Dairy product and method

Abstract

The invention provides a dairy product and a method. The present disclosure relates to a denatured whey protein composition. The denatured whey protein composition includes submicron particles having a volume weighted average diameter D (4, 3) of no more than 1 [mu] m and a substantially monomodal distribution, where at least 95 vol% of the particles are no more than 2 [mu] m. The present disclosure also relates to high protein beverages, food and nutritional compositions comprising such denatured whey protein compositions and methods for making such denatured whey protein compositions.

Description

[0001] This application is a divisional application with the same invention name as the parent case. The Chinese application number of the parent case is 202380048215.8, the international application number is PCT / IB2023 / 054315, and the application date is April 26, 2023.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Application No. 63 / 382,595 filed on November 7, 2022 and U.S. Provisional Application No. 63 / 363,914 filed on April 29, 2022, the entire contents of which are incorporated herein by reference. Technical Field

[0004] The present invention relates to denatured whey protein compositions comprising heat stable submicron particles, methods for preparing such compositions, and the use of such compositions in neutral ready-to-drink high protein beverages, high protein drinking yogurts and foods (including high protein snacks, egg white or nutritional bars), and set or stirred yogurts. Background Art

[0005] Ingredients with high protein content are used to prepare high protein foods and beverages. Desirable properties of ingredients with high protein content include heat stability to allow subsequent heat treatment to ensure product safety and extended shelf life, such as retort or ultra high temperature (UHT) processing, and suspendability with minimal or essentially no sedimentation, particularly for beverage applications.

[0006] Micronized whey protein concentrate (WPC) ingredients are known and used by the food industry to increase the protein content of a variety of applications including cultured and ready-to-drink beverages, and food products including bars and set or stirred yogurts.

[0007] Whey protein micronization is an advanced technology for producing whey protein particles. The size of whey protein particles is important in delivering the desired mouthfeel; particles of 0.1 to 3 μm provide a creamy mouthfeel, while particles of >3 μm cause a powdery and even gritty feel, and particles with a size of 0.1 μm produce a watery mouthfeel. In fact, it is known that particle sizes in the range of less than 0.1 μm contribute to a greasy taste, which is unpleasant if it is perceived as a primary tactile feature. In addition, it may be desirable to keep the amount of whey protein particles with a size below 0.5 μm to a minimum, because these smaller particles may provide an undesirable high viscosity for the product comprising them.

[0008] The main mechanism of whey protein particle formation involves two steps: first, whey protein unfolds during heating; second, the unfolded protein molecules aggregate mainly through disulfide bonds and hydrophobic interactions. Processing conditions such as temperature, heating time, pH and shear stress determine the reaction kinetics and physical and chemical properties of the particles.

[0009] Traditional micronization method comprises shearing or turbulence to limit the size of whey protein particles. However, when using high whey protein concentration to manufacture protein particles, micronization / aggregation reaction occurs very rapidly, which may be due to high molecular (protein) density and high collision efficiency. Although these technologies are effective in preventing the formation of very large microparticles / aggregates, the average size of the protein particles produced in the traditional micronization process is in the range of 1 micron to 10 microns, and most of the particles have a diameter greater than 1 micron. This particle size range is still easy to cause sedimentation or precipitation in liquid applications within the shelf life of the product, and causes undesirable instability. In addition, reducing the protein particle size while ensuring that all unfolded proteins form stable microparticles is technically unachievable.

[0010] Singer (US 4,734,287) prepared proteinaceous, water-dispersible colloids comprising substantially non-aggregated sweet whey protein coagulant particles having a mean diameter distribution of greater than about 0.1 μm to less than about 2.0 μm when dried, wherein less than about 2% of the total number of particles had a diameter exceeding 3.0 μm. According to Singer, these particles are heat unstable and will form fused aggregates when subjected to additional heat denaturation treatments.

[0011] McCarthy (US 5,350,590) sought to improve upon the composition provided by Singer and further demonstrated the heat stability of a commercial product manufactured according to Singer (designated SIMPLESSE 100D). McCarthy prepared whey "loosely bound" agglomerates comprising whey protein and casein suggesting that desolubilization of casein substantially prevented the formation of covalent bonds. McCarthy noted that agglomerates prepared without casein exhibited very poor heat stability. Figure 7 As shown, particles prepared only from whey protein were larger than 1 μm - even larger than 10 μm - and the peak of the particle distribution was larger than 1 μm, and in some cases larger than 10 μm.

[0012] Likewise, Huss and Spiegel (US 6,767,575) prepared denatured whey protein aggregates with an average aggregate size (median) between 1 μm and 4 μm starting from a relatively dilute protein stream (<3% w / v protein). In addition, most particles were larger than 1 μm, there were particles larger than 2 μm and even larger than 5 μm in size, and the peak of the particle distribution by volume was larger than 1 μm.

[0013] Villagran (US 6,605,311) utilizes similar conventional methods, starting from a 20 mass % whey protein solution, to produce a whey protein composition having a total protein content of less than 60%. According to Villagran, the resulting protein particles have a protein insolubility of about 80%, and a mean diameter particle size distribution range of about 0.1 μm to about 3.0 μm, wherein less than about 5% of the total number of particles have a diameter exceeding about 3.0 μm.

[0014] Arase (US20150181908) reported that the denatured protein product obtained by Villagran's 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. In fact, Arase reported that the denatured protein product obtained by Villagran's method showed visually recognizable gelation after heat treatment at 120°C in an autoclave for 15 minutes. Arase attempted to improve the product and use a feed with a lower protein mass percentage (12.5% ​​in Arase's working example). The average particle size of the resulting product was 0.38 to 0.7 μm, the protein insolubility was 43 to 47%, and it exhibited better stability than the product prepared by the Villagran method. However, the product prepared by the Arase method exhibited thermal instability because the average particle size increased to 0.5 to 3.5 μm when a 10% by mass protein solution was heated at 120°C for 15 minutes by an autoclave.

[0015] Havea (WO2010120199) provides a method for preparing a whey protein concentrate by providing a whey protein solution having a protein concentration of 15 to 50% (w / v) and a pH of 4.7 to 8.5; heating the solution to above 50°C for a period of time to denature the protein; and the heat treatment comprises heating the solution under turbulent conditions, such as a Reynolds number of at least 500.

[0016] Gulla (WO2013065014) and Cakir-Fuller (WO2020104954) reported that an exemplary heat-denatured whey protein concentrate prepared using the method described in Havea had a primary aggregate size D(4,3) for a 10% total solid protein solution of 1.70 μm, with good thermal stability, as indicated by 1% aggregate growth after heating the 10% total solid protein solution. Other exemplary heat-denatured whey protein concentrates prepared using the method described in Havea had a primary aggregate size D(4,3) of 1.62 to 2.50 μm for a 10% total solid protein solution and exhibited varying degrees of thermal stability.

[0017] Eriksen (WO2013117599) relates to the use of micronized whey protein in frozen dessert products. Eriksen exemplifies a micronized whey protein product in which at least about 10% of the particles are larger than 5 μm. For example, in Figure 2 In the Eriksen, a multimodal particle size distribution was shown with one peak between 0.1 and 1 μm and another between 1 and 10 μm, yielding a d of 0.097 μm. 10 , d of 0.316 μm 50 and d of 4.579 μm 90 .

[0018] Mikkelsen (WO2015059243) illustrates a method for producing a high-protein denatured whey protein composition, in which an aqueous solution containing sweet whey protein concentrate is prepared by dissolving whey protein concentrate in water to obtain a dry matter content of 16% and adjusting the pH to 6.4. Denaturation and micronization are carried out in a 6+6 scraped surface heat exchanger (SSHE), an APV shear agglomerator. After passing through a holding chamber (60 seconds), the product is cooled in the SSHE and then cooled to 10°C in a plate heat exchanger (PHE). During the heat treatment (80°C for 10 minutes), the protein denatures and forms particles with a size of 0.5 to 10 μm. The product suspension is pumped into a storage tank, and some of them are subsequently dried into a powder by spray drying. Mikkelsen reports that the resulting product suspension contains insoluble whey protein particles with a size range of 0.5 to 10 μm. Several additional patent applications from Arla Foods, including WO2018149869, WO2019110668 and WO2020187842, utilize the same process as outlined in Mikkelsen to obtain a denatured whey protein product comprising insoluble particles of denatured whey protein. Arla Foods sells microparticles WPC, including -YO8075, which is discussed in detail herein.

[0019] Burling (WO2005041677) prepared a whey protein gel suitable for stabilizing low-fat spreads and yogurts by cold gelation using only a small amount of whey protein. Burling noted that it is particularly important that the calcium concentration remain very low, the protein concentration remain relatively low and the pH remain at 7 or above during the heat treatment. In particular, pH>7 during the heating step promotes the formation of soluble aggregates or filaments with a size of 20 to 100 nm before gel formation.

[0020] A series of patent applications from Nestec, including WO2007110421, WO2007110422 and WO2007110423, relate to whey protein micelle powders produced in such a way that the micelles have an extremely sharp size distribution, such that more than 80% of the micelles produced have a size less 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 occurs at pH 6.0, but not at pH 6.8, which instead results in the formation of linear aggregates (i.e. acid gellable whey protein aggregates). It was found that the conversion yield of native whey protein to micelles decreases when the initial protein concentration is 12% or higher. The turbidity of a 3.4% protein solution measured at 500 nm was reported as 21 (Example 2), while the turbidity of a 4% whey protein micelle dispersion was reported as 80 at 500 nm (Examples 11-12).

[0021] The pasteurized liquid composition prepared by Minor (WO2009113845) includes 12g / 100mL or 16g / 100mL whey protein. The average particle size D(4,3) of the liquid composition obtained from static light scattering (Malvern Mastersizer 2000) falls between 3.7μm and 7.7μm before and after heat treatment. Example 4 of Minor shows that the average particle size D(4,3) of the whey protein composition obtained by static light scattering (Malvern Mastersizer 2000) is 0.48μm before spray cooking and 0.29μm after spray cooking.

[0022] 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 are also called whey protein micelles by Nestec (e.g., WO2007 / 110421), but their micellar properties are questionable. Nielsen emphasized that it is important to control the levels of Na, K and Ca ions to avoid gelation of the entire sample. Secondary heat treatment at UHT temperature without applying shear force resulted in gelation of the sample - heating at 150°C for 7 minutes and 30 seconds induced gelation of 14% and 16% protein content samples and heating at 150°C for 3 minutes and 10 seconds induced gelation of 20% protein content samples (and the 14% protein content sample was concentrated to 24%).

[0023] Dissanayake and Vasiljevic (J. Dairy Sci. 92, 1387-1397, 2009) prepared a whey protein composition by first heat treating a 10% (w / w) whey protein retentate sample at 90°C for 20 minutes and then applying dynamic high pressure shear (microfluidization) at 140 MPa using 1 or 5 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 a heat coagulation time (HCT) of 87.8 seconds for 1 pass and 102.5 seconds for 5 passes at 140°C.

[0024] The same group studied the functional properties of whey protein micronized at low pH (Dissanayake et al., J. Dairy Sci. 95, 1667-1679, 2012). The average particle size of whey protein micronized at low pH was about 100 nm. In particular, it was reported that the heat-treated, micronized, citric acid acidified (HTM-CA) and heat-treated, micronized, lactic acid acidified (HTM-LA) samples were 100 ± 6.3 nm and 101 ± 19.3 nm, respectively. Denatured micronized whey protein samples acidified with citric acid and lactic acid showed a thermal coagulation time (HCT) of more than 3 minutes at 140°C. However, the samples showed reduced thermal stability - the protein content retained in the supernatant after heating at 140°C for 10 seconds and subsequent centrifugation (12,000×g, 20°C) was only 19.7% and 10.9%, respectively, which may be caused by the specific acidulant effect.

[0025] While certain existing denatured whey protein compositions provide some resistance to gelling at 10% protein content in a standard heat coagulation time test (e.g., a HCT of 2 minutes or more at 140°C), 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.

[0026] Therefore, there remains a need for microparticulated whey protein compositions that are stable when exposed to a secondary heat treatment, in particular compositions that exhibit substantially no change in particle size distribution when exposed to a secondary heat treatment. Summary of the invention

[0027] The present disclosure relates to denatured whey protein compositions and methods of making and using such compositions.The denatured whey protein compositions include submicron-sized particles containing denatured whey protein.

[0028] For example, the particle size distribution of the denatured whey protein composition has some, 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 between about 1.0 μm and about 10.0 μm; a d of from about 0.5 to about 1.2 μm or from about 0.6 to about 1.0 μm; 50 ; d from about 0.9 to about 1.7 μm or from about 1.1 to about 1.5 μm 90; a volume weighted average diameter D(4,3) between about 0.6 and about 1.0 μm; and / or a substantially unimodal distribution such that at least 95% by volume or at least 98% by volume of the particles have a diameter of less than about 2.0 μm. Without wishing to be bound by theory, it is believed that at least the step of treating the whey protein solution and / or the whey protein retentate with an oxidant provides a denatured whey protein composition in which the reactivity of the denatured protein and the microparticles upon further heating is minimal, thereby making the microparticles relatively resistant to further growth of subsequent heat treatment. In fact, such denatured whey protein compositions are highly stable to secondary heating conditions and exhibit minimal or substantially no primary particle size growth after heating a 10% (w / w) protein content aqueous solution (at 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 residual variable 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 residual variable whey protein is less than 16%, or less than 14% or less than 12%. In certain embodiments, the denatured whey protein in the composition has a high degree of covalent bonding. For example, at least 60%, at least 70% or at least 80% of the beta-lactoglobulin in the denatured whey protein composition is covalently cross-linked to form a polymer (e.g., dimer, trimer, etc.). As another example, the ratio of covalent interaction to non-covalent interaction in the microparticle is at least 4 to 1. In addition, the high-protein liquid application prepared using the denatured whey protein composition, such as drinking yogurt or liquid nutritional composition, shows substantially no particle size change after heat treatment, and therefore shows minimal or substantially no sedimentation after long-term storage. Therefore, it is expected that the high-protein liquid application realized by the present technology provides excellent stability during the shelf life of the product.

[0029] In one aspect, the present disclosure provides a denatured whey protein composition comprising submicron particles having a particle size distribution as described herein. When exposed to a secondary heat treatment, the particle size distribution of the denatured whey protein composition is stable. For example, after heating a 10% (w / w) protein content aqueous solution at 120° C. for 15 minutes (e.g., in an autoclave or oil bath), the particle size distribution of the denatured whey protein composition is stable. 50 Maintained at about 0.6 to about 1.0 μm and / or d 90 Maintained at about 0.9 to about 1.7 μm.

[0030] The present disclosure provides a heat-stable denatured whey protein composition comprising at least 60% total protein on a dry weight basis and microparticles comprising denatured whey protein, wherein the microparticles have a particle size distribution comprising at least two characteristics selected from the group consisting of: (i) a volume weighted average diameter D(4,3) of not greater than about 1.0 μm, (ii) a d(4,3) of not greater than about 1.0 μm, 50 , (iii) a d of not greater than about 2.0 μm 90 , (iv) at least 92 volume % of the particles have a diameter less than about 2.0 μm, and (v) no more than 45 volume % of the particles have a diameter between about 1.0 and about 10.0 μm.

[0031] The present disclosure also provides liquid compositions, such as drinking yogurts, acidic beverages, neutral beverages or liquid nutritional compositions, which include the heat stable denatured whey protein compositions disclosed herein and food products, such as baked goods, bars, or set or stirred yogurts including the heat stable denatured whey protein compositions disclosed herein.

[0032] The present disclosure also provides a heat-treated, shelf-stable, high-protein liquid composition comprising at least 6% (w / v) whey protein, wherein the whey protein comprises microparticles comprising denatured whey protein; and wherein (i) the liquid composition has a d value of no greater than about 1.0 μm after applying a secondary heat treatment for microbial control. 50 and a d not greater than about 2.0 μm 90 (ii) the liquid composition has a temperature of 100s at 20°C -1 (iii) the liquid composition exhibits less than 10% sedimentation after storage at a temperature of about 20°C to about 25°C for 6 weeks; and / or (iv) the liquid composition exhibits less than 10% sedimentation after centrifugation at 1540 x g for 5 minutes.

[0033] The present disclosure also provides a liquid nutritional composition comprising at least 12% (w / v) total protein, such as 12% to 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 include a lipid component and / or a carbohydrate component; for example, 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, or 1 to 2 kcal / ml, or greater than 2 kcal / ml. In certain embodiments, the liquid nutritional composition has a caloric density of 1.5 to 2.5 kcal / ml. In certain embodiments, (i) the liquid nutritional composition has a d of not more than about 1.0 μm after secondary heat treatment for microbial control. 50 (ii) the liquid nutritional composition has a temperature of 100s at 20°C -1 (iii) the liquid nutritional composition exhibits less than 10% sedimentation after storage at a temperature of about 20°C to about 25°C for 6 weeks; and / or (iv) the liquid nutritional composition exhibits less than 10% sedimentation after centrifugation at 1540 x g for 5 minutes.

[0034] In one aspect, the present disclosure provides a denatured whey protein composition comprising microparticles, the microparticles comprising denatured whey protein, wherein at least 55% by volume of the microparticles have a diameter of about 0.2 μm to about 1.0 μm, and at least 95% by volume, at least 98% by volume, or at least 99% by volume of the microparticles have a diameter of less than about 2.0 μm; and wherein the denatured whey protein composition exhibits substantially no change in particle size distribution when exposed to a secondary heat treatment. In certain embodiments, after heating a 10% (w / w) protein content aqueous solution at 120° C. for 15 minutes (e.g., in an autoclave or oil bath), d 50 Maintained at about 0.6 to about 1.0 μm and / or d 90 Maintained at about 0.9 to about 1.7 μm.

[0035] In another aspect, the present disclosure provides a denatured whey protein composition comprising microparticles, the microparticles comprising denatured whey protein, wherein the microparticles have 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 have a diameter less than about 2.0 μm, and at least 99% by volume of the microparticles have a diameter less than about 3.0 μm; and wherein the denatured whey protein composition exhibits substantially no change in particle size distribution when exposed to a secondary heat treatment. In certain embodiments, after heating a 10% (w / w) protein content aqueous solution at 120° C. for 15 minutes (e.g., in an autoclave or oil bath), d 50 Maintained at about 0.6 to about 1.0 μm and / or d 90 Maintained at about 0.9 to about 1.7 μm.

[0036] In certain embodiments of any aspect disclosed herein, the stability of the particle size distribution is determined by evaluating the primary particle growth after (secondary) heat treatment of a 10% (w / w) protein content aqueous solution at 120°C for 15 minutes or at 140°C for 90 seconds (e.g., in an autoclave or oil bath).

[0037] In some such embodiments, a solution containing 10% (or 12%, or 14%, or 16%) (w / w) protein content of denatured whey protein exhibits minimal or substantially no change in particle size distribution when subjected to a primary particle growth test. For example, the primary particle growth test comprises a (secondary) heat treatment at 120° C. for 15 minutes (e.g., in an oil bath).

[0038] In some such embodiments, the d of the 10% (or 12%, or 14%, or 16%) (w / w) protein content solution is 50 There is essentially no increase after a primary particle growth test with heat treatment at 120°C for 15 minutes (eg, in an autoclave or oil bath).

[0039] In some such embodiments, after a primary particle growth test comprising heat treatment at 120°C for 15 minutes (e.g., in an autoclave or oil bath), the d 50 In some such embodiments, the d of the 10% (w / w) protein content aqueous solution after the primary particle growth test is not greater than about 1.0 μm. 50 From about 0.6 to about 1.0 μm.

[0040] In some such embodiments, the 10% (w / w) protein content aqueous solution is prepared before the primary particle growth test. 50 d after primary particle growth test 50The difference does not exceed 20%, 18%, 16%, 14%, 12%, 10%, 9%, 8%, 7%, 6% or 5%.

[0041] In some such embodiments, the d of the 10% (or 12%, or 14%, or 16%) (w / w) protein content solution is 90 There is essentially no increase after a primary particle growth test with heat treatment at 120°C for 15 minutes (eg, in an autoclave or oil bath).

[0042] In some such embodiments, after a primary particle growth test comprising heat treatment at 120°C for 15 minutes (e.g., in an autoclave or oil bath), the d 90 Less than about 2.0 μm. In some such embodiments, the d of the 10% (w / w) protein content aqueous solution after the primary particle growth test is 90 From about 0.9 to about 1.7 μm or from about 1.1 to about 1.5 μm.

[0043] In some such embodiments, the d of the 10% (w / w) protein content aqueous solution prior to the primary particle growth test is 90 After the primary particle growth test 90 The difference between the two parties is not more than 30%, not more than 25%, or not more than 20%.

[0044] In some such embodiments, the d of a 10% (w / w) protein content aqueous solution after a primary particle growth test comprising heat treatment at 120° C. for 15 minutes (e.g., in an autoclave or oil bath) is 50 is about 0.6 to about 1.0 μm, and the d of a 10% (w / w) protein content aqueous solution after the primary particle growth test 90 From about 0.9 to about 1.7 μm.

[0045] In certain embodiments of any of the aspects disclosed herein, the heat coagulation 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 containing 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.

