Denatured milk proteins and methods for their production

Abstract

The present disclosure includes denatured whey protein compositions. The compositions include at least 60 wt.% protein on a dry weight basis; less than 8 wt.% native glycomacropeptide (GMP) relative to the total weight of protein, greater than 2 wt.% enzyme hydrolyzed GMP; at least 8 wt.% proteolysis index; and greater than 50 wt.% denatured whey protein relative to the total weight of protein.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Patent Application No. 18 / 503,535, filed November 7, 2023, entitled “Denatured Milk Proteins, Methods of Manufacturing, and Protein Fortified Foods,” which is a continuation of U.S. Patent Application No. 18 / 097,003, filed January 13, 2023, entitled “Denatured Milk Proteins, Methods of Manufacturing, and Protein Fortified Foods,” each of which is incorporated herein by reference in its entirety. Technical Field

[0003] This technology relates to denatured whey protein compositions, including denatured whey proteins with low levels of natural glycomacropeptides. Background Technology

[0004] Milk-derived products (such as milk proteins, aldobionic products, and galactooligosaccharides) have become a major source of ingredients in a wide variety of foods and beverages. For example, milk-derived proteins have become a primary source of protein fortification in nutrition bars, sports drinks, and yogurt products. One source of milk protein is whey protein, a byproduct of cheese making. During cheese making, casein in milk forms cheese curds, while liquid whey is drained from the curds and transferred for further processing. In most cheese-making processes, liquid whey is a mixture of whey protein and a large amount of lactose and minerals, and this mixture undergoes additional purification to separate the whey protein from the lactose and minerals.

[0005] Whey protein obtained from cheese making also includes additional byproducts, such as cheese-making enzymes and the hydrolyzed proteins they produce. Natural glycomacropeptides (GMPs) are considered a lower-grade source of protein for muscle recovery because they contain fewer branched-chain amino acids (especially leucine), which stimulate muscle protein synthesis and are major components of muscle tissue after a period of intense exercise and resistance training. Furthermore, excessively high levels of native GMPs in whey protein compositions can lead to undesirable flavor and poor process incorporation, thus reducing the total amount of protein that can be fortified into the product. This technology can solve these problems and other challenges. Summary of the Invention

[0006] Typically, embodiments of this disclosure include denatured whey protein compositions. The composition comprises: at least 60% by weight of protein on a dry weight basis; less than 8% by weight of natural glycomacropeptide (GMP) and more than 2% by weight of enzymatically hydrolyzed GMP relative to the total weight of the protein; at least 8% by weight of the proteolysis index; and more than 50% by weight of denatured whey protein relative to the total weight of the protein.

[0007] In some embodiments, the denatured whey protein may comprise denatured enzyme-hydrolyzed cheese whey protein. In additional or alternative embodiments, the natural GMP is less than or about 7% by weight relative to the total weight of the protein. In a further embodiment, the denatured whey protein is characterized by a D50 particle size distribution value of less than or about 4.5 μm. In still more embodiments, the denatured whey protein is further characterized by a D10 particle size distribution value of less than or about 2.5 μm. In an additional embodiment, the denatured whey protein is further characterized by a D90 particle size distribution value of less than or about 8 μm. In a further embodiment, the denatured whey protein composition may comprise up to 7.0% by weight of fat on a dry weight basis. In an embodiment, the denatured whey protein composition may comprise at least 2% fat.

[0008] Embodiments of this technology also include a method for preparing a denatured whey protein composition. The method includes: filtering cheese whey from enzymatically coagulated milk to produce cheese whey residue and permeate; conjugating the cheese whey residue with one or more enzymes that selectively hydrolyze GMP in the cheese whey residue to form a GMP-reduced cheese whey residue composition; and heating the GMP-reduced cheese whey residue composition to form a denatured whey protein composition. The denatured whey protein composition may be characterized by comprising: at least 60% by weight of protein on a dry weight basis; less than 8% by weight of GMP and greater than 2% by weight of enzymatically hydrolyzed GMP relative to the total weight of protein; at least 8% by weight of proteolysis index; and greater than 50% by weight of denatured whey protein relative to the total weight of protein.

[0009] Embodiments of this technology also include a method for preparing a denatured whey protein composition. The method includes: filtering cheese whey from enzymatically coagulated milk to produce cheese whey residue and permeate; reducing the natural glycomacropeptides in the cheese whey residue to form a GMP-reduced cheese whey residue composition; and heating the GMP-reduced cheese whey residue composition to form a denatured whey protein composition. The denatured whey protein composition may be characterized by comprising: at least 60% by weight of protein on a dry weight basis; less than 11% by weight of GMP and greater than 2% by weight of enzymatically hydrolyzed GMP relative to the total weight of protein; at least 8% by weight of proteolysis index; and greater than 50% by weight of denatured whey protein relative to the total weight of protein.

[0010] In an additional embodiment, one or more enzymes selectively hydrolyzing GMP in cheese whey residues comprise at least one alkaline serine protease and at least one neutral protease. In an embodiment, GMP reduction comprises binding the cheese whey residues to one or more enzymes selectively hydrolyzing GMP in the cheese whey residues. In further embodiments, one or more enzymes selectively hydrolyzing GMP in cheese whey residues comprise at least one alkaline serine protease and at least one neutral protease. In a further embodiment, heating the GMP-reduced cheese whey residue composition also inactivates one or more enzymes selectively hydrolyzing GMP in the cheese whey residues. In an even further embodiment, the GMP-reduced cheese whey residue composition is heated to a temperature greater than or about 160°F. In further embodiments, heating the GMP-reduced cheese whey residue composition further comprises exposing the GMP-reduced cheese whey residue composition to high shear conditions. In a further embodiment, the denatured whey protein composition is characterized in that the weight ratio of natural GMP to total whey protein is less than or about 0.15. Additionally or alternatively, the denatured whey protein composition is characterized by a GMP of less than or about 11% by weight relative to the total protein weight. In further embodiments, the denatured whey protein composition is characterized by a GMP of less than or about 7% by weight relative to the total protein weight. In additional embodiments, the denatured whey protein composition is characterized by a fat content of up to 7% by weight on a dry weight basis. In a further embodiment, the denatured whey protein in the denatured whey protein composition is characterized by a D50 particle size distribution value of less than or about 5 μm.

[0011] Embodiments of this technology also include a denatured whey protein composition. This composition comprises: at least 60% by weight of protein on a dry weight basis; less than 11% by weight of natural GMP relative to the total weight of the protein; greater than 7% by weight of fat on a dry weight basis; a β-lactoglobulin to α-lactalbumin ratio greater than 5.00; and greater than 50% by weight of denatured whey protein relative to the total weight of the protein.

[0012] In some embodiments, the pre-denatured whey protein already comprises at least 30% by weight of denatured protein. In additional embodiments, the whey protein comprises at least 60% by weight of β-lactoglobulin relative to the total weight of the protein. Additionally or alternatively, in embodiments, the whey protein comprises up to 12% by weight of α-lactalbumin relative to the total weight of the protein. In further embodiments, the whey protein comprises a β-lactoglobulin to α-lactalbumin weight ratio greater than or about 7. In more embodiments, the native GMP is less than or about 10.5% by weight relative to the total weight of the protein. In further embodiments, the denatured whey protein is characterized by a D50 particle size distribution value of less than or about 0.3 μm. In still more embodiments, the denatured whey protein is further characterized by a D10 particle size distribution value of less than or about 0.1 μm. In additional embodiments, the denatured whey protein is further characterized by a D90 particle size distribution value of less than or about 1.0 μm.

