Dairy products and processing

Abstract

The present invention relates to a dairy protein composition, a method for preparing the same and uses thereof. In particular, the present invention relates to the use of the dairy protein composition for preparing high protein foods with low viscosity and / or hardness, including yoghurt.

Description

[Technical field]

[0001] The present invention relates to a dairy protein composition, a process for its preparation and its use. In particular, the present invention relates to the use of the dairy protein composition in the manufacture of low viscosity, high protein food products, including yoghurt. [Background technology]

[0002] High protein foods such as dairy products, sports drinks, medical drinks, cultured products, cheeses, etc. can be produced by adding ingredients with high protein content to a milk base or other composition. Desirable properties of high protein dairy ingredients include: ●Mild and milky taste, Concentration of protein, ●Thermal stability, • High nutritional value and / or ease of mixing with other ingredients in high protein foods.

[0003] High protein dairy ingredients include milk protein concentrates and isolates, whey protein concentrates and isolates, and caseinates.

[0004] In certain applications, it is desirable to achieve a low viscosity of a high protein food product while a high protein dairy ingredient provides a high protein content. Generally, the use of high protein dairy ingredients results in an unacceptably high viscosity in foods. Low viscosity can be achieved in foods by using microparticulated whey protein ingredients. However, while they do achieve the desired organoleptic properties, they do not provide the same casein:whey ratio as cow's milk, can have a strong protein or musty flavor profile, and are more expensive.

[0005] Therefore, there is a need for alternative dairy protein ingredients that have high protein content and provide low viscosity and clean flavor when used in food production.

[0006] It is an object of the present invention to provide improved or alternative dairy protein compositions, methods for their preparation and / or high protein food products comprising the dairy protein compositions, and / or at least to provide the public with a useful choice. Summary of the Invention

[0007] In one aspect, the invention generally provides a dairy protein composition comprising a dairy protein, the dairy protein composition comprising: a) comprising at least about 40% total protein by weight based on the dry matter in the composition; b) Milk proteins include casein; c) the total protein contains less than about 79% by weight of peptides having a molecular weight greater than about 20 kDa; d) The composition comprises: i. less than about 2 g calcium per 100 g total protein, and / or ii. Contains less than about 1.4 g calcium per 100 g dry matter in the composition.

[0008] In one aspect, the present invention provides a milk protein composition comprising a milk protein concentrate, a milk protein isolate, or a combination thereof, a) the composition comprises at least about 40% total protein by weight based on the dry matter in the composition; b) the total protein contains less than about 79% by weight of peptides having a molecular weight greater than about 20 kDa; c) The composition comprises i. less than about 2 g calcium per 100 g total protein, and / or ii. Contains less than about 1.4 g calcium per 100 g dry matter in the composition.

[0009] In one aspect, the present invention provides a dairy protein composition comprising a dairy protein concentrate, a dairy protein isolate, or a combination thereof, the dairy protein composition comprising: a) comprising at least about 40% total protein by weight based on the dry matter in the composition; b) Total protein is about 20 to about 79% by weight of a peptide having a molecular weight greater than about 20 kDa; about 15 to about 55% by weight of a peptide having a molecular weight of about 5 to about 20 kDa; about 2 to about 20% by weight of a peptide having a molecular weight of about 1 to about 5 kDa; about 2 to about 20% by weight of a peptide having a molecular weight of less than about 1 kDa; c) The composition comprises i. less than about 2 g calcium per 100 g total protein, and / or ii. Contains less than about 1.4 g calcium per 100 g dry matter in the composition.

[0010] In one aspect, the present invention provides a method for preparing a milk protein composition, the method comprising: a) providing an aqueous composition comprising milk protein, the aqueous composition comprising about 0.5 to about 20% by weight total protein; b) exposing the aqueous composition to the action of one or more proteolytic enzymes; c) inactivating the one or more proteolytic enzymes to produce a milk protein composition, where 1. The milk protein composition comprises at least about 40% total protein by weight based on the dry matter in the composition; 2. Milk proteins include casein, 3. The total protein contains less than about 79% by weight of peptides having a molecular weight greater than about 20 kDa; 4. The composition comprises: i. less than about 2 g calcium per 100 g total protein, and / or ii. Contains less than about 1.4 g calcium per 100 g dry matter in the composition.

[0011] In one aspect, the present invention provides a method for preparing a milk protein composition, the method comprising: a) providing an aqueous composition comprising a milk protein concentrate, a milk protein isolate, or a combination thereof, the composition comprising from about 0.5 to about 20% by weight total protein; b) exposing the aqueous composition to the action of one or more proteolytic enzymes; c) inactivating the one or more proteolytic enzymes to produce a milk protein composition, where 1. The milk protein composition comprises at least about 40% total protein by weight based on the dry matter in the composition; 2. The total protein contains less than about 79% by weight of peptides having a molecular weight greater than about 20 kDa; 3. The composition comprises: i. less than about 2 g calcium per 100 g total protein, and / or ii. Contains less than about 1.4 g calcium per 100 g dry matter in the composition.

[0012] In one aspect, the invention relates to a milk protein composition prepared by the method of the invention.

[0013] In one aspect, the present invention relates to a protein-containing food product comprising the dairy protein composition of the present invention.

[0014] In one aspect, the invention relates to the use of a dairy protein composition of the invention in the manufacture of a protein-containing food product.

[0015] In one aspect, the present invention relates to a protein-containing food product comprising a dairy protein composition of the present invention or a dairy protein composition prepared by a method of the present invention, wherein the food product is an acidified or fermented product.

[0016] In one aspect, the present invention relates to a protein-containing food product comprising a dairy protein composition of the present invention or a dairy protein composition prepared by a method of the present invention, wherein the protein-containing food product is yogurt.

[0017] In one aspect, the present invention provides a method for preparing a protein-containing food product, the method comprising: a) providing an aqueous composition comprising a milk protein composition of the present invention or a milk protein composition prepared by a method of the present invention; b) mixing with one or more additional additives to produce a protein-containing food product.

[0018] In one aspect, the present invention provides a method for producing an acidified protein-containing food product, comprising the steps of: a) providing an aqueous composition comprising a milk protein composition of the present invention or a milk protein composition prepared by a method of the present invention; b) acidifying the aqueous composition to produce an acidified protein-containing food product.

[0019] In one aspect, the present invention relates to a method for preparing a fermented protein-containing food product, the method comprising: a) i. a milk protein composition of the invention or a milk protein composition prepared by a method of the invention; ii. providing an aqueous composition comprising one or more media; b) incubating the aqueous composition for a period of time sufficient to produce a fermented protein-containing food product.

[0020] In one aspect, the present invention provides a method for preparing a bar, the method comprising: a) i. a milk protein composition of the invention or a milk protein composition prepared by a method of the invention; ii. providing a bar composition comprising one or more additional ingredients; b) forming the bar composition into a bar.

[0021] In one aspect, the present invention provides a method for preparing cheese, preferably processed cheese, comprising: a) i. a milk protein composition of the invention or a milk protein composition prepared by a method of the invention; ii. providing a cheese composition comprising one or more additional ingredients; b) heating the cheese composition to a temperature of at least about 70° C. to produce cheese.

[0022] In one aspect, the present invention provides a method for preparing a protein-containing food product, the method comprising: a) a milk protein composition comprising: i. at least about 40% total protein by weight based on dry matter in the composition; ii. providing a milk protein composition comprising less than about 2 g calcium per 100 g total protein and / or less than about 1.4 g calcium per 100 g dry matter; b) mixing the milk protein composition with one or more additional ingredients to produce an aqueous intermediate composition comprising about 0.5 to about 20% by weight total protein; c) exposing the aqueous intermediate composition to the action of one or more proteolytic enzymes; and d) inactivating the one or more proteolytic enzymes to produce the food product.

[0023] In one aspect, the present invention provides a method for preparing an acidified protein-containing food product, the method comprising: a) i. at least about 40% total protein by weight on dry matter of the composition; ii. providing a milk protein composition comprising less than about 2 g calcium per 100 g total protein and / or less than about 1.4 g calcium per 100 g dry matter; b) mixing the milk protein composition with one or more additional ingredients to produce an aqueous intermediate composition comprising about 0.5 to about 20% by weight total protein; c) exposing the aqueous intermediate composition to one or more proteases, and optionally inactivating the one or more proteases; d) acidifying the aqueous intermediate composition; producing an acidified protein-containing food product; Here, steps c) and d) are performed in the same order or in reverse order.

[0024] In one aspect, the present invention provides a method for preparing a fermented protein-containing food product, the method comprising: a) i. at least about 40% total protein by weight based on dry matter in the composition; ii. providing a milk protein composition comprising less than about 2 g calcium per 100 g total protein and / or less than about 1.4 g calcium per 100 g dry matter; b) mixing the milk protein composition with one or more additional ingredients to produce an aqueous intermediate composition comprising about 0.5 to about 20% by weight total protein; c) exposing the aqueous intermediate composition to one or more proteases, and optionally inactivating the one or more proteases; d) adding one or more media to the aqueous intermediate composition and incubating for a sufficient period of time; Producing a fermented protein-containing food product, Here, steps c) and d) are performed in the same order or in reverse order.

[0025] In one aspect, the present invention provides a method for preparing a protein-containing food product, the method comprising: a) a milk protein composition comprising a milk protein concentrate, a milk protein isolate, or a combination thereof, i. at least about 40% total protein by weight based on the dry matter in the composition; ii. providing a milk protein composition comprising less than about 2 g calcium per 100 g total protein and / or less than about 1.4 g calcium per 100 g dry matter; b) mixing the milk protein composition with one or more additional ingredients to produce an aqueous intermediate composition comprising about 0.5 to about 20% by weight total protein; c) exposing the aqueous intermediate composition to the action of one or more proteolytic enzymes; and d) optionally inactivating one or more proteolytic enzymes to produce a protein-containing food product.

[0026] In one aspect, the present invention provides a method for preparing an acidified protein-containing food product, the method comprising: a) a milk protein composition comprising a milk protein concentrate, a milk protein isolate, or a combination thereof, i. at least about 40% total protein by weight based on dry matter in the composition; ii. providing a milk protein composition comprising less than about 2 g calcium per 100 g total protein and / or less than about 1.4 g calcium per 100 g dry matter; b) mixing the milk protein composition with one or more additional ingredients to produce an aqueous intermediate composition comprising about 0.5 to about 20% by weight total protein; c) exposing the aqueous intermediate composition to one or more proteases, and optionally inactivating the one or more proteases; d) acidifying the aqueous intermediate composition producing an acidified protein-containing food product; Here, steps c) and d) are performed in the same order or in reverse order.

[0027] In one aspect, the present invention provides a method for preparing a fermented protein-containing food product, the method comprising: a) a milk protein composition comprising a milk protein concentrate, a milk protein isolate, or a combination thereof, i. at least about 40% total protein by weight based on dry matter in the composition; ii. providing a milk protein composition comprising less than about 2 g calcium per 100 g total protein and / or less than about 1.4 g calcium per 100 g dry matter; b) mixing the milk protein composition with one or more additional ingredients to produce an aqueous intermediate composition comprising about 0.5 to about 20% by weight total protein; c) exposing the aqueous intermediate composition to one or more proteases, and optionally inactivating the one or more proteases; d) adding one or more media to the aqueous intermediate composition and incubating for a sufficient period of time; Producing a fermented protein-containing food product. Here, steps c) and d) are performed in the same order or in reverse order.

[0028] The following embodiments may relate to any or all of the above aspects.

[0029] In various embodiments, the dairy protein composition is a dry composition, preferably a powder.

[0030] In various embodiments, the milk protein or milk protein composition may comprise or be provided from a milk protein concentrate (MPC), a milk protein isolate (MPI), caseinate, casein, casein co-precipitate, a retentate obtained by ultrafiltration or microfiltration of milk, or any combination of any two or more thereof. In various embodiments, the milk protein or milk protein composition may comprise a milk protein concentrate (MPC), a milk protein isolate (MPI), or a combination thereof. In various embodiments, the milk protein or milk protein composition may comprise a milk protein concentrate (MPC) or a milk protein isolate (MPI). In various embodiments, the milk protein may consist of a milk protein concentrate, a milk protein isolate, or a combination thereof.

[0031] In various embodiments, the retentate obtained by ultrafiltration or microfiltration of milk may comprise micellar casein concentrate.

[0032] In various embodiments, the total protein in the composition may comprise at least about 5, 10, 20, 30, 40, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 99% by weight casein, or the total protein in the composition may comprise 100% by weight casein. The various ranges may be between any two selected from these values, for example, the total protein may comprise about 5 to about 100, about 10 to about 100, about 40 to about 100, about 50 to about 100, about 60 to about 100, about 70 to about 100, about 75 to about 100, about 80 to about 100, about 5 to about 99, about 10 to about 99, about 40 to about 99, about 50 to about 99, about 60 to about 99, about 70 to about 99, about 75 to about 99, or about 80 to about 99 weight percent casein.

[0033] In various embodiments, milk proteins may include casein and whey proteins.

[0034] In various embodiments, the dairy protein composition may further comprise whey protein concentrate, or whey protein isolate, or a combination thereof.

[0035] In various embodiments, the total protein may contain less than about 79%, 78%, 77%, 76%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, or 40% by weight of peptides having a molecular weight greater than about 20 kDa.

[0036] In various embodiments, the total protein may contain from about 20 to about 79% by weight of peptides having a molecular weight greater than about 20 kDa.

[0037] In various embodiments, the total protein may contain about 15 to about 55% by weight of peptides having a molecular weight of about 5 to about 20 kDa.

[0038] In various embodiments, the total protein may contain less than about 30% by weight of peptides having a molecular weight of less than about 5 kDa.

[0039] In various embodiments, the total protein may contain from about 5 to about 30% by weight of peptides having a molecular weight of less than about 5 kDa.

[0040] In various embodiments, the total protein may contain less than about 20% by weight of peptides having a molecular weight of about 1 to about 5 kDa.

[0041] In various embodiments, the total protein may contain about 2 to about 20% by weight of peptides having a molecular weight of about 1 to about 5 kDa.

[0042] In various embodiments, the total protein may contain less than about 20% by weight of peptides having a molecular weight of less than about 1 kDa.

[0043] In various embodiments, the total protein may contain from about 2 to about 20% by weight of peptides having a molecular weight of less than about 1 kDa.