[0046] In certain embodiments of any aspect disclosed herein, (i) the denatured whey protein composition comprises 65% to 95%, 75% to 90% or 80% to 85% total protein on a dry weight basis; (ii) the residual denatured 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 interactions 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 cross-linked; (iv) β-lactoglobulin is no more than 80% or no more than 75% of the total protein in the denatured whey protein composition; (v) the denatured whey protein composition comprises at least 10% to 15% of the total protein in the denatured whey protein composition; (vi) the denatured whey protein composition has a lactose content of at most 10 wt%, at most 8 wt%, at most 6 wt%, or at most 4 wt%; (vii) the denatured whey protein composition has an ash content of at most 10 wt%, at most 8 wt%, at most 6 wt%, or at most 4 wt%; (viii) the denatured whey protein composition has a casein content of at most 5 wt%, at most 4 wt%, at most 3 wt%, at most 2 wt%, or at most 1 wt%; or a combination of any two or more or all of (ix) (i) to (viii).

[0047] The present disclosure also provides a method for preparing a denatured whey protein composition as described herein. The method includes (a) providing a whey protein solution or a whey protein retentate; (b) contacting the whey protein solution or the whey protein retentate with an oxidant; and (c) heat-treating the whey protein solution or the whey protein retentate under conditions allowing protein denaturation, wherein the total protein content of the whey protein solution or the whey protein retentate during step (c) is preferably at least 16% (w / w). In certain embodiments, the oxidant is a peroxide, such as hydrogen peroxide, and step (b) includes contacting the whey protein solution or the whey protein retentate with a peroxide and a peroxidase. In certain embodiments, the oxidant is optionally inactivated, removed or consumed before step (c), so that during the heat treatment of step (c), the whey protein solution or the whey protein retentate can be substantially free of an oxidant. In certain embodiments, during the heat treatment of step (c), the whey protein solution or whey protein retentate is substantially free of an oxidant, wherein no active steps are taken to inactivate, remove or consume the oxidant. For example, the oxidant may be present in an amount (e.g., <10 ppm) such that it is not desirable to remove or consume the oxidant. In certain embodiments, the heat treatment of step (c) comprises a temperature of at least 70°C.

[0048] The present disclosure also provides edible consumables, including foods and beverages, and liquid nutritional compositions comprising the denatured whey protein compositions described herein.

[0049] The present disclosure also provides a method for providing a nutritional supplement to a subject in need thereof. The method comprises enterally administering to the subject a denatured whey protein composition disclosed herein or a liquid nutritional composition comprising such a denatured whey protein composition.

[0050] The present disclosure also provides a liquid composition comprising a denatured whey protein composition, wherein the liquid composition exhibits minimal or substantially no change in particle size distribution after pasteurization or sterilization. In certain embodiments, the liquid composition has a d 50 and d less than 2 μm 90 In certain embodiments, the liquid composition exhibits substantially no sedimentation, maintains a relatively low viscosity, and exhibits a pleasant taste and mouthfeel without a powdery or gritty feel after storage (e.g., at a temperature of about 20°C to about 25°C) for 1, 3, 6, or 12 months.

[0051] It is an object of the present invention to provide: a denatured whey protein composition having improved heat stability; and / or a composition which exhibits minimal or substantially no primary particle size growth upon secondary heat treatment; and / or a product containing a denatured whey protein composition wherein the product has a particle size distribution which provides improved mouthfeel, sedimentation and / or shelf life; and / or to at least provide the public with a useful choice.

[0052] These and other objects of the present invention are described in the following paragraphs.These objects are considered to narrow the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 is a process flow diagram for preparing an exemplary denatured whey protein composition.

[0054] Figure 2 is a process flow diagram for making an exemplary liquid composition, ie, a high protein beverage, including a denatured whey protein composition.

[0055] Figure 3 is a process flow diagram for making an exemplary liquid composition, namely a drinking yogurt, including a denatured whey protein composition.

[0056] Figure 4 Shown is the particle size distribution of a high protein beverage prepared with powder A, powder G or powder H after heating at 120°C for 15 minutes.

[0057] Figure 5 Shown is the particle size distribution of a protein solution containing Powder A at 10%, 12%, 14% or 16% (w / w) protein after heating at 120°C for 15 minutes.

[0058] Figure 6 Shown are the thermal coagulation times of denatured whey protein compositions prepared at protein concentrations ranging from 10% to 20% (w / w).

[0059] Figure 7 Shown is the particle size distribution of high protein (15% (w / w) and 20% (w / w)) drinking yogurts comprising denatured whey protein concentrate at 1 week.

[0060] Figure 8 Shown are the thermal coagulation times (HCT) of denatured whey protein compositions prepared from samples treated with different peroxide dosage rates.

[0061] Fig. 9 Shown is the protein profile of samples analyzed by HPLC after treatment with enzymes and before heat treatment.

[0062] Fig.10 Shown is the particle size distribution of a 12% (w / v) low calorie high protein beverage before and after indirect UHT heating (143°C, 6 seconds). DETAILED DESCRIPTION

[0063] This detailed description is intended only to familiarize others skilled in the art with the invention, its principles and its practical applications so that others skilled in the art can modify and apply the invention in its various forms as they may best suit the requirements of a particular application. This specification and its specific embodiments are for illustrative purposes only. Therefore, the present invention is not limited to the embodiments described in this patent application and various modifications may be made.

[0064] A. Definition

[0065] As used in the specification and the appended claims, unless otherwise indicated, the following terms have the indicated meanings:

[0066] The term "about" generally refers to a range of values ​​that one of ordinary skill in the art would consider equivalent to the stated value (e.g., ±5 to 10% of the stated value). Ranges may be expressed herein as from "about" one particular value, and / or to "about" another particular value. When such a range is expressed, the range includes the recited values.

[0067] The term "denatured whey protein composition" refers to a composition comprising at least some denatured whey protein and preferably a significant amount of denatured whey protein. The composition may also contain non-denatured whey protein. However, the denatured whey protein composition preferably has an insolubility of at least 50%. Alternatively or additionally, the residual denatured whey protein in the composition is preferably less than 20%.

[0068] The term "dry weight basis" refers to the percentage of a substance in a composition or product after the water in the product has been removed. This can be calculated by applying a correction to allow for the water retained in the product.

[0069] The term "liquid nutritional composition" refers to an aqueous composition, which is preferably administered to the stomach of a subject by mouth or other means, usually by tube feeding. Such other means include nasogastric feeding and gastric feeding. The term "liquid nutritional composition" includes medical foods, enteral nutrition, foods for special medical purposes, liquid dietary substitutes and supplements. The liquid nutritional compositions described herein provide a significant amount of protein and carbohydrates, and lipids are also generally provided. 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 or have been treated for a disease or condition, be 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 but not limited to an athlete or an active elderly person, including people with specific nutritional needs.

[0070] The term "microparticles" refers to a population of insoluble aggregates comprising denatured whey proteins. Such aggregates may be formed via covalent (e.g., intermolecular sulfhydryl-disulfide exchange reactions) and / or non-covalent interactions (e.g., hydrophobic interactions). Typically, a single microparticle may have a particle size of about 0.1 μm to about 3.0 μm or larger (e.g., up to about 20 μm), and the particle population has a size distribution as further defined herein.

[0071] The term "non-dairy protein" refers to any protein that is not a dairy protein (ie, any protein that is not derived from animal milk). Non-dairy proteins include proteins of plant origin, fungal proteins, and algal proteins.

[0072] The term "non-whey protein" refers to any protein that is not whey protein. As used herein, the term "non-whey protein" includes casein and protein derived from one or more non-dairy sources.

[0073] The term "subject" includes humans and other primates and 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 requires nutritional support.

[0074] The term "substantially unhydrolyzed" refers to unhydrolyzed (intact) proteins and proteins with 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, the whey proteins and non-whey proteins used herein are substantially non-hydrolyzed.

[0075] The term "total protein" relates to the total amount of protein of a composition or product and can be determined, for example, using the Kjeldahl method and an appropriate nitrogen conversion factor for milk protein.

[0076] The term "whey protein concentrate" or "WPC" refers to a whey fraction from which lactose has been at least partially removed to increase the protein content to at least 20 wt%. In certain embodiments, WPC has at least 35 wt% of the total solids (TS) as whey protein, and preferably at least 60 wt%, at least 65 wt%, at least 70 wt%, at least 75 wt%, or at least 80 wt%. For the purposes of this specification, the term "WPC" includes whey protein isolate (WPI) when the context permits.

[0077] The term "whey protein isolate" as used herein refers to a composition consisting primarily of whey protein with negligible lipid and lactose content. Therefore, the preparation of WPI generally requires more rigorous separation methods, such as a combination of microfiltration and ultrafiltration or ion exchange chromatography. WPI is generally considered to refer to a composition in which at least 90% by weight of the solids are whey protein.

[0078] The term "yogurt" or "sour milk" refers to an acidic or fermented food or beverage product prepared from a dairy source and containing live microorganisms or chemical acidulants or both. The term "yogurt" or "sour milk" includes set or stirred yogurt and drinking yogurt, and also includes ambient yogurt.

[0079] B. Denatured Whey Protein Composition

[0080] Denatured whey protein compositions disclosed herein include submicron particles and can exhibit substantially no change in particle size distribution when exposed to secondary heat treatment. Such whey protein compositions are well suited for applications involving secondary heating conditions, such as high temperature pasteurization, ultra-high temperature (UHT) treatment, or retort heating for sterilization and microbial control. Denatured whey protein compositions disclosed herein can provide a thinner, less powdery mouthfeel than previously known compositions and exhibit minimal or substantially no grit or sandy feel.

[0081] 1. Particle size distribution characteristics

[0082] As discussed herein, the particle size distribution profile is based on the particle size distribution of a reconstituted liquid comprising the denatured whey protein composition disclosed herein.

[0083] In certain embodiments, the composition includes microparticles containing denatured whey protein, wherein the microparticles have a volume weighted average diameter D(4,3) of no more than about 1.0 μm. In some such embodiments, the volume weighted average diameter D(4,3) of the microparticles is 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% by volume, at least 93% by volume, at least 94% by volume, at least 95% by volume, at least 96% by volume, at least 97% by volume, at least 98% by volume, 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% by volume, at least 96% by volume, at least 97% by volume, at least 98% by volume, 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.

[0084] In certain embodiments, the composition includes microparticles containing 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% by volume, at least 86% by volume, at least 87% by volume, at least 88% by volume, 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% by volume, at least 96% by volume, at least 97% by volume, at least 98% by volume, 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 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 includes microparticles containing 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% by volume, at least 86% by volume, at least 87% by volume, at least 88% by volume, 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% of the microparticles by volume have a diameter of less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% of the microparticles by volume have a diameter of less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% of the microparticles by volume have a diameter of less than about 3.0 μm.

[0086] In certain embodiments, the composition includes microparticles containing 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 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% of the microparticles by volume have a diameter of less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% of the microparticles by volume have a diameter of less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% of the microparticles by volume have a diameter of less than about 3.0 μm.

[0087] In certain embodiments, the composition includes microparticles containing 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 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% of the microparticles by volume have a diameter of less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% of the microparticles by volume have a diameter of less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% of the microparticles by volume have a diameter of less than about 3.0 μm.

[0088] In certain embodiments, the composition includes microparticles containing 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% by volume, at least 86% by volume, at least 87% by volume, at least 88% by volume, 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% of the microparticles by volume have a diameter of less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% of the microparticles by volume have a diameter of less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% of the microparticles by volume have a diameter of less than about 3.0 μm.

[0089] In certain embodiments, the composition comprises microparticles comprising denatured whey protein, wherein the microparticles have a d of no more than about 1.2 μm, no more than about 1.1 μm, no more than about 1.0 μm, or 50 In some such embodiments, the microparticles have a d 50 In some embodiments, at least 85% by volume, at least 86% by volume, at least 87% by volume, at least 88% by volume, or at least 89% by volume of the microparticles have a diameter less than about 1.5 μm. In some embodiments, at least 91% by volume, at least 92% by volume, at least 93% by volume, at least 94% by volume, at least 95% by volume, at least 96% by volume, at least 97% by volume, at least 98% by volume, or at least 99% by volume of the microparticles have a diameter less than about 2.0 μm. In some embodiments, at least 98% by volume or at least 99% by volume of the microparticles have a diameter less than about 2.5 μm. In some embodiments, at least 99% by volume 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, wherein the microparticles have a d of no more than about 2.0 μm, no more than about 1.9 μm, or no more than about 1.8 μm. 90 In some such embodiments, the microparticles have a d 90 In some embodiments, at least 85% by volume, at least 86% by volume, at least 87% by volume, at least 88% by volume, or at least 89% by volume of the microparticles have a diameter less than about 1.5 μm. In some embodiments, at least 91% by volume, at least 92% by volume, at least 93% by volume, at least 94% by volume, at least 95% by volume, at least 96% by volume, at least 97% by volume, at least 98% by volume, or at least 99% by volume of the microparticles have a diameter less than about 2.0 μm. In some embodiments, at least 98% by volume 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 the microparticles have a d value 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 value 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 90In some such embodiments, at least 85%, at least 86%, at least 87%, at least 88%, or at least 89% of the microparticles by volume 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% of the microparticles by volume have a diameter of less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% of the microparticles by volume have a diameter of less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% of the microparticles by volume have a diameter of less than about 3.0 μm.

[0092] In certain embodiments, the composition comprises microparticles comprising denatured whey protein, wherein the microparticles have a d value 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 value 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 value 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% of the microparticles by volume 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% of the microparticles by volume have a diameter of less than about 2.0 μm. In some such embodiments, at least 98% or at least 99% of the microparticles by volume have a diameter of less than about 2.5 μm. In some such embodiments, at least 99% or at least 99.9% of the microparticles by volume have a diameter of less than about 3.0 μm.

[0093] In certain embodiments, the composition includes microparticles containing denatured whey protein, wherein no more than 45% by volume of the microparticles have a diameter of about 1.0 to about 10.0 μm. In some such embodiments, at least 85% by volume, at least 86% by volume, at least 87% by volume, at least 88% by volume, 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% by volume, at least 92% by volume, at least 93% by volume, at least 94% by volume, at least 95% by volume, at least 96% by volume, at least 97% by volume, at least 98% by volume, 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% by volume 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% by volume or at least 99.9% by volume of the microparticles have a diameter less than about 3.0 μm.

[0094] As used herein, a particle size of about 0.4 μm encompasses 0.35 μm to 0.44 μm, a particle size of about 0.5 μm encompasses 0.45 μm to 0.54 μm, a particle size of about 0.6 μm encompasses 0.55 μm to 0.64 μm, a particle size of about 0.7 μm encompasses 0.65 μm to 0.74 μm, a particle size of about 0.8 μm encompasses 0.75 μm to 0.84 μm, a particle size of about 0.9 μm encompasses 0.85 μm to 0.94 μm, and a particle size of about 1.0 μm encompasses 0.95 μm to 1.04 μm.

[0095] Methods for particle size are known in the art. For example, particle size can be measured using a Malvern Mastersizer 2000 (Malvern Instruments Ltd, Worcs, UK), wherein the refractive index of the particles is 1.46, and the refractive index of the solvent is 1.33. For example, in an exemplary embodiment, a suspension of 10% total solids (TS) is homogenized (e.g., 150 / 50 bar) at natural pH and the average particle size (characterized by D[4,3]) is determined using a Malvern Mastersizer 2000 (Malvern Instruments Ltd, Worcs, UK), wherein the refractive index of the particles is 1.46, and the refractive index of the solvent is 1.33, as described above.

[0096] 2. Stable particle size distribution

[0097] In certain embodiments, the particle size distribution of the denatured whey protein composition is substantially unchanged after a primary particle growth test comprising heat treatment at 120°C for 15 minutes (eg, in an autoclave or oil bath).

[0098] In certain embodiments, the denatured whey protein composition exhibits minimal or substantially 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 at 120° C. for 15 minutes (e.g., in an autoclave or oil bath). For example, a 10% (w / w) protein content aqueous solution containing a denatured whey protein composition exhibits minimal or substantially no change in particle size distribution after a primary particle growth test.

[0099] In some such embodiments, the d of the 10% (or 12%, or 14%, or 16%) (w / w) protein content solution is 50 There is essentially no increase after a primary particle growth test with heat treatment at 120°C for 15 minutes (eg, in an autoclave or oil bath).

[0100] In some such embodiments, after a primary particle growth test comprising heat treatment at 120°C for 15 minutes (e.g., in an autoclave or oil bath), the d 50 In some such embodiments, the d of the 10% (w / w) protein content aqueous solution after the primary particle growth test is not greater than about 1.0 μm. 50 From about 0.6 to about 1.0 μm.

[0101] In some such embodiments, the 10% (w / w) protein content aqueous solution is prepared before the primary particle growth test. 50 d after primary particle growth test 50 The difference does not exceed 20%, 18%, 16%, 14%, 12%, 10%, 9%, 8%, 7%, 6% or 5%.

[0102] In some such embodiments, the denatured whey protein compositions disclosed herein have a d value of about 0.040 for the denatured whey protein compositions when subjected to the primary particle growth test at a protein content of up to 16% (w / w). 50 is stable.

[0103] In some such embodiments, the d of the 10% (or 12%, or 14%, or 16%) (w / w) protein content solution is 90 There is essentially no increase after a primary particle growth test with heat treatment at 120°C for 15 minutes (eg, in an autoclave or oil bath).

[0104] In some such embodiments, after a primary particle growth test comprising heat treatment at 120°C for 15 minutes (e.g., in an autoclave or oil bath), the d 90is not greater than about 2.0 μm, not greater than about 1.9 μm, or not greater than about 1.8 μm. In some such embodiments, the d 90 From about 0.6 to about 2.0 μm, from about 0.7 to about 1.9 μm, from about 0.8 to about 1.8 μm, from about 0.9 to about 1.7 μm, from about 1.0 to about 1.6 μm, or from about 1.1 to about 1.5 μm.

[0105] In some such embodiments, the d of the 10% (w / w) protein content aqueous solution prior to the primary particle growth test is 90 d after primary particle growth test 90 The difference is no more than 30%, no more than 28%, no more than 26%, no more than 24%, no more than 22%, no more than 20%, no more than 18% or no more than 16%.

[0106] In some such embodiments, the denatured whey protein compositions disclosed herein have a d value of 0.0404 when subjected to the primary particle growth test at a protein content of up to 16% (w / w). 90 is stable.

[0107] In some such embodiments, the d of a 10% (w / w) protein content aqueous solution after a primary particle growth test comprising heat treatment at 120° C. for 15 minutes (e.g., in an autoclave or oil bath) is 50 is about 0.6 to about 1.0 μm, and the d of a 10% (w / w) protein content aqueous solution after the primary particle growth test 90 From about 1.1 to about 1.5 μm.

[0108] Methods for assessing primary particle growth are known in the art. In certain embodiments, primary particle growth is determined as described herein. Briefly, a 10% (w / w) protein solution (pH 6.8) is heated at 120° C. for 15 minutes (e.g., in an autoclave or oil bath). Particle size distribution can be assessed after heat treatment.

[0109] The stability of the particle size distribution of the denatured whey protein composition or the liquid composition comprising the denatured whey protein composition includes minimal or substantially no gelation, sedimentation or aggregation after the secondary heat treatment. The gelation of the liquid composition is considered to be a change of state from a liquid to a soft solid to a hard solid. If the solution no longer flows after heating, it is considered to have gelled.

[0110] In certain embodiments, the particle size distribution of the denatured whey protein composition is stable after sterilization and / or pasteurization (e.g., high temperature pasteurization, UHT treatment 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 treatment generally involves subjecting the composition to a temperature higher than 135°C, such as 135°C to 150°C to achieve sterilization. The typical holding time of UHT is 4 to 10 seconds (or longer). Generally, two variations of UHT treatment are used: direct or indirect. In a direct UHT heating system, steam is directly mixed with the liquid composition, while in an indirect UHT heating system, the liquid composition is heated by contacting with steam or superheated water on the entire metal surface of a heat exchanger (Zadow, 1986). Retort treatment generally involves subjecting the composition to a temperature between 110°C and 130°C in a sealed tank for 10 to 20 minutes to achieve sterilization.

[0111] 3. Thermal condensation time

[0112] In certain embodiments, the heat coagulation time (HCT) of a 10% (w / w) protein content aqueous solution containing a 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 a 10% (w / w) protein content aqueous solution containing a denatured whey protein composition is at least 14 minutes. 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.

[0113] Methods for determining the heat setting time (HCT) are known in the art. The HCT method involves sealing 1 mL of a sample in a glass tube, which is clamped on a platform and placed in a silicone oil bath, which is thermostatically controlled at 140° C. with a defined shaking rate. The length of time between placing the container in the oil bath and the onset of visible aggregate formation is defined as HCT (Singh H & Creamer LK (1992), Determination of heat stability, In: Advanced Dairy Chemistry edFox PF Elsevier).

[0114] Applicants believe, without wishing to be bound by any theory and based on experience including that described herein, that any liquid composition having a heat setting time of less than 60 seconds has a high risk of extensive fouling and plugging of the UHT heating equipment, while any liquid composition having a HCT of 65 to 80 seconds at 140° C. has a potential risk of fouling. As described herein, liquid compositions having a heat setting time above 80 seconds are considered stable to UHT heating treatment at 140° C. for 5 seconds. Alternatively or additionally, samples having a HCT of less than 3 minutes have a high risk of gelling and agglomeration in the distillation pot after heating at 121° C. in an oil bath.

[0115] In some such embodiments, the denatured whey protein compositions disclosed herein exhibit minimal or substantially no gelling or coagulation at 140°C after 2 minutes at a protein content of up to 16% (w / w).