[0013] Embodiments of this technology include protein-fortified food products. Embodiments include food products containing at least 3% by weight of total protein. In one embodiment, the protein comprises a denatured whey protein composition containing less than 8% by weight of natural glycomacropeptides and more than 2% by weight of enzymatically hydrolyzed glycomacropeptides relative to the total weight of the whey protein composition.

[0014] Embodiments of this technology also include protein-fortified food products. The food products contain at least 3% by weight of total protein. This protein includes a denatured whey protein composition comprising: less than 11% by weight of natural glycomacropeptides, greater than 7% by weight of fat, and a ratio of β-lactoglobulin to α-lactalbumin greater than 5.00 relative to the total weight of the whey protein composition. Attached Figure Description

[0015] A further understanding of the nature and advantages of selected embodiments of the present technology can be achieved by referring to the remainder of the specification and accompanying drawings, in which similar reference numerals may be used in several drawings to denote similar components. In some cases, a sublabel is associated with a reference numeral and follows a hyphen to indicate one of a plurality of similar components. When a reference numeral without an existing sublabel is mentioned, it is intended to refer to all such plurality of similar components.

[0016] Figure 1 Selected operations in a formation method according to some embodiments of the present technology are shown;

[0017] Figure 2A It is a graph depicting the particle size distribution of an undenatured whey protein composition (WPC) with 80% protein content on a dry basis.

[0018] Figure 2B It is a graph depicting the particle size distribution of the denatured whey protein composition according to embodiments of the present disclosure;

[0019] Figure 2C It is a comparison Figure 2A and 2B A graph showing the particle size distribution;

[0020] Figure 3A This is a graph showing the capillary electrophoresis curve of an undenatured whey protein composition (WPC) with 80% protein content on a dry basis.

[0021] Figure 3B It is a graph showing the capillary electrophoretic characteristics of the denatured whey protein composition according to embodiments of the present disclosure;

[0022] Figure 4A It is a graph depicting the particle size distribution of the undenatured high-fat whey protein composition;

[0023] Figure 4B It is a graph depicting the particle size distribution of the denatured high-fat whey protein composition according to embodiments of the present disclosure.

[0024] Figure 4C It is a comparison Figure 4A and 4B The particle size distribution curve; and

[0025] Figure 5 This is an illustration of the sample according to Example 3.

[0026] Several figures are included as illustrations. It should be understood that these figures are for illustrative purposes and are not considered to be to scale unless specifically stated otherwise. Furthermore, as illustrations, the figures are provided to aid understanding and may not include all aspects or information compared to a true representation, and may include exaggerated material for illustrative purposes.

[0027] In the drawings, similar parts and / or features may have the same numerical reference labels. Furthermore, various components of the same type can be distinguished by using letters to differentiate similar components and / or features after the reference label. If only the first numerical reference label is used in the specification, the description applies to any similar parts and / or features having the same first numerical reference label, regardless of the letter suffix. Detailed Implementation

[0028] Whey proteins (as described herein) are actually a collection of different proteins, primarily including β-lactoglobulin (β-Lg) and α-lactalbumin (α-La), as well as glycomacropeptide (GMP, sometimes called cGMP or CMP), which cleaves native κ-casein in milk via papain activity, causing the milk to coagulate into cheese curds. Depending on the purification process and degree of purification, whey protein concentrate (WPC) can be formed by concentrating whey proteins to a protein content of 25-90% by weight of the total solids, or whey protein isolate (WPI) can be formed by concentrating whey proteins to a protein content of 90-99% by weight of the total solids.

[0029] Whey protein obtained from cheese making also includes additional byproducts, such as cheese-making enzymes and the hydrolyzed proteins they produce. For example, from enzymatically coagulated milk... * The conventional composition of obtained whey typically contains:

[0030]

[0031] * 1Walstra P, Wouters JTM, Geurts TJ. Milk Components, Dairy Science and Technology. 2nd edition, CRC Press; 2006: Chapter 2.

[0032] 2. Fogeding EA, Luck P, Vardhanabhuti B. Encyclopedia of Dairy Sciences. Second Edition, Elsevier Ltd.; 2011: Whey Protein Products.

[0033] Hydrolyzed proteins include glycomacropeptides (GMPs) from the hydrolysis of κ-casein, thus the resulting para-κ-casein can form a major component of cheese curd. Smaller, more soluble GMPs are carried away along with whey proteins and can constitute 13-20% by weight of the protein present in the whey protein fraction on a dry weight basis. Unfortunately, GMPs are considered a inferior source of protein for muscle recovery because they contain fewer branched-chain amino acids, which stimulate muscle protein synthesis and are a major component of muscle tissue after a period of intense exercise and resistance training.

[0034] Furthermore, a high natural GMP content in whey protein compositions can lead to undesirable flavors and poor processing combinations. Specifically, on a dry weight basis, a high natural GMP content (e.g., 12% by weight or more) in the presence of proteins in whey protein fractions increases artificial flavors, off-flavors, and / or cardboard flavors commonly found in liquid whey, dry whey, dairy protein concentrates, and milk, while reducing natural milk flavor. Additionally, natural whey proteins can interact with water in food or beverage products, negatively increasing the viscosity of the product. This interaction is problematic because it limits the amount of whey protein composition that can be incorporated, reducing the total amount of fortified protein in the product.

[0035] This technology overcomes these problems by providing denatured whey protein compositions, such as powdered or denatured whey protein compositions, that exhibit a high total protein weight percentage, a reduction attributable to natural GMP. Specifically, this technology surprisingly discovers that by carefully processing whey protein compositions to selectively reduce GMP levels without hydrolyzing the remaining proteins in the whey protein composition (e.g., β-lactoglobulin (β-Lg) and α-lactalbumin (α-La)), denatured whey protein compositions with a low natural GMP percentage and high protein content can be provided. It has been observed that such denatured whey protein compositions do not exhibit the negative effects associated with compositions containing a high natural GMP weight percentage.

[0036] For example, compositions according to one or more embodiments of this disclosure can exhibit lower processing viscosity and reduced cardboard flavor and more milk flavor. Specifically, this disclosure surprisingly finds that when natural GMP is enzymatically reduced in a whey protein composition, the processing viscosity decreases, such that the viscosity is about 10% lower than that of a composition in which natural GMP is not enzymatically reduced, for example, about 20%, for example, about 30%, for example, about 40%, for example, about 50% lower than that of a composition without enzymatic reduction. For example, in the compositions discussed herein without enzymatic reduction, the processing viscosity of the composition can be greater than 200 centipoise, for example, from 201 centipoise to 500 centipoise. Conversely, compositions according to the present technology exhibit less than 200 centipoise, for example, about 175 centipoise or less, for example, about 150 centipoise or less, for example, about 125 centipoise or less, for example, about 100 centipoise or less, for example, about 75 centipoise or less, for example, about 50 centipoise or less, for example, as low as about 25 centipoise, or any range or value between them. In some embodiments, the compositions according to the present technology may also have the ability to reduce bound water. Therefore, the compositions according to the present technology may be uniquely suited to fortify aqueous foods to a higher level than previously thought achievable with whey protein compositions, because of the observed lower increase in viscosity (typically associated with these formulations), which is at least in part due to reduced interaction with water.

[0037] Figure 1 Exemplary operations in method 100 according to some embodiments of the present technology are illustrated; the method can be performed in various processing devices known in the art. Method 100 may include a number of optional operations, which may or may not be specifically associated with some embodiments of the method according to the present technology. For example, many operations are described to provide a wider range of composition formations, but are not critical to the technology or may be performed by easily understood alternative methods.