[0044] In various embodiments, the total protein is a) about 20 to about 79% by weight of a peptide having a molecular weight greater than about 20 kDa; b) about 15 to about 54% by weight of a peptide having a molecular weight of about 5 to about 20 kDa; c) about 2 to about 20% by weight of a peptide having a molecular weight of about 1 to about 5 kDa; d) about 2 to about 20% by weight of a peptide having a molecular weight of less than about 1 kDa; or e) Any combination of any two or more of a) to d) may be included.

[0045] In various embodiments, the total protein is a) about 20 to about 79% by weight of a peptide having a molecular weight greater than about 20 kDa; b) about 15 to about 54% by weight of a peptide having a molecular weight of about 5 to about 20 kDa; c) about 2 to about 17% by weight of a peptide having a molecular weight of about 1 to about 5 kDa; d) about 2 to about 20% by weight of a peptide having a molecular weight of less than about 1 kDa; or e) Any combination of any two or more of a) to d) is also acceptable.

[0046] In various embodiments, the total protein is a) about 35 to about 65% of peptides having a molecular weight greater than about 20 kDa; b) about 25 to about 50% by weight of a peptide having a molecular weight of about 5 to about 20 kDa; c) about 4 to about 12% by weight of a peptide having a molecular weight of about 1 to about 5 kDa; d) about 2 to about 6% by weight of a peptide having a molecular weight of less than about 1 kDa; or e) Any combination of any two or more of a) to d) may be included.

[0047] In various embodiments, the milk proteins may be at least partially hydrolyzed. In various embodiments, the degree of hydrolysis of the total protein may be less than about 10%, less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 1%, less than about 0.9%, less than about 0.8%, less than about 0.75%, less than about 0.7%, less than about 0.65%, less than about 0.5%, less than about 0.25%, less than about 2%, less than about 0.15%, or less than about 0.1%.

[0048] In various embodiments, the dairy protein composition may comprise from about 50% to about 90% total protein by weight based on the dry matter in the composition.

[0049] In various embodiments, the protein-containing food product may be a liquid nutritional composition, a beverage, ice cream, an acidified product, a fermented product, buttermilk, cheese, processed cheese, a cheese analog, quark, a pudding, a frozen dessert, a coffee whitener, a gel, a bar, or a baked good.

[0050] In various embodiments, the fermented product may be yogurt, milk, kefir, skyr, petit swiss, ambient yogurt, fermented milk drink, smoothie, or sour cream. In various embodiments, the yogurt is drinking yogurt, set yogurt, Greek-style yogurt, or stirred yogurt.

[0051] In various embodiments, the acidified product can be an acidified milk drink, yogurt, cheese, processed cheese, cheese analogs, or buttermilk.

[0052] In various embodiments, the beverage may be a dairy drink, an acidic drink, a juice, a smoothie, or a sports drink. In various embodiments, the dairy drink may be a liquid nutritional composition, low-lactose milk, flavored milk, or fortified milk.

[0053] In various embodiments, the food product may include a dairy base, which may include one or more of skim milk, skim milk powder, whey protein concentrate, whey protein isolate, whole milk powder, whole milk, lactose, standardized milk protein concentrate, caseinate, cream, anhydrous milk fat, and fat-filled milk powder.

[0054] In various embodiments, the food product may include one or more lipids, carbohydrates, proteins, flavors, vitamins, minerals, dairy products, water, food additives, colors, fruit preparations, or combinations of any two or more of these ingredients.

[0055] In various embodiments, the protein-containing food product has reduced hardness and / or viscosity compared to a control food product having the same ingredient composition, casein and protein content as the food product of the present invention, except that it does not contain the dairy protein composition of the present invention.

[0056] In various embodiments, the food product is a solid or solidified gel having a reduced firmness of about 40 to about 80% compared to a control solid or solidified gel, hi other embodiments, the food product is a semi-solid or liquid food product and exhibits a reduced viscosity of about 40 to about 99% compared to a control semi-solid or liquid food product.

[0057] In various embodiments, the mouthfeel (e.g., hardness or thickness) is reduced and / or there is negligible or no increase in objectionable flavors (e.g., bitterness or saltiness) compared to a control food having the same ingredient composition, casein and protein content as the food product of the present invention, except that it does not include the milk protein composition of the present invention.

[0058] In various embodiments, the aqueous composition may contain less than about 2.5 g calcium per 100 g casein.

[0059] In various embodiments, the method of preparing a milk protein composition may include subjecting the milk protein composition to ion exchange chromatography, calcium chelation and / or ultrafiltration under acidic conditions to produce a wet milk protein composition, hi various embodiments, the calcium in the composition is reduced by at least about 40% by weight, or from about 40% to about 99% by weight.

[0060] In various embodiments, the method of preparing the dairy protein composition further comprises drying the dairy protein composition to form a powdered (dry) dairy protein composition.

[0061] In various embodiments, the proteolytic enzyme may comprise one or more proteases. In various embodiments, the protease is an endopeptidase.

[0062] In various embodiments, the protease may be a metalloprotein endopeptidase or a serine endopeptidase. In various embodiments, the metalloprotein endopeptidase may be a zinc endopeptidase.

[0063] In various embodiments, the proteolytic enzyme may include chymotrypsin, trypsin, pepsin, papain, bacillolysin, pancreatin, bromelain, carboxypeptidase, or a combination of any two or more thereof.

[0064] In various embodiments, the proteolytic enzyme may have optimal activity at a pH between about pH6 and about pH11.

[0065] In various embodiments, the proteolytic enzyme is derived from a Bacillus genus, such as Bacillus amyloliquefaciens, an Aspergillus genus, such as Aspergillus oryzae, or a Fusarium genus.

[0066] In various embodiments, the method comprises: a) a temperature of about 0°C to about 85°C; b) About 30 seconds to about 48 hours, c) a pH of about pH 6 to about pH 8, or d) may include a step of exposing the aqueous composition to the action of one or more proteases in any combination of two or more of a) to c).

[0067] In various embodiments, the method may include exposing the aqueous composition to the action of one or more proteolytic enzymes at a pH of about pH 4 to about pH 11, about pH 6 to about pH 11, about pH 6 to about pH 10, about pH 6 to about pH 9, or about pH 6 to about pH 8.

[0068] In various embodiments, the method may include adding one or more food grade acids or acidic substances, which may be selected from glucono-delta-lactone (GDL), lactic acid, citric acid, malic acid, acetic acid, tartaric acid, fumaric acid, hydrochloric acid, phosphoric acid, sulfuric acid, and combinations of any two or more thereof.

[0069] In various embodiments, the medium is a bacterial medium, which is selected from the group consisting of Lactobacillus, Streptococcus, Leuconostoc, Lactococcus, Lactobacillus subsp. Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus, or Lactobacillus casei.

[0070] In various embodiments, acidification or incubation of the aqueous intermediate composition inactivates one or more proteolytic enzymes.

[0071] In various embodiments, the yogurt comprises: a) Hardness of about 300g to about 8000g, b) a breaking force of about 10 to about 500 g; or c) A yogurt containing (a) and (b).

[0072] In various embodiments, the yogurt is 50s -1 The viscosity of the stirred yogurt is about 1 to about 4000 mPa·s.

[0073] In various embodiments, the yogurt is a drinking yogurt having a pourable uniform consistency.

[0074] Any of the embodiments or preferences described herein may be relevant alone or in combination with any one or more of the aspects described herein, unless otherwise stated or indicated.

[0075] The invention may also be broadly described as consisting of any or all combinations of any two or more of the parts, elements and features individually or collectively referred to or indicated in the specification of this application. Where specific integers that have known equivalents in the art to which the invention pertains are referred to herein, such known equivalents are deemed to be incorporated herein as if individually set forth.

[0076] Reference to a range of numerical values ​​disclosed herein (e.g., 1 to 10) also explicitly defines all rational numbers within that range (e.g., 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9, and 10), and ranges of rational numbers within that range (e.g., 2-8, 1.5-5.5, and 3.1-4.7), and therefore all subranges of all ranges. What is disclosed herein is hereby expressly disclosed. These are merely examples of what is specifically intended, and all possible combinations of numerical values ​​between the lowest and highest values ​​recited are considered to be expressly disclosed in this application in the same manner.

[0077] References are made herein to external sources, including patents and other documents, generally for the purpose of providing a context for discussing features of the invention. Unless otherwise stated, reference to such sources should not be construed as an admission that such sources are prior art or form part of the common general knowledge in the art in any jurisdiction. [Brief description of the drawings]

[0078] The invention will now be described, by way of example only, with reference to the accompanying drawings in which: [Figure 1] 1 is a flow diagram illustrating an exemplary process for producing the dairy protein compositions of the present invention. [Diagram 2] 1 is a flow chart illustrating an exemplary method for producing a protein-containing food product of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0079] The present invention relates to low calcium (calcium deficient) partially hydrolyzed dairy protein compositions and methods for their production. The present invention also relates to food products, such as acidic and fermented foods, comprising the dairy protein compositions and methods for producing such foods. The present invention also relates to food products prepared using calcium deficient MPC, where partial hydrolysis of the dairy proteins is carried out "on-line" during food preparation.

[0080] Dairy protein compositions and / or food products prepared using the methods described herein have reduced viscosity and / or firmness, while maintaining desirable texture, flavor and other properties, compared to known food products that contain alternative dairy protein ingredients. 1.Definition

[0081] The term "milk protein concentrate (MPC)" refers to a milk protein product in which 40% or more of the weight of the non-fat solids is protein or 70% or more, 80% or more, 85% or more of the weight of the non-fat solids is protein and the weight ratio of casein to whey protein is between about 95:10 and about 50:50, preferably between 90:10 and 80:20. MPCs with a milk protein content of 90% or more are sometimes referred to as milk protein isolates (MPIs). When MPCs are mentioned, MPIs should be considered to be included where applicable in the context. Milk protein concentrates may also include modified MPCs, such as calcium-deficient MPCs or other counterion modified MPCs. Such concentrates are known to those skilled in the art and are often expressed as "MPC" followed by the percentage of dry matter of the milk protein. For example, MPC70 is an MPC that uses 70% of the dry matter as milk protein.

[0082] The phrase "calcium depleted" is used herein to refer to compositions such as milk protein concentrates (MPCs) in which the concentration of calcium bound to casein is reduced and is lower than the concentration of calcium bound to casein in a corresponding non-deficient composition. Such compositions may also be depleted in other divalent cations, such that the concentration of divalent cations (e.g., magnesium) bound to casein is lower compared to the corresponding non-deficient composition. Similarly, calcium in casein refers to bound calcium, which is calcium bound to casein.

[0083] As used herein, the term "partially hydrolyzed" refers to milk proteins that have been acted upon by one or more proteolytic enzymes.

[0084] "Casein" refers to a compound of casein and metal ions, produced by acid precipitation of casein and redissolving with a base containing the metal ion. Hydroxide solutions containing sodium, potassium or ammonium can be used to produce sodium caseinate, potassium caseinate or ammonium caseinate. Suitable descriptions of caseinates and methods of producing caseinates as used herein can be found in Fox & McSweeney, 2003 and Dairy Processing Handbook, 2003.

[0085] Here, "food" refers to compositions including food and beverages for human or animal consumption. Consumption occurs through eating and drinking. In various embodiments, the food provided herein meets food safety standards required by the U.S. Food and Drug Administration (FDA), the U.S. Department of Agriculture, the European Food Safety Authority, and / or other state or local food regulatory agencies. The term includes compositions that can be combined with or added to other ingredients to make a composition that can be ingested by humans or animals.

[0086] The term "liquid nutritional composition" refers to an aqueous composition that is preferably administered orally or perorally. Alternatively, the liquid nutritional composition may be administered by other means, such as tube feeding into the patient's stomach, including nasogastric feeding and gastric feeding. Liquid nutritional compositions include "medical foods," "enteral nutrients," "foods for special medical purposes," liquid meal replacements, and supplements. Liquid nutritional compositions provide high amounts of protein, carbohydrates, and normal fats, and optionally vitamins and minerals. In an exemplary embodiment, the liquid nutritional composition provides a balanced diet.

[0087] As used herein, a "peptide" refers to a compound consisting of two or more amino acids linked in a chain form via a bond between the carboxyl group of one amino acid and the amino group of the adjacent amino acid. The term includes peptides and proteins of any length or molecular weight, including peptides or proteins containing two or more amino acids, for example, peptides or proteins containing 2-250, 2-300, or 2-400 amino acids, or peptides or proteins with molecular weights from less than 1 kDa to more than 20 kDa. The term includes peptide or protein fragments cleaved from a longer peptide or protein by hydrolysis, and unhydrolyzed or "intact" peptides or proteins.

[0088] As used herein, the term "milk protein" refers to the value calculated using a milk protein conversion factor from the percentage of nitrogen in a sample using the following formula: % Milk Total Protein = % Nitrogen x 6.38 Conniff, editor, 1997. § 33.2.11 AOAC Official Method 991.20 Nitrogen in Milk (Total). Official Methods of Analysis of AOAC International. Sixteenth Edition and Third Revision, Volume 2, AOAC International. Gaithersburg, Maryland (Chapter 33, p. 11)

[0089] As used herein, the term "total protein" refers to all protein from any source or ingredient present in the composition. "Total milk protein" refers to all of the milk-derived proteins in the composition, particularly the caseins and whey proteins.

[0090] As used herein, the term "intact casein" refers to casein in a milk protein composition that does not undergo substantial hydrolysis after being subjected to the action of proteolytic enzymes.

[0091] The term "comprises" as used herein means "comprises at least partially." In interpreting statements herein that include this term, the feature preceded by the term must be present in each statement, although other features may also be present. Related terms such as "comprises" and "included" are to be interpreted in the same manner. 2. Milk Protein Composition

[0092] In one aspect, the invention generally provides dairy protein compositions comprising dairy proteins, where the dairy protein compositions comprise dairy proteins. a) the composition comprises at least about 40% total protein by weight based on the dry matter in the composition; b) Milk proteins include casein; c) the total protein contains less than about 79% by weight of peptides having a molecular weight greater than about 20 kDa; d) The composition comprises: i. less than about 2 g calcium per 100 g total protein, and / or ii. Contains less than about 1.4 g calcium per 100 g dry matter in the composition.

[0093] In various embodiments, the milk protein may comprise a milk protein concentrate (MPC), a milk protein isolate (MPI), caseinate, casein, casein co-precipitate, a retentate obtained by ultrafiltration or microfiltration of milk, or any combination of any two or more thereof. In various embodiments, the milk protein may comprise a milk protein concentrate (MPC), a milk protein isolate (MPI), or a combination thereof. In various embodiments, the milk protein may comprise a milk protein concentrate (MPC) or a milk protein isolate (MPI).