[0116] In certain embodiments, the HCT of 12% (w / w) protein content solution containing a denatured whey protein composition is at least 2 minutes. In some such embodiments, the HCT of 12% (w / w) protein content solution containing a denatured whey protein composition is at least 8 minutes. In some such embodiments, the HCT of 14% (w / w) protein content solution containing a denatured whey protein composition is at least 4 minutes. In some such embodiments, the HCT of 16% (w / w) protein content solution containing a denatured whey protein composition is at least 2 minutes.

[0117] 4. High Protein Content

[0118] In certain embodiments, the denatured whey protein composition comprises 60% to 95% by weight of total protein based on dry weight. In some such embodiments, the denatured whey protein composition comprises 65% to 95% by weight of total protein based on dry weight. In some such embodiments, the denatured whey protein composition comprises 70% to 95% by weight of total protein based on dry weight. In some such embodiments, the denatured whey protein composition comprises 75% to 90% by weight of total protein based on dry weight. In some such embodiments, the denatured whey protein composition comprises 80% to 85% by weight of total protein based on dry weight.

[0119] In certain embodiments, the denatured whey protein composition comprises at least 60 wt%, at least 65 wt%, at least 70 wt%, at least 75 wt%, or at least 80 wt% total protein based on dry weight. In some such embodiments, the denatured whey protein composition comprises at least 70 wt%, at least 75 wt%, or at least 80 wt% total protein based on dry weight.

[0120] 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 calculates protein concentration by multiplying the total nitrogen result by the nitrogen conversion factor for milk protein, 6.38.

[0121] Optionally, non-protein nitrogen (NPN) is also determined, for example, by: ISO 8968-4 / IDF 020-4 - Milk - Determination of nitrogen content - Part 4: Determination of non-protein nitrogen content and true protein content of samples calculated as (m 总氮 -m NPN )*6.38. Unless otherwise specified herein, total protein content is not reported with reference to NPN.

[0122] 5. High variability

[0123] Whey proteins include bovine serum albumin (BSA), α-lactalbumin, β-lactoglobulin, lactoferrin, immunoglobulins and other small proteins. Cheese whey-based ingredients may contain additional proteins, such as glycomacropeptide (GMP) and Peptone 5 (pp5), which are casein-related proteins. Heat treatment of whey or WPC leads to denaturation of BSA, α-lactalbumin, β-lactoglobulin, lactoferrin, immunoglobulins. In contrast, GMP and pp5 are non-denaturable.

[0124] As described herein, acid casein whey lacking GMP can be used as a source of whey protein. For example, an exemplary ingredient can be prepared using 60% cheese WPC and 40% acid WPC, which will provide a higher level of heat-denatured protein than a corresponding ingredient prepared from cheese WPC alone.

[0125] Methods for determining the degree of protein denaturation are known in the art. The exemplary method used herein relies on HPLC (Elgar et al. (2000) J Chromatography A, 878, 183-196); other methods suitable include methods relying on Agilent 2100 Bioanalyzer ((Agilent Technologies, Inc. 2000, 2001-2007, Waldbronn, Germany) and microfluidic chips, and methods utilizing 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).

[0126] The denatured whey protein composition may be characterized by measuring the residual denatured protein as a proportion of the total protein (TN x 6.38) according to the following formula:

[0127]

[0128] Soluble whey protein was determined using reverse phase HPLC as described above (Elgar et al., 2000) and expressed as grams protein / 100 grams powder.

[0129] Variable whey protein was measured as Σ(bovine serum albumin + α-lactalbumin + β-lactoglobulin + lactoferrin + immunoglobulins).

[0130] In certain embodiments, the residual variable 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 residual variable whey protein is less than 16%, or less than 14% or less than 12%.

[0131] For an exemplary denatured whey protein composition, the proportion of denatured protein that has been denatured can be estimated according to the following formula:

[0132]

[0133] Denatured whey protein compositions can also be characterized by insolubility. For example, a suspension containing a denatured whey protein composition in NaCl is treated with acetic acid and centrifuged. The total protein content of the supernatant and the suspension that was not centrifuged is determined, and the insoluble percentage of the sample can be determined as follows:

[0134]

[0135] In certain embodiments, the denatured whey protein composition is at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% insoluble.

[0136] 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-denatured whey protein in the composition is present in a denatured state. In some such embodiments, at least 60% of the heat-denatured whey protein is present in a denatured state, at least 70% of the heat-denatured whey protein is present in a denatured state, or at least 80% of the heat-denatured whey protein is present in a denatured state.

[0137] Denatured whey protein compositions can also be characterized according to the ratio of non-denatured protein (e.g., GMP) to residual denatured 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.

[0138] 6. Covalent / non-covalent bonds

[0139] Both covalent and non-covalent interactions can participate in the formation of whey protein microparticles. Covalent interactions involve electron sharing 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 provide irreversible aggregation of whey proteins.

[0140] 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, making it highly reactive in covalent interactions. BSA contains 17 SS bonds and one SH group, also making it capable of initiating covalent aggregation. α-Lactalbumin contains 4 SS bonds but no free SH groups to act as a starting point for covalent aggregation reactions.

[0141] In certain embodiments, the denatured whey protein in the composition has a high degree of covalent bonding.

[0142] 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 with another protein (e.g., another β-lactoglobulin molecule and / or BSA and / or α-lactalbumin) to form a multimer (e.g., dimer, trimer, etc.). In some such embodiments, at least 85% of the β-lactoglobulin in the denatured whey protein composition is covalently cross-linked with another protein.

[0143] As another example, the ratio of covalent interactions to non-covalent interactions in the microparticle is at least 2 to 1, at least 3 to 1, or at least 4 to 1.

[0144] 7. Other characteristics of the denatured whey protein composition

[0145] In certain embodiments, alpha-lactalbumin does not exceed 60%, does not exceed 50%, or does not exceed 40% of the total protein in the denatured whey protein composition.

[0146] In certain embodiments, the denatured whey protein composition includes about 5% to about 30% alpha-lactalbumin, about 10% to about 25% alpha-lactalbumin, or about 15% to about 20% alpha-lactalbumin, relative to the total protein in the denatured whey protein composition.

[0147] In certain embodiments, the beta-lactoglobulin does not exceed 80%, does not exceed 75%, or does not exceed 70% of the total protein in the denatured whey protein composition.

[0148] In certain embodiments, the denatured whey protein composition includes 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 relative to the total protein in the denatured whey protein composition.

[0149] In certain embodiments, the denatured whey protein composition has a ratio of alpha-lactalbumin to beta-lactoglobulin of about 1:15 to about 10:1, about 1:10 to about 5:1, or about 1:5 to about 1:1.

[0150] In certain embodiments, the denatured whey protein composition exhibits significant turbidity when in aqueous solution. For example, a 3.4% (w / w) protein content aqueous solution containing a denatured whey protein composition exhibits a turbidity value of at least 50 absorption units measured at 500nm and 20°C. In some such embodiments, a 3.4% (w / w) protein content aqueous solution containing a denatured whey protein composition exhibits a turbidity value of at least 75 or at least 100 absorption units measured at 500nm. As another example, a 4% (w / w) protein content aqueous solution containing a denatured whey protein composition exhibits a turbidity value of at least 90 absorption units measured at 500nm. In some such embodiments, a 4% (w / w) protein content aqueous solution containing a denatured whey protein composition exhibits a turbidity value of at least 100, at least 110 or at least 120 absorption units measured at 500nm.

[0151] To assess turbidity, it may be necessary to further dilute the aqueous solution. For example, a 3.4% (w / w) protein content aqueous solution is 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 this case, the dilution factor can be used to obtain the absorbance units of the original solution.

[0152] 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, such as from about 30 mM to about 150 mM. The monovalent metal cations include Na + and K + .

[0153] In certain embodiments, the denatured whey protein composition comprises less than about 10 wt%, less than about 9 wt%, less than about 8 wt%, less than about 7 wt%, less than about 6 wt%, less than about 5 wt%, less than about 4 wt%, less than about 3 wt%, less than about 2 wt%, less than about 1 wt%, or less than about 0.1 wt% lactose. In some such embodiments, the denatured whey protein composition has a lactose content of at most 10 wt%, at most 8 wt%, at most 6 wt%, or at most 4 wt%.

[0154] 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.

[0155] In certain embodiments, the denatured whey protein composition has been treated or is derived from a source (eg, WPC) that has been treated to reduce the lactose concentration, for example, by hydrolysis of lactose.

[0156] 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.

[0157] In certain embodiments, the denatured whey protein composition comprises less than about 20 wt%, less than about 15 wt%, less than about 10 wt%, less than about 9 wt%, less than about 8 wt%, less than about 7 wt%, less than about 6 wt%, less than about 5 wt%, less than about 4 wt%, less than about 3 wt%, less than about 2 wt%, less than about 1 wt%, or less than about 0.1 wt% fat. In some such embodiments, the denatured whey protein composition has a fat content of at most 10 wt%, at most 8 wt%, at most 6 wt%, or at most 4 wt%.

[0158] In certain embodiments, the denatured whey protein composition has 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.

[0159] In certain embodiments, the denatured whey protein composition comprises less than about 12 wt%, less than about 11 wt%, less than about 10 wt%, less than about 9 wt%, less than about 8 wt%, less than about 7 wt%, less than about 6 wt%, less than about 5 wt%, or less than about 4 wt% ash. In some such embodiments, the denatured whey protein composition has an ash content of at most 10 wt%, at most 8 wt%, at most 6 wt%, or at most 4 wt%.

[0160] In certain embodiments, the denatured whey protein composition has a whey protein to ash ratio of 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, or at least 20 to 1.

[0161] In certain embodiments, the denatured whey protein composition comprises casein 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. In some such embodiments, the denatured whey protein composition has a casein content of at most 5 % by weight, at most 4 % by weight, at most 3 % by weight, at most 2 % by weight or at most 1 % by weight. In some such embodiments, the denatured whey protein composition is substantially free of casein.

[0162] In certain embodiments, the denatured whey protein composition has a whey protein to casein 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, or at least 35 to 1.

[0163] Each feature of the denatured whey protein composition described herein can be combined with any other feature or features, unless clearly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous can be combined with any other feature or features indicated as being preferred or advantageous.

[0164] By way of example and not limitation, in certain embodiments, the denatured whey protein composition comprises microparticles containing denatured whey protein and has a particle size distribution that is substantially stable after secondary heat treatment. The particle size distribution of the denatured whey protein composition has some, 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 between about 1.0 μm and about 10.0 μm; from about 0.5 to about 1.2 μm or from about 0.6 to about 1.0 μm; 50 ; d from about 0.9 to about 1.7 μm or from about 1.1 to about 1.5 μm 90; a volume-weighted average diameter D(4,3) between about 0.6 and about 1.0 μm; and / or a substantially unimodal distribution such that at least 95% by volume or at least 98% by volume of the particles have a diameter less than about 2.0 μm.

[0165] After a primary particle growth test comprising heat treatment at 120°C for 15 minutes (e.g., in an autoclave or oil bath), some and preferably all of the aforementioned particle size distribution characteristics remain substantially the same. For example, the d 50 Not more than about 1.0 μm, d of 10% (w / w) protein content aqueous solution after primary particle growth test 90 is not greater than about 2.0 μm, not greater than about 1.9 μm, or not greater than about 1.8 μm. In some such embodiments, the d of the 10% (w / w) protein content aqueous solution after the primary particle growth test is 50 The primary particle growth test results in a 10% (w / w) protein content aqueous solution with a d value of about 0.6 to about 1.0 μm. 90 From about 1.1 to about 1.5 μm.

[0166] In some such embodiments, the denatured whey protein composition has a substantially stable particle size distribution and exhibits minimal or substantially no primary particle size growth after heating a 10% (w / w) protein content aqueous solution (at pH 6.8) at 120° C. for 15 minutes, and further has the following properties or a combination of properties:

[0167] (i) the denatured whey protein composition comprises from 65 wt% to 95 wt%, from 75 wt% to 90 wt%, or from 80 wt% to 85 wt% total protein on a dry weight basis;

[0168] (ii) the residual denatured 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%;

[0169] (iii) the ratio of covalent interactions 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 cross-linked;

[0170] (iv) β-lactoglobulin is no more than 80% or no more than 75% of the total protein in the denatured whey protein composition;

[0171] (v) the denatured whey protein composition has a lactose content of at most 10 wt%, at most 8 wt%, at most 6 wt% or at most 4 wt%;

[0172] (vi) 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%, up to 6%, or up to 4% by weight;

[0173] (vii) the denatured whey protein composition has an ash content of at most 10 wt%, at most 8 wt%, at most 6 wt%, or at most 4 wt%;

[0174] (viii) the denatured whey protein composition has a casein content of at most 5 wt%, at most 4 wt%, at most 3 wt%, at most 2 wt% or at most 1 wt%; or

[0175] (ix) A combination of any two or more or all of (i) to (viii).

[0176] In certain embodiments, the denatured whey protein composition is heat stable (e.g., a 10% (w / w) protein content aqueous solution exhibits a HCT of at least 3, at least 6, at least 9, 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.

[0177] C. Whey protein source

[0178] The whey protein source for the denatured whey protein composition can be any source providing one or more whey proteins, such as sweet whey or acid casein whey or milk whey (obtained from the microfiltration of skim milk as the permeate phase) or a combination thereof. Exemplary sources of whey protein include, but are not limited to, rennet whey or cheese whey, lactic acid whey, inorganic acid whey, casein whey and microfiltration skim milk whey. In certain embodiments, the whey protein source for the denatured whey protein composition is whey protein concentrate (WPC) or whey protein isolate (WPI). The whey protein composition, including WPC and WPI, can be derived from acid casein whey or cheese whey. Alternative sources of whey protein for the denatured whey protein composition include enriched compositions containing separate whey proteins (e.g., compositions rich in beta-lactoglobulin or compositions rich in alpha-lactalbumin).

[0179] WPC is rich in whey proteins, but also contains other components such as lipids, lactose, minerals / ash, and in the case of cheese whey based WPC, glycomacropeptide (GMP), an immutable casein-related non-globular protein.A typical process for producing whey protein concentrate utilizes membrane filtration.

[0180] WPC is usually described as a percentage of whey protein (w / w) in "WPC". For example, WPC80 is a WPC with 80% whey protein by weight.

[0181] The whey protein may be derived from any mammalian species, such as cows, sheep, goats, horses, buffalo, deer and camels. Preferably, the whey protein is bovine.

[0182] In certain embodiments, denatured whey protein may be obtained according to methods described in US 6,767,575 (Huss & Spiegel), US 2006 / 0204643 (Merrill et al.), US 4,734,827 (Singer et al.), US 5,494,696 (Holst et al.), US 2012 / 0114795 (Havea et al.; also published as WO 2010 / 120199), EP 0 412 590 (Hakkaart et al.), and US 5,188,842 (Visser et al.; also published as EP 0 347 237). Each method of preparing a denatured whey protein component will impart different properties, so anyone using the present invention should select a protein component to best suit their method.

[0183] In certain embodiments, the whey protein source is available as a powder, preferably a WPC or WPI powder.

[0184] The denatured whey protein composition can be prepared by a mixture of cheese and / or acid WPC and / or WPI or by a mixture of protein. In certain embodiments, the whey protein source is or includes WPC and / or WPI. In certain embodiments, the whey protein source is or includes a blend of WPC and / or WPI and optionally one or more ingredients including whey and / or non-whey proteins.

[0185] In certain embodiments, the whey protein source is cheese whey or WPC prepared by cheese whey. In some such embodiments, coloring agent is used in cheese preparation process. For example, cheese manufacturers can use annatto to provide orange-yellow color for colored cheese such as cheddar cheese, Leicester cheese, Gloucester cheese etc. The annatto coloring agent is derived from the seed of Bixa orellana (annatto), which is a natural shrub in Central America. Seed contains carotenoid pigments, including bixin, norbixin and orelline. In some such embodiments, the whey protein source includes coloring agent.

[0186] D. Manufacturing method

[0187] In at least one aspect, the present disclosure provides methods for making a denatured whey protein composition. Exemplary methods for making a denatured whey protein composition are described below and are described in Figure 1 Suitable modifications of the methods for achieving the denatured whey protein compositions described herein will be apparent to those skilled in the art.

[0188] In certain embodiments, the method comprises (a) providing a whey protein solution or a whey protein retentate; (b) contacting the whey protein solution or the whey protein retentate with an oxidant; and (c) heat treating the whey protein solution or the whey protein retentate under conditions that allow the desired protein denaturation to occur, wherein the total protein content of the whey protein solution or the whey protein retentate is preferably at least 16% (w / w). In some such embodiments, the denaturation heat treatment is carried out at a temperature greater than 70° C. under conditions of mechanical shear and / or turbulence, as known in the art.

[0189] In certain embodiments, the oxidizing agent is a peroxide, such as hydrogen peroxide, and step (b) comprises contacting the whey protein solution or the whey protein retentate with the peroxide and a peroxidase.

[0190] In certain embodiments, the amount of oxidant is less than 300 ppm, such as from about 5 to about 250 ppm, such as 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.

[0191] In certain embodiments, the amount of oxidant is less than 1200×10 -6 kg / kg whey protein, such as about 20×10 -6 kg / kg whey protein to about 900×10 -6 kg / kg whey protein, about 40×10 -6 kg / kg whey protein to about 800×10 -6 kg / kg whey protein, or about 60×10 -6 kg / kg whey protein to about 700×10 -6 kg / kg whey protein, such as about 80×10 -6 kg / kg whey protein to about 600×10 -6 kg / kg whey protein, such as about 100×10 -6 kg / kg whey protein to about 400×10 -6 kg / kg whey protein.

[0192] In certain embodiments, the amount of oxidant is less than 2000×10 -6 kg / kg variable whey protein, such as about 30×10 -6 kg / kg variable whey protein to about 1400×10 -6 kg / kg variable whey protein, about 60×10 -6 kg / kg variable whey protein to about 1200×10 -6 kg / kg variable whey protein, or about 90×10 -6 kg / kg variable whey protein to about 1000×10 -6 kg / kg variable whey protein, such as about 120×10 -6 kg / kg variable whey protein to about 800×10 - 6 kg / kg variable whey protein, such as about 150×10 -6 kg / kg variable whey protein to about 600×10 -6 kg / kg variable whey protein.

[0193] In certain embodiments, the amount of oxidant is less than 2400×10 -6 kg / kg β-lactoglobulin protein, such as about 40×10 -6 kg / kg β-lactoglobulin protein to about 1800×10 -6 kg / kgβ-lactoglobulin protein, about 80×10 -6 kg / kg β-lactoglobulin protein to about 1600×10 -6 kg / kg β-lactoglobulin protein, or about 120 × 10 -6 kg / kg β-lactoglobulin protein to about 1400×10 -6 kg / kg β-lactoglobulin protein, such as about 160×10 -6 kg / kg β-lactoglobulin protein to about 1200×10 -6 kg / kg β-lactoglobulin protein, such as about 200×10 -6 kg / kg β-lactoglobulin protein to about 800×10 -6 kg / kg β-lactoglobulin protein.

[0194] In certain embodiments, the molar ratio of oxidant to β-lactoglobulin protein is less than 2, e.g., 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.

[0195] In some such embodiments, the whey protein solution or whey protein retentate includes a whey protein source such as WPC and / or WPI. In some such embodiments, the whey protein source includes a colorant.

[0196] In certain embodiments, the oxidizing agent is a peroxide, such as hydrogen peroxide, and step (b) comprises contacting the whey protein solution or the whey protein retentate with the peroxide and a peroxidase.

[0197] In certain embodiments, a certain amount of oxidant is added in one step. More preferably and in order to avoid catalyst inhibition and deactivation, the total amount of peroxide is added in more than one step, for example in 2, 3, 4, 5 or more steps. More preferably, the amount of peroxide is added continuously. One skilled in the art can determine the optimal rate of oxidant addition as a function of catalyst / enzyme activity and protein concentration in the whey solution.

[0198] In certain embodiments, the oxidant is optionally inactivated, removed or consumed before step (c), so that during the heat treatment of step (c), the whey protein solution or the whey protein retentate may be substantially free of the oxidant. In certain embodiments, during the heat treatment of step (c), the whey protein solution or the whey protein retentate is substantially free of the oxidant, wherein no active step is taken to inactivate, remove or consume the oxidant. For example, the oxidant may be present in an amount (e.g., <10ppm) that does not wish to remove or consume the oxidant.

[0199] In certain embodiments, the denaturing heat treatment step is carried out at a temperature above 70°C and under conditions of high shear stress / force, which can be generated by increasing the slurry viscosity, or the wall shear rate, or changing the flow pattern to turbulent, or applying mechanical shear, or a combination thereof.

[0200] In an exemplary embodiment, Figure 1 As shown, the 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 and optionally in combination with a peroxidase); and subjecting the whey protein retentate to a heat treatment step that allows protein denaturation to occur (preferably, when no detectable oxidizing agent is left).

[0201] like Figure 1 As shown, cheese whey collected after cheese making is clarified and heated (pasteurized) to provide microbial control and also to inactivate any remaining rennet and starter cultures from cheese making.

[0202] General conditions for thermalization (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 a batch hold process), about 72°C (about 161°F) for 15 seconds (also known as a high temperature short time (HTST) process), about 89°C (about 192°F) for 1 second, or any alternative thermal and non-thermal treatments that have an equivalent bactericidal effect to the above treatments.

[0203] like Figure 1 In some 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 embodiments, the whey protein retentate has a TS content of at least 18%, at least 20%, at least 22% or at least 24%.

