 1 to 5.

13. The composition according to claim 12, wherein the composition is (i) a liquid composition, for example, a drinking yogurt, a liquid nutritional composition (for example, a medical food), or a high-protein beverage such as a neutral or acidic high-protein beverage, or (ii) a food product, for example, a baked food product, a bar, or a set-type yogurt or a stirred-type yogurt.

14. The composition is a food product, - The food product is a set-type yogurt, and the set-type yogurt optionally contains about 6% to about 20% (w / v) total protein, and optionally at least 50% (w / w) of the total protein is derived from the denatured whey protein composition, and the set-type yogurt has reduced hardness and / or reduced volume-weighted average particle size compared to a control set-type yogurt product having the same component composition and the same protein content (provided that the control set-type yogurt product does not contain the denatured whey protein composition described in any one of claims 1 to 5). - The food product is a stirred yogurt, and the stirred yogurt optionally contains about 6% to about 20% (w / v) total protein, and optionally at least 50% (w / w) of the total protein is derived from the denatured whey protein composition, and the stirred yogurt has a lower viscosity and / or a lower volume-weighted average particle size compared to a control stirred yogurt product having the same component composition and the same protein content (provided that the control stirred yogurt product does not contain the denatured whey protein composition described in any one of claims 1 to 5). The composition according to claim 13.

15. A method for preparing a heat-stable denatured whey protein composition, the method comprising: (a) providing an aqueous whey protein solution or a whey protein retaining solution; (b) contacting the aqueous whey protein solution or the whey protein retaining solution with an oxidizing agent in combination with an enzyme catalyst or a chemical catalyst, preferably a catalyst such as a peroxidase enzyme; and (c) subjecting the whey protein solution or the whey protein retaining solution to heat treatment under high shear stress / force conditions, wherein the total protein content of the aqueous whey protein solution or the whey protein retaining solution in step (c) is preferably at least 16% (w / w), and the high shear stress / force is optionally an increase in serum phase viscosity, 1000 s －１ A method generated by a wall shear rate exceeding 2000, a turbulent flow pattern of at least 2000 Re, the application of mechanical shear, or a combination thereof.

16. The method according to claim 15, wherein the oxidizing agent is hydrogen peroxide or benzoyl peroxide, and / or the catalyst is a peroxidase enzyme.

17. The method according to claim 15 or 16, wherein the oxidizing agent is present in an amount of less than 300 ppm or less than 200 ppm in step (b), or the molar ratio of the oxidizing agent to β-lactoglobulin is less than 2, preferably less than 1, or about 0.1 to about 0.

85.

18. A heat-stable denatured whey protein composition prepared by the method described in claim 15 or 16.

19. A heat-treated, room-temperature storable high-protein liquid composition containing at least 6% (w / v) whey protein, The aforementioned whey protein includes fine particles containing denatured whey protein, (i) The liquid composition has d particles of about 1.0 μm or less after a secondary heat treatment applied for microbial control. ５０ and / or d of about 2.0 μm or less ９０ (ii) The liquid composition has 100s －１ (iii) The liquid composition has a viscosity of less than 400 mPa·s or less than 200 mPa·s as measured at 20°C, and the liquid composition exhibits a sedimentation of less than 10% after being stored at a temperature of about 20°C to about 25°C over the shelf life of the liquid composition, and / or (iv) the liquid composition exhibits a sedimentation of less than 10% after being centrifuged at 1540 × g for 5 minutes. A heat-treated, room-temperature storable high-protein liquid composition.

20. The heat-treated, room-temperature storable liquid composition according to claim 19, wherein the liquid composition comprises at least 8%, at least 10%, at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) of denatured whey protein.

21. The heat-treated, room-temperature storable liquid composition according to claim 19 or 20, wherein the liquid composition is a liquid nutritional composition used to provide nutrition to an object in need of nutrition, and the liquid composition has an energy density of at least 0.5, at least 1.0, at least 1.5, or at least 2.0 kcal / ml.

22. The heat-treated, shelf-stable liquid composition according to claim 19 or 20, further comprising other dairy proteins and / or non-dairy proteins.

23. A liquid nutritional composition comprising fine particles containing denatured whey protein, At least 95% of the total protein in the liquid nutritional composition is denatured whey protein. The liquid nutritional composition optionally further comprises a lipid component and / or a carbohydrate component. Liquid nutritional composition.

24. The liquid nutritional composition according to claim 23, wherein substantially all of the total protein in the liquid nutritional composition is denatured whey protein.

25. The liquid nutritional composition according to claim 23 or 24, wherein the liquid nutritional composition contains at least 12% (w / v) total protein.

26. The liquid nutritional composition according to claim 23 or 24, wherein the liquid nutritional composition contains 12% (w / v) or less of total protein.

27. (i) The liquid composition has d particles of about 1.0 μm or less after a secondary heat treatment applied for microbial control. ５０ (ii) The liquid composition is at 20°C for 100 seconds. －１ The liquid nutritional composition according to claim 23 or 24, having a viscosity of less than 400 mPa·s or less than 200 mPa·s as measured by (iii) the liquid composition exhibits a sedimentation of less than 10% after being stored at a temperature of about 20°C to about 25°C for at least 3 months, and / or (iv) the liquid composition exhibits a sedimentation of less than 10% after being centrifuged at 1540 × g for 5 minutes.

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