[0038] The whey composition raw materials used herein can be obtained from cheese making processes, whey protein concentrates, whey protein isolates (e.g., from milk), or combinations thereof. In some embodiments, the whey raw material can be produced during the cheese-making process, and when the cheese-making process uses rennet (e.g., chymosin), it can be referred to as "sweet whey," and when acid is used to form curd, it can be referred to as "acidic whey." The pH of sweet whey is typically in the range of about 5.6 to 6.6, while the pH of acidic whey is typically in the range of 4.3 to 4.6. While any suitable whey raw material can be used, in some embodiments, the whey raw material used to form the denatured whey protein composition is "sweet whey," and thus can be considered as filtered from lactose and minerals.

[0039] For example, in some embodiments, the whey protein raw material may be a whey protein residue filtered from sweet whey (102), which has a protein composition of 25% or higher on a dry weight basis, such as about 50% or higher, such as about 60% or higher, such as about 70% or higher, such as about 80% or higher, or any range or value between them. In embodiments, the whey protein residue may be concentrated using ultrafiltration or other methods known in the art. Nevertheless, in some embodiments, the whey protein residue may be diluted prior to processing, for example by mixing with water to form a whey protein raw material with a protein concentration in solution of about 30% or lower, such as about 18% or lower, such as about 16% or lower, such as about 15% or lower, such as about 14% or lower, or any range or value between them.

[0040] In some embodiments, the natural GMP level of 103 whey protein raw material can be reduced using one or more enzymes that selectively reduce natural GMP levels without hydrolyzing β-lactoglobulin and / or α-lactalbumin. GMP can also be reduced by using expensive chromatographic systems or by mixing natural whey protein isolates purified directly from unprocessed milk with whey protein concentrates from cheese production. In embodiments, the one or more enzymes can be one or more proteases. Sources of the one or more proteases can include microbial, fungal, plant, and / or animal sources. For example, one or more proteases can be derived from fungi of the genus *Aspergillus*, bacteria of the genus *Bacillus* (e.g., *Bacillus subtilis*) and / or animals (e.g., trypsin, chymotrypsin, etc.), and other sources. However, in some embodiments, the one or more proteases include acidic proteases, neutral proteases, alkaline proteases, or combinations thereof. Additionally or alternatively, the protease can be an endopeptide, an exopeptide, or a combination thereof. Therefore, in some embodiments, one or more proteases may include aspartic proteases, serine proteases, cysteine ​​proteases, or combinations thereof. Nevertheless, in one embodiment, one or more proteases may include serine proteases, such as basic serine proteases, alone or in combination with at least one neutral protease.

[0041] Although one or more enzymes are selected, the selected one or more enzymes may be added to the whey protein raw material in an amount of about 0.001% by weight or greater based on the weight of the total protein in the composition, for example, about 0.0025% by weight or greater, for example, about 0.005% by weight or greater, for example, about 0.0075% by weight or greater, for example, about 0.01% by weight or greater, or any range or value between them. It should be understood that the foregoing range may refer to the total amount of enzymes contained in the whey protein raw material, or to the amount of each enzyme added to the whey protein raw material.

[0042] However, in one aspect, the amount of one or more enzymes added is chosen to hydrolyze at least about 10% by weight or more of the natural GMP present in the whey protein raw material, for example, about 15% by weight or more, for example, about 20% by weight or more, for example, about 25% by weight or more, for example, about 35% by weight or more, for example, about 40% by weight or more, for example, about 45% by weight or more, for example, about 50% by weight or more, for example, about 60% by weight or more, for example, about 65% by weight or more, for example, about 70% by weight or more, for example, about 72.5% by weight or more, for example, about 75% by weight or more, for example, about 80% by weight or more, for example, about 85% by weight or more, or any range or value between them. In other words, in some embodiments, according to any one or more of the above percentages, the denatured whey protein composition according to the present technology may have a reduced amount of natural GMP compared to the whey protein raw material.

[0043] Regardless of the amount of one or more enzymes chosen, a GMP-selective enzyme is added to the whey protein raw material to hydrolyze the natural GMP in the whey protein composition. In some embodiments, the hydrolysis phase may last for about 72 hours or less, such as about 60 hours or less, such as about 48 hours or less, such as about 36 hours or less, such as about 24 hours or less, such as about 12 hours or less, such as about 10 hours or less, such as about 8 hours or less, such as about 5 hours or more, or any range or value in between. Hydrolysis may occur at temperatures of about 60°F or lower, such as about 55°F or lower, such as about 50°F or lower, such as about 45°F or lower, or any range or value in between.

[0044] Nevertheless, in some embodiments, the natural GMP level 103 of the whey protein feedstock can be reduced by utilizing a high-fat feedstock. By using whey protein retentates from a microfiltration process (in which a portion of the GMP permeates through the microfiltration membrane), the whey protein feedstock can provide the desired low natural GMP level and high denatured whey protein level. Therefore, in some embodiments of this technology, based on the dry weight of the whey protein feedstock, a high-fat feedstock with a fat content of about 7% or more can be utilized, for example, about 8% or more, for example, about 9% or more, for example, about 10% or more, for example, about 11% or more, for example, about 12% or more, for example, about 13% or more, for example, about 14% or more, for example, about 15% or more, for example, about 16% or more, for example, about 17% or more, for example, about 18% or more, for example, about 19% or more, for example, about 20% or more, for example, about 20.5% or more, or any range or value between them.

[0045] In some embodiments, the low natural GMP and high fat levels discussed above in high-fat feedstocks can be obtained or improved by utilizing microfiltration membranes (e.g., microfiltration membranes with pore sizes of about 0.5 micrometers or smaller, such as about 0.4 micrometers or smaller, such as about 0.3 micrometers or smaller, or such as about 0.08 micrometers or larger, or any range therebetween). That is, in some embodiments, the microfiltration membrane process can be selected to retain denatured whey proteins and fats while allowing some or all of the natural proteins (including natural GMPs) to pass through to the permeate side. It should be understood that in some embodiments, other filtration methods can be used to provide high-fat, low-natural GMP feedstocks.

[0046] However, in embodiments using microfiltration membranes, it was surprisingly found that the ratio of β-lactoglobulin to α-lactalbumin increased. That is, not wanting to be bound by theory, β-lactoglobulin may be more prone to early denaturation than α-lactalbumin. Therefore, β-lactoglobulin may be retained by the membrane to a greater extent, while a higher proportion of α-lactalbumin passes through with the permeate. In some embodiments, whey protein feedstock compositions according to the present technology exhibit a β-lactoglobulin to α-lactalbumin ratio of about 2.75 or greater, for example, about 3 or greater, for example, about 3.5 or greater, for example, about 4 or greater, for example, about 4.5 or greater, for example, about 5 or greater, for example, about 5.5 or greater, for example, about 6 or greater, for example, about 6.5 or greater, for example, about 7 or greater, for example, about 7.5 or greater, or any range or value between them.

[0047] For example, in some embodiments, the whey protein raw material composition may include about 65% by weight of β-lactoglobulin or more, such as about 67.5% by weight or more, such as about 70% by weight or more, such as about 72.5% by weight or more, such as about 75% by weight or more, or any range or value between them. Additionally or alternatively, in embodiments, the whey protein raw material composition may contain about 15% by weight or less of α-lactalbumin, such as about 12.5% ​​by weight or less, such as about 10% by weight or less, such as about 7.5% by weight or less, or any range or value between them.