[0094] In one aspect, the present invention provides a milk protein composition comprising a milk protein concentrate, a milk protein isolate, or a combination thereof, a) the composition comprises at least about 40% total protein by weight based on the dry matter in the composition; b) the total protein contains less than about 79% by weight of peptides having a molecular weight greater than about 20 kDa; c) The composition comprises i. less than about 2 g calcium per 100 g total protein, and / or ii. Contains less than about 1.4 g calcium per 100 g dry matter in the composition.

[0095] In various embodiments, the composition comprises at least about 40%, about 45%, about 50%, about 60%, about 70%, about 80%, about 90%, or at least about 95% total protein by weight based on dry matter in the composition, and various ranges can be selected from any two of these ranges. In various embodiments, the composition may comprise about 40 to about 99%, about 40 to about 90%, or about 40 to about 80% total protein by weight based on dry matter in the composition.

[0096] In various embodiments, the milk protein or milk protein composition may contain at least about 10, 20, 30, 40, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99 or 100% by weight of casein based on dry matter, and the weight of casein can be selected from various ranges selected from between any of the following values ​​based on dry matter: about 10 to about 100, about 40 to about 100, about 50 to about 100, about 60 to about 100, about 70 to about 100, about 75 to about 100, or about 80 to about 100, about 10 to about 90, about 40 to about 90, about 50 to about 90, about 60 to about 90, about 70 to about 90, about 75 to about 90, or about 80 to about 90% by weight.

[0097] In various embodiments, the milk protein or milk protein composition may comprise whey protein. In various embodiments, the milk protein or milk protein composition may comprise about 1 to about 50%, about 1 to about 40%, about 1 to about 30%, or about 1 to about 20% by weight of whey protein based on dry matter.

[0098] In various embodiments, whey protein may comprise or be provided by a component that comprises whey protein concentrate, whey protein isolate, or a combination thereof.Other suitable whey protein sources known in the art can be used.For example, in some embodiments, whey protein may comprise or be provided by a component that comprises whey liquid, such as cheese whey or acid whey.

[0099] The calcium-deficient milk proteins (particularly casein) present in the composition of the invention are subjected to the action of proteolytic enzymes to achieve partial hydrolysis, which results in a molecular weight distribution associated with the advantageous characteristics described herein.

[0100] In various embodiments, the total protein may contain less than about 79%, 78%, 77%, 76%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, or 40% by weight of peptides having a molecular weight greater than about 20 kDa.

[0101] In various embodiments, the total protein may comprise 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, or 70 to about 79% by weight of peptides having a molecular weight greater than about 20 kDa, and various ranges may be between any two of these values, such as, for example, about 25 to about 79%, about 30 to about 79%, about 35 to about 79%, about 40 to about 79%, about 50 to about 79%, about 20 to about 75%, about 25 to about 75%. , about 30 to about 75%, about 35 to about 75%, about 40 to about 75%, about 50 to about 75%, about 20 to about 70%, about 25 to about 70%, about 30 to about 70%, about 35 to about 70%, about 40 to about 70%, about 50 to about 70%, about 20 to about 65%, about 25 to about 65%, about 30 to about 65%, about 35 to about 65%, about 40 to about 65%, about 50 to about 65%, about 20 to about 60%, about 30 to about 60%, about 40 to about 60%, about 20 to about 55%, about 30 to about 5%, or about 40 to about 60%.

[0102] In various embodiments, the total protein may comprise about 15, 20, 25, 30, 35, 40, 45, 50, or about 55% by weight of peptides having a molecular weight of about 5 to about 20 kDa, and various ranges may be between any two of these values, for example, about 15 to about 55%, about 15 to about 50%, about 15 to about 45%, about 15 to about 40%, about 15 to about 35%, 20 to about 50%, about 20 to about 45%, about 20 to about 40%, about 20 to about 35%, 25 to about 50%, about 25 to about 45%, about 25 to about 40%, about 25 to about 35%, 30 to about 50%, about 30 to about 45%, or about 30 to about 40% by weight.

[0103] In various embodiments, the total protein may contain less than about 30% peptides having a molecular weight less than about 5 kDa, or less than about 28%, less than about 25%, less than about 24%, less than about 23%, less than about 22%, less than about 21%, or less than about 20% peptides having a molecular weight less than about 5 kDa.

[0104] In various embodiments, the total protein may contain about 5%, 10%, 15%, 20%, 21%, 22%, 23%, 24%, or 25 to about 30% peptides having a molecular weight of less than about 5 kDa, and various ranges may be between any two of these values, such as various ranges such as about 5 to about 25%, or about 10 to about 25%, or about 15 to about 25%, or about 20 to about 25%, about 5 to about 22%, about 5 to about 21%, or about 5 to about 20%, or about 10 to about 20%, or about 15 to about 20%, or about 5 to about 15%, or about 10 to about 15% by weight.

[0105] In various embodiments, the total protein may contain less than about 20% peptides having a molecular weight in the range of about 1 to about 5 kDa, or less than about 18%, less than about 16%, less than about 14%, or less than about 12% peptides having a molecular weight in the range of about 1 to about 5 kDa.

[0106] In various embodiments, the total milk protein may contain about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18% to about 20% peptides having a molecular weight in the range of about 1 to about 5 kDa, and various ranges may be between any two of these values, for example, about 2 to about 18%, about 2 to about 16%, about 2 to about 15%, about 2 to about 14%, about 2 to about 12%, about 2 to about 10%, about 2 to about 9%, about 3 to about 20%, about 3 to about 18%, about 3 to about 16%, about 3 to about 15%, about 3 to about 14%, or about 3 to about 12%, about 3 to about 10%, about 3 to about 9%, about 4 to about 20%, about 4 to about 18%, about 4 to about 16%, about 4 to about 15%, about 4 to about 14%, or about 4 to about 12%, about 4 to about 10%, about 4 to about 9%, about 5 to about 20%, 5 to about 18%, about 5 to about 16%, about 5 to about 15%, about 5 to about 14%, or about 5 to about 12%, about 5 to about 10%, or about 5 to about 9% by weight.

[0107] In various embodiments, the total protein may contain less than about 20% peptides having a molecular weight less than about 1 kDa, or less than about 15%, less than about 10%, or less than about 9% peptides having a molecular weight less than about 1 kDa.

[0108] In various embodiments, the total protein may contain 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15-20% by weight of peptides having a molecular weight of less than about 1 kDa, and various ranges may be between any two of these values, such as about 2 to about 20%, about 2 to about 15%, about 2 to about 10%, about 2 to about 9%, about 2 to about 8%, about 2 to about 7%, about 2 to about 6%, about 2 to about 5%, about 3 to about 20%, about 3 to about 15%, about 3 to about 10%, about 3 to about 9%, about 3 to about 8%, about 3 to about 7%, about 3 to about 6%, about 3 to about 5%, about 5 to about 20%, about 5 to about 15%, or about 5 to about 10% by weight.

[0109] In various embodiments, the total protein may have a peptide molecular weight distribution corresponding to the following molecular weight distribution: a) about 20%, 25%, 30%, 35%, 40%, 45%, or 50 to about 79% of peptides having a molecular weight greater than about 20 kDa; b) about 15, 20, 25, 30, 35, 40, 45, 50, or about 55% by weight of a peptide having a molecular weight of about 5 to about 20 kDa; c) about 2%, 3%, 4%, 5%, 10%, 12%, 14%, 16%, 18 to about 20% by weight of a peptide having a molecular weight of about 1 to about 5 kDa, and d) about 2%, 3%, 4%, 5%, 10%, 15 to about 20% by weight of peptides having a molecular weight of less than about 1 kDa.

[0110] In various embodiments, the total protein may have a peptide molecular weight distribution corresponding to the following molecular weight distribution: a) about 20 to about 79% of peptides having a molecular weight greater than about 20 kDa; b) about 15 to about 54% by weight of a peptide having a molecular weight of about 5 to about 20 kDa; c) about 2 to about 17% by weight of a peptide having a molecular weight of about 1 to about 5 kDa, and d) about 2 to about 20% by weight of a peptide having a molecular weight of less than about 1 kDa.

[0111] In various embodiments, the total protein is a) less than about 79%, 78%, 77%, 76%, 75%, 70%, 65%, 60%, 55%, 50%, 45% or 40% by weight of peptides having a molecular weight greater than about 20 kDa; b) It may contain about 15 to about 55%, about 15 to about 50%, about 15 to about 45%, about 15 to about 40%, about 15 to about 35%, 20 to about 50%, about 20 to about 45%, about 20 to about 40%, about 20 to about 35%, 25 to about 50%, about 25 to about 45%, about 25 to about 40%, about 25 to about 35%, 30 to about 50%, about 30 to about 45%, or about 30 to about 40% by weight.

[0112] In various embodiments, the total protein is a) less than about 79%, 78%, 77%, 76%, 75%, 70%, 65%, 60%, 55%, 50%, 45% or 40% of peptides having a molecular weight greater than about 20 kDa; b) may contain less than about 30%, less than about 28%, less than about 25%, or less than about 20% peptides having a molecular weight of less than about 5 kDa.

[0113] In various embodiments, the total protein is a) less than about 79%, 78%, 77%, 76%, 75%, 70%, 65%, 60%, 55%, 50%, 45% or 40% of peptides having a molecular weight greater than about 20 kDa; b) It may contain less than about 20%, less than about 18%, less than about 16%, less than about 14%, or less than about 12% peptides having a molecular weight in the range of about 1 to about 5 kDa.

[0114] In various embodiments, the total protein is a) less than about 79%, 78%, 77%, 76%, 75%, 70%, 65%, 60%, 55%, 50%, 45% or 40% of peptides having a molecular weight greater than about 20 kDa b) may contain less than about 20%, less than about 15%, or less than about 10% peptides having a molecular weight of less than about 1 kDa.

[0115] The molecular weight distribution of the peptides present in the composition of the present invention can be measured by the following method. XL Column (2 x 30 cm) and TSKSW XLProteins / peptides are separated using a size-exclusion HPLC method using guard columns and maintaining all columns at 30 °C (TOSOH Corporation). Samples are dissolved in a mobile phase buffer consisting of 0.1 M potassium phosphate pH 6.0, 0.3 M potassium chloride and 6 M urea to a final protein concentration of 2-4 mg / mL. Reference standards (glutathione (reduced, 307 Da), insulin B chain (oxidized, 3,496 Da), myoglobin (16,952 Da), carbonic anhydrase (28,982 Da), glycerylaldehyde-3-phosphate dehydrogenase (35,688 Da)) are introduced before and after each sample set. Samples are run for a total of 70 min at a flow rate of 0.45 ml / min using an injection volume of 50 μL of sample solution between 2 and 4 mg / ml. By monitoring the absorbance at 220 nanometers, , to detect proteins and peptides. The retention times of the standards are fitted to a quadratic curve and the resulting equation is used to calculate retention times corresponding to four molecular weight ranges, namely >20 kDa, 5-20 kDa, 1-5 kDa, and <1 kDa. For the chromatograms of the hydrolysate samples, the area under the curve for each molecular weight range is calculated and expressed as a percentage of the total protein / peptide material. Other suitable methods for determining molecular weight distribution are well known and obvious to those skilled in the art.

[0116] In various embodiments, the milk protein may be partially hydrolyzed. In various embodiments, the degree of hydrolysis of the milk protein is less than about 10%, less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, less than about 1%, less than about 0.75%, less than about 0.5%, less than about 0.25%, less than about 0.2%, less than about 0.15%, or less than about 0.1%, and suitable ranges may be selected from these values, such as about 0.1 to about 10%, about 0.1 to about 5%, or about 0.1 to about 3%.

[0117] The degree of hydrolysis is measured using the o-phthalaldehyde OPA method first described by Church et al. 1983 (J Dairy Sci, 66(6), 1219-1227), but with modified reducing agents reported by Frister et al. 1988 (Fessenhuis, J. Analytical Chemistry, 330, 631-633).

[0118] Briefly, samples are diluted in water to a level where the absorbance is within the range of the standard absorbance. Glycine is diluted in water to obtain a standard curve of 0.25-1.00 mol / mL glycine. 40 mg OPA (dissolved in 1 mL methanol or ethanol) is added to 25 mL of 100 mM sodium tetraborate and 2.5 mL of 20% (wt / wt) SDS, and the volume is adjusted to 50 mL with water. 100 mg of N,N-dimethyl-2-mercaptoammonium chloride is added instead of β-mercaptoethanol. Three milliliters of the prepared MOPA reagent are added to 0.4 milliliters of samples and standards in colorimetric dishes, and the absorbance is read accurately for 2 minutes using a spectrophotometer set at 340 nm. The degree of hydrolysis of the samples is calculated using the standard curve by calculating the number of amino groups and then the percentage of the theoretical total number of peptide bonds per gram of protein. Lysine side chains are taken into account in the calculation.

[0119] In various embodiments, the dairy protein composition comprises less than about 2 g calcium per 100 g total protein, or less than about 1.9, 1.8, 1.6, 1.5, 1.4, 1.2, 1, 0.8, 0.75, 0.6, 0.5, 0.4, 0.3, 0.2 or 0.1 g calcium per 100 g total protein, with various ranges between any two of these values, e.g., about 0.1 to about 2, about 0.5 to about 2, about 1 to about 2, about 0.1 to about 1.5, about 0.5 to about 1.5, or about 1 to about 1.5 g calcium per 100 g total protein.

[0120] In various embodiments, the dairy protein composition contains less than about 1.4 g calcium per 100 g dry matter, or less than about 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, 0.5, or 0.4 g calcium per 100 g dry matter.

[0121] Calcium content can be measured using inductively coupled plasma optical emission spectroscopy. Improved wet digestion procedures and ICP-OES measurements were performed according to the methods described in Methods for the Determination of Metals and Inorganic Chemicals in Environmental Samples. Method 200.2: Sample Preparation Procedures for the Spectrochemical Determination of Total Recoverable Elements. Environmental Systems Monitoring Laboratory, Environmental Protection Agency, Cincinnati, Ohio, 1994.

[0122] In various embodiments, the dairy protein composition comprises less than 65, 60, 55, or about 50 grams of intact casein per 100 grams of composition dry matter.