[0204] In certain embodiments, filtering step is carried out to provide a retentate with about 65 % by weight to about 95 % by weight total protein. In some such embodiments, the total protein content of the whey protein retentate is about 75 % by weight to about 90 % by weight or about 80 % by weight to about 85 % by weight. In some such embodiments, the total protein content of the whey protein retentate is at least 65 % by weight, at least 70 % by weight, at least 75 % by weight or at least 80 % by weight.

[0205] At this stage, particularly during thermalization, 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). Without wishing to be bound by theory, uncontrolled formation of denatured aggregates may result in the formation of larger than desired aggregates during the thermal denaturation step.

[0206] In some such embodiments, lactalbumin solution and / or lactalbumin retentate have low-level denaturation before the denaturation heat treatment step.For example, before the denaturation heat treatment step, the denaturation level of lactalbumin solution and / or lactalbumin retentate can be less than 10%, such as about 3% to about 8%.Or, before the denaturation heat treatment step, the denaturation level of lactalbumin solution and / or lactalbumin retentate can be less than 9%, less than 8%, less than 7%, less than 6% or less than 5%.In some such embodiments, before the denaturation heat treatment step, the denaturation level of lactalbumin solution and / or lactalbumin retentate is about 5%, about 4%, about 3%, about 2% or about 1%.

[0207] like Figure 1As shown, the retentate (and / or whey protein solution) is contacted with an oxidant. It should be understood that the time for treating the whey protein stream with an oxidant can vary. For example, a diluted whey protein composition (i.e. before UF / DF) is contacted with an oxidant.

[0208] In some such embodiments, the oxidant is any food grade oxidant. In some such embodiments, the oxidant is oxygen (O2), ozone, peroxides, including alkyl hydroperoxides, superoxide, peroxynitrite, peroxydisulfuric acid or lactoperoxidase.

[0209] In some such embodiments, the oxidant is a peroxide, preferably an organic peroxide. Exemplary peroxides include, but are not limited to, metal peroxides (e.g., alkali metal peroxides such as sodium peroxide; alkaline earth metal peroxides, such as magnesium peroxide or calcium peroxide; or transition metal peroxides such as zinc peroxide), hydrogen peroxide, and benzoyl peroxide. In some such embodiments, the oxidant is a perborate such as sodium perborate.

[0210] In some such embodiments, the oxidizing agent is benzoyl peroxide or hydrogen peroxide. Hydrogen peroxide is available in various concentrations and purities. In one embodiment, the hydrogen peroxide is food grade.

[0211] In some such embodiments, the oxidant is used with a catalyst, which may be an enzyme catalyst or a chemical catalyst. In some such embodiments, the oxidant is used with a catalyst enzyme such as a microbial peroxidase. Exemplary fungal peroxidases are (DSM Food Specialties). Other exemplary microbial peroxidases 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.

[0212] In some such embodiments, the oxidant is added over a period of time. For example, an initial amount of the oxidant may be added, and then subsequent amounts of the oxidant may be added as the initial amount is consumed. Thus, the total amount of peroxide may be added in more than one step, such as in 2, 3, 4, 5 or more steps, or continuously over a period of time.

[0213] Exemplary oxidizing agents, including hydrogen peroxide, have been previously used by the dairy industry to bleach whey.

[0214] Hydrogen peroxide (H2O2) is a clear, colorless liquid with a slightly pungent odor. Hydrogen peroxide is one of two bleaching agents currently approved in the United States for bleaching whey to decolorize whey compositions made from cheese containing annatto.

[0215] Previous reports provide contradictory information about the effect of hydrogen peroxide on whey protein denaturation. Kramer et al., J. Agric. Food Chem, 65, 10258-10269 (2017) reported that when low levels of hydrogen peroxide (e.g., 500 μM, i.e., a 5:1 molar ratio compared to protein) were present during the heating phase, enhanced aggregation of β-lactoglobulin (β-Lg) alone and a mixture of β-Lg and α-lactalbumin (α-La) was observed. Similarly, Kang et al., J. Dairy Sci., 93, 3891-3901 (2010) reported that hydrogen peroxide concentrations above the legal limit (500 ppm) increased the amount of whey protein denaturation. On the other hand, there are several reports that hydrogen peroxide reduces the denaturation of whey proteins (see, e.g., Fish and Mickelsen, Journal of Dairy Science, 50(7), 1045-1048 (1967), noting that treatment of skim milk with hydrogen peroxide before preheating at 89°C for 30 minutes reduces the denaturation of acid whey proteins; Sutariya and Patel, Food Chemistry, 223, 114-120 (2017), indicating that the addition of hydrogen peroxide to whey protein isolate prevents whey protein denaturation and aggregation; see also US2016 / 0235082, Figure 8 ). In addition, Bechtle (US 3,818,109) reported that a combination of mild heating (e.g., a temperature of 60°C to 70°C) and hydrogen peroxide treatment avoided any appreciable denaturation of whey proteins while achieving sterilization of the culture medium. It is noteworthy that these reports appear to describe the application of heat to samples containing hydrogen peroxide.

[0216] On the other hand, in certain embodiments, the method for making a denatured whey protein composition disclosed herein provides for removing or consuming an oxidant prior to the denaturation heat treatment step. Thus, in some such embodiments, the oxidant is consumed prior to subjecting the whey protein solution or whey protein retentate to a denaturation heat treatment step under conditions that allow protein denaturation to occur. Thus, during the denaturation heat treatment step, the whey protein solution or whey protein retentate may be substantially free of an oxidant.

[0217] In certain embodiments, the oxidant is used together with a catalyst enzyme such as a microbial peroxidase, and the amounts of the oxidant and the peroxidase are such that the oxidant is completely consumed by the enzyme. Alternatively, when the oxidant is hydrogen peroxide, catalase can be used to catalyze the decomposition of hydrogen peroxide into water and oxygen. In certain embodiments, catalase is not used.

[0218] The whey protein solution or whey protein retentate may be sampled to confirm that it is substantially free of oxidizing agents. For example, the whey protein solution or whey protein retentate may be sampled and tested for detectable peroxide using colorimetric test strips available from Merck / MilliporeSigma.

[0219] In some such embodiments, the method further comprises adjusting the pH of the whey protein solution or the whey protein retentate before the denaturing heat treatment step. In some such embodiments, the pH of the whey protein solution or the whey protein retentate before 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 whey protein solution or the whey protein retentate before the denaturing heat treatment step is about 5.8 to about 7.0, about 6.0 to about 6.8, or about 6.2 to about 6.6.

[0220] like Figure 1 As shown, the whey protein solution or the whey protein retentate is subjected to heat treatment. The heat treatment is performed to impart the desired denaturation.

[0221] In certain embodiments, the denaturing heat treatment step comprises a high wall shear rate, even under laminar flow, for example at least 1000 s -1 The whey protein solution or whey protein retentate is heated to greater than about 70° C. at a wall shear rate of about 1000 s -1 To about 10000s -1 , or about 1500s -1 To about 5000s -1 , or about 1500s -1 To about 4000s -1 or about 2000s -1 To about 3000s -1 .

[0222] In certain embodiments, the denaturing heat treatment step comprises heating the whey protein solution or whey protein retentate to greater than about 70° C. under turbulent conditions (e.g., 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.

[0223] Turbulent flow is defined as having sufficient mass flow rate in the heating tube to provide a Reynolds number of at least 2000, known as Re. This Reynolds number is characteristic of turbulent flow and is known in the field of fluid dynamics. The determination of Re depends on the mass velocity of the fluid and its highest viscosity at the target heating temperature, which is defined as the nominal viscosity determined from measurements of the pressure drop along a known length of a horizontal pipe of known uniform circular cross-section at a known flow rate of the fluid heat treated at uniform temperature prior to drying using the Hangen-Poiseuille equation. In order to calculate the Re for a given process, the formula for Newtonian fluids can be used, which uses the highest viscosity of the heat treated fluid at the target temperature. This means that the Re will be higher at all other viscosities (i.e., lower viscosities) and will fall into the turbulent region.

[0224] In certain embodiments, the denaturing heat treatment step comprises heating the whey protein solution or whey protein retentate to greater than about 50° C. in combination with mechanical shear. In some such embodiments, a homogenizer, colloid mill, high pressure pump, scraped surface heat exchanger, high shear mixer, etc. is used to mix the solution or break up aggregates during or after formation.

[0225] In certain embodiments, the denaturation heat treatment step comprises heating the whey protein solution or whey protein retentate to a temperature of about 70° C. to about 90° C. at pH 6 to pH 7 while under conditions of sufficiently high shear stress / force resulting from increased serum viscosity, high wall shear rate, turbulence, mechanical shear, or a combination of two or more. In some such embodiments, the denaturation heat treatment step comprises heating the whey protein solution or whey protein retentate to a temperature of about 80° C. to about 85° C. at pH 6.3 to pH 6.7 while under conditions of sufficiently high shear stress / force resulting from increased serum viscosity, high wall shear rate, turbulence, mechanical shear, or a combination of two or more.

[0226] Known factors such as heating temperature and pH may affect protein aggregation. However, as reported herein, variations within defined ranges of temperature and pH did not significantly affect particle size, indicating that the methods disclosed herein are robust and reproducible.

[0227] In certain embodiments, the whey protein solution or whey protein retentate is substantially free of oxidizing agents during the heat treatment step that allows protein denaturation to occur. For example, the oxidizing agent may be inactivated, consumed and / or removed prior to the heat treatment step.

[0228] One of the main mechanisms of protein aggregation is believed to be the formation of intermolecular disulfides involving sulfhydryl-disulfide interchange. Without wishing to be bound by theory, it is believed that pretreatment with an oxidant affects aggregation pathways and reaction kinetics by promoting intermolecular sulfhydryl-disulfide interchange. Without wishing to be bound by theory, it is believed that the microparticles disclosed herein are primarily disulfides bonded to low levels of thiols. Additionally or independently, limiting the pre-denaturation of whey protein (denaturation level prior to the denaturation heat treatment step) can also enable the formation of small particles.

[0229] After the denaturation heat treatment step, the denatured whey protein composition may be dried, or the denatured whey protein composition may be directly used to prepare a high protein product without drying.

[0230] In certain embodiments, the denatured whey protein composition is not subjected to a particle size reduction or particle size selection procedure such as further microfiltration to achieve the particle size distribution described herein.

[0231] E. Application and usage

[0232] In at least one aspect, the present disclosure provides ingredients for preparing edible consumables, beverages, or liquid nutritional compositions such as medical foods. In some such embodiments, the ingredients are provided in liquid or dry (powder) form or a blend thereof.

[0233] Examples of suitable edible consumables include, but are not limited to, baked products (e.g., muffins, cookies, brownies, etc.), puffed and / or extruded snack products, protein fortified snacks, confectionery products (including chocolates, gels, ice cream, caramels, snack bars, spreads, sauces, dressings), dairy products (including drinkable yogurts (including ambient yogurts), stirred yogurts, and cheeses), food additives (such as protein sprays), and dietary supplement products (including daily tablets).

[0234] An exemplary edible consumable is a solid set gel, such as set yogurt. Set yogurt prepared using the denatured whey protein composition provided herein can exhibit reduced hardness compared to a control yogurt having the same ingredient composition and protein content, except that the control yogurt does not include the denatured whey protein composition provided herein.

[0235] Another exemplary edible consumable is a semi-solid food, such as stirred yogurt. Stirred yogurt prepared using the denatured whey protein composition provided herein can exhibit reduced viscosity compared to a control yogurt having the same ingredient composition and protein content, except that the control yogurt does not include the denatured whey protein composition provided herein.

[0236] In certain embodiments, the set or stirred yogurt exhibits reduced in-mouth texture (or reduced grittiness) and / or exhibits negligible or no increase in undesirable flavors compared to an appropriate control yogurt.

[0237] The example of suitable beverage includes but is not limited to culture and ready-to-drink (RTD) beverage and aqueous liquid (such as concentrate) that can be diluted and can be reconstructed to provide the powder of such beverage.In certain embodiments, beverage is drinking type yogurt (including environmental drinking type yogurt), milk, milk powder, sports supplement including dairy and non-dairy base sports supplement, fruit juice or formula (such as infant formula, follow-up formula or growth formula) in powder or liquid form.In certain embodiments, beverage is neutral high protein beverage.In certain embodiments, beverage is acidic high protein beverage.

[0238] An exemplary beverage is a drinking yogurt. Drinking yogurt is a fermented milk beverage with low viscosity. Drinking yogurt may be ambient drinking yogurt.

[0239] Another exemplary beverage is a sports drink. Sports drinks typically include a high level of protein, have a low fat content and are supplemented with vitamins and minerals, flavorings, sweeteners, stabilizers and / or salt.

[0240] In certain embodiments, the denatured whey protein composition disclosed herein is used as a component for preparing a nutritional composition, preferably a liquid nutritional composition (e.g., medical food). As an example only, US federal law and Food and Drug Administration regulations define medical food as "a food formulated to be consumed 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, are established by medical evaluation" (§5 (b) of the 1988 Orphan Drug Act (Orphan Drug Act) (21 U.S.C. 360 ee (b) (3)) and FDA regulations 21 CFR 101.9 (j) (8)). In some such embodiments, the liquid nutritional composition also includes fat and / or carbohydrate components.

[0241] In certain embodiments, the denatured whey protein compositions disclosed herein are formulated into nutritional compositions by wet blending. In some such embodiments, the nutritional composition is a wet blend nutritional composition, such as a wet blend meal replacement composition or a wet blend infant formula.

[0242] The denatured whey protein compositions disclosed herein can be used to prepare powdered or ready-to-mix compositions, particularly wet blended compositions, wet blended dietary replacement compositions and / or wet blended infant formulas. Wet blended compositions are compositions prepared using a wet blending step, wherein two or more ingredients are combined into an aqueous solution and optionally subsequently dried into a powder. For example, wet blending may be required for ready-to-mix nutritional compositions that require a wet blending step to incorporate a fat source into the composition.

[0243] The denatured whey protein compositions disclosed herein are particularly useful in wet blending applications because the low viscosity of the compositions can enable high total solids content to be achieved during the evaporation process. When the solids content during the evaporation step is high, less water must be removed during spray drying, which reduces energy usage accordingly. In addition, the heat-stabilized denatured whey protein compositions disclosed herein can reduce line scaling during evaporation and drying, which reduces production downtime accordingly, and increases plant throughput and product yield.

[0244] The method for preparing the wet blend composition is known to those skilled in the art. In short, the method for preparing the wet blend composition may include the step of combining two or more components to form an aqueous solution. The aqueous solution may optionally be subjected to one or more additional steps, such as homogenization, standardization, heat treatment (such as pasteurization and / or UHT treatment), homogenization, evaporation and / or spray drying. The aqueous solution may include a heat-stable protein composition of the present invention and one or more additional ingredients, optionally selected from one or more lipids, one or more carbohydrates, one or more additional ingredients as described herein, or any combination of these.

[0245] In at least one aspect, the present disclosure provides a method for providing nutritional support to a subject, comprising administering to the subject enterally a nutritional composition comprising a denatured whey protein composition disclosed herein. In certain embodiments, the subject is a subject in need of nutritional support. Therefore, the nutritional composition described herein is used in a method for providing nutritional support to a subject in need thereof. 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 can be oral or via a tube (e.g., nasogastric or gastric feeding).

[0246] 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 the rate of glycogen resynthesis, regulate blood glucose levels, increase the insulin response to elevated blood glucose concentrations, reduce satiety, reduce satiety, increase food intake, increase calorie intake, improve glucose metabolism, increase recovery rate after surgery, increase rehabilitation efficacy prior to surgery or chemotherapy, increase recovery rate after injury, increase recovery rate after exercise, increase athletic performance, and / or provide nutrition. Enteral administration can be oral or via a tube (e.g., nasogastric or gastric).

[0247] In certain embodiments, the nutritional compositions comprising the denatured whey protein compositions disclosed herein are used to maintain or increase muscle protein synthesis, maintain or increase muscle mass, prevent or increase loss of muscle mass, 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 the insulin response to elevated blood glucose concentrations, reduce satiety, reduce fullness, increase food intake, increase calorie intake, improve glucose metabolism, increase the rate of recovery after surgery, increase preconditioning prior to surgery or chemotherapy, increase the rate of recovery after injury, increase the rate of recovery after exercise, increase athletic performance, and / or provide nutrition.

[0248] F. High Protein Liquid Composition

[0249] 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 or a drinkable yogurt, or an acidic or neutral pH liquid nutritional composition, each of which is further discussed in 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 6% to 30% (w / v) denatured whey protein, such as 7% to 28% (w / v) denatured whey protein, 8% to 26% (w / v) denatured whey protein or 10% to 24% (w / v) denatured whey protein.

[0250] 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 includes other milk proteins and / or non-milk proteins. In other such embodiments, the high-protein liquid composition includes whey proteins from two or more sources, such as heat-denatured whey protein compositions, non-denatured whey protein compositions, whey protein hydrolysates, or a composition (such as MPC) comprising both whey and casein, and the total whey protein in the composition is the sum of the total whey proteins present in the heat-denatured whey protein compositions, non-denatured whey protein compositions, whey protein hydrolysates and / or MPC.

[0251] 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 the liquid composition is the sum of all proteins contributed by all protein-containing ingredients in the composition. For example, in embodiments where the liquid composition comprises a heat-denatured whey protein composition and other non-dairy and / or dairy proteins derived from, for example, 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 20% (w / v), at least 21% (w / v), at least 22% (w / v) total protein. In some such embodiments, the liquid composition comprises 6% to 30% (w / v) total protein, for example 7% to 28% (w / v) total protein, 8% to 26% (w / v) total protein or 10% to 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.

[0252] 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 total protein in the high-protein liquid composition is casein. In some such embodiments, at most 5%, at most 4%, at most 3%, at most 2%, at most 1% or at most 0.1% of 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 includes a total protein that is no more than 12% (w / v), for example, 5% (w / v), 6% (w / v), 7% (w / v), 8% (w / v), 9% (w / v), 10% (w / v) or 11% (w / v) of total protein. In some such embodiments, at least 90% or at least 95% of total protein in the high-protein liquid composition is denatured whey protein. In some such embodiments, substantially all total proteins in the high-protein liquid composition are denatured whey proteins.

[0253] In certain embodiments, the high-protein liquid composition comprises at least 12% (w / v) total protein, for example 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 12% to 18% (w / v) total protein. In some such embodiments, at least 90% or at least 95% of total protein in the high-protein liquid composition is denatured whey protein. In some such embodiments, substantially all total proteins in the high-protein liquid composition are denatured whey proteins.

[0254] 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% to 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% to 20% (w / v) total protein is less than 1 μm.

[0255] In certain embodiments, the liquid composition is sterilized and / or pasteurized in a manner suitable for commercial application. 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 for 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 comprises a d(4,3) of less than 1 μm, ... 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 comprises d(4,3) less than 1 μm, d(4,3) less than 1 μm, 50 and d less than 3 μm 90 .

[0256] The lethal effect of high temperature on microorganisms depends on temperature and holding time, and it is well known that the time required to kill the same number of microorganisms decreases when the temperature is increased. The time taken to reduce a specific amount of the initial number of microorganisms at a specific temperature is generally referred to as the "F value". The F0 value is the equivalent time in minutes for a specified temperature that gives the same thermal lethality as at 121°C. As known in the relevant art, acidic products generally require lower F0 values ​​than products processed at neutral pH to achieve commercial sterility.

[0257] In certain embodiments, the liquid composition is shelf-stable after sterilization and / or pasteurization. In some such embodiments, when stored for at least 2 months, at least 3 months, at least 6 months, or at least 12 months aseptically packaged at a temperature of about 20°C to about 25°C for a long time, the liquid composition shows minimal or substantially no sedimentation, gelation, or aggregation, and negligible bacterial growth. In some such embodiments, the liquid composition shows minimal or substantially no powdery or gritty feel. Therefore, for example, the liquid composition can be a heat-treated, shelf-stable liquid composition.

[0258] In certain embodiments, the liquid composition exhibits minimal or substantially no settling under conditions that facilitate settling.

[0259] In certain embodiments, in addition to denatured whey protein, the high-protein liquid composition also includes non-whey and / or non-milk proteins. Exemplary sources of non-whey proteins include, but are not limited to, skim milk powder (SMP) or whole milk powder or milk protein concentrate (MPC), or milk protein isolate (MPI) or micellar casein concentrate (MCC). As another example, casein can be included in the high-protein liquid composition in the form of sodium caseinate or potassium caseinate or calcium caseinate or magnesium caseinate.

[0260] In certain embodiments, the high protein liquid composition comprises one or more, two or more, or three or more non-dairy proteins. Suitable non-dairy proteins included in the liquid composition include algae protein, fungal protein (e.g., mycoprotein), vegetable protein, and animal protein and hydrolyzed forms thereof. In some such embodiments, the liquid composition comprises soy protein, rice protein, and / or pea protein.

[0261] In addition, the high protein liquid composition may include whey proteins from two or more sources. For example, the liquid composition may include a mixture of WPC and WPI, one or both of which have been thermally denatured. As another example, the liquid composition may include a mixture of WPC prepared in different ways or with different properties.

[0262] In addition to the methods disclosed herein, exemplary methods for preparing heat-denatured whey protein compositions suitable for liquid compositions are provided in PCT International Applications PCT / NZ2007 / 000059 (published as WO2007 / 108709) and PCT / NZ2010 / 000072 (published as WO2010 / 120199) and PCT International Application PCT / IB2012 / 056103 (published as WO2013 / 065014), each of which is incorporated herein by reference in its entirety.