[0048] Although methods for reducing natural GMP are used, the raw materials for which natural GMP is reduced can undergo denaturation as known in the art 104. For example, in some embodiments, the whey protein composition can be heated to a temperature of about 176°F or higher, such as about 140°F to about 300°F, such as about 160°F to about 210°F, such as about 170°F to about 200°F, or any range or value between them, wherein the heating converts at least a portion of the starting whey protein into denatured whey protein. Advantageously, if used, this heating also denatures any enzymes. While heating, the slurry can be mixed or stirred to reduce the aggregation level of the denatured whey protein. The slurry can be mixed and heated for about 1 second to about 120 seconds, such as about 2.5 seconds to about 105 seconds, such as about 5 seconds to about 90 seconds, or any range or value between both.

[0049] During or after heating, the denatured whey protein composition may be subjected to mechanical shear conditions 105. In some embodiments, mechanical shear conditions may further denature the whey protein and / or inactivate one or more enzymes, or may reduce aggregates that form with whey protein denaturation. Mechanical shear conditions as used herein generally refer to conditions in which at least about 1000 seconds are applied. -1 High shear conditions, such as applying shear forces greater than or approximately 10,000 s. -1 Shear force, for example, applied for greater than or approximately 50,000 seconds -1 Shear force, for example, applied for greater than or approximately 100,000 seconds -1 Shear force, applied for up to 500,000 seconds -1 Shear force. In some embodiments, the denatured whey protein composition is typically sheared for about 0.1 to 120 seconds at a temperature of about 120 to 300°F using a high-shear mixer, colloid mill, or swept-surface heat exchanger.

[0050] However, it should be understood that in some embodiments, at least a portion of the protein in the whey protein composition is denatured before heating. For example, in some embodiments, the whey protein raw material may have about 5% by weight or more of denatured protein based on the total weight of the protein in the whey protein raw material, such as about 20% by weight or more, such as about 25% by weight or more, such as about 30% by weight or more, such as about 35% by weight or more, such as about 40% by weight or more, or any range or value between them.

[0051] Nevertheless, after heating, or in the denatured whey protein composition of this technology, the final protein comprises about 45% by weight or more denatured whey protein relative to the total weight of the protein in the whey protein composition, for example about 50% by weight or more, for example about 55% by weight or more, for example about 60% by weight or more, for example about 65% by weight or more, for example about 70% by weight or more, for example about 75% by weight or more, for example about 77.5% by weight or more, for example about 80% by weight or more, for example about 85% by weight or more, for example about 90% by weight or more, or any range or value between them.

[0052] As discussed above, in some embodiments, the denatured whey protein composition may be a highly denatured whey protein composition, and thus may contain about 50% by weight or more of protein on a dry weight basis, such as about 55% by weight or more, such as about 60% by weight or more, such as about 65% by weight or more, such as about 70% by weight or more, such as about 75% by weight or more, such as about 80% by weight or more, or any range or value between them.

[0053] Despite the final composition of the protein, the denatured whey protein composition can optionally be cooled and concentrated after heating and shearing, followed by drying at 106°C to produce a powdered or denatured whey protein composition. The drying process can include processes such as spray drying, heating, and evaporation. As will be discussed in more detail below, the denatured whey protein composition can then be packaged or added directly to other ingredients to prepare a food or beverage composition.

[0054] Regardless of the method used to reduce natural GMP, the denatured whey protein composition according to this technology may contain less than 12% by weight of natural GMP based on the total weight of the protein in the denatured whey protein composition, such as about 11% by weight or less, such as about 10% by weight or less, such as about 9% by weight or less, such as about 8% by weight or less, such as about 7% by weight or less, such as about 6.5% by weight or less, such as about 6% by weight or less, such as about 5.9% by weight or less, such as about 5% by weight or less, such as about 4% by weight or less, such as about 3% by weight or less, such as about 2% by weight or less, such as about 1% by weight or less, or any range or value between them.

[0055] Therefore, in some embodiments, the denatured whey protein composition may exhibit a natural GMP to denatured whey protein weight ratio of about 0.15 or less, such as about 0.125 or less, such as about 0.1 or less, such as about 0.09 or less, such as about 0.085 or less, or any range or value in between.

[0056] Furthermore, as can be understood from the above, during the hydrolysis of GMP in whey protein raw materials, hydrolyzed GMP is formed and can be advantageously retained in the denatured whey protein composition based on the desired end use. That is, the present technology has found that denatured GMP (GMP that has undergone at least one hydrolysis reaction, also referred to herein as enzymatically hydrolyzed GMP) does not exhibit the same side effects (e.g., cardboard flavor / artificial flavor) as natural GMP. Therefore, in some embodiments, the denatured whey protein composition may contain about 1% by weight or more of denatured GMP based on the total weight of protein in the denatured whey protein component, for example, about 2% by weight or more, for example, about 3% by weight or more, for example, about 4% by weight or more, for example, about 5% by weight or more, for example, about 6% by weight or more, for example, about 7% by weight or more, for example, about 8% by weight or more, for example, about 9% by weight or more, or any range or value between them.

[0057] Similarly, in some embodiments, due to the action of one or more enzymes on natural GMP, the denatured whey protein composition may have an increased proteolysis index. The proteolysis index is a measure of the increase in non-protein nitrogen (NPN) relative to the total Kjeldahl nitrogen (TKN) of a sample, and its determination method is described in the examples below. That is, as proteins are broken down by enzymatic activity into primary amino acids or small peptides soluble in trichloroacetic acid (referred to as non-protein nitrogen), the proteolysis index (PI) in the sample increases, and thus can serve as an indicator of protein hydrolysis products. Therefore, in some embodiments, the denatured whey protein composition according to the present technology may have a proteolysis index of about 6% by weight or greater, for example, about 7% by weight or greater, for example, about 8% by weight or greater, for example, about 9% by weight or greater, for example, about 10% by weight or greater, for example, about 12.5% ​​by weight or greater, for example, about 15% by weight or greater, for example, about 17.5% by weight or greater, for example, about 20% by weight or less, for example, about 22.5% by weight or greater, for example, about 25% by weight or greater, or any range or value between them.

[0058] Furthermore, while in some embodiments discussed above, the denatured whey protein composition may be considered "high in fat" and has a fat content of about 7% or more, for example, about 8% or more, for example, about 9% or more, for example, about 10% or more, for example, about 11% or more, for example, about 12% or more, for example, about 13% or more, for example, about 14% or more, for example, about 15% or more, for example, about 16% or more, for example, about 17% or more, for example, about 18% or more, for example, about 19% or more, for example, about 20% or more, for example, about 20.5% or more, or any range or value between them, in some embodiments, the denatured whey protein composition contains less than 7% by weight of fat, for example, about 6.5% by weight or less, for example, about 6% by weight or less, for example, about 2% by weight or more, or any range or value between them.

[0059] Nevertheless, the particles of the denatured whey protein composition according to the present technology may have an average particle size of about 0.001 μm to about 11 μm, for example about 0.005 μm to about 9 μm, for example about 0.01 μm to about 7 μm, for example about 0.015 μm to about 5 μm, or any range or value between them.

[0060] Surprisingly, this technology has discovered that by forming the low-natural-GMP denatured whey protein compositions discussed herein, a narrow particle size distribution of whey proteins can be obtained, which can further improve the flavor properties of the denatured whey protein compositions of this technology. For example, the particles can have a D90 particle size distribution value of about 8 μm or less, such as about 7 μm or less, such as about 6 μm or less, such as about 5 μm or less, such as about 4 μm or less, such as about 3 μm or less, such as about 2 μm or less, such as about 1.75 μm or less, such as about 1.5 μm or less, such as about 1.25 μm or less, or any range or value between them. The D90 particle size distribution value is the particle diameter in which 90% of the sample mass contains particles of this size or smaller.