[0123] Intact caseins and hydrolysis products can be identified and quantified using a variety of methods including SDS-PAGE (Laemmli UK: Cleavage of structural proteins during phage T4 head assembly. Cleavage of structural proteins during head assembly of bacteriophage T4. Nature. 1970, 227: 680-685.10.1038 / 227680a0) and microfluidic SDS electrophoresis techniques (SG Anema, 2009 Separation and quantification of milk proteins using a "lab-on-a-chip" microfluidic SDS electrophoresis technique. International Journal of Dairy Science, Vol. 19, No. 4, April 2009, pp. 198-204), size exclusion HPLC and reversed-phase HPLC (Maurmayr et al. (2013)), which are used to characterize the abundance of α-casein in milk. s1 -, α s2-, β-, κ-casein, α-whey protein, β-lactoglobulin and lactoferrin were measured and quantified (Agricultural Science Introduction 78,(3),201-205.). SDS-PAGE and size-exclusion HPLC techniques separate proteins and peptides by mass, whereas RP-HPLC separates proteins and peptides based on hydrophobicity / hydrophilicity. These methods can determine the reduction of intact casein in hydrolyzed material, as long as the intact casein in the starting material can be compared. Using identical protein content, the reduction of intact casein appears as a reduction in the protein band intensity in the various electropherograms, allowing comparison of hydrolyzed and unhydrolyzed material. If the initial casein content of the material is known, these techniques are useful to show the reduction of intact casein in the product.

[0124] In cases where the casein content in the raw material is unknown, the HPLC-MS peptide spectrum method can be used to measure hydrolyzed casein and whey proteins. If marker casein and whey peptides are synthesized as standards, absolute quantification is possible, which is routinely done (McGrath (2016) Proteomic characterization of the thermally induced hydrolysis of sodium caseinate. International Journal of Dairy Science 53, 51-59). This allows for an estimation of the complete casein reduction and the casein and whey content provided in the starting material.

[0125] In various embodiments, the dairy partial composition is in at least a partial form, e.g., the dairy protein composition is a powder. In other embodiments, the dairy protein composition is in liquid form. In some embodiments, the dairy protein composition comprises a liquid that has been reconstituted from a powder composition.

[0126] In various embodiments, the composition may include an inactivated proteolytic enzyme. 3. Method for producing milk protein composition

[0127] A method for producing a calcium-deficient, partially hydrolyzed dairy protein composition is disclosed. raw materials

[0128] Any suitable dairy protein source can be used to prepare the dairy protein compositions according to the methods disclosed herein.

[0129] In various embodiments, the dairy protein may comprise a milk protein concentrate (MPC), a milk protein isolate (MPI), caseinate, casein, casein co-precipitate, or any combination of any two or more thereof. In various embodiments, the dairy protein may comprise a milk protein concentrate (MPC), a milk protein isolate (MPI), or a combination thereof. In various embodiments, the dairy protein may comprise a milk protein concentrate (MPC) or a milk protein isolate (MPI).

[0130] Generally, MPCs are prepared by invoking an ultrafiltration process to prepare a logistics rich in casein and whey protein. In another embodiment, a milk protein concentrate may be prepared by mixing a skim milk stream with a whey protein concentrate stream, treating the skim milk stream or the combined stream with cation exchange, and optionally concentrating or drying. Suitable MPCs for use herein may be prepared from a mixture of MPCs.

[0131] In various embodiments, the caseinate may be sodium caseinate, ammonium caseinate, potassium caseinate, or a combination of any two or more thereof.

[0132] Casein co-precipitates contain casein and whey and may be obtained by a combination of heating and acidification to obtain a coagulum, followed by processing and drying of the coagulum. In addition to calcium co-precipitates, casein co-precipitates prepared by methods known to those skilled in the art are suitable.

[0133] In various embodiments, milk proteins can include or be provided from the retentate obtained by ultrafiltration or microfiltration of milk.

[0134] In various embodiments, the milk protein concentrate may be prepared by a process that includes ultrafiltration of fresh liquid milk and percolation to produce a retentate.

[0135] The milk protein may be provided in the form of calcium-deficient MPC. Calcium-deficient MPC is an MPC that has a lower calcium content than the corresponding non-calcium-deficient MPC. These products also typically have a lower content of other divalent cations, such as magnesium, compared to the corresponding non-consumable products. The calcium-deficient MPC is preferably dried to a moisture content of less than 6%, or a water activity level that is favorable for storage of the dried ingredient for several months without undue deterioration.

[0136] The preferred MPC of the present invention has calcium engineered by cation exchange method.The production and application of these calcium-deficient MPCs have been previously disclosed in US Patent 7,157,108, published PCT application WO2008 / 026940, and published US patent application 2010 / 0021595.These documents are fully incorporated herein by reference.Other methods for preparing calcium-deficient MPCs will be apparent to those skilled in the art.

[0137] In other embodiments, the milk protein may be provided in the form of a non-calcium deficient milk source such as MPC or MPI. The milk protein may be subjected to a calcium depletion step before or after proteolysis to reduce the calcium content.

[0138] Aqueous compositions comprising dairy protein may be prepared from a powdered or liquid dairy protein source, such as powdered or liquid MPC or retentate. Aqueous compositions comprising dairy protein may be formed by mixing two or more source materials together to obtain desired properties of the starting composition, such as protein and calcium levels.

[0139] In various embodiments, the aqueous composition contains less than about 2.5 grams of calcium per 100 grams of casein, or less than about 2.4, 2.2, 2.0, 1.8, 1.6, 1.4, 1.2, 1.0, or 0.8 grams of calcium per 100 grams of casein. In various embodiments, the aqueous composition contains from about 0.8 to about 2.5 grams, or from about 1.0 to about 2.5 grams, or from about 1.4 to about 2.5 grams, or from about 2.0 to about 2.5 grams, or from about 0.8 to about 2.0 grams, or from about 1.0 to about 2.0 grams, or from about 1.4 to about 2.0 grams.

[0140] In various embodiments, the method may include subjecting the milk protein composition to ion exchange chromatography, calcium chelation, and mixing with carbon dioxide, followed by filtration and / or ultrafiltration under acidic conditions to reduce the calcium in the composition by at least about 40, 50, 60, 70, 80, 90, 95, or 99% by weight, or to reduce the calcium in the composition by about 40 to about 99%, about 50 to about 99%, about 60 to about 99%, about 70 to about 95%, or about 40 to about 95%, or about 50 to about 95%, about 60 to about 95%, or about 40 to about 90%, or about 50 to about 90%, or about 60 to about 90%, or about 70 to about 90% by weight to produce an aqueous milk protein composition.

[0141] In various embodiments, ion exchange chromatography may involve exchanging calcium in the milk protein composition for sodium, potassium, or a combination thereof.

[0142] In various embodiments, the aqueous composition may comprise at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 15, 16, 18, or at least about 20% total protein by weight based on the dry matter in the composition, and various ranges may be between any two of these values, such as from about 2 to about 20, 4 to about 20, 5 to about 20, 8 to about 20, 2 to about 18, 4 to about 18, 5 to about 18, or about 8 to about 20% by weight.

[0143] The dairy protein compositions of the present invention may be used directly in the preparation of protein-containing food products, or may be packaged aseptically in liquid form or dried and packaged in powder form. Protein Degradation

[0144] In various embodiments, the method of the present invention includes exposing the aqueous composition to one or more proteases to at least partially hydrolyze the milk proteins in the composition. As used herein, unless otherwise defined, "subjected to the action of one or more proteases" or analogues means that at least a portion of the peptides in the milk proteins are hydrolyzed, resulting in at least a portion of the milk proteins in the composition being hydrolyzed. To achieve at least partial hydrolysis, the conditions for the proteases must be optimized, e.g., temperature, pH and treatment time must be adjusted to achieve the proteolytic activity of the enzyme.

[0145] Any proteolytic enzyme that achieves at least partial hydrolysis of at least a portion of the milk proteins can be used.

[0146] In various embodiments, the protease may include one or more proteases. In various embodiments, the one or more proteases belong to Enzyme Commission (EC) class 3.4.21 (serine proteases), 3.4.24 (metalloendopeptidases), or 3.4.17 (carboxypeptidases). In various embodiments, the one or more proteases are subtilisins, serine proteases, acid proteases, or neutral proteases. Examples of suitable protease enzymes are described in WO2016164096A1. In various embodiments, the protease is an endopeptidase.

[0147] In various embodiments, the one or more proteases may belong to one or more of Enzyme Commission (EC) classes 3.4.21 (serine endopeptidases), 3.4.22 (cysteine ​​endopeptidases), 3.4.24 (metalloendopeptidases), and 3.4.17 (metallocarboxypeptidases). In various embodiments, the one or more proteases may be subtilisins, serine proteases, acid proteases, alkaline proteases, or neutral proteases.

[0148] In various embodiments, the one or more proteases are selected from the group consisting of EC class 3.4.17, 3.4.21, 3.4.22, 3.4.23.1, 3.4.23.2, 3.4.23.3, 3.4.23.5, 3.4.23.12, 3.4.23.15, 3.4.23.16, 3.4.23.17, 3.4.23.19, 3.4.23.20, 3.4.23.21, 3.4.23.22, 3.4.23.23, 3.4.23.24, 3.4.23.25, 3.4.23.26, 3.4.23.28, 3.4.23.29, The class may belong to one or more of 3.4.23.30, 3.4.23.31, 3.4.23.32, 3.4.23.34, 3.4.23.35, 3.4.23.36, 3.4.23.38, 3.4.23.39, 3.4.23.40, 3.4.23.41, 3.4.23.42, 3.4.23.43, 3.4.23.44, 3.4.23.45, 3.4.23.46, 3.4.23.47, 3.4.23.48, 3.4.23.49, 3.4.23.50, 3.4.23.51, 3.4.23.52, or 3.4.24. In various embodiments, the one or more proteases are selected from the group consisting of EC class 3.4.21, 3.4.22, 3.4.23.1, 3.4.23.2, 3.4.23.3, 3.4.23.5, 3.4.23.12, 3.4.23.15, 3.4.23.16, 3.4.23.17, 3.4.23.19, 3.4.23.20, 3.4.23.21, 3.4.23.22, 3.4.23.23, 3.4.23.24, 3.4.23.25, 3.4.23.26, 3.4.23.28, 3.4.23.29, 3.4.23.30, 3.4.23.31, 3.4.23.32, 3.4.23.33, 3.4.23.34, 3.4.23.35, 3.4.23.36, 3.4.23.37, 3.4.23.38, 3.4.23.39, 3.4.23.40, 3.4.23.41, 3.4.23.42, 3.4.23.43, 3.4.23.44, 3.4.23.45, 3.4.23.46, 3.4.23.47, 3.4.23.48, 3.4.23.49, 3.4.24 ... .4.23.30, 3.4.23.31, 3.4.23.32, 3.4.23.34, 3.4.23.35, 3.4.23.36, 3.4.23.38, 3.4.23.39, 3.4.23.40, 3.4.23.41, 3.4.23.42, 3.4.23.43, 3.4.23.44, 3.4.23.45, 3.4.23.46, 3.4.23.47, 3.4.23.48, 3.4.23.49, 3.4.23.50, 3.4.23.51, 3.4.23.52, or 3.4.2.

[0149] In various embodiments, the proteolytic enzyme may comprise one or more proteases. In various embodiments, the protease is an endopeptidase.

[0150] In various embodiments, the protease may be a metalloprotein endopeptidase or a serine endopeptidase. In various embodiments, the metalloprotein endopeptidase may be a zinc endopeptidase.

[0151] In various embodiments, the proteolytic enzyme may include chymotrypsin, trypsin, pepsin, papain, bacillolysin, pancreatin, bromelain, carboxypeptidase, or a combination of any two or more thereof.

[0152] In various embodiments, the protease is not an aspartic endopeptidase. In various embodiments, the protease does not belong to the EC class 3.4.23.18.

[0153] In various embodiments, the proteolytic enzyme may have optimal activity at a pH between about pH6 and about pH11.

[0154] A proteolytic enzyme is considered to have optimal activity under particular conditions (e.g., a particular pH range or temperature) if the activity of the enzyme under the particular conditions is at least 60%, at least 65%, or at least 70%, or at least 75%, or at least 80%, or at least 85%, or at least 90%, or at least 95% of the maximum reaction rate of the enzyme's catalysis under those conditions.

[0155] In various embodiments, the proteolytic enzyme is derived from Bacillus, Fusarium, or plant material.

[0156] In various embodiments, the protease may be derived from a Bacillus species, e.g., Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus stearothermophilus, Bacillus licheniformis, Aspergillus species, e.g., Aspergillus oryzae, or Fusarium species. In various embodiments, the protease may be derived from a Bacillus species, e.g., Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus stearothermophilus, Bacillus licheniformis, or Fusarium species. In various embodiments, the protease may be derived from a Bacillus species, e.g., Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus stearothermophilus, Bacillus licheniformis, or Fusarium species.

[0157] In various embodiments, the proteolytic enzymes may be derived from plant materials, such as fruit or plant materials. In various embodiments, the proteolytic enzymes may be derived from fruit materials, such as pineapple or papaya. Examples of such enzymes include bromelain and papain.

[0158] In various embodiments, the method may include exposing the aqueous composition to one or more proteases at a temperature of about 0 to about 85°C, about 0 to about 50°C, about 0 to about 40°C, about 0 to about 30°C, about 0 to about 20°C, about 1 to about 85°C, about 1 to about 50°C, about 1 to about 40°C, about 1 to about 30°C, or about 1 to about 20°C.

[0159] In various embodiments, the method may include exposing the aqueous composition to the action of one or more proteases for from about 30 seconds to about 48 hours, from 1 minute to about 48 hours, from 1 minute to about 24 hours, from 1 minute to about 12 hours, from 1 minute to about 6 hours, from 2 minutes to about 24 hours, from 2 minutes to about 12 hours, from 2 minutes to about 6 hours, from 10 minutes to about 24 hours, from 10 minutes to about 12 hours, from 10 minutes to about 6 hours, from 20 minutes to about 24 hours, from 20 minutes to about 12 hours, or from about 20 minutes to about 6 hours.

[0160] The pH corresponding to the casein isoelectric point (pI) (pH 4.6) is preferably avoided before or after protein hydrolysis. At the pI of casein, casein may cause undesired aggregation in milk proteins. Thus, in various embodiments, the pH of the aqueous composition may be greater than pH 4.7, 4.8, 4.9 or pH 5. In various embodiments, the pH of the aqueous composition may be about pH 4.7 to about pH 8, about pH 4.8 to about pH 8, about pH 5 to about pH 8, about pH 5.5 to about pH 8, or about pH 6 to about pH 8.