[0263] In certain embodiments, one or more protein components, such as heat-denatured whey protein compositions or high-protein liquid compositions, can be treated to reduce lactose content. In some such embodiments, the protein component or high-protein liquid composition is treated with an enzyme such as beta-galactosidase, or filtered to remove lactose. Suitable enzyme treatment and filtration protocols to reduce lactose content will be apparent to those skilled in the art.

[0264] 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 at most 10% (w / v), at most 8% (w / v), at most 6% (w / v), or at most 4% (w / v).

[0265] In some embodiments, the high-protein liquid composition comprises a lipid component. In some such embodiments, the lipid component exists with an amount of 30% (w / v) at the most. In some embodiments, the high-protein liquid composition comprises a carbohydrate component. In some such embodiments, the carbohydrate component exists with an amount of 30% (w / v) at the most. In some embodiments, the high-protein liquid composition has a caloric density that is no more than 1kcal / ml, 1 to 2kcal / ml or greater than 2kcal / ml.

[0266] In certain embodiments, the high protein liquid composition includes at least one monovalent cation and / or at least one divalent metal cation. In some such embodiments, the high protein liquid composition includes at least 30 mg / 100 mL, at least 50 mg / 100 mL, or at least 100 mg / 100 mL of divalent metal cations.

[0267] Each feature of high protein liquid composition as described herein can be combined with any other one or more features, unless clearly indicated to the contrary. In particular, any feature indicated as preferred or advantageous can be combined with any other one or more features indicated as preferred or advantageous. An exemplary liquid composition is a liquid nutritional composition. The liquid nutritional composition can be provided with enteral nutrition by oral administration or by tube administration (e.g., nasogastric or gastric feeding). In addition to protein, exemplary liquid nutritional compositions also contain a certain level of fat and / or carbohydrate and combination to obtain a caloric value of at least 0.5kcal / g or kcal / ml. For example, the liquid nutritional composition can have a thermal density of up to 3kcal / g or higher. So far, it is difficult to achieve such a high thermal density with low viscosity and enough albumen.

[0268] In certain embodiments, the liquid nutritional composition also includes a lipid component. The lipid component may include vegetable lipids or animal (e.g., cattle, pigs, etc.) lipids, including milk lipids and fish oils. In some such embodiments, vegetable oils are suitable because they are easy to prepare and saturated fatty acid content is relatively low. Exemplary vegetable oils include canola (rapeseed) oil, corn oil, sunflower oil, olive oil or soybean oil.

[0269] 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 g of lipid per 100 mL of the composition, and various ranges can be selected between these values, such as about 0.01 to about 30, about 0.1 to about 30, about 1 to about 30, 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 of lipid per 100 mL of the composition.

[0270] In certain embodiments, the liquid nutritional composition also includes a carbohydrate component. The carbohydrate component may include 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, oligosaccharides of glucose. Suitable indigestible carbohydrates include, but are not limited to, oligofructose, inulin and galacto-oligosaccharides.

[0271] 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, or at least about 45 g of carbohydrate per 100 mL of the composition, and various ranges can be selected between these values, such as about 0.01 to about 4, about 0.01 to about 45, about 0.1 to about 45, about 1 to about 45, about 4 to about 45, about 5 to about 50, about 6 to about 65, about 7 to about 80, about 8 to about 90, about 9 to about 100 g of carbohydrate per 100 mL of the composition. 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 carbohydrates.

[0272] In certain embodiments, the liquid nutritional composition also includes a stabilizer or emulsifier. Suitable emulsifiers include lecithin, monoglycerides and diglycerides, polyglycerol esters, milk phospholipids, citric acid esters (CITREM) and diacetyl tartaric acid esters (DATEM) of monoglycerides and diglycerides. These emulsifiers can be added in an amount of about 0.003g to about 0.06g per gram of lipid. Suitable stabilizers include but are not limited to carrageenan, gellan gum, pectin, guar gum, locust bean gum, xanthan gum, carboxymethyl cellulose and microcrystalline cellulose and combinations thereof. Those skilled in the art will recognize that, in addition to those listed above, many different gum forms are suitable for liquid compositions disclosed herein.

[0273] In certain embodiments, the liquid nutritional composition further comprises a source of amino acids, amino acid precursors, amino acid metabolites, or any combination of any two or more thereof, preferably free amino acids, amino acid precursors or amino acid metabolites.

[0274] In certain embodiments, the liquid nutritional composition also includes vitamins and / or minerals required for long-term maintenance of patient nutrition, and small amounts of components such as antioxidants, flavoring agents and / or coloring agents. The amount of vitamins and / or minerals used in the liquid nutritional composition is a typical amount of a dietary replacement product known to those skilled in the art. The micro-nutrition requirements of various subgroups of a population are also known. The recommended daily requirements of vitamins and minerals can be specified for various population subgroups. See, for example, dietary reference intakes (DRIs): dietary allowances and adequate intakes recommended by the United States National Academy of Sciences, the Institute of Medicine, and the Food and Nutrition Board (2010).

[0275] In certain embodiments, the liquid nutritional composition is a neutral pH liquid nutritional composition including at least 12% (w / v) total protein, such as 12% to 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. Therefore, in certain embodiments, the neutral pH liquid nutritional composition may include 12% to 18% (w / v) total protein, wherein substantially all of the total protein comes from denatured whey protein. In some such embodiments, the neutral pH liquid nutritional composition includes a level and combination of fat and / or carbohydrates reaching 1 to 3 kcal / ml, such as 1.5 to 2.5 kcal / ml of caloric density. An exemplary neutral pH liquid nutritional composition includes 12% (w / v) total protein, wherein substantially all of the total protein comes from denatured whey protein and has a caloric density of 1.60 kcal / ml. Another exemplary neutral pH liquid nutritional composition includes 15% (w / v) total protein, wherein substantially all of the total protein comes from denatured whey protein and has a caloric density of 2.25 kcal / ml.

[0276] Another exemplary liquid composition is a high protein drinking yogurt. Methods of making liquid compositions such as drinking yogurt are well known in the art.

[0277] Figure 3 An exemplary method for preparing a drinking yogurt is shown. In the exemplary method, ingredients (e.g., skim milk powder and whey protein composition) are combined 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, for example, to 38°C or 42°C. The mixture is then inoculated with a bacterial starter culture. The mixture is then fermented until at least a pH of less than 4.6 is reached, followed by cooling to 20°C. The gel so formed is broken and sheared, then packaged and cooled to 4°C.

[0278] In certain embodiments, the high protein drinking yogurt comprises 15% total protein, wherein the same drinking yogurt comprises at least 12% denatured whey protein (i.e., at least 80% (w / w) of the total protein comes from denatured whey protein). In certain embodiments, the high protein drinking yogurt comprises 20% total protein, wherein the same drinking yogurt comprises at least 17% denatured whey protein (i.e., at least 85% (w / w) of the total protein comes from denatured whey protein).

[0279] In certain embodiments, the high protein drinking yogurt has been sterilized and / or pasteurized. For example, the drinking yogurt may have been sterilized.

[0280] In certain embodiments, the average particle size of the drinking yogurt (characterized by D(4,3)) 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 of the drinking yogurt (characterized by D(4,3)) is about 0.6 μm to about 1.5 μm, or about 0.8 μm to about 1.3 μm, or about 0.9 μm to about 1.2 μm. In some such embodiments, the average particle size of the drinking yogurt (characterized by D(4,3)) in which at least 75%, at least 80%, or at least 85% of the total protein of the drinking yogurt comes from the denatured whey protein composition is about 0.6 μm to about 1.5 μm, or about 0.8 μm to about 1.3 μm, or about 0.9 μm to about 1.2 μm.

[0281] In certain embodiments, the drinking yogurt has a 50 Less than 2.0 μm, less than 1.5 μm or less than 1.0 μm. In some such embodiments, the d 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. In some such embodiments, the d 50 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, wherein at least 75%, at least 80% or at least 85% of the total protein of the drinking yogurt comes from the denatured whey protein composition.

[0282] In certain embodiments, the drinking yogurt has a 90 Less than 3.0 μm, less than 2.5 μm or less than 2.0 μm. In some such embodiments, the d90 of the drinking yogurt is from about 1.3 μm to about 2.5 μm, or from about 1.4 μm to about 2.4 μm, or from about 1.5 μm to about 2.3 μm. In some such embodiments, the d90 of the drinking yogurt is from about 1.3 μm to about 2.5 μm, or from about 1.4 μm to about 2.4 μm, or from about 1.5 μm to about 2.3 μm, wherein at least 75%, at least 80%, or at least 85% of the total protein of the drinking yogurt comes from the denatured whey protein composition.

[0283] In certain embodiments, the high-protein drinking yogurt shows minimal or substantially no sedimentation during the shelf life of the product, for example up to 6 weeks for frozen drinking yogurt, and up to 6 to 9 months for ambient drinking yogurt. For example, the high-protein drinking yogurt shows minimal or substantially no sedimentation during the shelf life of the product (e.g., up to 6 weeks). In some such embodiments, the high-protein drinking yogurt shows less than 20% sedimentation during the shelf life of the product. In some such embodiments, the high-protein drinking yogurt shows less than 15%, less than 12%, less than 10%, less than 8% or less than 6% sedimentation during the shelf life of the product. In some such embodiments, the high-protein drinking yogurt shows about 2% to about 6% or about 3% to about 5% sedimentation during the shelf life of the product.

[0284] As another example, centrifugation test can be used to evaluate sedimentation. Therefore, in some such embodiments, high protein drinking yogurt shows less than 20% sedimentation after centrifugation for 5 minutes at 1540xg. In some such embodiments, the liquid composition shows 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 shows about 2% to about 6% or about 3% to about 5% sedimentation after centrifugation.

[0285] 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 viscosity of the high-protein drinking yogurt comprising 10% to 20% (w / v) total protein is 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.

[0286] The viscosity of the sample can be measured by methods known in the art. For example, one method involves using a MCR302 rheometer (Anton-Paar) and subjecting the sample to a pre-shear (e.g., 300 s -1 for 1 min), followed by a shear rate sweep (e.g., from 0.001-398 s -1 , viscosity at 50s -1 and / or 100s -1The apparent viscosity can be measured at 10°C using a Haake viscometer (Haake Mess-Technik, Gmbh & Co., Karlsruhe, Germany). The shear rate is increased from 0 to 120 s over a period of 180 s. -1 , then reduce the shear to 0s over a period of 30 seconds -1 To measure the viscosity. The apparent viscosity is 50s -1 obtained at a shear rate of .

[0287] The compositions, methods, and uses described herein will be better understood by reference to the following exemplary embodiments and examples, which are included as illustrations of the scope of the invention and not as limitations.

[0288] G. High protein coagulated and stirred yogurt

[0289] In at least one aspect, the present disclosure provides a high-protein food comprising denatured whey protein. The high-protein food can be a yogurt, such as a set yogurt or a stirred yogurt, each of which is discussed in further detail herein. Methods for making set and stirred yogurts are generally known in the art.

[0290] In certain embodiments, high-protein food comprises at least 6% (w / w) denatured whey protein. In some such embodiments, high-protein food 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, food comprises 6% to 30% (w / w) denatured whey protein, for example 7% to 25% (w / w) denatured whey protein, 8% to 20% (w / w) denatured whey protein or 10% to 15% (w / w) denatured whey protein.

[0291] 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 is denatured whey protein. In some such embodiments, substantially all of the total protein in the high-protein food is denatured whey protein. In other such embodiments, the high-protein food includes other milk proteins and / or non-milk proteins. In other such embodiments, the high-protein food includes whey proteins from two or more sources, such as heat-denatured whey protein compositions, non-denatured whey protein compositions, whey protein hydrolysates, or a composition (such as MPC) comprising both whey and casein, and the total whey protein in the composition is the sum of the total whey proteins present in the heat-denatured whey protein compositions, non-denatured whey protein compositions, whey protein hydrolysates and / or MPC.

[0292] In certain embodiments, high protein food comprises at least 5%, at least 10%, at least 15% or at least 20% (w / w) total protein. The total protein in food is the sum of all proteins contributed by all protein-containing ingredients in the product. For example, in the embodiment that food comprises heat-denatured whey protein composition and other non-milk and / or milk protein derived from for example skim milk powder (SMP) or milk protein concentrate (MPC), the total protein in food is the sum of the total protein present in heat-denatured whey protein composition and SMP and / or MPC. In some such embodiments, food 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 comprises 6% to 30% (w / w) total protein, for example 8% to 25% (w / w) total protein or 10% to 15% (w / w) total protein. In some such embodiments, the high-protein food 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.

[0293] In certain embodiments, food has been sterilized and / or pasteurized in a manner suitable for commercial use. In some such embodiments, sterilization and / or pasteurization include thermal (i.e. heating) treatment. In other such embodiments, sterilization and / or pasteurization include non-thermal treatment.

[0294] In certain embodiments, the high protein food comprises a solidified gel, such as a solidified yogurt. In some such embodiments, the solidified 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 containing denatured whey protein.

[0295] 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. In some such embodiments, the set gel exhibits a breaking force of less than 50 g, less than 45 g or less than 40 g.

[0296] In certain embodiments, the high protein food is a semi-solid food, such as stirred yogurt. In some such embodiments, the particle size distribution of the semi-solid food after sterilization or pasteurization includes a particle size distribution of less than 1 μm. 50 and d less than 5 μm 90 In some such embodiments, the semi-solid food 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.

[0297] In some such embodiments, the semi-solid food has a temperature of 50 s at 20°C. -1 A viscosity of less than 500 mPa.s or less than 400 mPa.s measured under .

[0298] H. Exemplary Embodiments

[0299] In one aspect, the present disclosure provides a heat stable denatured whey protein composition comprising microparticles comprising denatured whey protein, wherein at least 50% by volume or at least 55% by volume of the particles have a diameter of 0.2 μm to 1.0 μm.

[0300] In another aspect, the present disclosure provides a heat stable denatured whey protein composition comprising microparticles comprising denatured whey protein, wherein the microparticles have a volume weighted average diameter D(4,3) of no more than about 1.0 μm and at least 95% by volume of the particles have a diameter less than about 2.0 μm.

[0301] In certain embodiments of any aspect disclosed herein, the present disclosure provides a heat stable denatured whey protein composition comprising microparticles, the microparticles comprising denatured whey protein, wherein the microparticles exhibit minimal or substantially no primary particle size growth after heating a 10% (w / w) protein content aqueous solution (at pH 6.8) at 120° C. for 15 minutes. In some such embodiments, after a primary particle growth test comprising heat treatment at 120° C. for 15 minutes, the d 50 No greater than about 1.0 μm, preferably about 0.6 to about 1.0 μm. In some such embodiments, after a primary particle growth test comprising heat treatment at 120° C. for 15 minutes, the d 90 Less than about 2.0 μm, preferably about 0.9 to about 1.7 μm or about 1.1 to about 1.5 μm.

[0302] 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 degree of hydrolysis of the whey protein is less than about 5%. In some such embodiments, the whey protein is substantially non-hydrolyzed.

[0303] 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.

[0304] 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 total protein.

[0305] In certain embodiments of any of the aspects disclosed herein, the residual denatured whey protein in the denatured whey protein composition is less than 16%, less than 14%, or less than 12%.

[0306] 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%.

[0307] In certain embodiments of any of the aspects disclosed herein, the heat stable denatured whey protein composition has a lactose content of at most 10%, at most 8%, at most 6%, or at most 4% by weight.

[0308] In certain embodiments of any of the aspects disclosed herein, the heat stable denatured whey protein composition has a fat content of at most 20%, at most 18%, at most 16%, at most 14%, at most 12%, at most 10%, at most 8% or at most 6%.

[0309] In certain embodiments of any of the aspects disclosed herein, the heat stable denatured whey protein composition has an ash content of at most 10%, at most 8%, at most 6%, or at most 4% by weight.

[0310] In certain embodiments of any of the aspects disclosed herein, the heat stable denatured whey protein composition comprises less than about 10% casein by weight.

[0311] In certain embodiments of any of the aspects disclosed herein, the heat stable denatured whey protein composition is substantially free of casein.

[0312] Exemplary embodiments 1. A heat stable denatured whey protein composition comprising at least 60% total protein on a dry weight basis and microparticles comprising denatured whey protein, wherein the microparticles have a particle size distribution comprising at least two characteristics selected from the group consisting of: (i) a volume weighted average diameter D(4,3) of not greater than about 1.0 μm, (ii) a d(4,3) of not greater than about 1.0 μm. 50 , (iii) a d of not greater than about 2.0 μm 90 , (iv) at least 92 volume % of the particles have a diameter less than about 2.0 μm, and (v) no greater than 45 volume % of the particles have a diameter between about 1.0 and about 10.0 μm.

[0313] Exemplary embodiment 2. The heat-stable denatured whey protein composition of exemplary embodiment 1, wherein the particle size distribution comprises (i) a volume weighted average diameter D(4,3) of not more than about 1.0 μm, for example, from about 0.6 to about 1.0 μm, and (iii) a volume weighted average diameter D(4,3) of not more than about 2.0 μm, for example, from about 0.9 to about 1.7 μm. 90 .

[0314] Exemplary embodiment 3. The heat stable denatured whey protein composition of exemplary embodiment 1, wherein the particle size distribution includes (ii) a particle size distribution of no greater than about 1.0 μm, such as about 0.6 to about 1.0 μm. 50 , and (iii) a d of no greater than about 2.0 μm, such as about 0.9 to about 1.7 μm 90 .

[0315] 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 no more than about 1.0 μm, such as about 0.6 to about 1.0 μm.

[0316] 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.

[0317] 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 particles having a diameter of about 1.0 to about 10.0 μm.

[0318] Exemplary embodiment 7. The heat-stable denatured whey protein composition of any one of exemplary embodiments 1-6, wherein the particle size distribution is substantially unchanged after a 10% (w / w) protein content aqueous solution comprising the denatured whey protein composition is heated at 120°C for 15 minutes.

[0319] Exemplary embodiment 8. The heat-stable denatured whey protein composition of any one of exemplary embodiments 1-7, wherein after heating a 10% (w / w) protein content aqueous solution comprising the denatured whey protein composition at 120° C. for 15 minutes, d 50 From about 0.6 to about 1.0 μm.

[0320] Exemplary embodiment 9. The heat-stable denatured whey protein composition of any one of exemplary embodiments 1-8, wherein after heating a 10% (w / w) protein content aqueous solution comprising the denatured whey protein composition at 120° C. for 15 minutes, d 90 No greater than about 2.0 μm, such as about 0.9 to about 1.7 μm.

[0321] Exemplary embodiment 10. A heat-stable denatured whey protein composition according to any one of exemplary embodiments 1-9, wherein the volume-weighted average diameter D(4,3) is no greater than about 1.0 μm, such as about 0.6 to about 1.0 μm, after heating a 10% (w / w) protein content aqueous solution comprising the denatured whey protein composition at 120°C for 15 minutes.

[0322] Exemplary embodiment 11. A heat stable denatured whey protein composition according to any one of exemplary embodiments 1-10, wherein a 10% (w / w) protein content aqueous solution of the denatured whey protein composition has 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.

[0323] Exemplary embodiment 12. The heat stable denatured whey protein composition of any of exemplary embodiments 1-11, wherein the composition comprises less than about 10% by weight casein.

[0324] Exemplary embodiment 13. The heat stable denatured whey protein composition of any of exemplary embodiments 1-12, wherein the composition is substantially free of casein.

[0325] 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% β-lactoglobulin, from about 35% to about 75% β-lactoglobulin, or from about 40% to about 65% β-lactoglobulin relative to the total protein in the composition.

[0326] Exemplary embodiment 15. The heat stable denatured whey protein composition of any of exemplary embodiments 1-14, wherein the composition comprises at least 70% (w / w) whey protein relative to the total protein, at least 80% (w / w) whey protein relative to the total protein, or at least 90% (w / w) whey protein relative to the total protein.

[0327] 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%.

[0328] 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%.

[0329] Exemplary embodiment 18. A liquid composition comprising the heat stable denatured whey protein composition of any one of exemplary embodiments 1-17.

[0330] Exemplary embodiment 19. The liquid composition of exemplary embodiment 18, wherein the liquid composition is a drinking yogurt.

[0331] Exemplary embodiment 20. A food product comprising the heat stable denatured whey protein composition of any one of exemplary embodiments 1-17.

[0332] Exemplary embodiment 21. The food product of Exemplary embodiment 20, wherein the food product is a baked good, a bar, or set or stirred yogurt.

[0333] Exemplary embodiment 22. The food of exemplary embodiment 20, wherein the food is a set yogurt, the set yogurt optionally comprising about 6% to about 20% (w / v) total protein, optionally, at least 50% (w / w) of the total protein coming from the denatured whey protein composition, wherein the set yogurt exhibits reduced hardness and / or reduced volume-weighted average particle size compared to a control set yogurt product having the same ingredient composition and the same protein content, the difference being that the control set yogurt product does not include the denatured whey protein composition described in any one of exemplary embodiments 1-17.

[0334] Exemplary embodiment 23. The food of exemplary embodiment 20, wherein the food is a stirred yogurt, the stirred yogurt optionally comprising from about 6% to about 20% (w / v) total protein, optionally at least 50% (w / w) of the total protein coming from the denatured whey protein composition, wherein the stirred yogurt exhibits reduced viscosity and / or reduced volume-weighted average particle size compared to a control stirred yogurt product having the same ingredient composition and the same protein content, except that the control stirred yogurt product does not include the denatured whey protein composition of any one of exemplary embodiments 1-17.