[0061] In addition, the particles may have a D50 particle size distribution value of about 5 micrometers or less, such as about 4.5 μm or less, such as about 4 μm or less, such as about 3.5 μm or less, such as about 3 μm or less, such as about 2.5 μm or less, such as about 2 μm or less, such as about 1.5 μm or less, such as about 1 μm or less, such as about 0.75 μm or less, such as about 0.5 μm or less, such as about 0.3 μm or less, or any range or value between them. The D50 particle size distribution value is the particle diameter in which 50% of the sample mass contains particles of that size or less.

[0062] Additionally or alternatively, the particles may have a D10 particle size distribution value of about 3 μm or less, such as about 2.5 μm or less, such as about 2 μm or less, such as about 1.5 μm or less, such as about 1 μm or less, such as about 0.5 μm or less, such as about 0.4 μm or less, such as about 0.3 μm or less, such as about 0.2 μm or less, such as about 0.1 μm or less, such as about 0.05 μm or less, or any range or value between them. The D10 particle size distribution value is the particle diameter in which 10% of the sample mass contains particles of that size or less.

[0063] As discussed above, denatured whey protein compositions can be packaged in a denatured form or incorporated into food or beverage products to produce fortified foods and / or beverages. Suitable food and beverage products may include protein bars, granola bars, yogurt, drinkable yogurt, pudding products, ready-to-drink beverages, ready-to-mix beverage powders, baked goods, medical nutrition products, nutritional products, meat products, cheese, butter, cereal products, cream cheese, dairy products, etc.

[0064] As merely an example of the methods and products for fortifying products using denatured whey protein compositions discussed herein, yogurt compositions known in the art can be formed. For example, a scoopable or drinkable yogurt milk, which may contain whole milk, skim milk, or combinations thereof fortified with a denatured whey protein composition according to the present technology, can be sent to a fermentation tank where a yogurt culture is added to produce a yogurt mixture. The yogurt mixture can then be mixed with other ingredients or flavorings and packaged.

[0065] The viscosity of the fortified food and / or beverage products discussed herein (the methods thereof will be discussed in more detail in the examples) can be about 500 centipoise (cP) or less, for example about 400 centipoise or less, for example about 300 centipoise or less, for example about 250 centipoise or less, for example about 200 centipoise or less, for example about 150 centipoise or less, for example about 100 centipoise or less, for example about 50 centipoise or less, or any range or value between them.

[0066] Furthermore, the fortified food and / or beverage products discussed herein may be fortified to a protein level greater than 3% or about 8% by weight or more based on the weight of the food and / or beverage product, for example about 8.5% by weight or more, for example about 9% by weight or more, for example about 9.5% by weight or more, for example about 10% by weight or more, for example about 12.5% ​​by weight or more, for example about 15% by weight or more, for example about 17.5% by weight or more, for example about 20% by weight or more, for example about 22.5% by weight or more, for example about 25% by weight or more, for example about 27.5% by weight or more, for example about 30% by weight or more, or any range or value between them.

[0067] Furthermore, certain implementations of this disclosure can be better understood through the following examples, which are non-limiting and exemplary in nature.

[0068] Example

[0069] Test methods and procedures

[0070] Quantitative analysis of glycomacropeptide (GMP), α-lactalbumin (α-LA), and β-lactoglobulin (β-LG)

[0071] Quantitative analysis in GMP was performed using the Beckman capillary electrophoresis system. The capillaries were DOV-1701OH Deactivated TSP standard FS tubes (600 mm x 50 μm) with 100 x 800 μm slit openings, and were regenerated / activated with 0.1 N HCl solution.

[0072] Preparation of sample solution:

[0073] - Prepare a 1% protein solution for each sample using deionized (DI) water.

[0074] - Vortex the sample until it is homogeneous.

[0075] - Allow the powder sample to hydrate for at least 30 minutes, then continue with sample preparation.

[0076] - Sample buffer was prepared using 0.0787 g DTT (threo-1,4-dimercapto-2,3-butanediol) and 30 g reducing buffer. The reducing buffer consisted of 0.5 g / L methyl hydroxypropyl cellulose, 167 mM Tris, 42 mM 3-morpholinopropanesulfonic acid, and 67 mM disodium ethylenediaminetetraacetate in 8 M urea solution.

[0077] - After the sample is hydrated, mix the sample buffer and the sample in a 1:1 ratio to a total of 4 mL.

[0078] - Vortex the sample and let it stand for one hour.

[0079] - Filter the sample into the glass vial using a PVDF 0.22μm syringe filter. Cap the vial.

[0080] The sample solution was injected for 10 seconds at 3.4 kPa. Separation was performed at 45°C with a voltage of 25 kV (initially increased from 0 to 25 kV over 3 minutes). Milk proteins were detected at 214 nm. The amount of each protein component was determined by comparing the peak area of ​​each component with the total peak area. In this example:

[0081]

[0082]

[0083] Fat quantification

[0084] Fat composition was determined by a Mojonnier improvement to the Roese-Gottlieb procedure (reference method AOAC 989.05) for fat extraction. The Roese-Gottlieb procedure uses ether to extract fat from dairy products.

[0085] calculate:

[0086]

[0087] in:

[0088] - Fat weight: (Weight of the sample beaker after extraction) - (Weight of the empty beaker).

[0089] Blank: The blank analysis value must be subtracted from the obtained fat weight.

[0090] - Sample weight: The weight of the sample before the extraction procedure.

[0091] Quantitative analysis of protein using total Kjeldahl nitrogen (TKN)

[0092] Protein analysis using Kjeldahl nitrogen (refer to method AOAC 991.20) is used to determine the weight percentage of nitrogen-containing compounds. Sulfuric acid digests proteins and other nitrogen-containing compounds, converting nitrogen to ammonium sulfate. A catalyst is used to increase the reaction rate and raise the boiling point of sulfuric acid. The amount of nitrogen associated with proteins and soluble nitrogen is obtained by titrating ammonia with standard hydrochloric acid.

[0093] Determination of TKN (crude total protein)

[0094]

[0095] in:

[0096] -ΔmL = Amount of standardized HCl added to the sample - Amount added to the blank sample, in mL (usually calculated by the distillation apparatus and given as ΔmL).

[0097] -N = the exact normality of standardized HCl from the Certificate of Analysis, in meq / mL.

[0098] -14.007 = the formulation weight of nitrogen, in mg / meq.

[0099] The constant 6.38 represents the grams of dairy protein per gram of nitrogen. Other constants for ammonium sulfate are 4.7218 and for L-tryptophan, it is 7.2904.

[0100] -W = Sample size in grams.

[0101] - Divide by 10, the result is grams per 100 grams.

[0102] - Results are expressed as a percentage of protein weight (grams per 100 grams of sample).

[0103] Determination of NPN (non-protein nitrogen)

[0104] Non-protein nitrogen (NPN) consists of urea, ammonia, free amino acids, creatine, uric acid, peptides, and amino alcohols of phospholipids (Ruska and Jonkus, 2014), and is soluble in trichloroacetic acid (TCA). Generally, more than 30 amino acids are sufficient to classify a peptide as a protein, although this is not a hard and fast rule. Peptides consisting of fewer than 30 amino acids may be present in the TCA-soluble fraction and classified as NPN. This method uses the addition of trichloroacetic acid (TCA) to precipitate proteins. Proteins are filtered off, and the NPN in the filtrate is measured. The amount of NPN can then be determined.