[0161] In various embodiments, the method may include exposing the aqueous composition to one or more proteases at a pH of about pH 4.7 to about pH 8. In various embodiments, the method may include exposing the aqueous composition to the action of one or more proteases at a pH of about pH 6 to about pH 8.

[0162] In various embodiments, the method may include exposing the aqueous composition to the action of one or more proteases for about 30 seconds to about 48 hours at a temperature of about 0 to about 85° C. and a pH of about pH 4.7 to about pH 8. In various embodiments, the method may include exposing the aqueous composition to the action of one or more proteases for about 30 seconds to about 48 hours at a temperature of about 0 to about 85° C. and a pH of about pH 6 to about pH 8.

[0163] In various embodiments, the aqueous composition to which the protease is added is heated or incubated at a temperature for a period of time to achieve the degree of hydrolysis and / or molecular weight distribution of the milk protein required. As will be appreciated by those skilled in the art, the exact temperature and period of time will vary depending on the protease used. To achieve the desired protein hydrolysis, other conditions may be required, such as the presence of a specific cation and / or a specific pH to achieve optimal activity of the enzyme. Those skilled in the art can easily determine the appropriate conditions for a given protease using manufacturer information and basic testing without undue experimentation.

[0164] Suitable methods for inactivating one or more proteases will be apparent to the skilled artisan. In various embodiments, the method may include inactivating one or more proteases by one or more of the following inactivation methods: Heating at a temperature of at least about 70° C. for at least about 15 seconds; Change the pH to below pH4 or above pH11. Changing the solvent conditions (e.g., increasing ionic strength by adding salt) Use of enzyme inhibitors (e.g. EDTA), Evaporation and drying, Use of ultrafiltration membranes, immobilization on an inert support (e.g. Roehm Eupergit, carrageenan particles, chitosan particles, or any other suitable material) for use in stirred tank or fixed bed reactors, or in membrane or hollow fiber reactors; Using pulsed electric fields, and / or Use ultrasonic treatment.

[0165] In various embodiments, the method may include exposing the composition to one or more proteolytic enzymes to reduce intact casein in the composition by at least about 5%, about 10%, about 12%, about 15%, or at least about 18% by weight, with various ranges being between any two of these values, such as from about 5 to about 18%, or from about 10 to about 18%, or from about 12 to about 18%, or from about 15 to about 18%, or from about 5 to about 15%, or from about 10 to about 15%, or from about 12 to about 15%, or from about 5 to about 12%, or from about 10 to about 12%, or from about 5 to about 10%.

[0166] In various embodiments, the method may further include drying the dairy protein composition. Any suitable method of one skilled in the art may be used, including concentrating in an evaporator and / or dryer. In various embodiments, the dairy protein composition may be dried to form a powder. 4. Protein-rich foods

[0167] In another aspect, the present invention relates to a protein-containing food product comprising the composition of the present invention.

[0168] In another aspect, the present invention relates to the use of the composition of the present invention in the manufacture of a protein-containing food product.

[0169] In various embodiments, the protein-containing food product may be a liquid nutritional composition, a beverage, ice cream, an acidified product, a fermented product, buttermilk, cheese, processed cheese, a cheese analog, quark, a pudding, a frozen dessert, a coffee whitener, a gel, a bar, or a baked good.

[0170] In various embodiments, the fermented product may be yogurt, milk, kefir, skyr, petit swiss, ambient yogurt, fermented milk drink, smoothie, or sour cream. In various embodiments, the yogurt is drinking yogurt, set yogurt, Greek-style yogurt, or stirred yogurt.

[0171] In various embodiments, the fermented product may be yogurt, milk, kefir, skyr, petit suisse, ambient yogurt, fermented milk drink, smoothie, fromage frais, mascarpone, creme fraiche, or sour cream. In various embodiments, the yogurt may be drinking yogurt, set yogurt, Greek yogurt, filtered yogurt, or stirred yogurt.

[0172] In various embodiments, the acidified product can be an acidified milk drink, yogurt, cheese, processed cheese, cheese analogs, or buttermilk.

[0173] In various embodiments, the beverage may be a dairy drink, an acidic drink, a juice, a smoothie, or a sports drink. In various embodiments, the dairy drink may be a liquid nutritional composition, low-lactose milk, flavored milk, or fortified milk.

[0174] In various embodiments, the processed cheese may be a processed cheese spread, a "slice on slice" processed cheese, a processed cheese "lollipop," individually wrapped processed cheese slices, processed cheese triangles, processed cream cheese, processed cheese sauce, or a processed cheese block.

[0175] In various embodiments, the pH of the protein-containing food product may be from about pH3 to about pH8.

[0176] In various embodiments, food-safe acidic or alkaline additives can be used to adjust the pH of the protein-containing food. In various embodiments, the pH of the protein-containing food can be adjusted to about pH 3 to about pH 8, for example, about pH 4 to about pH 7, about pH 4 to about pH 6.8, about pH 5 to about pH 7, or about pH 5 to about pH 6.8. In various embodiments, the pH of the protein-containing food can be adjusted to about pH 6.8.

[0177] After calibration with standards of pH 4, 7, and 10 (Pronalys, LabServ), the pH can be measured by equilibrating the sample to 25°C and measuring with a pH probe (EC620132, Thermo Scientific). The pH can also be measured using a MeterLab PHM250 ion analyzer (Radiometer, Copenhagen). Other methods of measuring pH will be apparent to the skilled practitioner.

[0178] Food products prepared using the dairy protein compositions and / or methods may have reduced hardness and / or viscosity compared to a control food having the same ingredient composition, casein and protein content as the food product of the invention, except that the control food does not include a dairy protein composition of the invention and / or is not prepared by a method of the invention. In various embodiments, the food product is a solid or solidified gel having a reduced hardness of about 40 to about 80% compared to a control solid or solidified gel. In other embodiments, the food product is a semi-solid or liquid food product and exhibits a reduced viscosity of about 40 to about 99% compared to a control semi-solid or liquid food product.

[0179] Food products prepared using the dairy protein compositions and / or methods of the present invention exhibit reduced in-mouth texture (e.g., hardness or thickness) and / or negligible or no increase in undesirable flavors (e.g., bitterness or saltiness) compared to a control food product having the same ingredient composition, casein and protein content as the present invention, but lacking the dairy protein composition of the present invention.

[0180] In various embodiments, a protein-containing food product may comprise the dairy protein composition of the present invention and at least one lipid source.In various embodiments, a protein-containing food product may comprise the dairy protein composition of the present invention and at least one carbohydrate source.

[0181] In various embodiments, a protein-containing food product may be prepared by a process comprising providing a dairy protein composition of the present invention and mixing with at least one lipid source and at least one carbohydrate source.

[0182] In various embodiments, a protein-containing food product may comprise the dairy protein composition of the present invention, at least one lipid source, and at least one carbohydrate source.

[0183] In various embodiments, the protein-containing food product may contain at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 30%, 40%, or at least about 50% by weight total protein. In various embodiments, the protein-containing food product may contain from about 1 to about 50% by weight total protein, with useful ranges being between any two of these values ​​(e.g., from about 1 to about 40%, or from about 1 to about 30%, or from about 1 to about 20%, or from about 1 to about 16%, 1 to about 15%, 1 to about 14%, or from about 1 to about 12%, or from about 1 to about 10%, or from about 2 to about 50%, or from about 2 to about 40%, or from about 2 to about 30%, or from about 2 to about 20%, or from about 2 to about 10%). 6%, 2 to about 15%, 2 to about 14%, or about 2 to about 12%, or about 2 to about 10%, about 4 to about 50%, or about 4 to about 40%, or about 4 to about 30%, or about 4 to about 20%, or about 4 to about 16%, 4 to about 15%, 4 to about 14%, or about 4 to about 12%, or about 4 to about 10%, about 5 to about 50%, or about 5 to about 40%, or about 5 to about 30%, or about 5 to about 20%, or about 5 to about 16%, 5 to about 15%, 5 to about 14%, or about 5 to about 12%, or about 5 to about 10%).

[0184] In various embodiments, the protein-containing food may contain at least about 0.1% lipid by weight, such as about 0.1%, about 0.2%, about 0.5%, about 1%, about 3%, about 5%, about 10%, etc. In various embodiments, the protein-containing food may contain from about 0.1 to about 40% lipid by weight, with useful ranges being between any two of these values ​​(e.g., from about 0.1 to about 40%, or from about 0.5 to about 40%, or from about 1 to about 40%, or from about 3 to about 40%, or from about 5 to about 40%, or from about 10 to about 40%, or from about 15 to about 40%, or from about 20 to about 40%, or from about 0.1 to about 35%, or from about 0.5 to about 35%, or from about 1 to about 35%, or from about 3 to about 35%, or or about 5 to about 35%, or about 10 to about 35%, or about 15 to about 35%, or about 20 to about 35%, or about 0.1 to about 30%, or about 0.5 to about 30%, or about 1 to about 30%, or about 3 to about 30%, or about 5 to about 30%, or about 10 to about 30%, or about 15 to about 30%, or about 20 to about 30%, or about 0.1 to about 20%, or about 0.5 to about 20%, or about 1 to about 20%, or about 3 to about 20%, or about 5 to about 20%, or about 10 to about 20%, or about 15 to about 20%).

[0185] In various embodiments, the protein-containing food may contain at least about 0.1% by weight carbohydrate, e.g., about 0.1%, or about 0.5%, or about 1%, or about 3%, or about 5%, or about 10% by weight. In various embodiments, the protein-containing food may contain from about 0.1 to about 40% by weight carbohydrate, with useful ranges being between any two of these values ​​(e.g., from about 0.1 to about 40%, or from about 0.5 to about 40%, or from about 1 to about 40%, or from about 3 to about 40%, or from about 5 to about 40%, or from about 10 to about 40%, or from about 15 to about 40%, or from about 20 to about 40%, or from about 0.1 to about 35%, or from about 0.5 to about 35%, or from about 1 to about 35%, or from about 3 to about 35%, or is about 5 to about 35%, or about 10 to about 35%, or about 15 to about 35%, or about 20 to about 35%, or about 0.1 to about 30%, or about 0.5 to about 30%, or about 1 to about 30%, or about 3 to about 30%, or about 5 to about 30%, or about 10 to about 30%, or about 15 to about 30%, or about 20 to about 30%, or about 0.1 to about 20%, or about 0.5 to about 20%, or about 1 to about 20%, or about 3 to about 20%, or about 5 to about 20%, or about 10 to about 20%, or about 15 to about 20%).

[0186] In various embodiments, the protein-containing food product may contain at least about 15 mg / 100 g calcium.

[0187] In various embodiments, the protein-containing food product may contain at least about 40 mg / 100 g calcium.

[0188] In various embodiments, the food of the present invention is an acidic or fermented food in which the hardness and / or viscosity of the food of the present invention is reduced compared to a control food having the same ingredient composition and the same casein and protein content as the food of the present invention, except that it does not contain the milk protein composition of the present invention. For solid coagulated gel foods, the hardness of the fermented food is reduced by about 40 to about 80%. For semi-solid or liquid foods, the viscosity of the fermented food is reduced by about 40 to about 99%.

[0189] In various embodiments, the food products of the invention are acidified or fermented foods that exhibit reduced mouth texture (e.g., hardness or thickness) and / or negligible or no increased objectionable flavor (e.g., bitterness or saltiness) compared to a control food having the same ingredient composition, casein and protein content as the food products of the invention, except that it does not include the milk protein composition of the invention.

[0190] For the avoidance of doubt, the control food contains the same ingredients in the same relative amounts, and the same casein, total protein, lipid and / or carbohydrate content, as the food of the invention to which it is being compared. Additional Ingredients

[0191] In various embodiments, the one or more additional ingredients may be lipids, carbohydrates, proteins, flavors, vitamins, minerals, dairy products, water, food additives, colors, fruit preparations, or combinations of any two or more of these ingredients.

[0192] In various embodiments, the lipid may be a vegetable lipid or an animal lipid, including dairy lipid. Vegetable oil is generally representative due to its ease of incorporation and low content of saturated fatty acids. Examples of vegetable oil include canola (rapeseed) oil, corn oil, sunflower oil, olive oil, soybean oil, or hydrogenated vegetable oil.

[0193] In various embodiments, the dairy lipids include cream, butter, shortening, anhydrous milk fat (AMF), buttermilk, its hydrolyzed products, combinations of hydrolyzed and / or non-hydrolyzed compositions, one or more stages of hard milk fat extracts from the milk fat fraction, including hard (H), soft-hard (SH) and soft-soft-hard (SSH) extracts, one or more stages of soft milk fat extracts from the milk fat fraction, including soft (S), soft-soft (SS) and soft-soft-soft (SSS) extracts, combinations of hard milk fat extracts, combinations of soft milk fat extracts, any combination of hard milk types or more. These compositions can be obtained from whole milk or colostrum, and any derivative of whole milk or colostrum, including cream, cultured cream, and whey cream (milk lipids obtained from whey, including acid whey or cheese whey, preferably cheese whey). Cultured cream is cream from whole milk or colostrum fermented with acid-producing microorganisms, preferably lactic acid bacteria.

[0194] In various embodiments, the vegetable oil can be coconut oil, corn oil, cottonseed oil, canola oil, rapeseed oil, olive oil, palm oil, peanut oil, safflower oil, safflower oil, sesame oil, soybean oil, sunflower oil, hazelnut oil, almond oil, cashew oil, macadamia nut oil, pecan nut oil, pistachio oil, walnut oil, oil from melon and gourd seeds, pumpkin seed oil, apricot oil, glycerin, avocado oil, flaxseed oil, linseed oil, grapeseed oil, hemp oil, linseed oil, rice bran oil, wheat germ oil, or any combination of two or more thereof.In some embodiments, the vegetable oil can be hydrogenated coconut oil.

[0195] In various embodiments, the carbohydrate may include monosaccharides, disaccharides, oligosaccharides, and polysaccharides, as well as mixtures thereof, including sugars, sucrose, and sucralose. Many of these are commercially available starches, modified starches, maltodextrins (3-20 glucose equivalents (DE)) or corn syrup (>20 DE) for long chain carbohydrates. It may also include non-digestible carbohydrates, such as fructooligosaccharides, inulin, and galactooligosaccharides. In various embodiments, the carbohydrate may include polyhydric alcohols, such as, for example, selected from the group consisting of glycerol (glycerin), maltitol, erythritol, sorbitol, and any combination of any two or more of these.