[0335] Exemplary embodiment 24. A nutritional composition comprising the heat stable denatured whey protein composition of any one of exemplary embodiments 1-17.

[0336] Exemplary embodiment 25. A liquid nutritional composition comprising the heat stable denatured whey protein composition of any one of exemplary embodiments 1-17.

[0337] Exemplary embodiment 26. A method of providing nutrition to a subject in need thereof, the method comprising administering the nutritional composition of Exemplary embodiment 24 or the liquid nutritional composition of Exemplary embodiment 25 to the subject.

[0338] 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.

[0339] Exemplary embodiment 28. A method for preparing a heat-stable denatured whey protein composition, the method comprising: (a) providing a whey protein solution or a whey protein retentate; (b) contacting the whey protein solution or the whey protein retentate with an oxidizing agent; and (c) heat treating the whey protein solution or the whey protein retentate under conditions of high shear stress / force, wherein the total protein content of the whey protein solution or the whey protein retentate in step (c) is preferably at least 16% (w / w); wherein the high shear stress / force is optionally generated by an increased slurry phase viscosity, a wall shear rate above 1000 s-1, a turbulent flow mode with a Re of at least 2000, application of mechanical shear or a combination thereof.

[0340] Exemplary embodiment 29. The method of exemplary embodiment 28, wherein the oxidizing agent is hydrogen peroxide or benzoyl peroxide.

[0341] Exemplary embodiment 30. The method of exemplary embodiment 29, wherein step (b) further comprises contacting the whey protein solution or the whey protein retentate with a combination of an oxidizing agent and a catalyst (eg, an enzyme catalyst or a chemical catalyst, preferably peroxidase).

[0342] Exemplary embodiment 31. The method of any of exemplary embodiments 28-30, wherein the oxidant is present in step (b) in an amount of less than 300 ppm or less than 200 ppm.

[0343] Exemplary embodiment 32. The method of any one of exemplary embodiments 28-30, wherein the oxidant is less than 1200×10 -6 kg / kg whey protein, less than 2000×10 -6 kg / kg variable whey protein or less than 2400×10 -6 An amount of kg / kg of β-lactoglobulin protein is present in step (b).

[0344] 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).

[0345] Exemplary embodiment 34. The method of any one of exemplary embodiments 28-33, wherein the whey protein solution or whey protein retentate has a low level of denaturation prior to step (c).

[0346] Exemplary embodiment 35. The method of exemplary embodiment 34, wherein the denaturation level of the whey protein solution or the whey protein retentate prior to step (c) is less than 10%, such as from about 3% to about 8%.

[0347] 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.

[0348] Exemplary embodiment 37. A heat stable denatured whey protein composition prepared by the method of any one of exemplary embodiments 28-35.

[0349] Exemplary embodiment 38. A method for preparing a liquid composition, the method comprising combining the heat-stable denatured whey protein composition of any 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 microbial activity.

[0350] Exemplary embodiment 39. A heat-treated, shelf-stable, high-protein liquid composition comprising at least 6% (w / v) whey protein, wherein the whey protein comprises microparticles comprising denatured whey protein; and wherein (i) the liquid composition has a d of no greater than about 1.0 μm after a secondary heat treatment for microbial control. 50 and / or a d of not greater than about 2.0 μm 90 (ii) the liquid composition has a temperature of 100s at 20°C -1 (iii) the liquid composition exhibits less than 10% sedimentation after storage of the liquid composition at a temperature of about 20°C to about 25°C for the shelf life; and / or (iv) the liquid composition exhibits less than 10% sedimentation after centrifugation at 1540 x g for 5 minutes.

[0351] Exemplary embodiment 40. 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, wherein the liquid composition or the liquid nutritional composition or the heat-treated shelf-stable liquid composition:

[0352] ·Having a d of no greater than about 1.0 μm 50 and a d not greater than about 2.0 μm 90 ;

[0353] comprising at least 12%, at least 14%, at least 16%, at least 18%, or at least 20% (w / v) total protein; and / or

[0354] • 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.

[0355] 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.

[0356] 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.

[0357] Exemplary embodiment 43. The heat-treated shelf-stable liquid composition of exemplary embodiment 42, wherein the drinking yogurt has a shelf-stable liquid composition of 100% by volume at 20°C for 50 s. -1 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 measured at room temperature.

[0358] 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.

[0359] Exemplary embodiment 45. The heat-treated shelf-stable liquid composition of exemplary embodiment 39, wherein the liquid composition is an acidic beverage.

[0360] 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.

[0361] 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.

[0362] Exemplary embodiment 48. The heat-treated shelf-stable liquid composition of exemplary embodiment 47, wherein the acidic beverage has a shelf-stable liquid composition having ... -1 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 measured at room temperature.

[0363] 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.

[0364] Exemplary embodiment 50. The heat-treated shelf-stable liquid composition of exemplary embodiment 39, wherein the liquid composition is a neutral beverage.

[0365] 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.

[0366] Exemplary embodiment 52. A 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.

[0367] Exemplary embodiment 53. The heat-treated shelf-stable liquid composition of exemplary embodiment 39, wherein the liquid composition is a liquid nutritional composition for providing nutrition to a subject in need thereof.

[0368] 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, such as Ca++.

[0369] Exemplary embodiment 55. A 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.

[0370] 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.

[0371] Exemplary embodiment 57. The heat-treated shelf-stable liquid composition of any one of exemplary embodiments 53 to 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.

[0372] Exemplary embodiment 58. The heat-treated shelf-stable liquid composition of any one of exemplary embodiments 39-57, further comprising a non-whey protein, preferably selected from casein, caseinates, micellar casein isolates, and mixtures thereof.

[0373] Exemplary embodiment 59. A liquid nutritional composition comprising at least 6% (w / v) whey protein, wherein the whey protein comprises microparticles comprising denatured whey protein; and wherein the liquid nutritional composition further comprises a lipid component, wherein the lipid component is optionally present in an amount of up to 30% (w / v), a carbohydrate component, wherein the carbohydrate component is optionally present in an amount of up to 30% (w / v), at least one monovalent cation, and 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 the divalent metal cation.

[0374] 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.

[0375] Exemplary embodiment 61. A method of providing nutrition to a subject in need thereof, the method comprising administering to the subject the liquid nutritional composition of any one of exemplary embodiments 25, 53-57, or 59-60.

[0376] Exemplary embodiment 62. A heat-treated high-protein coagulated gel, the coagulated 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; and wherein the coagulated gel (i) exhibits a hardness of less than 1000 g.sec and / or (ii) a breaking force of less than 50 g.

[0377] Exemplary embodiment 63. The heat-treated high-protein coagulated gel of exemplary embodiment 62, wherein the coagulated gel comprises yogurt.

[0378] Exemplary embodiment 64. The heat-treated high-protein coagulated gel of exemplary embodiment 62 or exemplary embodiment 63, wherein the coagulated gel comprises at least 12%, at least 14%, at least 16%, at least 18% or at least 20% (w / w) whey protein.

[0379] Exemplary embodiment 65. A heat-treated high protein semi-solid food, the semi-solid food 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; and wherein the semi-solid food has a temperature of 50s at 20°C. -1 A viscosity of less than 1000 mPa.s, less than 800 mPa.s, less than 600 mPa.s or less than 400 mPa.s measured under reduced pressure.

[0380] Exemplary embodiment 66. The heat-treated high-protein semi-solid food of Exemplary embodiment 65, wherein the semi-solid food comprises stirred yogurt.

[0381] Exemplary embodiment 67. The heat-treated high-protein semi-solid food of exemplary embodiment 65 or exemplary embodiment 66, wherein the semi-solid food comprises at least 12%, at least 14%, at least 16%, at least 18% or at least 20% (w / w) whey protein.

[0382] Exemplary embodiment 68. A liquid nutritional composition comprising microparticles comprising denatured whey protein, wherein the liquid nutritional composition comprises no more than 12% (w / v) total protein and at least 95% of the total protein in the liquid nutritional composition is denatured whey protein; and wherein the liquid nutritional composition optionally also comprises a lipid component and / or a carbohydrate component.

[0383] 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.

[0384] Exemplary embodiment 70. The liquid nutritional composition of Exemplary embodiment 68 or Exemplary embodiment 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).

[0385] Exemplary embodiment 71. The liquid nutritional composition of any one of exemplary embodiments 68-70, 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 the divalent metal cation.

[0386] 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.

[0387] 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 to 2 kcal / ml.

[0388] Exemplary embodiment 74. The liquid nutritional composition of any one of exemplary embodiments 68-71, wherein the liquid nutritional composition has a caloric density greater than 2 kcal / ml.

[0389] Exemplary embodiment 75. The liquid nutritional composition of any one of exemplary embodiments 68-71, wherein the liquid nutritional composition has a caloric density of 1.5 to 2.5 kcal / ml.

[0390] Exemplary embodiment 76. The liquid nutritional composition of any one of exemplary embodiments 68-75, wherein (i) the liquid composition has a d value of no greater than about 1.0 μm after secondary heat treatment for microbial control. 50 ; (ii) the liquid composition has a viscosity of less than 400 mPa.s or less than 200 mPa.s measured at 100 s-1 at 20°C; (iii) the liquid composition exhibits less than 10% sedimentation after storage at a temperature of about 20°C to about 25°C for at least 3 months; and / or (iv) the liquid composition exhibits less than 10% sedimentation after centrifugation at 1540 x g for 5 minutes.

[0391] Exemplary embodiment 77. A liquid nutritional composition comprising microparticles containing denatured whey protein, wherein the liquid nutritional composition comprises at least 12% (w / v) total protein and at least 95% of the total protein in the liquid nutritional composition is denatured whey protein; wherein the liquid nutritional composition further comprises a lipid component and / or a carbohydrate component.

[0392] 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.

[0393] Exemplary embodiment 79. The liquid nutritional composition of Exemplary embodiment 77 or Exemplary embodiment 78, wherein the liquid nutritional composition has a neutral pH.

[0394] Exemplary embodiment 80. The liquid nutritional composition of any one of exemplary embodiments 77-79, wherein said liquid nutritional composition comprises 12% to 18% (w / v) total protein, wherein substantially all of said total protein is from denatured whey protein.

[0395] 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).

[0396] Exemplary embodiment 82. The liquid nutritional composition of any one of exemplary embodiments 77-81, 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 the divalent metal cation.

[0397] 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.

[0398] 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 to 2 kcal / ml.

[0399] Exemplary embodiment 85. The liquid nutritional composition of any one of exemplary embodiments 77-82, wherein the liquid nutritional composition has a caloric density greater than 2 kcal / ml.

[0400] 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 to 2.5 kcal / ml.

[0401] Exemplary embodiment 87. The liquid nutritional composition of any one of exemplary embodiments 77-86, wherein (i) the liquid composition has a d value of no greater than about 1.0 μm after secondary heat treatment for microbial control. 50 (ii) the liquid composition has a temperature of 100s at 20°C -1(iii) the liquid composition exhibits less than 10% sedimentation after storage at a temperature of about 20°C to about 25°C for at least 3 months; and / or (iv) the liquid composition exhibits less than 10% sedimentation after centrifugation at 1540 x g for 5 minutes.

[0402] I. Embodiment

[0403] In order to demonstrate the practice of the subject matter disclosed herein, the following examples were prepared and tested. However, the examples should not be construed as limiting the scope of the invention.

[0404] method

[0405] Preparation and evaluation of denatured whey protein powder

[0406] All powders were recombined to 10% (w / w) protein concentration using ambient (~20°C) deionized water with a magnetic stirrer. The protein powder was gradually added to the water under vortex mixing conditions, carefully avoiding any foaming. Once all powders were dispersed, hydration was continued for 30 minutes under continuous stirring. The solution was homogenized at 150 / 50 bar and tested for primary particle size, primary particle size growth, thermal coagulation time, insolubility and denaturation as described below.

[0407] The primary particle size of the reconstituted powder was determined using a Malvern Mastersizer 2000 or Malvern Mastersizer 3000 (Malvern Instruments Ltd, Worcs, UK), with a refractive index of 1.46 for the particles and 1.33 for the solvent. The volume-weighted average particle size d[4,3], D 50 and D 90 Percentile value of the value.

[0408] The volume % of particles within a defined size range is determined by taking the sum of the relevant size classes of the raw data reported by the Mastersizer software.

[0409] Primary particle size growth was determined after heating a 10% protein solution (pH 6.8) in a silicone oil bath at 120°C for 15 minutes. A 5 mL aliquot of the 10% protein solution was placed in an 8 mL glass vial with a closed cap and placed in a shaking unit inserted in an oil bath at 120°C while gently shaking the sample during the 15 minute heating period. The sample was immediately cooled by plunging into ice water and the particle size analysis was performed as described above.

[0410] Thermal coagulation time: A 1 ml aliquot of a 10% protein (w / w) solution (pH 6.8) was placed in a glass vial, which was clamped on a platform and placed in a silicone oil bath thermostatically controlled at 140° C. with a gentle shaking rate. The length of time (minutes) that elapsed between the placement of the container in the oil bath and the onset of visible aggregate formation was defined as the thermal coagulation time (HCT).

[0411] Insoluble (%) = 7.5 g of denatured whey protein powder was dissolved in 192.5 g of 0.1 M sodium chloride solution by mixing for 30 minutes using a magnetic stirrer. The pH of a 50 g portion of the suspension was adjusted to pH 4.6 using 15% acetic acid (the amount of acetic acid added was recorded for the dilution correction factor). 40 g of the suspension was transferred to a 50 ml centrifuge tube and centrifuged (Beckman) at 7250 g for 20 minutes. The total nitrogen content was tested by the Kjeldahl method and the total protein in the supernatant of the solution was analyzed (ISO 8968-1 / 2 | IDF 020-1 / 2 - Milk - Determination of nitrogen content - Part 1 / 2: Determination of nitrogen content using the Kjeldahl method). The total protein content was calculated by multiplying the total nitrogen result by a conversion factor of 6.38. Another portion of the suspension (not centrifuged) was also analyzed for total protein.

[0412]

[0413] Denatured and soluble proteins were analyzed based on the method described by Elgar et al. (2000) J Chromatography A, 878, 183-196.

[0414] The powders were characterized by measuring the residual variable protein as a proportion of the total protein according to the following formula:

[0415]

[0416] Soluble whey protein was determined using reverse phase HPLC and is reported as g protein / 100 g powder.

[0417] Variable whey protein was measured as ∑ bovine serum albumin + α-lactalbumin + β-lactoglobulin + lactoferrin + immunoglobulins.

[0418] The denaturation % of a denaturable protein can be calculated based on the following formula:

[0419]

[0420] The percentage of covalently cross-linked β-lactoglobulin was determined using a 2100 Agilent Bioanalyzer system (Agilent, USA) utilizing microfluidic SDS electrophoresis using the method of Anema (2009) with modifications (SG Anema, 2009, “Lab on a chip” microfluidic SDS electrophoresis for separation and quantification of milk proteins. International Dairy Journal, Vol. 19, No. 4, pp. 198-204).

[0421] The proportions of residual native protein, covalently aggregated protein and non-covalently aggregated protein were determined by dissolving the denatured whey protein composition in a buffer that dissociates the different bonds and measuring the change in soluble protein after a centrifugation step.

[0422] To determine the residual native protein content, the samples before and after heating were dissolved / or diluted to 10% (w / w) protein in water. A 300 mg aliquot of the sample was dissolved in 1.2 mL of acetate buffer (0.2 M, pH 4.35).

[0423] Prepare 0.1M phosphate buffer (pH=6.7) prepared from 0.0281mol NaH2PO4.2H2O and 0.0218mol Na2HPO4; add urea to a final concentration of 8M, and add sodium dodecyl sulfate (SDS) to a final concentration of 2% (w / v). Dissolve a 300mg aliquot in 1.2mL phosphate / urea / SDS buffer (PSU buffer).

[0424] The prepared samples were shaken using a test tube shaker for 1 hour.

[0425] The samples were centrifuged at 20,000 x g for 1 hour at 25°C.

[0426] Carefully take a 100 μL aliquot of the supernatant and run using a bioanalyzer system and prepared according to the method of Anema. The area under the curve of the protein peak is automatically integrated by software (2100 Bioanalyzer Expert Software package). The peak area is used to calculate the binding type based on the sample preparation as described above.

[0427] in accordance with

[0428] The acetate buffer used in this experiment precipitates any denatured whey protein or residual casein in the solution. Unheated samples are expected to have low levels of denatured protein. Heated samples are expected to have high levels of denaturation. The residual native protein in the heated samples is calculated as the proportion of the unheated feed native protein.

[0429]

[0430] Urea can break hydrophobic bonds and SDS can break non-covalent bonds such as hydrogen and hydrophobic bonds. Heated samples added to phosphate buffer containing urea and SDS will cause solubilization of non-covalent bonds, thus allowing calculation of non-covalently aggregated proteins compared to total soluble proteins in the unheated solution, taking into account the residual native-like proteins.

[0431]

[0432] Therefore, covalently aggregated protein can be calculated as the remaining portion of protein not solubilized by PSU buffer.

[0433] Covalently aggregated protein = 1 - (residual native-like protein - non-covalently aggregated protein)

[0434] Preparation and analysis of high protein liquid nutritional compositions and / or beverages

[0435] Figure 2 A flow chart showing the overall process of producing a high protein beverage.

[0436] Neutral high protein drink

[0437] The required proteins, carbohydrates, minerals and stabilizers are dry mixed and hydrated in water heated to 55°C for 60 minutes, and defoamers are added if necessary. Oil is added to the mixture. The pH is adjusted to about pH 6.8 ± 0.1 using 5% KOH. The mixture is homogenized at 55°C, 150 / 50 bar. Beverages can be sterilized in two ways:

[0438] (1) The homogenized mixture was filled into a distillation pot and heated at 120°C for 15 minutes.

[0439] (2) The homogenized mixture is UHT sterilized at 145°C for 4 seconds and then homogenized at 150 / 50 bar. The product is aseptically packaged.

[0440] Acidic high protein drinks

[0441] The same process is followed as for the neutral beverage; however, a pectin solution is prepared at 80°C and added to the water before adding protein, carbohydrates, minerals and stabilizers. The pH is adjusted to pH 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 packaged aseptically.

[0442] Unless otherwise stated, a rheometer such as the Anton Paar instrument was used, using a cup and hammer assembly, at 100 s. -1The viscosity of the formulation is measured at a shear rate of 20° C. at 20° C. It should be understood that other methods of measuring or estimating viscosity are well known in the art and may be used where appropriate.

[0443] The particle size distribution of the beverage was measured in the same manner as described for primary particle size, and volume % analysis of the particles was performed in the same manner as above.

[0444] Sedimentation Test: After appropriate heat treatment of the beverage, the product is stored at 25°C for shelf life testing at 1 month, 3 months, 6 months or 12 months. To test for sedimentation, the product is inverted 3 times and poured out carefully. The container is placed upside down for 30 minutes; the weight of the container plus the sediment is recorded; the amount of sediment is calculated by subtracting the weight of the empty container. The sedimentation % is then calculated by the ratio of the sedimentation weight to the total weight of the product in the container.

[0445] Preparation and analysis of high-protein drinking yogurt, set and stirred yogurt

[0446] Figure 3 A flow chart of a yogurt manufacturing process is shown. Denatured whey protein particle concentrate, skim milk powder (SMP) and optional MPC are mixed in 10°C water for another 60 minutes using an overhead stirrer. The composition is kept overnight at 4°C. The mixture is preheated to 60°C and then homogenized at 150 / 50 bar. The composition is then heated to 95°C and kept for 6 minutes, 80°C for 20 minutes, or equivalently 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 yogurt. Drinking and / or stirred yogurt is cooled to 20°C and processed to break the gel. The yogurt is loaded into a container and cooled to about 4°C. Immediately after inoculation, the coagulated yogurt is loaded into a tank and fermented to produce a coagulated yogurt.

[0447] The pH was measured at about 10° C. using a pH meter (model pHM210; Radiometer Analytical SAS, France) calibrated with pH 7.0 and pH 4.0 buffers before use. The pH of each sample was measured at least twice on day 7 from separate tanks.

[0448] The apparent viscosity was measured at 10° C. using a Haake viscometer (Haake Mess-Technik, Gmbh & Co., Karlsruhe, Germany). The shear rate was increased from 0 to 120 s over a period of 180 s. -1 , then reduce the shear to 0s over a period of 30s -1 To measure viscosity. Record at 50s -1Apparent viscosity at shear rate. Tests were performed in duplicate on day 7 from separate sample jars.

[0449] The sedimentation via centrifugation was determined by centrifuging a sample of drinking yogurt at 3000 rpm (1540 x g) for 5 minutes at a temperature of 10-15°C using a Sorvall Evolution RC centrifuge (rotor F13S-14x 50cY, rotor code C=SLA600TC angle rotor). The supernatant was carefully removed and the sides of the tube were dried. The sedimentation as a percentage of the total weight of the original sample in the tube was calculated as % sedimentation.

[0450] Sedimentation was determined by carefully pouring the drinking yogurt from the container to allow the liquid to drain out. Sedimentation was determined by measuring the height of the sediment in the bottle and the height of the yogurt in the bottle and calculating the sediment as a percentage of the total sample. Tests were performed in duplicate from separate sample jars after 1 week and 6 weeks of storage.