[0105] Sample preparation

[0106] A. Mix the sample thoroughly.

[0107] B. Peel a 150 mL beaker or a 4 oz snap-cap. For powder or solid samples, add 10-15 glass beads to the beaker or snap-cap before peeling.

[0108] C. Based on the table below, transfer an appropriate number of samples to peeled beakers. Record the sample weight to the nearest 0.0001 g. For powder or solid samples, add 20 mL of deionized water and shake to prepare the sample.

[0109] D. Using a graduated cylinder or automatic dispenser, add 15 mL of 33% TCA to the sample.

[0110] E. Place the beaker or spring-loaded lid back onto the balance.

[0111] F. Add deionized water to the sample until the total mass of the sample, TCA solution, and added water is approximately 50 g. Record the gross weight.

[0112] G. Mix well and let the solution stand for 10 minutes.

[0113] H. Filter with slow filter paper and pour into a clean spring-loaded lid or beaker.

[0114] I. Transfer approximately 7-10g of filtrate to a Kjeldahl digestion tube. Record the exact mass of the added filtrate.

[0115] The J.TKN (Total Kjeldahl Nitrogen) method is used to determine nitrogen (N) in the filtrate for the following calculations:

[0116]

[0117] ·mL = The amount of standardized HCl required to titrate the sample distillate.

[0118] • N = Precise normality (meq / mL) of standardized HCl from a batch analysis certificate (COA) or as determined in the laboratory.

[0119] 14.007 = the weight of nitrogen in the formulation, in mg / meq.

[0120] 6.38 is a factor that converts nitrogen into dairy protein.

[0121] W = the number of grams of sample filtrate used.

[0122] • A = the weight of the sample used (in grams)

[0123] B = Total grams of sample solution

[0124] ·10 Convert the results to grams per 100 grams.

[0125] Determination of true protein

[0126] True protein is a measure of total nitrogen (TKN) (sometimes called crude protein) after deducting non-protein nitrogen (NPN). True protein = TKN - NPN, expressed as the weight percentage of true protein (grams of true protein per 100 grams of sample).

[0127] Determination of protein hydrolysis index (PI)

[0128] The protein hydrolysis index is a measure of the increase in non-protein nitrogen (NPN) relative to the total Kjeldahl nitrogen (TKN) of a sample. The protein hydrolysis index (PI) in a sample increases as proteins are broken down by enzymatic activity into major amino acids or small peptides soluble in trichloroacetic acid (referred to as non-protein nitrogen).

[0129]

[0130] The protein hydrolysis index is expressed as the NPN weight percentage of total crude protein (TKN) (grams of NPN per 100 grams of total crude protein).

[0131] Determination of denatured whey protein

[0132] The denatured whey protein (DWP) method measures whey proteins that have undergone secondary and tertiary structural disruption and possible destruction, making the protein insoluble in environments where it is normally soluble in its native form.

[0133] Sample preparation

[0134] - Prepare a sample solution containing approximately 1.2% (w / w) protein. Adjust the pH to 6.8 using 0.1N hydrochloric acid or 0.1N sodium hydroxide.

[0135] - After pH adjustment, record the sample weight and the final solution weight.

[0136] -The volume was divided into two parts for testing-

[0137] -10ml for total TKN. Test TKN. This will be the percentage of total TKN.

[0138] -25g of the DWP fraction in the beaker is used for the following processing.

[0139] Determination of denatured whey protein fractions (pH 4.6)

[0140] Add approximately 10 mL of distilled water to the beaker containing the DWP fraction.

[0141] Place the small magnetic stir bar into the beaker. Place the beaker on the stirring plate and insert the pH probe and temperature compensator into the solution. Turn on the stirrer.

[0142] - Adjust the pH to 4.60 ± 0.02 by adding 1 normal HCl (for coarse adjustment) and 0.1 normal HCl (for fine adjustment).

[0143] Place the beaker on the balance. Adjust the weight of the solution to approximately 50 grams using distilled water (e.g., tare weight + 50 grams). Record the weight to the nearest 0.0001 g.

[0144] Let the solution stand at room temperature for one hour. Vortex the contents of the beaker for 10-15 seconds to mix. Transfer the contents to a centrifuge tube.

[0145] Place the test tubes in the centrifuge, ensuring they are counterbalanced. Centrifuge at 10°C and 10,000 rpm for 15 minutes. Do not use the brakes to accelerate or decelerate.

[0146] - Without transferring any precipitate, transfer approximately 10 mL of the supernatant to the identified culture tube. This is the undenatured whey protein fraction. Test the TKN in this supernatant. This is "% TKN in the undenatured whey protein fraction".

[0147] Calculate denatured whey protein (DWP):

[0148]

[0149] Determination of particle size distribution

[0150] Particle size distribution was determined using a Malvern Mastersizer 3000 with Hydro EV. The method parameters were: particle refractive index 1.46, particle absorptivity index 0.0001, dispersant refractive index 1.33, and water as the dispersant. The Mie scattering model was used for the analysis.

[0151] Example 1

[0152] A glycomacropeptide (GMP)-reduced denatured whey protein composition was prepared using whey protein composition (WPC), which is a sweet whey protein composition with a dry weight content greater than 25%. The process begins with a WPC retentate (80% protein on a dry basis), which is concentrated using ultrafiltration (UF) of pasteurized sweet whey separated from cheese curd. The 80% WPC retentate (WPC80) is transferred to a temperature-controlled storage tank and maintained at a low temperature of approximately 45°F or lower, where it is mixed with water to form a 14% protein solution. The storage tank contains 5,000 lbs of whey protein mixture in batches, of which 700 lbs of protein from the 80% WPC retentate (retentate) is mixed with water to form a diluted whey protein solution. The diluted whey protein solution is maintained at approximately 45°F or lower until the protease is added.

[0153] One or more enzymes are added, in which case one or more proteases initiate the incubation phase of protein hydrolysis, selectively hydrolyzing GMPs in the whey protein mixture while leaving other whey proteins (such as α-lactalbumin and β-lactoglobulin) essentially unhydrolyzed. In this experiment, the proteases comprise at least one basic serine protease and at least one neutral protease. In this example, one or more proteases are added to the whey protein mixture at a level of about 0.001% by weight to about 5% by weight or about 0.012% by weight or less of the total protein in the substrate. For a 5000-pound batch, approximately 38 grams of enzymes are added.

[0154] The hydrolysis phase, under stirring at ≤45°F, lasts for at least 5 hours, but can last 72 hours or longer, without hydrolyzing the major whey proteins (i.e., α-lactalbumin and β-lactoglobulin). Subsequently, the enzymatically treated whey protein mixture is heated to 190°F for 6 seconds (operating range 176–195°F for 5–90 seconds) while undergoing mechanical shearing to achieve enzyme inactivation and whey protein denaturation. Following heating and mechanical shearing, the resulting denatured whey protein mixture is optionally cooled (e.g., 50°F) and concentrated (e.g., by evaporation or nanofiltration), then spray-dried to produce a powdered, GMP-reduced denatured whey protein composition.

[0155] like Figures 2A to 2C The micronized denatured whey protein composition, as described, has a particle size distribution (characterized by D90) of less than or about 1.7 micrometers compared to the whey protein raw material, and exhibits a narrower range than the raw material.