[0196] In various embodiments, the protein may be a dairy protein or a non-dairy protein.In various embodiments, the protein may be milk, whey, casein, caseinate, egg, egg white, egg yolk, vegetable, plant, alfalfa, clover, pea, legume, kidney bean, soybean, kidney bean, lupin, cocoa, horn bean, nut, peanut, rye, grain, whole grain, rice, hemp, wheat gluten, fungal or algae protein, protein concentrate thereof, protein isolate thereof, hydrolysate thereof, or any combination of two or more thereof.

[0197] In various embodiments, the protein may be a protein powder. The protein powder may be any of the protein sources described. The protein powder may be non-agglomerated, agglomerated, roll-pressed, freeze-dried, drum-dried, spray-dried, or foam-spray-dried protein powder. In various embodiments, the protein powder comprises whey protein concentrate (WPC) or whey protein isolate (WPI). In various embodiments, the protein powder comprises whole milk powder, skim milk powder, or milk protein concentrate (MPC).

[0198] In various embodiments, the one or more additional ingredients may be flavorings, including but not limited to sweeteners, natural flavors, natural flavors, artificial flavors, herbs, and flavorings.

[0199] In some embodiments, the one or more additional ingredients may include nuts and / or seeds.

[0200] In various embodiments, one or more additional ingredients may be vitamins. The vitamins may include fat-soluble or water-soluble vitamins. Suitable vitamins include, but are not limited to, vitamin C, vitamin A, vitamin E, vitamin B12, vitamin K, riboflavin, niacin, vitamin D, vitamin B6, folic acid, pyridoxine, thiamine, pantothenic acid, and biotin. Forms of vitamins may include salts of vitamins, derivatives of vitamins, compounds with the same or similar activity as vitamins, and metabolites of vitamins.

[0201] In various embodiments, the one or more additional components may be minerals, including but not limited to chloride, sodium, calcium, iron, chromium, copper, iodine, zinc, magnesium, phosphorus, potassium, chromium. Suitable forms of the minerals include soluble mineral salts, sparingly soluble mineral salts, insoluble mineral salts, non-reactive minerals such as chelated minerals, mineral complexes, carbonyl minerals, reduced minerals, and combinations thereof.

[0202] In various embodiments, the food product may contain at least about 10, 20, 25, 30, 40, 50, 60, 70, 75, 80, 90, or 100% of the Recommended Daily Intake (RDI) of vitamins and minerals, such as 100 milliliters, 250 milliliters, 500 milliliters, or 1 liter, as set by European (FSMP) or USDRA regulations.

[0203] In various embodiments, one or more additional ingredients may be dairy products. In some embodiments, the dairy products may be selected from the group consisting of milk powder protein concentrate, skim milk powder, whole milk powder, whey protein concentrate, whey protein isolate, caseinate, milk fat, cream, curd casein, cheese or cream cheese. In various embodiments, one or more additional ingredients may be other dairy products such as milk powder protein concentrate, skim milk powder, whole milk powder, whey protein concentrate, whey protein isolate, caseinate, milk fat or cream.

[0204] In various embodiments, the one or more additional ingredients may be food additives, including but not limited to gelling enzymes, antifoaming agents, stabilizers, emulsifiers, preservatives, fiber, probiotics, antioxidants, flavor enhancers, colors, acidity regulators, or emulsifying salts. In various embodiments, the one or more additional ingredients may be food additives, including but not limited to rennet, antifoaming agents, stabilizers, emulsifiers, preservatives, fiber, probiotics, antioxidants, flavor enhancers, colors, acidity regulators. Potassium sorbate in acidified products is a useful preservative.

[0205] In various embodiments, one or more additional components may be a stabilizer or emulsifier. Useful emulsifiers include lecithin, mono- and diglycerides, polyglycerol esters, milk phospholipids, citrate esters (Cytrem), polysorbate 60, glyceryl monostearate, and datum. Useful stabilizers include carrageenan, gellan gum, pectin, guar gum, locust bean gum, carboxymethylcellulose, alginate, agar, oat gum, tragacanth gum, acacia gum, xanthan gum, karaya gum, tara gum, starch, modified starch, and microcrystalline cellulose, gelatin, or combinations thereof. Useful stabilizers include carrageenan, gellan gum, pectin, guar gum, locust bean gum, carboxymethylcellulose, alginate, agar, oat gum, tragacanth gum, acacia gum, xanthan gum, karaya gum, tara gum, starch, modified starch and microcrystalline cellulose, or combinations thereof. In addition to those listed above, those skilled in the art will recognize that many different gel forms are suitable for use in the compositions disclosed herein.

[0206] In various embodiments, the one or more additional components may be salts or acidity adjusters such as sodium chloride, potassium chloride, ethylenediaminetetraacetic acid (EDTA) salts, lactic acid, acetic acid, citric acid, potassium hydroxide, phosphates such as dipotassium phosphate, disodium phosphate, citrates such as disodium citrate, dipotassium citrate, tripotassium citrate, etc. The citrates may be selected from the group consisting of disodium citrate, dipotassium citrate, tripotassium citrate, and trisodium citrate. In some embodiments, the phosphates may be selected from the group consisting of dipotassium phosphate, disodium phosphate, n-phosphates, diphosphates, and polyphosphates.

[0207] In various embodiments, the one or more additional components may be a source of amino acids, amino acid precursors or amino acid metabolites, or any combination of any two or more thereof, preferably free amino acids, amino acid precursors or amino acid metabolites.

[0208] The method of combining one or more additional ingredients with the milk protein concentrate to produce the protein-containing food product will depend on the protein-containing food product being formed. These methods are known to those skilled in the art. 5. Yogurt

[0209] The dairy protein compositions of the present invention are particularly useful in the production of yogurt.

[0210] In various embodiments, the protein-containing food product is a yogurt. In various embodiments, the yogurt is a set yogurt or a stirred yogurt. In various embodiments, the stirred yogurt is a drinking yogurt.

[0211] In various embodiments, the protein-containing food product is room temperature yogurt. By fermenting the room temperature yogurt and then heat-treating it, it is possible to provide yogurt that has excellent shelf life (e.g., no significant microbial growth) even when stored at room temperature for a long period of time.

[0212] In various embodiments, the pH of the yogurt is less than about pH 4.7.

[0213] In various embodiments, the pH of the yogurt is less than about pH 4.6.

[0214] In various embodiments, the yogurt has a titratable acidity (TA) of about 0.9 to about 2.0 (equivalent lactic acid %). In other embodiments, the yogurt has a titratable acidity (TA) of about 0.6 to about 2.0 (equivalent lactic acid %). The method for measuring TA is to heat the food sample to 20-25°C and mix to ensure homogeneity. Weigh 10 g of sample into a glass beaker and mix with 10 mL of distilled water and stir continuously using a magnetic stirrer. Place a pH electrode in the sample solution and titrate to pH 8.30 with 0.1 M sodium hydroxide. Record the volume (mL) of NaOH and calculate the titratable acidity as lactic acid equivalents using the following formula: TA = (NaOH volume (mL) x 0.9) / (yogurt mass (g))

[0215] In various embodiments, the hardness of the set yogurt is about 300 to about 8000 g.s., or about 500 to about 8000, or about 1000 to about 8000, or about 2000 to about 8000, or about 3000 to about 8000, or about 4000, to about 8000, or about 5000 to about 8000, or about 5000 to about 7000, or about 1000 to about 7000, or about 2000 to about 7000, or about 3000 to about 7000, or about 4000, to about 7000, or about 5000 to about 7000, or about 500 to about 5000, or about 1000 to about 5000, or about 2000 to about 5000, or about 3000 to about 5000 g.s.

[0216] In various embodiments, the breaking force of the set yogurt is about 10 to about 500, or about 20 to about 500, or about 50 to about 500, or about 100 to about 500, or about 200 to about 500, or about 300 to about 500, or about 20 to about 400, or about 50 to about 400, or about 100 to about 400, or about 200 to about 400, or about 300 to about 400, or about 20 to about 300, or about 50 to about 300, or about 100 to about 300, or about 200 to about 300 g.

[0217] Breaking force and strength can be evaluated using a Stable Micro Systems TAHD Plus Texture Analyzer. A 1.27 cm organic glass cylinder is used for one compression test. The initial force to break the surface of the set yogurt is recorded as the breaking force (g) and the area under the curve as hardness (gs).

[0218] In various embodiments, the stirred yogurt is stirred for 50 s -1The viscosity at is less than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 17, 18, 20, 25, 30, 40, 50, 100, 200, 250, 500, 750, 1000, 1250, 1500, 1750, 2000, 2500, 3000, or about 4000 mPa.s, and the various ranges may be between any two of these values, for example, from about 1 to about 4000, from about 1 to about 2000, from about 1 to about 1000, from about 1 to about 500, about 1 to about 200, about 1 to about 100, about 1 to about 50, about 1 to about 30, about 1 to about 20, 5 to about 4000, about 5 to about 2000, about 5 to about 1000, about 5 to about 500, about 5 to about 200, about 5 to about 100, about 5 to about 50, about 5 to about 30, about 5 to about 20, 10 to about 4000, about 10 to about 2000, about 10 to about 1000, about 10 to about 500, about 10 to about 200, about 10 to about 100, about 10 to about 50, about 10 to about 30, or about 10 to about 20 mPa.

[0219] Viscosity can be measured using a Haake VT500 or Viscotester IQ Viscometer. Refrigerate the sample (4°C) and set the Haake water bath to 10°C. Stir the sample gently before testing to achieve a uniform consistency. Depending on the thickness of the sample, use an MV1 or SV DIN rotor and configure using a cup and pendulum. Fill the cup with the yogurt sample to latitude, ensuring no bubbles are present. Screw the rotor onto the instrument and zero, place the pendulum into the cup and fix it in place inside the cup. Apply a shear rate scan from 0 to 1201 / s and hold for 50 seconds. -1 The apparent viscosity is reported as

[0220] In various embodiments, the drinking yogurt has a uniform pourable consistency. -1The viscosity at is less than about 1, 2, 3, 4, 5, 7, 8, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 125, 150, 200, 300, 400, 500, 600, 700 or 800 mPa.s, and various ranges may be between any two of these values, for example, about 0 to 800 mPa.s. about 800, about 0 to about 500, about 0 to about 200, about 0 to about 100, about 0 to about 50, about 0 to about 30, about 2 to about 800, about 2 to about 500, about 2 to about 200, about 2 to about 100, about 2 to about 50, about 2 to about 30, about 5 to about 800, about 5 to about 500, about 5 to about 200, about 5 to about 100, about 5 to about 50, or about 5 to about 30 mPa.s.

[0221] The sensory quality of the set yogurts can be evaluated using the following sensory procedure: The yogurt set is placed in transparent sample cups displaying a random three-digit blind code. The samples are stored at 4°C and submitted to a team of experts immediately after being removed from the cooler. The sensory evaluation is carried out by a group of eight experts familiar with the yogurt tasting experience. The yogurt samples are evaluated by participants describing texture, flavor attributes and intensity. A consensus approach is used to collate the attributes that best describe each sample.

[0222] Flavor can be evaluated using a trained sensory panel. For example, samples are evaluated by trained sensory panelists (n=8-12) utilizing the sensory vocabulary created during the attribute generation session. One attribute generation session and one training and calibration session are held prior to the two consensus profiling evaluation sessions. In consensus profiling, trained sensory panel members collaborate to agree on an intensity rating (on a 0-150 mm line scale) for each sensory attribute, rather than providing two or three independent ratings. All samples are evaluated in two parts, one set per course, and one sample in each course. All samples are evaluated at room temperature (approximately 18-20°C) and sampled under white light in clear sample cups labeled with random three-digit code labels. Samples are presented to team members in a random order. Other sensory panels or alternative methods of evaluating flavor will be apparent to those skilled in the art.

[0223] In various embodiments, the yogurt has an acceptable bitterness. Within the 150-point (mm) range, the attribution threshold of a typical consumer is approximately 10-15 points (depending on the individual). A score below 4.5 in the example suggests that some group members may not notice the bitterness. Therefore, it is assumed that the bitterness level detected by the formal group will be acceptable to some consumers. A reference value of 0.01% caffeine in filtered water was set as the threshold intensity, 0.03% as weak intensity, and 0.06% as medium intensity.

[0224] Particle size distribution can be determined using a Malvern Mastersizer 2000. Deionized water (refractive index (RI) = 1.33) is used to disperse the sample, and the refractive index of milk fat (RI = 1.46) is used as the dispersed phase. Drops of sample are added until a shading value of 10-15% is obtained. Particle size is reported as surface weighted mean diameter (D[3,2]) and volume weighted mean diameter (D[4,3]).

[0225] In various embodiments, the D[3,2] particle size distribution of the acidic dairy beverage is less than about 15 μm, or about 14 μm, or about 13 μm, or about 12 μm, or about 11 μm, or about 10 μm.

[0226] The viscosity of acidified dairy beverages can be measured with a Brookfield DV2T Viscometer. Samples are lightly stirred prior to testing to achieve a uniform consistency. Spindle numbers S-61, S-62, S-63, S-64 are used at a constant speed of 30 or 60 rpm, depending on the viscosity of the sample. Tests are performed at ambient temperature. Viscosity is taken 60 seconds after the start of the test or until a constant value is reached. Viscosity is expressed in millipascal-seconds (mPa.s). 6. Protein Bars

[0227] The dairy protein compositions described herein are particularly useful in producing protein bars. The dairy protein compositions of the present invention are useful in producing protein bars having high protein content while maintaining acceptable breaking force, water activity and flavor. The artisan will recognize that a protein bar will have adequate breaking force, ensure adequate bar texture and adequate water activity, and limit dry yeast and mold growth.

[0228] In various embodiments, the protein-containing food product is a protein bar.

[0229] In various embodiments, the breaking force of the protein stick is from about 500g to about 10,000g, or from about 500g to about 9,000g, or from about 500g to about 8,000g, or from about 500g to about 7,000g, or from about 500g to about 6,000g, or from about 500g to about 6,000g, or from about 500g to about 4,000g, or from about 500g to about 3,000g, 1,000g to about 10,000g, or from about 1,000g to about 9,000g, or from about 1,000g to about 8,000g, or from about 1,000g to about 7,000g. g, or about 1,000 g to about 6,000 g, or about 1,000 g to about 6,000 g, or about 1,000 g to about 4,000 g, or about 1,000 g to about 3,000 g, 2,000 g to about 10,000 g, or about 2,000 g to about 9,000 g, or about 2,000 g to about 8,000 g, or about 2,000 g to about 7,000 g, or about 2,000 g to about 6,000 g, or about 2,000 g to about 6,000 g, or about 2,000 g to about 4,000 g, or about 2,000 g to about 3,000 g.