[0451] Phase separation was determined by measuring the height of the clear layer formed on top of the yogurt and the height of the yogurt in the bottle and calculating the phase separation as a percentage of the total sample. Tests were performed in duplicate from separate sample jars after 1 and 6 weeks of storage.

[0452] The breaking force and hardness were evaluated using a TAHD Plus Texture Analyser from Stable Micro Systems, Godalming, UK. A 27 cm Perspex cylinder was used in a compression test; the initial breaking force of the set yoghurt surface was recorded as the breaking force (g) and the area under the curve was recorded as the hardness (gs).

[0453] The sensory attributes of foods were evaluated using the following sensory procedure. Foods were presented in clear sample cups labeled with a random 3-digit blind code. Samples were presented to the panel at a temperature appropriate for the specific food. The sensory evaluation was conducted by a panel of 8 expert panelists who were highly experienced in tasting specific foods. Food samples were evaluated with participants describing texture and flavor attributes and intensity. A consensus method was used to best collate the attributes describing each sample.

[0454] Figure 2 is an exemplary process flow diagram for making an exemplary liquid composition including denatured whey protein concentrate, namely a high protein beverage.

[0455] Figure 3 is an exemplary process flow diagram for making an exemplary liquid composition, namely a drinking yogurt, including a denatured whey protein composition.

[0456] Example 1A: Preparation of the denatured whey protein particle composition of the present invention

[0457] Use standard commercial ultrafiltration / diafiltration technology that the cheese whey of annatto coloring is concentrated to the protein concentration of about 22-24% (w / w).Table 1 shows the specific conditions for producing powder A, B, C and D from colored whey.By pre-loading the retentate silo with fungal peroxidase MaxiBright and adding the hydrogen peroxide that amounts to 50-80ppm, the whey retentate of coloring is decolorized.Select the ratio of hydrogen peroxide to retentate, hydrogen peroxide addition rate and hydrogen peroxide to add the mixing holding time of retentate in silo after retentate is added, to provide the best improvement to the variability whey protein composition, provide expected whiteness simultaneously and prevent the formation of peculiar smell.This step is completed at the temperature of<10 ℃.Before further processing, retentate is sampled and uses Merck peroxide indicator test strip (MQuant peroxide test strip, 0.5-25mg / L H2O2) test detectable peroxide, and do not detect peroxide.

[0458] The pH of the whey protein concentrate was adjusted as described in Table 1 using a 2% (w / v) sodium hydroxide solution and preheated to 54°C using a heat exchanger heated by hot water. Subsequently, a denatured whey protein composition was prepared according to the method described in WO2010120199. The whey protein concentrate was fed to two identical single-tube high-pressure steam-heated shell-and-tube heat exchangers in series at a sufficiently high flow rate using a high-pressure pump with a delivery pressure of 250-350 bar to achieve a Reynolds number of ≥2100. As shown in Table 1, the concentrate left the first high-pressure heater at 71-73°C and left the second high-pressure heater at 80-85°C.

[0459] After coming out of the second heater, the heat treated whey protein concentrate passes through a holding pipe and a pipe to the nozzle group at the top of the spray dryer, which has a similar pipe specification to the high pressure heater, which means that the turbulent flow is maintained. The length of the holding pipe is selected in such a way that the combined pipe to the nozzle group provides an additional 20s residence time for the heated flow before spray drying, where the temperature loss of the pipe from the outlet of the second high pressure heater until the nozzle group is <1°C. This means that no additional mechanical shear inducing device is used during heating and after the heater-reactor system and before spray drying.

[0460] In the spray dryer, the heat treated concentrate is delivered to a set of three nozzles and atomized into a droplet spray at a pressure greater than 200 bar. An inlet hot air temperature of 210-220°C and an outlet chamber temperature of 60-70°C are used. The powder is further dried and then cooled in a vibrating fluidized bed, after which the material is sieved and packaged to produce a powder with <5% moisture. According to the method described in WO2010120199, several uncolored whey protein concentrates are also processed without the addition of hydrogen peroxide and peroxidase (powders E, F, G) under turbulent flow with a Reynolds number of ≥2100. A commercially available micronized WPC (Nutrilac YO-8075) from Arla Foods (powder H) was obtained and a comparative test was performed using the method described in the method section.

[0461] Table 1. Comparison of process variables for producing denatured whey compositions

[0462]

[0463] Table 2. Typical overall composition (dry weight basis) and protein distribution of whey protein concentrates before heating.

[0464] Composition Annatto Colored Whey Uncolored whey Protein (w / w) 84.9 83.75 Fat (w / w) 5.78 5.3 Lactose (w / w) 5.66 5.75 Ash content (w / w) 2.62 3.0 Ca(mg / 100g) 390 395 Na(mg / 100g) 160 227 K(mg / 100g) 620 609 Soluble GMP / Total soluble protein (%) 20.3 21.5 Soluble pp5 / total soluble protein (%) 3.2 4.8 Soluble α-lac / total soluble protein (%) 13.7 14.7 Soluble BSA / Total soluble protein (%) 1.07 1.7 Soluble β-lg / total soluble protein (%) 57.6 52.6 Soluble IgG / total soluble protein (%) 4.1 4.6

[0465] Table 3. Comparison of characteristics of reconstituted denatured whey protein concentrate powders

[0466]

[0467] Example 1B: Turbidity of denatured whey protein concentrate powder after reconstitution

[0468] The turbidity of the denatured whey protein concentrate was determined by reconstituting a powder prepared according to Powder C (described 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) so that the reading was within the measurement range of the spectrophotometer. The absorbance of the 0.025% protein solution at 500 nm at about 20° C. was 0.857. Taking into account the dilution factor, the turbidity of the 0.1% protein solution was 3.4 absorbance units.

[0469] Example 2: Preparation of 9% protein (w / v) shelf-stable beverage

[0470] This example describes the preparation of an exemplary high protein liquid beverage comprising a powder (Powder A) using a method according to the present invention compared to the comparative powders (Powders F, G, H) described in Example 1. The beverage was prepared under neutral conditions according to the procedure in the Methods section. The recipe for preparing the beverage is shown in Table 4 and the nutritional composition of the beverage is summarized in Table 5.

[0471] Table 4. Formulations of high protein liquid beverages including different denatured whey protein concentrates

[0472] Ingredients (g / 100g) Powder A Powder F Powder G Powder H water 87.66 90.14 87.66 88.67 Powder A 10.70 Powder F 11.02 Powder G 10.70 Powder H 10.60 Canola Oil 0.18 0.18 0.18 0.18 Sodium chloride 0.06 0.06 0.06 0.06 Tripotassium citrate monohydrate 0.060 0.060 0.060 0.060 Gellan Gum DAIDupont 0.040 0.040 0.040 0.040 Novagel 1518 0.190 0.190 0.190 0.190 Defoaming agent 0.01 0.01 0.01 0.01 5% Potassium Hydroxide 0.8 0.8 0.8 0.8

[0473] Table 5. Nutritional composition of high protein liquid beverages including different denatured whey protein concentrates

[0474] Components Nutritional composition Protein (g / 100ml) 9.0 Protein from denatured WPC (g / 100ml) 9.0 Fat (g / 100ml) 0.8 Carbohydrate (g / 100ml) 1.0 Ash content (g / 100ml) 0.5 Total solids (g / 100ml) 11.9 Energy (kcal / ml) 0.5 Sodium (mg / 100ml) 93 Potassium (mg / 100ml) 65 Calcium (mg / 100ml) 32

[0475] Table 6 shows the properties of high protein beverages prepared using different denatured whey protein concentrates. When compared to the properties of powder A summarized in Example 1, Table 3, the high protein beverage including powder A showed minimal changes in particle size characteristics after heating at 120°C for 15 minutes. The high protein beverage prepared with powder A had a uniform consistency and a smooth appearance that was very thin and pourable. Powder F caused the high protein beverage to gel, forming large aggregates with a granular appearance in solution during heat treatment and therefore showed significant instability to sterilization conditions. Powder G and powder H exhibited significant particle growth due to heat treatment, which resulted in an unpleasant gritty / powdery mouthfeel. In Figure 4 The difference in particle size distribution after sterilization can be clearly seen in Figure 2. Figure 4 Shown is the particle size distribution of a high protein beverage prepared with powder A, powder G or powder H after heating at 120°C for 15 minutes.

[0476] Table 6. Beverage properties after distillation heat treatment

[0477]

[0478] Example 3: Primary Particle Size Growth

[0479] This example shows the particle size of a denatured whey protein concentrate of the invention and a comparative denatured whey protein concentrate after heating at 120°C for 15 minutes.

[0480] All powders were prepared according to the reconstitution method described in the method, however, 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 method.

[0481] Table 7 shows the D 50 and D 90 The values ​​can be compared with the corresponding powders in Example 1, Table 3, which show the particle size before heating at 120°C for 15 minutes. The appearance of the denatured whey protein composition after heating is also described in the table as liquid, viscous liquid or gel state.

[0482] Table 7. Change in particle size of denatured whey protein compositions after heating at 120°C for 15 minutes.50 and D 90 Reported in μm.

[0483]

[0484] For solutions containing up to 16% (w / w) protein and 18% (w / w) protein, the powder of the present invention showed no significant change in particle size after heat treatment at 120°C for 15 minutes and remained liquid. The comparative composition gelled at a protein content of 10% (w / w) or 12% (w / w) protein after the same heat treatment. The denatured whey protein composition of the present invention exhibits excellent heat stability even at higher protein concentrations and exhibits minimal or no change in particle size distribution compared to the comparative denatured whey protein composition. Figure 5 Shown is the particle size distribution of protein solutions of Powder A containing 10% to 16% (w / w) protein after heating at 120°C for 15 minutes.

[0485] After heat treatment, D 50 Less than 1μm and D 90 A particle size distribution of less than 2 μm is an important feature to achieve good shelf life stability, minimal settling, low viscosity and good mouth feel properties such as no powdery or gritty feel. This is representative of all powders of the present invention.

[0486] Figure 5 Shown is the particle size distribution of a protein solution containing Powder A at 10%, 12%, 14% or 16% (w / w) protein after heating at 120°C for 15 minutes.

[0487] Example 4: Thermal coagulation time at high protein concentration and use of the denatured whey protein concentrate of the present invention in a high protein liquid nutritional composition

[0488] This example shows the heat coagulation time (at 140°C) of a protein solution containing a denatured whey protein concentrate according to the invention and a comparative whey protein concentrate of high protein concentration.

[0489] All compositions were prepared by the recombinant method described in the methods, however, protein concentrations of 12, 14, 16, 18% and 20% (w / w) protein were also prepared. Heating was performed according to the thermal clotting time method described in the methods.

[0490] Figure 6The denatured whey protein compositions of the present invention are shown to be significantly more thermally stable than comparative denatured whey protein compositions. Based on the inventors' experience, it is known that samples take at least 1 minute to 1.5 minutes to reach 140°C, and therefore, any sample that does not show a thermal coagulation time greater than 1 minute is unlikely to be stable during UHT heat treatment. All denatured whey protein compositions of the present invention have a thermal coagulation time of 1 minute or greater at up to 20% (w / w) protein. When prepared at a concentration of 14% (w / w) or greater, powders G and H exhibit a thermal coagulation time of less than 1 minute; when prepared at a concentration of 10% (w / w) or greater, powder F exhibits a thermal coagulation time of less than 1 minute.

[0491] Figure 6 : Thermal coagulation time of denatured whey protein compositions prepared at protein concentrations ranging from 10% to 20% (w / w).

[0492] As a next step, the particle size distribution of protein solutions of the inventive powder C containing 10%-18% (w / w) protein was measured after heating at 140°C for 90 seconds and immediate cooling. The values ​​in Table 8 can be compared with the corresponding powders in Example 1, Table 3, which shows the particle size before heating. Powder C not only remains liquid after such heating conditions, but also shows minimal particle size changes at protein concentrations up to 18% (w / w) protein when heated at 140°C for 90 seconds.

[0493] Table 8. Particle size distribution of powder C after heating at 140°C for 90 seconds. 50 and D 90 Reported in μm. The appearance of the composition after heating is described.

[0494]

[0495] Based on these findings, several high protein liquid nutritional compositions were prepared using the powder of the present invention (Powder A) as described in Tables 9, 10, 11. Since the comparative denatured whey protein concentrate at a concentration of 14% (w / w) or higher was not heat stable at 140°C, these samples were not further tested in high protein nutritional compositions. Beverages were prepared according to the procedure shown in the Methods section.

[0496] Table 9. Formulations of neutral and acidic pH high protein liquid beverages including powder A

[0497] Ingredients (g / 100g) Recipe 1 Recipe 2 Recipe 3 water 80.86 69.92 78.41 Powder A 16.99 15.909 19.05 Canola Oil 0.62 6.10 0.4 sugar 0.6 2.0 0.64 Maltodextrin T400 5.0 Phosphatidylcholine (Metarin P IP) 0.10 0.13 0.40 Tripotassium citrate monohydrate 0.060 0.06 0.032 MCC / CMC stabilizer (Novagel 1518) 0.10 0.10 Gellan Gum 0.025 Carrageenan 0.04 Pectin YM 115 0.056 Sodium chloride 0.03 Trisodium citrate dihydrate 0.075 Calcium carbonate 0.209 Defoaming agent 0.01 0.01 0.01 5% Potassium Hydroxide 0.6 0.45 15% hydrochloric acid 1.0 Product pH 7.0 7.0 4.0 UHT process conditions (temperature, time) 145℃, 4 seconds 145℃, 4 seconds 110℃, 4 seconds

[0498] Table 10. Nutritional compositions of pH neutral and acidic high protein liquid beverages including various protein contents and compositions

[0499]

[0500] Table 11. Beverage properties of pH neutral and acidic high protein liquid beverages with various protein contents and compositions after UHT heat treatment

[0501] Recipe 1 Recipe 2 Recipe 3 <![CDATA[Volume-weighted particle size d 4,3 (μm)]]> 0.71 0.68 0.71 <![CDATA[D 50 (μm)]]> 0.64 0.46 0.67 <![CDATA[D 90 (μm)]]> 1.21 1.13 1.16 % Particle Volume <1 μm 77.9 85.2 78.8 % Particle Volume 1-10μm 22.1 14.7 21.2 % Particle Volume > 2 μm 0.6 3.4 0.0 <![CDATA[Viscosity (mPa.s at 100 s -1 ))]]> 7.9 11.8 9.7

[0502] Additional high protein liquid nutritional compositions were prepared using Powder A as described in Tables 12, 13, and 14.

[0503] Table 12. Formulation of neutral pH high protein liquid beverage including powder A

[0504] Ingredients (g / 100g) Recipe 4 Recipe 5 water 70.12 58.04 Powder A 15.91 18.10 Canola Oil 6.10 8.51 sugar 2.00 2.0 Maltodextrin T400 5.00 12.40 Phosphatidylcholine (Metarin P IP) 0.13 0.13 Tripotassium citrate monohydrate 0.06 0.06 MCC / CMC stabilizer (Novagel 1518) 0.10 0.10 Dimagnesium phosphate trihydrate 0.033 0.065 Carrageenan 0.04 0.04 Defoaming agent 0.01 0.01 Sodium chloride 0.049 Potassium chloride 0.095 5% Potassium Hydroxide 0.45 0.45 Product pH 6.76 6.75 UHT process conditions (temperature, time) 146℃, 6 seconds 146℃, 6 seconds

[0505] Table 13. Nutritional composition of pH neutral high protein liquid beverages including various protein contents and compositions

[0506]

[0507] Table 14. Beverage properties of pH neutral high protein liquid beverages with various protein contents and compositions after UHT heat treatment

[0508] Recipe 4 Recipe 5 <![CDATA[Volume-weighted particle size d 4,3 (μm)]]> 0.692 2.74 <![CDATA[D 50 (μm)]]> 0.587 0.75 <![CDATA[D 90 (μm)]]> 1.09 7.74 % Particle Volume <1 μm 91.33 62.1 % Particle Volume 1-10μm 8.66 37.92 <![CDATA[Viscosity at time 0 (mPa.s at 100 s -1 ))]]> 19.9 81.8 <![CDATA[Viscosity at 1 month (mPa.s at 100 s -1 ))]]> 21.9 60.9 <![CDATA[Viscosity at 3 months (mPa.s at 100 s -1 ))]]> 21.9 <![CDATA[Viscosity at 6 months (mPa.s at 100 s -1 ))]]> 24.6 88.8 Percentage of sediment at 1 month (%) 0.5 0.74 Sediment percentage at 3 months (%) 0.36 Sediment percentage at 6 months (%) 0.70 0.72

[0509] Powder A showed minimal particle size growth after UHT heat treatment when used to prepare pH neutral and acidic beverages with different compositions. The particle size of the beverages after heating can be compared with Example 1 in Table 3. Powder A showed very low viscosity for high protein beverages after UHT heat treatment and had a uniform consistency throughout with no signs of particles or separation in the packaging.

[0510] Example 5: Use of the improved denatured whey protein particle composition of the present invention in high-protein drinkable yogurt

[0511] This example describes the preparation of a number of exemplary high protein drinking yogurts comprising powders B and C using the method according to the invention in comparison with other denatured whey protein concentrates. Table 15 shows the nutritional composition and properties of yogurts comprising denatured whey protein concentrates.

[0512] Flow chart of yogurt manufacturing process Figure 3As shown. Denatured whey protein concentrate and skim milk powder (SMP) were reconstituted in water to produce an aqueous composition comprising 15 wt% protein, 12 wt% protein from a denatured whey protein particle composition or 20 wt% protein, 17 wt% from a denatured whey protein particle composition. For each batch, 9.2% (w / w) SMP was reconstituted and filled 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. Balance to 100% was reached with additional water.

[0513] Table 15. Composition and properties of high protein drinking yogurts containing denatured whey protein concentrate (all compositions had acceptable flavor)

[0514]

[0515] When powders B and C were used to prepare 15% (w / w) protein drinking yogurt, the yogurt showed minimal changes in particle size properties after one week of preparation compared to the powder properties in Example 1, Table 3. Comparative powders E, F and G showed particle size growth one week after yogurt manufacturing. Yogurt prepared with powders B and C also showed lower sedimentation (%) than the comparative yogurt. Similarly, at the end of the shelf life, 6 weeks after preparation, the sediment (%) remained low and no phase separation was observed for powders B and C. The overall appearance of the yogurt was smooth and uniform in appearance and was very thin and pourable, and had no powdery or granular texture. In contrast, powders E and F showed a thicker texture with a powdery / gritty mouthfeel. At the end of the 6-week shelf life, there was significant sediment formation in the drinking yogurt prepared with powders E, F and G. The sediment was a very solid gel layer that could not be redispersed back into the drinking yogurt by shaking.

[0516] When powder C was used to prepare 20% (w / w) protein drinking yogurt, it showed minimal change in particle size properties compared to the powder properties in Example 1, Table 3, one week after yogurt production. The yogurt was smooth and uniform, and very thin and pourable. Powders F and G were also used to prepare 20% (w / w) protein drinking yogurt; however, they were found to be unsuitable for use during heat treatment at 85°C for 15 minutes because the samples resulted in the formation of large gelled particles and lumps, and the viscosity increased significantly. The samples had a granular texture that would cause clogging of processing equipment.

[0517] Figure 7 The particle size distribution of 15% (w / w) and 20% (w / w) protein yoghurts 1 week after production is shown, which clearly shows the smaller particle size distribution of the denatured whey protein concentrate of the present invention.

[0518] Figure 7: Particle size distribution of high protein (15% (w / w) and 20% (w / w)) drinking yogurts comprising denatured whey protein concentrate at 1 week.

[0519] Example 6: Use of improved denatured whey protein concentrate in high protein groups and stirred yogurt

[0520] This example describes the preparation of exemplary high protein groups and stirred yogurts including the denatured whey protein composition of the present invention, Powder C, compared to Powder F. The recipes, compositions and yogurt properties of the set and stirred yogurts are summarized in Table 16. Figure 3 Yogurt was prepared with some modifications as stated in the methods.

[0521] Table 16. Recipes, composition and properties of set and stirred yogurts using Powder C and Powder F mixed with skimmed milk powder and MPC.

[0522]

[0523] The stirred yogurt prepared with powder C showed lower viscosity compared to powder F; the yogurt prepared with powder C was soft, spoonable, and had a shiny and smooth appearance, while the yogurt prepared with powder F had a rougher texture. The set yogurt prepared with powder C showed lower breaking force and hardness compared to powder F.

[0524] Example 7: Use of improved denatured whey protein concentrate in protein bars

[0525] An exemplary manner showing the use of denatured whey protein concentrate in protein bars is described.

[0526] Prepare protein bars by mixing maltodextrin and powder C. Combine glucose syrup, glycerine and water and heat to 50-55°C. Add the glucose syrup and glycerine mixture to the maltodextrin and protein mixture, then add the oil and lecithin mixture. Heat the confectionery fat and lecithin until the fat is melted. Use a Hobart mixer (model N-50) to mix the mixture at speed 1 for 90 seconds, then scrape down the bowl. Continue mixing (speed 2) until a uniform mass is obtained.

[0527] The mixture was poured and spread evenly into a bar frame (~16 mm deep) and rolled out to make it flush with the frame, cutting off any excess. The mixture was left overnight to cure. The mixture was unwound from the frame and cut into 30 mm x 100 mm bars. The bars were packaged in foil bags for storage until use. The composition and breaking force (g) properties of the protein bars are described in Table 17.