[0156] Figure 2A The particle size of the undenatured (feed) material was depicted, ranging from 0.011 to 9.86 micrometers. Following GMP enzymatic hydrolysis and subsequent enzyme inactivation and whey protein denaturation, the particle size decreased to a range of 0.017 to 4.03 micrometers. Figure 2B As shown. Figure 2C Provides a comparative overlay plot of particle size distribution of whey protein raw material and sample 1.

[0157] The weight level of natural GMP in total protein of the whey protein composition raw material and the micronized denatured whey protein composition was determined by capillary electrophoresis (CE) and defined as the peak area of ​​natural GMP divided by the peak area of ​​total protein on the CE plot, such as... Figure 3A and 3B As shown.

[0158] As discussed above, denatured whey protein (DWP) and the proteolysis index (PI) (defined as non-protein nitrogen divided by total nitrogen) were determined by wet chemical analysis. Table 1 illustrates the reduction of natural GMP from the typical cheese whey level of 12-25% (20.52% in this case) to below 8%, specifically 5.84%. Similarly, as GMP hydrolysis occurs, the proteolysis index increases accordingly, as natural GMP proteins are broken down into smaller fragments, leading to an increase in the amount of non-protein nitrogen. Figure 3A and 3B It was shown that although GMP was significantly reduced during hydrolysis, α-lactalbumin (a-La) and β-lactoglobulin (b-Lg) remained essentially unhydrolyzed due to controlled hydrolysis, further illustrated by the minimal reduction (<10%) in the β-lactoglobulin to α-lactalbumin ratio (Table 1). The increase in denatured whey protein was a direct result of the time and temperature treatment during enzyme inactivation.

[0159] Table 1:

[0160]

[0161] *- Denatured whey protein %

[0162] ^- Fat percentage on a dry basis

[0163] 1-Protein hydrolysis index

[0164] 2- Protein percentage on dry basis

[0165] Example 2

[0166] The feedstock for the high-fat whey protein composition (hfWPC) is obtained via microfiltration. The feed composition from the retentate side of the microfiltration process is 75.48% protein and 17.66% fat on a dry basis. Water is added to adjust the high-fat whey protein composition feed to 14% protein (operating range 10-24%).

[0167] Protein denaturation was achieved by preheating the feed to 130°F (operating range 120-150°F), then heating to a denaturation temperature of 176-195°F and holding for 5-90 seconds, while simultaneously mechanically shearing to control particle size during heating. After the heating and shearing process, the denatured whey protein composition was cooled to below 50°F and dried into powder by spray drying to form Sample 2.

[0168] The total solids, fat, protein, denatured whey protein, and particle size distribution of the denatured high-fat whey protein composition were analyzed, and the results are shown in Table 2 and... Figures 4A-4C As shown, this is the result of the high-fat whey protein composition before (feeding) and after the denaturation-shearing process. Figure 4A The particle size range of high-fat WPC feedstock was described as 0.011–11.2 micrometers. After denaturation and shearing processes, the particle size decreased to the range of 0.011–2.75 micrometers. Figure 4B As shown. Figure 4C A comparative overlay of particle size distributions for both high-fat WPC feedstock and modified high-fat WPC is provided.

[0169] As described above and shown in Table 1, this technique reveals that the high-fat WPC formed in this study exhibits different chemical properties compared to WPC 80. Compared to the starting feed WPC 80, the high-fat WPC's starting material has lower GMP (10.5% vs 20.5%), higher denatured whey protein (DWP) (45.10% vs 17.27%), higher dry-basis fat (17.66% vs 6.35%), and a higher β-lactoglobulin to α-lactalbumin ratio (7.65% vs 3.37%). The production of high-fat WPC utilizes both ultrafiltration and microfiltration, while the production of WPC 80 utilizes only ultrafiltration. One possible explanation is that, due to the pore size of the microfiltration membrane, most of the DWP and fat are retained in the retentate, while some GMP, α-lactalbumin, β-lactoglobulin, and other whey proteins permeate to the permeate side. It is believed that β-lactoglobulin is more prone to denaturation during milk and subsequent sweet whey pasteurization compared to α-lactalbumin, thus the retained protein contains more denatured β-lactoglobulin than α-lactalbumin. Therefore, the ratio of β-lactoglobulin to α-lactalbumin in high-fat WPC is higher than that in WPC80. Interestingly, despite differences in DWP values ​​in the starting feed, the ultimately denatured high-fat WPC (hfWPC) and denatured WPC80 achieved similar particle size distributions and denatured whey protein (DWP) values ​​(for comparison). Figure 2C and Figure 4C (and Tables 1 and 2).

[0170] Table 2:

[0171]

[0172] Example 3

[0173] A denatured whey protein composition was used to fortify drinkable yogurt applications to observe its functional benefits in managing the viscosity of high-protein products.

[0174] The sample according to Example 3 was prepared using the following yogurt preparation method:

[0175] 1. Mix the powder and liquid components of the basic formulation (Table 3) to obtain a homogeneous solution.

[0176] 2. Pasteurize at 80-85℃ (176-185°F) for 30 minutes.

[0177] 3. Cool to 42°C (108°F).

[0178] 4. Inoculate with 0.02% of the starter culture (CHR Mild 2.0)

[0179] 5. Maintain the temperature at 42℃ (108°F) during fermentation.

[0180] 6. Stop the fermentation process when the pH value reaches 4.6.

[0181] 7. Store at 4℃ (39°F).

[0182] Table 3

[0183]

[0184] The protein content, total solids percentage, and viscosity of the samples and controls were then analyzed, and the results are shown in Table 4.

[0185] Table 4

[0186] Drinkable yogurt—fortified with the following substances protein% Total solids % Viscosity (cP) Comparison (without WPC) 10.00 18.00 >25000 Standard GMP undenatured WPC80 (control) 8.32 14.42 4,040 GMP reduces denaturation of WPC80 10.08 16.54 90 GMP-reduced undenatured HfWPC (control) 9.94 16.93 860 GMP reduces denaturation of HfWPC 10.03 17.06 80 Reference for commercially available products 7.54 14.02 580

[0187] *Brookfield viscometer, using a No. 6 rotor, at 100 rpm and 42°F.

[0188] Consumers may be interested in different levels of protein fortification. Examples of protein fortification in drinkable yogurt are shown in Tables 3 and 4. Figure 5 As shown in the figure, compared to commercially available products with a protein level of 7.54%, the drinkable yogurt fortified to 10% protein using the GMP-reduced denatured WPC80 composition and the GMP-reduced denatured hfWPC according to this technology has a preferred lower viscosity. Therefore, this technology exhibits a significant reduction in viscosity, even at higher fortification levels. The reduced viscosity benefits the consumer experience due to the increased protein content per serving and the ease of consumption of the product.

[0189] As shown, increasing protein content in yogurt applications is challenging. Yogurt viscosity typically increases with increasing protein content. This is because proteins commonly used to fortify foods (such as casein and undenatured whey protein) bind to and interact with water, resulting in a higher viscosity texture. The benefit of using denatured whey compositions according to this technology in products such as yogurt is fortifying protein content while maintaining an acceptable viscosity. This is shown in Table 4, which compares viscosity data for yogurt fortified with a control (without added WPC), conventional GMP undenatured WPC80, and GMP-reduced denatured WPC80, demonstrating the practical solution to viscosity issues offered by GMP-reduced denatured whey protein compositions. As shown, the control yogurt (without added WPC) is very viscous, with a viscosity >25000 cP, making it unpouringable and unsuitable as a drinkable yogurt product. Similarly, the drinkable yogurt fortified with conventional GMP undenatured WPC80 has a viscosity of 4040 cP, which is still too viscous for a drinkable yogurt product and is unacceptable. On the other hand, yogurt fortified with GMP-reduced modified WPC80 according to this technology exhibits an acceptable viscosity of 90 cP for pouring and as a drinkable yogurt product. Furthermore, compared to a commercial reference viscosity of 580 cP, GMP-reduced modified WPC80 demonstrates highly preferred performance.