[0230] The breaking force (g) of the bars can be assessed using a TAHD Plus Texture Analyser from Stable Micro Systems, Godalmin, UK. Texture measurements are performed by penetration. Force is measured over a set penetration depth of 12mm. A 5mm stainless steel cylindrical probe is pushed into the rod to a depth of 12mm at a constant speed of 1mm / s and withdrawn at a speed of 10mm / s. The relationship of force (g) of the probe movement versus time (s) is measured. Three compressions are applied to the surface of each rod sample. Two bars were assessed for each sample. Samples are removed from storage at 20°C and texture measurements are made at 20°C in a temperature controlled room.

[0231] In various embodiments, the water activity of the protein bar is less than about 0.65, less than about 0.6, less than about 0.55, or less than about 0.5.

[0232] Water activity can be measured in the following way: Water activity analysis is performed using an Aqua Lab Dew Point Moisture Analyzer 4TE DUO (Meter, Pullum, WA, USA). Standard solutions are measured to verify calibration, and then samples are measured directly.

[0233] The sensory attributes of protein bars can be evaluated using the following sensory procedure: The bars are placed in transparent sample cups labeled with a random three-digit blind code. The sensory evaluation is performed by a team of at least seven experts familiar with bar tasting. The warm-up samples compared to the control are tasted first and the panel is calibrated in terms of taste and texture qualities using a 0 to 7 point scale, where 0 indicates no difference from the control and 7 indicates an extreme difference from the control. The protein samples are rated by participants describing the nature and intensity of their texture and flavor compared to the control sample.

[0234] Can measure bar color using ColorFlex EZ (HunterLab). Calibrates using standard black and white tiles. Reports color using L*, A*, b* color spaces.

[0235] In one aspect, the present invention provides a method of preparing a bar, the method comprising: a) i. a milk protein composition of the invention or a milk protein composition prepared by a method of the invention; ii. providing a bar composition comprising one or more additional ingredients; b) forming the bar composition into a bar.

[0236] In various embodiments, the method includes the step of heating and / or mixing the bar composition.

[0237] In various embodiments, the method includes forming the bar composition into a bar by molding the composition and / or extruding the composition. In various embodiments, the bar composition is molded or extruded and then cut into bars.

[0238] cheese

[0239] The dairy protein compositions of the present invention are particularly useful in the production of cheese, particularly processed cheese. The dairy protein compositions of the present invention are useful for producing processed cheese having a high protein content while maintaining acceptable firmness, yield stress, meltability and flavor.

[0240] In various embodiments, the protein-containing food product is a processed cheese. In various embodiments, the processed cheese may be a processed cheese spread, a "slice-on-slice" processed cheese, a processed cheese "lollipop", individually wrapped processed cheese slices, processed cheese triangles, processed cream cheese, processed cheese sauce, or a processed cheese block.

[0241] In various embodiments, when the processed cheese is separately packaged processed cheese slices or "slice on slice" processed cheese, the processed cheese has a hardness of about 6N to about 13N, or about 6N to about 12N, or about 7N to about 15N, or about 7N to about 14N, or about 7N to about 13N, or about 7N to about 12N.

[0242] Hardness can be assessed by penetration testing using a TAHD Plus Texture Analyzer from Stable Micro Systems. A 6mm diameter stainless steel probe is inserted 10mm into the sample at a speed of 1mm / s. Each product sample is tested five times at different locations on the packaged product. The peak force measured is recorded as hardness (N). The average of the results is reported.

[0243] Yield stress can be assessed using a Brookfield rotational viscometer with a four-sided stainless steel blade (6 mm diameter, 12 mm height) inserted into the sample to a depth of 19 mm and rotated at 0.5 rpm for 30 seconds. The sample height is at least 30 mm. The product is tested three times at different locations for each sample. The average results are reported as stress (Pa).

[0244] In various embodiments, the process cheese has a yield stress of less than 111000, 10500, 10000, 9500, 9000, 8500, 8000, 7500, 7000, 6500, 6000, 5500, 5000, 4500, 4000, 3500, 3000, 2500, 2000, 1500, 1000 or 500 Pa, with various ranges within these values. It may be between any two selected from the above, for example, about 500 to about 11000 Pa, about 2000 to about 10000 Pa, about 2000 to about 9000 Pa, about 2000 to about 7500 Pa, about 2000 to about 6000 Pa, about 3000 to about 10000 Pa, about 3000 to about 9000 Pa, about 3000 to about 7500 Pa, or about 3000 to about 6000 Pa.

[0245] In one aspect, the present invention provides a method for preparing cheese, preferably processed cheese, comprising: a) i. a milk protein composition of the invention or a milk protein composition prepared by a method of the invention; ii. providing a cheese composition comprising one or more additional ingredients; b) heating the cheese composition to a temperature of at least about 70° C. to produce cheese.

[0246] In various embodiments, the method includes mixing the composition. 7. Food Preparation Methods

[0247] In another aspect, the present invention provides a method for preparing a protein-containing food product, the method comprising: a) providing a milk protein composition of the present invention or a milk protein composition prepared by the method of the present invention, and a milk protein composition of the present invention; b) mixing with one or more additional ingredients to produce a protein-containing food product.

[0248] In various embodiments, a protein-containing food product can be prepared by a method comprising providing a dairy protein composition of the present invention and mixing with at least one lipid source. In various embodiments, a protein-containing food product can be prepared by a method comprising providing a dairy protein composition of the present invention and mixing with at least one carbohydrate source. In various embodiments, a protein-containing food product can be prepared by a method comprising providing a dairy protein composition of the present invention and mixing with at least one lipid source and at least one carbohydrate source. Other steps of preparation and packaging of the protein-containing food product will depend on the product being manufactured and will be known to those skilled in the art.

[0249] In various embodiments, the method may include providing an aqueous composition comprising a milk protein composition. In some embodiments, the method may include reconstituting a milk powder protein composition, and optionally combining with one or more additional dry ingredients, to produce the aqueous composition. 8. Methods for producing acidic or fermented foods

[0250] In another aspect, the invention relates to a method for producing an acidified protein-containing food product, the method comprising the steps of providing a dairy protein composition of the invention or produced by a method of the invention, and acidifying the composition to produce an acidified protein-containing food product.

[0251] In another aspect, the present invention relates to a method for preparing a fermented protein-containing food product, the food product comprising: a) This method is i. a milk protein composition of the invention or a milk protein composition prepared by a method of the invention; ii. providing a composition comprising one or more media; b) incubating the composition for a period of time sufficient to produce a fermented protein-containing food product.

[0252] The composition can be heat treated prior to acidification or fermentation to reduce pathogens or other microorganisms present in the mixture and to induce β-lactoglobulin-k-casein interactions to alter the texture of the yogurt. Heat treatments used include pasteurization (72°C for 15 s or 63°C for 30 min), pasteurization (85°C for 20-30 min or 90-95°C for 5 min), sterilization (110°C for 30 min or 130°C for 40 s) or UHT (135-145°C for 1-5 s; e.g. 145°C for 1-2 s).

[0253] In various embodiments, the acidification step comprises adding one or more food grade acids or acidic substances, hi various embodiments, the food grade acids or acidic substances are selected from the group consisting of gluconic acid delta lactone (GDL), lactic acid, citric acid, malic acid, acetic acid, tartaric acid, fumaric acid, phosphoric acid, hydrochloric acid, sulfuric acid, or a combination of any two or more thereof.

[0254] In various embodiments, the acidification or fermentation step includes adding one or more culture media. In various embodiments, the culture media is a microbial culture media, preferably a bacterial culture media. In various embodiments, the culture media is selected from the group consisting of Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus casei, or Lactobacillus, Streptococcus, Leuconostoc, Lactococcus or Bifidobacterium.

[0255] In various embodiments, the method of the present invention includes incubating the composition for a sufficient time to produce a fermented protein-containing food product. A sufficient time may be the time necessary to produce a desired hardness, viscosity and / or acidity. One of skill in the art can readily determine the time required to produce a fermented product based on the particular medium used, the culture conditions (e.g., temperature), the properties of the water composition (e.g., protein content, pH), and the desired attributes of the final product (e.g., acidity, hardness, viscosity).

[0256] In various embodiments, the acidified or fermented food product may be smoothed or sheared to break down the gel structure. Smoothing provides a pourable consistency. Equipment for such smoothing may be anything that imparts shear to the product, including back pressure valves, rotor stator shear pumps, homogenizers, or on-line screens or filters. 9. How to produce food by online protein hydrolysis

[0257] In another aspect, the invention relates generally to a method of producing a protein-containing food product comprising the steps of providing a composition comprising calcium-deficient milk protein and one or more additional ingredients, and proteolyzing the milk protein in the composition to produce the protein-containing food product.

[0258] The methods of the present invention are useful for producing any of the protein-containing food products disclosed herein.

[0259] In various embodiments, the method comprises: a) at least about 40% total protein by weight based on the dry matter in the composition; b) providing a milk protein composition comprising a milk protein, preferably a milk protein composition comprising a milk protein concentrate, a milk protein isolate, or a combination thereof, comprising less than about 2 g calcium per 100 g total protein, and / or less than about 1.4 g calcium per 100 g dry matter in the composition.

[0260] In one aspect, the invention provides a protein-containing food product prepared by the above method.

[0261] In various embodiments, the milk protein or milk protein composition may comprise a milk protein concentrate (MPC), a milk protein isolate (MPI), caseinate, casein, casein co-precipitate, a retentate obtained by ultrafiltration or microfiltration of milk, or any combination of any two or more thereof. In various embodiments, the milk protein may comprise a milk protein concentrate (MPC), a milk protein isolate (MPI), or a combination thereof. In various embodiments, the milk protein may comprise a milk protein concentrate (MPC) or a milk protein isolate (MPI).

[0262] In various embodiments, the milk protein composition comprises at least about 40%, about 45%, about 50%, about 60%, about 70%, about 80%, about 90%, or at least about 95% total protein by weight based on dry matter in the composition, and various ranges may be between any two of these values. In various embodiments, the composition may comprise about 40 to about 99%, about 40 to about 90%, or about 40 to about 80% total protein by weight based on dry matter in the composition.

[0263] In various embodiments, the milk protein or milk protein composition may comprise at least about 10, 20, 30, 40, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99 or 100% casein by weight on a dry matter basis, and various ranges may be between any two of these values, for example, about 10 to about 100%, about 40 to about 100, about 50 to about 100, about 60 to about 100, about 70 to about 100, about 75 to about 100, or about 80 to about 100%, about 10 to about 90, about 40 to about 90, about 50 to about 90, about 60 to about 90, about 70 to about 90, about 75 to about 90, or about 80 to about 90% by weight.

[0264] In various embodiments, the milk protein or milk protein composition may comprise whey protein. In various embodiments, the milk protein or milk protein composition may comprise about 1 to about 50% by weight, about 1 to about 40% by weight, about 1 to about 30% by weight, or about 1 to about 20% by weight of whey protein based on dry matter.

[0265] In various embodiments, the whey protein may comprise or be provided from ingredients including whey protein concentrate, whey protein isolate, or combinations thereof. Other suitable whey protein sources known in the art may be used.

[0266] In various embodiments, the dairy protein composition comprises less than about 2 g calcium per 100 g total protein, or less than about 1.9, 1.8, 1.6, 1.5, 1.4, 1.2, 1, 0.8, 0.75, 0.6, 0.5, 0.4, 0.3, 0.2 or 0.1 g calcium per 100 g total protein, with various ranges between any two of these values, e.g., about 0.1 to about 2, about 0.5 to about 2, about 1 to about 2, about 0.1 to about 1.5, about 0.5 to about 1.5, or about 1 to about 1.5 g.

[0267] In various embodiments, the dairy protein composition contains less than about 1.4 g calcium per 100 g dry matter, or less than about 1.3, 1.2, 1.1, 1.0, 0.9, 0.8, 0.7, 0.6, 0.5, or 0.4 g calcium per 100 g dry matter.

[0268] The method further includes mixing the milk protein composition with one or more additional ingredients to produce an intermediate aqueous composition comprising from about 0.5 to about 20% total protein by weight.

[0269] In various embodiments, the aqueous intermediate composition may comprise at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 15, 16, 18, or at least about 20% total protein by weight based on the dry matter in the composition, and various ranges may be between any two of these values, such as from about 2 to about 20, 4 to about 20, 5 to about 20, 8 to about 20, 2 to about 18, 4 to about 18, 5 to about 18, or about 8 to about 20% by weight.

[0270] The one or more additional components may include any component described herein, in particular any component disclosed in any of paragraphs

[0177] to

[0193] .

[0271] In various embodiments, the one or more additional ingredients may include nonfat or skim milk powder.

[0272] In various embodiments, the method includes exposing the aqueous intermediate composition to the action of one or more proteases. Any of the proteases described herein can be used under any of the conditions described herein, such as the enzymes and conditions described in any of paragraphs

[0141] to

[0155] or in the embodiments.

[0273] In some embodiments, the method includes a step of inactivating one or more proteases. Any method of inactivating proteases described herein can be used. In other embodiments, no specific inactivation is required, and one or more proteases are inactivated by a subsequent processing step in preparing a protein-containing food, for example, by heating the composition with one or more media to produce a fermented food, or by producing an acidified food, or by producing an acidified composition.

[0274] In various embodiments, the method may include adjusting the pH to a desired pH to obtain optimal activity of one or more proteolytic enzymes or to improve the solubility of one or more components in the aqueous intermediate composition.

[0275] In various embodiments, the total protein may have a peptide molecular weight distribution corresponding to the following molecular weight distribution: a) about 20%, 25%, 30%, 35%, 40%, 45%, or 50 to about 79% of peptides having a molecular weight greater than about 20 kDa; b) about 15, 20, 25, 30, 35, 40, 45, 50, or about 55% by weight of a peptide having a molecular weight of about 5 to about 20 kDa; c) about 2%, 3%, 4%, 5%, 10%, 12%, 14%, 16%, 18 to about 20% by weight of a peptide having a molecular weight of about 1 to about 5 kDa, and d) about 2%, 3%, 4%, 5%, 10%, 15 to about 20% by weight of peptides having a molecular weight of less than about 1 kDa.