[0528] The breaking force (g) of the bars was evaluated using a TAHD Plus Texture Analyser from Stable Micro Systems in Godalming, UK. Texture measurements were performed by penetration. Force was measured at a set penetration depth of 12 mm. A 5 mm stainless steel cylindrical probe was pushed into the bar at a constant rate of 1 mm / s to a depth of 12 mm and then withdrawn at a rate of 10 mm / s. The force (g) versus time (s) for the probe to move was measured. Three compressions were performed on the surface of each bar sample. Two bars were evaluated for each sample. The samples were removed from 20°C storage and texture measurements were performed in a controlled room at 20°C.

[0529] Table 17. Formulation, composition and properties of protein bars using powder C

[0530]

[0531] Powder C produced acceptable protein bars in terms of texture. The sensory properties of the bars were acceptable in terms of texture attributes, including hardness, firmness, stickiness, and graininess. We detected no off-flavor attributes in the protein bars. The hardness of the protein bars after 1 month of storage was 2310 g, and a hardness of less than about 4000 g after 12 months of storage was suitable. The water activity of the bars after 1 month of storage was 0.51, which is below the microbial stability limit of 0.65. The results indicate that the denatured whey protein concentrate is suitable for protein bars.

[0532] Example 8: Effect of addition rate on improving thermal stability of denatured whey protein particle composition

[0533] Uncolored cheese whey solutions were prepared by reconstructing cheese whey protein (WPC 80) powder until 20-22% (w / w) protein concentration. The composition of WPC 80 powder is summarized in Table 18. The solution was homogenized at 200 / 50 bar and then divided into four (4) batches. The pH of three batches was adjusted to 6.2 using a 1M concentration of KOH:NaOH (50:50) blend, and one sample was adjusted to pH 5.3 with 6M HCl. pH 5.3 is close to the optimal pH for the maximum activity of MaxiBright, which allows peroxide to be added at a higher rate without inhibiting enzyme activity. Enough equal amounts of commercially available MaxiBright solutions were added to all four samples, and the samples were mixed at 4°C in a cooler using an overhead stirrer at 150rpm. The samples named as follows 1) control - no peroxide added, 2) pH 6.2-75 ppm peroxide added in one step, 3) pH 6.2-75 ppm peroxide added incrementally at a rate of 10 ppm / hr, and 4) pH 5.3-75 ppm peroxide added incrementally at a rate of 30 ppm / hr. 75 ppm peroxide is equivalent to 0.375 moles of H2O2 / mole of β-lactoglobulin protein. The enzyme dairy bleaching activity according to the COA from DSM is 5000 U / g. After completing the peroxide dosing and ensuring that there is no peroxide remaining in the solution, the pH of sample 4 is adjusted to 6.2 using the aforementioned KOH:NaOH blend before heating. Subsequently, in order to produce a heat-stable denatured whey protein particle composition, a combination of a scraped surface heat exchanger and a steam-heated pilot-scale tubular heat exchanger is used. The sample is preheated to 55°C in the scraped surface heat exchanger and then heated to 85°C in the steam-heated tubular heat exchanger. Adjust flow rate to achieve 1800s in tubular heat exchanger -1 The sample temperature was maintained at 85°C for an additional 20 seconds using a holding tube with tube specifications similar to those of a steam-heated tubular heat exchanger to provide the residence time required to achieve the desired HCT. The samples were then cooled, collected, and tested for HCT as described in Section 3 - Thermal Condensation Time. All samples were tested for protein profile before and after heat treatment as described in Section 5 - High Denaturation.

[0534] Figure 8 : Thermal coagulation time of denatured whey protein compositions prepared from samples treated with different peroxide dosage rates.

[0535] Fig. 9 : The protein profile of the samples after enzyme treatment and before heat treatment was analyzed by HPLC, including glycomacropeptide (gmp), Peaks for peptone 5 (pp5), α-lactalbumin (alac), lactoferrin (Lf), BSA, β-lactoglobulin (blac), and immunoglobulin (igg).

[0536] Table 18. Composition of WPC 80 powder

[0537] Moisture content (g / 100g powder) 4.5 Ash content (g / 100g powder) 2.63 Carbohydrate (g / 100g powder) 6.8 Fat (g / 100g powder) 5.6 Protein content (g / 100g powder 80.5 β-Lg (% protein) 52.9 α-Lac (% protein) 13.2 GMP (% protein) 23.2 pp5 (% protein) 4.9 Lactoferrin (% protein) 0.2 BSA (% protein) 1.5 IGG (% protein) 4 Ca(mg / 100g powder) 386 K(mg / 100g powder) 567 Na(mg / 100g powder) 115

[0538] Example 9: Use of the improved denatured whey protein particle composition of the present invention in a low calorie high protein liquid nutritional composition at neutral pH

[0539] This example describes the preparation of a neutral pH low calorie high protein beverage comprising the Powder A denatured whey protein composition of the present invention. The formulation, composition and properties of the neutral pH low calorie high protein beverage are summarized in Table 19. Figure 2 Beverages were prepared at a protein level of 12% (w / v) with some modifications under UHT conditions. Indirect UHT conditions of 143°C for 6 seconds were used.

[0540] Table 19. Formulation of neutral pH low calorie high protein liquid beverage including Powder A.

[0541] Ingredients (g / 100g) Recipe 6 water 82.81 Powder A 15.40 Canola Oil 0.59 Sodium chloride 0.01 Tripotassium Citrate 0.22 Trisodium Citrate 0.10 Potassium chloride 0.08 Dimagnesium phosphate 0.18 Choline chloride 0.03 Mineral and vitamin premixes 0.06 Tricalcium phosphate 0.09 Gellan Gum 0.04 Cellulose Gum 0.20 Lecithin 0.01 Defoaming agent 0.01 5% Potassium Hydroxide 0.18 pH 6.8 UHT process conditions (temperature, time) 143℃, last 6s

[0542] Table 20. Nutritional composition of a neutral pH low calorie high protein liquid beverage including powder A

[0543] Components Recipe 6 Protein (g / 100ml) 12.4 Protein from denatured WPC (g / 100ml) 12.4 Fat (g / 100ml) 1.6 Ash content (g / 100ml) 1.2 Sodium (mg / 100ml) 77 Potassium (mg / 100ml) 218 Calcium (mg / 100ml) 108 Phosphorus (mg / 100ml) 102 Iron (mg / 100ml) 1.1 Magnesium (mg / 100ml) 38.3 Zinc (mg / 100ml) 1.0 Manganese (mg / 100ml) 0.24 Selenium (μg / 100ml) 9.4 Copper (mg / 100ml) 0.13 Chloride (mg / 100ml) 63 Iodine (μg / 100ml) 19 Vitamin A (%RE) (μg / 100ml) 167 Vitamin C (mg / 100ml) 19 Vitamin D (μg / 100ml) 1.7 Vitamin E (mg / 100ml) 1.2 Vitamin K (μg / 100ml) 5.7 Thiamine (mg / 100ml) 0.35 Riboflavin (mg / 100ml) 0.26 Niacin (mg / 100ml) 2.1 Vitamin B6 (mg / 100ml) 0.30 Biotin(μg / 100ml) 5.3 Folic acid (μg / 100ml) 36 Vitamin B12 (μg / 100ml) 0.19 Vitamin B5 (mg / 100ml) 0.91 Choline (mg / 100ml) 35 Carbohydrate (g / 100ml) 1.7 Energy (Cal / 100ml) 72

[0544] Table 21. Beverage properties of neutral pH low calorie high protein liquid beverages before UHT heat treatment.

[0545] Recipe 6 <![CDATA[Volume weighted particle size d 4,3 (μm)]]> 0.85 <![CDATA[D 50 (μm)]]> 0.57 % Particle Volume <1 μm 93 % Particle Volume 1-10μm 6.3 % Particle Volume > 2 μm 3.1 <![CDATA[Viscosity (mPa·s at 100 s -1 ))]]> 42

[0546] Table 22. Beverage properties of neutral pH low calorie high protein liquid beverages including Powder A after indirect UHT heat treatment.

[0547] Recipe 6 <![CDATA[Volume-weighted particle size d 4,3 (μm)]]> 1.3 <![CDATA[D 50 (μm)]]> 0.65 <![CDATA[D 90 (μm)]]> 3.15 % Particle Volume <1 μm 76 % Particle Volume 1-10μm 24 % Particle Volume > 2 μm 15 <![CDATA[Viscosity at time 0 (mPa.s at 100 s -1 ))]]> 31 <![CDATA[Viscosity at 1 month (mPa.s at 100 s -1 ))]]> 37 Viscosity at 3 months (mPa.s at 100s-1) 50 Viscosity at 6 months (mPa.s at 100s-1) 54

[0548] Fig.10 : Particle size distribution of a 12% (w / v) low-calorie, high-protein beverage before and after indirect UHT heating (143°C for 6 s).

[0549] From the above results, it can be seen that powder A showed the smallest particle size growth in formulation 6 beverage after indirect UHT heat treatment. -1 The viscosity was less than 100 mPa.s during shelf life storage. The formulation 6 beverage showed a uniform consistency with no separation and visible protein particles or aggregates in the packaging.

[0550] Example 10: Use of improved denatured whey protein concentrate in high protein biscuits

[0551] This example describes the preparation of high protein biscuits including the Powder C denatured whey protein composition of the present invention. High protein (-20%) biscuits were prepared based on the recipe and method of Cooper et al., J. Food Sci., 49(2), 376-379 (1984) to compare the performance of the denatured whey protein composition of the present invention with another insoluble whey protein ingredient, lactalbumin, based on equivalent protein. The recipe of the high protein biscuit is summarized in Table 23.

[0552] Table 23. High protein biscuit recipe

[0553] Ingredients (weight, g) Powder C Whey protein butter 122 122 Sugar (seasoning bottle) 100 100 Powder C 90 - <![CDATA[NZMP SureProtein TM Whey Protein 825]]> - 80 Flour, regular (10% protein), 200 200 Baking powder (double acting) 6.4 6.4 water 71 85

[0554] Butter at 20°C was mixed in a Hobart mixer (Model N50, Hobart Corporation, US) with a flat beater at speed 2 for 30 s. Sugar was added over 30 s; the bowl was scraped down and mixed for an additional 60 s. The egg white ingredients were added at speed 1 over the next 60 s; scraped down and mixed for an additional 60 s at speed 2. Sifted flour and baking powder were added at speed 1 over 60 s. Water (55 g) was added at speed 1 over 60 s. If the dough was too brittle and dry and did not form a cohesive mass, additional water was added while mixing. The dough was rolled into a 5 mm thickness and cut into 50 mm diameter discs. The discs were placed on a metal tray lined with baking paper and baked at 180°C for 12.5 minutes until the bottom surface was brown. The baked cookies were cooled on a wire rack for 30 minutes before evaluation. The experiment was repeated.

[0555] The biscuits were evaluated subjectively for color; surface appearance (smooth to rough); initial texture (appearance when broken in half: closed to open texture); crispness (easy to break when bitten); residual mouthfeel (fine to powdery; powdery, amount of residue). The spread ratio (width / thickness) was determined using 6 baked biscuits according to the AACC method (American Association of Cereal Chemists (1984) Approved Methods of the American Association of Cereal Chemists, Vol. 1 (Eighth Edition), Method 10-50D, Revised 1986. Baking quality of cookie flour. St Paul, Minnesota, US). The evaluation of the baked biscuits can be seen in Table 24.

[0556] Table 24. Characteristics of high protein biscuits

[0557] Features Powder C Comparator: Whey protein high protein biscuits color golden brown Light beige Surface appearance Smooth, some slight cracks All cookies are smooth and slightly cracked Internal texture Open, similar to shortbread Open, similar to shortbread Crisp A crispy bite A little crispy Residual taste Good solubility Powdery, slightly sandy taste Width (W), mm 49.0,49.2,49.5,49.5 49.1,49.3；49.0,48.7 Thickness (T), mm 10.8,10.5；10.8,10.5 9.5,9.5；10.1,9.8 Spread ratio = W / T 4.6±0.1 5.0±0.1

[0558] The biscuits containing Powder C (of Example 1) as the protein source were browner, crispier and had a better mouthfeel than the comparative ingredient.

[0559] Example 11: Use of improved denatured whey protein concentrate in high-protein ambient yogurt

[0560] This example describes the preparation of an exemplary high protein ambient yogurt comprising a denatured whey protein composition of the present invention compared to Powder F. The recipe and composition of the ambient yogurt are summarized in Table X. Figure 3 , yogurt was prepared by modification. Amidated low methoxyl (LMA) pectin and gellan gum were added during reconstitution of dry ingredients, and cream was added during the last 10 minutes of reconstitution. After stirring to break the gel, ambient yogurt was heated at 75°C for 30 seconds and packaged, and then stored at ambient temperature.

[0561] Table 25. Recipe and composition of ambient yogurt using powder C and powder F mixed with skimmed milk powder.

[0562]

[0563] The pH, flavor, viscosity, particle size distribution, visible sediment at the end of shelf life (%), phase separation at the end of shelf life (%), and sediment after centrifugation of the resulting ambient yogurts were evaluated using the methods described herein.

[0564] It is expected that the pH of the yogurt product will be acceptable.The viscosity and sedimentation of ambient yogurt prepared with powder C are expected to remain low during shelf life and have acceptable organoleptic properties.

[0565] It should be understood that the foregoing detailed description and the accompanying examples are merely illustrative and should not be considered as limiting the scope of the invention, which is limited only by the appended claims and their equivalents. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications may be made without departing from the spirit and scope of the invention, including but not limited to those involving chemical structures, substituents, derivatives, intermediates, syntheses, formulations or methods, or any combination of such changes and modifications of the uses of the invention.

[0566] All references cited above (patents and non-patents) are incorporated into this patent application by reference. The discussion of these references is intended only to summarize the assertions made by their authors. It is not admitted that any reference (or part of any reference) is relevant prior art (or prior art). Applicants reserve the right to question the accuracy and relevance of the references cited.

Claims

1. A heat stable denatured whey protein composition comprising at least 60% total protein on a dry weight basis and microparticles comprising denatured whey protein, wherein the microparticles have a particle size distribution comprising at least two characteristics selected from the group consisting of: (i) a volume weighted average diameter D(4,3) of not greater than about 1.0 μm, (ii) a d(4,3) of not greater than about 1.0 μm 50 , (iii) a d of not more than about 2.0 μm 90 , (iv) at least 92 volume percent of the particles have a diameter less than about 2.0 μm, and (v) no more than 45 volume percent of the particles have a diameter of about 1.0 to about 10.0 μm.

2. The heat stable denatured whey protein composition according to claim 1, wherein the particle size distribution comprises: (i) a volume weighted average diameter D(4,3) of not greater than about 1.0 μm, such as from about 0.6 to about 1.0 μm, and (iii) a volume weighted average diameter D(4,3) of not greater than about 2.0 μm, such as from about 0.9 to about 1.7 μm. 90 ;or (ii) a d of no greater than about 1.0 μm, such as about 0.6 to about 1.0 μm 50 , and (iii) a d of no greater than about 2.0 μm, such as about 0.9 to about 1.7 μm 90 .

3. A heat stable denatured whey protein composition according to claim 1 or claim 2, wherein the particle size distribution comprises (iv) at least 92%, at least 95% or at least 98% by volume of the particles having a diameter less than about 2.0 μm and / or (v) no more than 45% by volume of the microparticles having a diameter of about 1.0 to about 10.0 μm.

4. A heat stable denatured whey protein composition comprising at least 60% total protein on a dry weight basis and microparticles comprising denatured whey protein, wherein after heating a 10% (w / w) protein content aqueous solution comprising the denatured whey protein composition at 120°C for 15 minutes, 50 From about 0.6 to about 1.0 μm; wherein after heating a 10% (w / w) protein content aqueous solution comprising the denatured whey protein composition at 120°C for 15 minutes, 90 Not greater than about 2.0 μm, such as about 0.9 to about 1.7 μm; wherein after heating a 10% (w / w) protein content aqueous solution comprising the denatured whey protein composition at 120°C for 15 minutes, the volume weighted average diameter D(4,3) is no greater than about 1.0 μm, such as from about 0.6 to about 1.0 μm; and / or A 10% (w / w) protein content aqueous solution of the denatured whey protein composition has 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.

5. The heat stable denatured whey protein composition of claim 4, wherein the microparticles have a particle size distribution comprising at least two characteristics selected from the group consisting of: (i) a volume weighted mean diameter D(4,3) of no greater than about 1.0 μm, (ii) a d(4,3) of no greater than about 1.0 μm. 50 , (iii) a d of not more than about 2.0 μm 90 , (iv) at least 92 volume % of the particles have a diameter less than about 2.0 μm, and (v) no more than 45 volume % of the particles have a diameter of about 1.0 to about 10.0 μm.

6. A 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 casein, preferably wherein the composition is substantially free of casein.

7. A heat stable denatured whey protein composition according to any one of claims 1 to 6, wherein the composition comprises from about 30% to about 80% β-lactoglobulin, from about 35% to about 75% β-lactoglobulin, or from about 40% to about 65% β-lactoglobulin relative to the total protein in the composition.

8. A heat stable denatured whey protein composition according to any one of claims 1 to 7, wherein the composition comprises at least 70% (w / w) whey protein relative to the total protein, at least 80% (w / w) whey protein relative to the total protein, or at least 90% (w / w) whey protein relative to the total protein.

9. The heat stable denatured whey protein composition according to any one of claims 1 to 8, wherein the residual denatured whey protein in the denatured whey protein composition is less than 16%.

10. A heat stable denatured whey protein composition according to any one of claims 1 to 9, wherein a 3.4% (w / w) protein content aqueous solution containing the heat stable denatured whey protein composition exhibits a turbidity value of at least 50 absorbance units measured at 500 nm at about 20°C, preferably a turbidity value of at least 100 absorbance units, and / or a 4% (w / w) protein content aqueous solution containing the denatured whey protein composition exhibits a turbidity value of at least 90 absorbance units measured at 500 nm at about 20°C, preferably a turbidity value of at least 100 absorbance units.

11. The heat stable denatured whey protein composition according to any one of claims 1 to 10, 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 11.

13. The composition of claim 12, wherein the composition is (i) a liquid composition, such as a drinkable yogurt, a liquid nutritional composition (such as a medical food), or a high-protein beverage, such as a neutral or acidic high-protein beverage, or (ii) a food, such as a baked good, a bar, or a set or stirred yogurt.

14. The composition according to claim 13, wherein the composition is a food, wherein The food product is a set yogurt, the set yogurt optionally comprising from about 6% to about 20% (w / v) total protein, optionally at least 50% (w / w) of the total protein coming from the denatured whey protein composition, wherein the set yogurt exhibits reduced hardness and / or reduced volume weighted average particle size compared to a control set yogurt product having the same ingredient composition and the same protein content, except that the control set yogurt product does not include the denatured whey protein composition according to any one of claims 1 to 11; or The food product is a stirred yogurt, optionally comprising from about 6% to about 20% (w / v) total protein, optionally at least 50% (w / w) of the total protein coming from the denatured whey protein composition, wherein the stirred yogurt exhibits reduced viscosity and / or reduced volume weighted average particle size compared to a control stirred yogurt product having the same ingredient composition and the same protein content, except that the control stirred yogurt product does not include the denatured whey protein composition according to any one of claims 1-11.

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

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

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

85.

18. A heat stable denatured whey protein composition prepared by the method according to any one of claims 15 to 17.

19. A heat-treated, shelf-stable high protein liquid composition comprising at least 6% (w / v) whey protein, wherein the whey protein comprises microparticles containing denatured whey protein; and wherein (i) the liquid composition has a d value of no greater than about 1.0 μm after secondary heat treatment for microbial control; 50 and / or a d of not greater than about 2.0 μm 90 (ii) the liquid composition has a temperature of 100s at 20°C -1 (iii) the liquid composition exhibits less than 10% sedimentation after storage of the liquid composition at a temperature of about 20°C to about 25°C for the shelf life; and / or (iv) the liquid composition exhibits less than 10% sedimentation after centrifugation at 1540 x g for 5 minutes.

20. The heat-treated shelf-stable liquid composition of 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) denatured whey protein.

21. The heat-treated shelf-stable liquid composition of claim 19 or claim 20, wherein the liquid composition is a liquid nutritional composition for providing nutrition to a subject in need thereof, 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. A heat treated shelf-stable liquid composition according to any one of claims 19 to 21 further comprising other dairy and / or non-dairy proteins.

23. A liquid nutritional composition comprising microparticles comprising denatured whey protein, wherein at least 95% of the total protein in the liquid nutritional composition is denatured whey protein; Wherein the liquid nutritional composition optionally further comprises a lipid component and / or a carbohydrate component.

24. The liquid nutritional composition of 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 claim 24, wherein the liquid nutritional composition comprises at least 12% (w / v) total protein.

26. The liquid nutritional composition according to claim 23 or claim 24, wherein the liquid nutritional composition comprises no more than 12% (w / v) total protein.

27. The liquid nutritional composition according to any one of claims 23 to 26, wherein (i) the liquid composition has a d value of no greater than about 1.0 μm after secondary heat treatment for microbial control. 50 (ii) the liquid composition has a temperature of 100s at 20°C -1 (iii) the liquid composition exhibits less than 10% sedimentation after storage at a temperature of about 20°C to about 25°C for at least 3 months; and / or (iv) the liquid composition exhibits less than 10% sedimentation after centrifugation at 1540 x g for 5 minutes.

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