[0190] Table 4 shows viscosity data for the control (without WPC), GMP-reduced undenatured hfWPC, and GMP-reduced denatured hfWPC. This example demonstrates the practical solution that GMP-reduced denatured whey protein compositions offer for products with the desired low viscosity. As the data shows, the control yogurt (without WPC) is very viscous, with a viscosity >25000 cP, making it unpouringable and unsuitable as a drinkable yogurt product. In contrast, drinkable yogurt fortified with conventional GMP-reduced undenatured hfWPC exhibits a lower viscosity of 860 cP, which is still too viscous for a drinkable yogurt product. On the other hand, yogurt fortified with GMP-reduced denatured hfWPC according to this technology exhibits an acceptable viscosity of 80 cP for pouring and as a drinkable yogurt product.

[0191] Example 4

[0192] A denatured whey protein composition was used to fortify scoopable yogurt applications to observe functional benefits in managing the viscosity of high-protein products.

[0193] Table 5:

[0194]

[0195] The protein content, total solids percentage, and viscosity of the samples and controls were then analyzed, and the results are shown in Table 6.

[0196] Table 6

[0197]

[0198] Tables 5 and 6 show examples of protein-fortified scoopable yogurts. The data indicate that the GMP-reduced denatured whey protein composition according to this disclosure exhibits a greater leverage effect in increasing the protein content of scoopable yogurts without a corresponding increase in viscosity or a change in acceptable texture. The viscosity of a commercial reference scoopable yogurt containing 10% protein is 11,000 cP. When the protein content is increased to 14.7% using GMP-reduced undenatured hfWPC, the viscosity is 63,000 cP, which is too high for a scoopable yogurt product. However, when GMP-reduced denatured hfWPC or GMP-reduced denatured WPC80 according to this technology is used to increase the protein content to 14.7%, the viscosity is 9,000 to 12,000 cP, equivalent to a commercially available reference yogurt with a protein content of 10%. This demonstrates the advantage of using the GMP-reduced denatured whey protein composition according to this technology to achieve an increase in the protein content of scoopable yogurt while maintaining viscosity. This improvement also enhances the consumer experience due to the more familiar viscosity and texture.

[0199] Example 5

[0200] Samples of undenatured whey protein concentrate, GMP-reduced WPC 80, undenatured high-fat WPC, and denatured WPC (80% protein on a dry basis) were prepared for descriptive flavor analysis. Whey protein was rehydrated at 10% (w / v). Products were dispensed into lidded soufflé cups with 3-digit codes and evaluated. The beverages were evaluated in duplicate by a seven-member trained panel using established sensory language for whey protein. Descriptive flavor analysis employed a 0-15 scale and Spectrum. TM Methods (Meilgaard et al. 1999; Drake and Civille 2003). Paper ballots were used. Each panel member evaluated each product in duplicate during different sessions. Data were analyzed using Fisher's minimum significance difference (LSD) as a post-hoc test via general linear model ANOVA (SAS version 9.1, Cary, NC).

[0201] Table 7 summarizes the analysis of the samples by the trained team. An unexpected observation was that, according to this technique, reducing GMP and denaturing whey proteins resulted in a significant reduction in cardboard flavor, which is generally considered an off-flavor and present in many dairy proteins. Furthermore, the reduction of GMP led to an improvement in milk flavor, which is considered a beneficial flavor property in products suitable for dairy applications.

[0202] When evaluating GMP-reduced denatured high-fat whey protein compositions, similar flavor observations further confirmed the results of GMP-reduced denatured WPC. Similarly, it was observed that lower GMP, combined with protein denaturation, directionally led to a reduction in cardboard flavor and a significant increase in milk flavor.

[0203] Table 7

[0204]

[0205] Aroma and flavor intensity were scored on a general intensity scale of 0 to 15 (spectral method, Meilgaard et al., 1999). ND - Not detected.

[0206] The mean values ​​differed in columns followed by different letters (p<0.05). No unlisted attributes were detected in these samples.

Claims

1. A denatured whey protein composition comprising: At least 60% by weight of whey protein on a dry weight basis; Relative to the total weight of proteins in the denatured whey protein composition, less than 8% by weight of natural glycomacropeptide (GMP) and more than 2% by weight of enzymatically hydrolyzed GMP; At least 8.0% by weight of protein hydrolysis index; and The denatured whey protein comprises more than 50% by weight of the total protein in the denatured whey protein composition.

2. The denatured whey protein composition of claim 1, wherein the denatured whey protein comprises denatured enzyme-hydrolyzed cheese whey protein.

3. The denatured whey protein composition of claim 2, wherein the natural GMP is less than or equal to 7 by weight relative to the total weight of the proteins in the denatured whey protein composition.

4. The denatured whey protein composition of claim 1, wherein the denatured whey protein is characterized in that the D50 particle size distribution value is less than or equal to 4.5 µm.

5. The denatured whey protein composition of claim 4, wherein the denatured whey protein is further characterized in that the D10 particle size distribution value is less than or equal to 2.5 µm.

6. The denatured whey protein composition of claim 4, wherein the denatured whey protein is further characterized in that the D90 particle size distribution value is less than or equal to 8.0 µm.

7. The denatured whey protein composition of claim 1, further comprising up to 7.0% by weight of fat on a dry weight basis.

8. A method for preparing a denatured whey protein composition, the method comprising: Cheese whey from enzymatically coagulated milk is filtered into cheese whey retentate and permeate; and To reduce the natural glycomacropeptides (GMPs) in the cheese whey retentate to form a GMP-reduced cheese whey retentate composition; and Heating the GMP-reduced cheese whey retentate composition to form a denatured whey protein composition. The denatured whey protein composition is characterized by: At least 60% by weight of whey protein on a dry weight basis; Relative to the total weight of proteins in the denatured whey protein composition, less than 8% by weight of natural GMP and more than 2% by weight of enzymatically hydrolyzed GMP; At least 8.0% by weight of protein hydrolysis index; as well as The denatured whey protein comprises more than 50% by weight of the total protein in the denatured whey protein composition.

9. The method of claim 8, wherein the reduction of natural GMP in the cheese whey retrieval comprises combining the cheese whey retrieval with one or more enzymes that selectively hydrolyze GMP in the cheese whey retrieval to form a GMP-reduced cheese whey retrieval composition, wherein the one or more enzymes that selectively hydrolyze GMP in the cheese whey retrieval comprise at least one alkaline serine protease and at least one neutral protease, or wherein the reduction of GMP occurs by microfiltration of the cheese whey retrieval.

10. The method of claim 9, wherein heating of the GMP-reduced cheese whey retentate composition also inactivates one or more enzymes that selectively hydrolyze GMP in the cheese whey retentate.

11. The method of claim 8, wherein the GMP-reduced cheese whey retentate composition is heated to a temperature greater than or equal to 160°F.

12. The method of claim 8, wherein heating of the GMP-reduced cheese whey retentate composition further comprises exposing the GMP-reduced cheese whey retentate composition to high shear conditions.

13. The method of claim 8, wherein the denatured whey protein composition is characterized in that the weight ratio of natural GMP to total whey protein is less than or equal to 0.

15.

14. The method of claim 8, wherein the GMP-reduced cheese whey retentate already contains at least 30% by weight of denatured protein prior to heating.

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