[0276] In various embodiments, the total protein may have a peptide molecular weight distribution corresponding to the following molecular weight distribution: a) about 20 to about 79% of peptides having a molecular weight greater than about 20 kDa; b) Contains about 15 to about 54% by weight of peptides having a molecular weight of about 5 to about 20 kDa. c) about 2 to about 17% by weight of a peptide having a molecular weight of about 1 to about 5 kDa, and d) about 2 to about 20% by weight of a peptide having a molecular weight of less than about 1 kDa.

[0277] In various embodiments, the total protein is a) less than about 79%, 78%, 77%, 76%, 75%, 70%, 65%, 60%, 55%, 50%, 45% or 40% of peptides having a molecular weight greater than about 20 kDa; b) about 15 to about 55%, about 15 to about 50%, about 15 to about 45%, about 15 to about 40%, about 15 to about 35%, 20 to about 50%, about 20 to about 45%, about 20 to about 40%, about 20 to about 35%, 25 to about 50%, about 25 to about 45%, about 25 to about 40%, about 25 to about 35%, 30 to about 50%, about 30 to about 45%, or about 30 to about 40%.

[0278] In various embodiments, the total protein is a) less than about 79%, 78%, 77%, 76%, 75%, 70%, 65%, 60%, 55%, 50%, 45% or 40% of peptides having a molecular weight greater than about 20 kDa; b) containing less than about 30%, less than about 28%, less than about 25%, or less than about 20% peptides having a molecular weight of less than about 5 kDa.

[0279] In various embodiments, the total protein is a) less than about 79%, 78%, 77%, 76%, 75%, 70%, 65%, 60%, 55%, 50%, 45% or 40% of peptides having a molecular weight greater than about 20 kDa; b) Contains less than about 20%, less than about 18%, less than about 16%, less than about 14%, or less than about 12% peptides having a molecular weight in the range of about 1 to about 5 kDa.

[0280] In various embodiments, the total protein is a) less than about 79%, 78%, 77%, 76%, 75%, 70%, 65%, 60%, 55%, 50%, 45% or 40% of peptides having a molecular weight greater than about 20 kDa; b) containing less than about 20%, less than about 15%, or less than about 10% peptides having a molecular weight of less than about 1 kDa.

[0281] In various embodiments, the protein-containing food product comprises: a) less than about 3, 2.75, 2.5, 2.25, 2, 2.1, 2, 1.9, 1.8, 1.5, 1, or 0.5 grams of calcium per 100 grams of casein in the food, or from about 0.5 to about 3 grams of calcium per 100 grams of casein in the food; b) less than about 3, 2.75, 2.5, 2.25, 2, 2.1, 2, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, 1.2, 1.1, or 1 g of calcium per 100 g of total protein in the food, or from about 1 to about 3 g of calcium per 100 g of total protein in the food, and / or c) Contains less than about 2, 1.75, 1.5, 1.4, 1.3, 1.2, 1.1, 1, 0.9, 0.8, 0.5 g of calcium, or from about 0.5 to about 2 g of calcium per 100 g of dry matter in the food.

[0282] In various embodiments, the protein-containing food is an acidic food. The acidic food may be an acidic food as described herein.

[0283] In one embodiment, the method of the invention comprises exposing the aqueous intermediate composition to one or more proteases, optionally inactivating the one or more proteases, and then acidifying the aqueous intermediate composition to produce an acidified protein-containing food product. In another embodiment, the protein hydrolysis and acidification steps are reversed to include acidifying the aqueous intermediate composition, then exposing the aqueous intermediate composition to one or more proteases, and optionally inactivating the one or more proteases to produce an acidified protein-containing food product.

[0284] In various embodiments, the protein-containing food is a fermented food. The fermented food may be any of the fermented foods described herein.

[0285] In one embodiment, the method comprises exposing the aqueous intermediate composition to one or more proteases, optionally inactivating the one or more proteases, then adding one or more media to the aqueous intermediate composition and incubating for a sufficient time to produce a fermented protein-containing food product. In another embodiment, the protein hydrolysis and fermentation steps are reversed, and the method comprises adding one or more media to the aqueous intermediate composition and incubating for a sufficient time, after which exposing the aqueous intermediate composition to the action of one or more proteases, and optionally inactivating the one or more proteases to produce a fermented protein-containing food product.

[0286] In various embodiments, the method includes adding a casein-containing composition or a whey-containing composition or a mixture thereof to the milk protein composition or the aqueous intermediate composition and adjusting the ratio of casein to whey to a desired level. In various embodiments, the adjustment can be made before proteolysis, after proteolysis, before inactivation of proteolytic enzymes, after inactivation of proteolytic enzymes, before or after addition of one or more media, before or after culturing the aqueous intermediate composition to produce a fermentation product, or before or after acidification.

[0287] In various embodiments, the method may further include the step of adding one or more additional components. In various embodiments, one or more other components may be added before proteolysis, after proteolysis, before the proteolytic enzymes are inactivated, before the proteolytic enzymes are inactivated, after the proteolytic enzymes are inactivated, before or after adding one or more media, before or after culturing the aqueous intermediate composition to produce a fermentation product, or before or after acidification. For example, it may be desirable to add certain components after proteolysis to avoid adverse effects of the proteolytic enzymes on these components. EXAMPLES

[0288] 1. Example 1

[0289] This example describes the preparation of a dairy protein composition of the present invention.

[0290] MPC with reduced calcium was prepared according to the method described in US Pat. No. 7,157,108.

[0291] As shown in Table 1, the following MPC powders were reconstituted in water at 25-55°C for 1 hour using an overhead stirrer to produce an aqueous composition containing 10% by weight of protein. MPC A (protein 69.9%, calcium 2180mg per 100g powder) MPC B (70% protein, 326mg calcium per 100g powder) MPC C (50% protein, 401mg calcium per 100g powder)

[0292] Additionally, a fresh MPI retentate having reduced calcium levels was prepared according to the method described in US Pat. No. 7,157,108.

[0293] As shown in Table 1, the following MPI powders were reconstituted in water at 25-55°C for 1 hour using an overhead stirrer to produce aqueous compositions containing approximately 6.61-6.82% protein by weight. MPI A (protein 6.82%, calcium 2720mg per 100g protein) MPI B (protein 6.95%, calcium 82mg per 100g protein)

[0294] The composition was cooled to 5° C. and the appropriate amount of DuPont Food Pro PNL enzyme (as recommended by the manufacturer) was added.

[0295] The composition was incubated at 5° C. and neutral pH for 30 minutes to allow protein hydrolysis to occur.

[0296] The composition was then heated to 90°C for 8-10 minutes and held for 5 minutes to inactivate the enzymes before being cooled to ambient temperature.

[0297] A summary of the MPC and MPI, including components, composition, hydrolysis conditions, and property descriptions of the partial hydrolysis products, is shown in Table 1. [Table 1-1] [Table 1-2]

[0298] 2. Example 2

[0299] This example describes the preparation of a dairy protein composition of the present invention.

[0300] The following component powder combinations were reconstituted in water at 25-55°C with an overhead stirrer for 1 hour to produce aqueous compositions containing 6.9% or 10% by weight protein: MPC B (protein 70%, calcium 326mg per 100g MPC) MPC D (protein 82.4%, calcium 270mg per 100g MPC) MPC E (protein 70%, calcium 328mg per 100g MPC)

[0301] The composition was cooled to 5° C. or heated to 45° C. and the appropriate amount (as per the manufacturer's recommendations) of one of the following enzymes was added: DuPont Food Pro PNL DSM MaxiPro Serine proteases from Fusarium sp.

[0302] The composition was incubated at temperature and pH to allow hydrolysis of the proteins to occur.

[0303] The compositions were then heated to inactivate the enzymes and then cooled to ambient temperature and, for some samples, dried to form a powder.

[0304] An overview of the MPC, including components, composition, hydrolysis conditions, and characteristics of the partial hydrolysis products, is shown in Table 2. [Table 2-1] [Table 2-2]

[0305] Example 2A

[0306] According to Example 2, an MPC composition was prepared.

[0307] The composition was cooled and an appropriate amount of one of the following enzymes was added: DuPont Food Pro PNL Biocatalyst Promod TM 523MDP

[0308] The composition was incubated at temperature and pH to allow hydrolysis of the proteins to occur.

[0309] The composition was then heated to inactivate the enzyme and then cooled to ambient temperature.

[0310] A summary of the MPC including components, composition, hydrolysis conditions, and characteristics of the partially hydrolyzed products is given in Table 2A below. [Table 3]

[0311] 3. Example 3

[0312] This example describes the preparation of a milk protein composition of the invention and the reduction of complete casein using a method from Anema (SG Anema, "Lab on a Chip" Separation and Quantification of Milk Proteins Using Microfluidic SDS Electrophoresis Technology. International Journal of Dairy Science, Vol. 19, No. 4, April 2009, pp. 198-204).

[0313] MPCD (82.4% protein, 270 mg calcium per 100 g MPC) powder was reconstituted in water at 25-55°C for 1 hour using an overhead stirrer to produce an aqueous composition containing 6.9% protein by weight.

[0314] The composition was heated to 45° C. and the appropriate amount of DuPont Food Pro PNL (as per the manufacturer's recommendations) was added.

[0315] The composition was incubated at temperature and pH to allow hydrolysis of the proteins to occur.

[0316] The composition was then heated to 90° C. for 30 seconds and held at 90° C. for 5 minutes to inactivate the enzymes, then cooled to ambient temperature and dried to a powder.

[0317] A summary of the MPC, including components, composition, hydrolysis conditions, and characteristics of the partial hydrolysis products, is shown in Table 3. [Table 4]

[0318] The milk protein composition of the present invention in Table 3 and the starting material MPC D (not hydrolyzed) were dissolved exactly to 3.5% protein. To analyze the complete casein reduction in the samples, a 2100 Amgelen Bioanalyzer System (Amgelen, USA) was used and the samples were prepared based on the Anema method. The area under the curve of the casein band was automatically integrated by the software (2100 Bioanalyzer Expert package). The band area, which shows the reduction rate compared to MPC D, was calculated.

[0319] The starting material is known to have a casein:whey ratio of approximately 80:20, allowing confirmation of complete casein reduction based on g protein / 100g total weight. Using the [Bioanalyzer] microfluidic chip, it has previously been found that the band intensities for BSA, LF, and IgG are very low and may be too low for accurate quantification. This therefore impacts the casein to whey ratio, resulting in an over-representation of casein when compared to the total protein content. The Protein 80 kit used in this method claims to separate proteins with molecular weights ranging from ~5 to 80 kDa, so small peptides and larger proteins may not be detected. These factors result in a casein to whey ratio greater than the commonly accepted 80:20. Due to these factors, we hypothesize that the casein to whey ratio reported using Bioanalyzer data is likely skewed, and therefore the expected casein content was calculated for the control MPCD (82.4g protein / 100g total weight x 0.8 casein). The bands are well distinguished and captured within the molecular weight range so that changes in the detection of whey proteins do not affect the detection and quantification of casein. [Table 5]

[0320] 4. Example 4

[0321] This example describes the use of a dairy protein composition of the invention in the production of a high protein (10% by weight) set yoghurt.

[0322] Skim milk powder was reconstituted in water and the MPC composition prepared as described in Example 2 was added at ambient temperature.

[0323] The mixture was heated at 85°C for 15 minutes and then cooled to 43°C.

[0324] The mixture was inoculated with a starter (Chr Hansen YF-L702).

[0325] The mixture was placed in a pot.

[0326] The mixture was incubated at 43°C for approximately 9-16 hours until the final pH was approximately 4.6 to form yogurt.

[0327] The yogurt was stored refrigerated at 4°C.

[0328] Using the method of the present invention, the pH, breaking force and firmness of the yoghurt were evaluated. The organoleptic properties of the yoghurt were also evaluated. The composition and properties of the yogurt are shown in Table 5. [Table 6]

[0329] Example 4A

[0330] This example describes the use of a dairy protein composition of the invention in the production of a high protein (10% by weight) set yoghurt.

[0331] A set yogurt was prepared as in Example 4 using a dairy protein composition prepared as in Example 2A.

[0332] The composition and properties of the yogurt are shown in Table 5A. [Table 7]

[0333] Example 4B

[0334] This example describes the use of a dairy protein composition of the invention in the production of a high protein (12% by weight) set yoghurt.

[0335] The MPC prepared as in Example 3 was used to prepare set yogurt as in Example 4 above.

[0336] The composition and properties of the yogurt are shown in Table 5B. [Table 8]

[0337] 5. Example 5

[0338] This example describes the preparation of stirred yoghurt containing a dairy protein composition according to the present invention.

[0339] The stirred yoghurt of the present invention was prepared by the following method. 1. The MPC composition prepared in Example 2 was added to water to redissolve the powder and mixed thoroughly. 2. The yogurt mixture was subjected to a conventional yogurt milk heat treatment. 3. The mixture was cooled to 43°C. 4. Add the starter and stir until completely combined. 5. The inoculated mixture was incubated at 43° C. until the pH was about 4.6 (about 14 hours). 6. The yogurt was cooled to 20°C and sheared to break down the gel and produce stirred yogurt. 7. The yogurt was packaged and stored at 4°C.

[0340] The ingredients, composition, production conditions, and properties of stirred yogurt are summarized in Table 6.

[0341] Reference samples were prepared containing non-partially hydrolyzed MPC. Reference sample 1 (Reference 1) was prepared using MPC A. Reference sample 2 (Reference 2) was prepared using MPC B. [Table 9]

[0342] 6. Example 6

[0343] In this example, the sensory evaluation of the stirred yogurt produced in Example 5 is described.

[0344] An expert sensory panel (n=12) used quantitative descriptive sensory analysis to capture the sensory qualities of the yogurt. Sensory attributes were defined and intensity was scored on a fixed unstructured 150 mm line scale ranging from absent to strong. All data was collected using CompuSense 20® and analyzed using Minitab 18®. Yogurt was stored at 4°C until evaluation and gently stirred as well between the second samplings to ensure samples were stable. All samples were placed in clear sample cups at 20°C and identified with a random three-digit code under a red light. Panelists chose the mouthwash they would use, including water, soda water, plain water crackers, apple, and cucumber. [Table 10]

[0345] Row values ​​with different superscript letters indicate significant differences (P<0.05) between samples.

[0346] Sensory evaluation of yogurt prepared using the present invention (Sample I) showed that it is possible to produce a very thin, drinkable yogurt without affecting the flavor properties. Sample I was evaluated as not being too thick. Sample I has a similar bitterness level to the reference yogurt and a similar saltiness to the reference yogurt.

[0347] 7. Example 7

[0348] This example illustrates the preparation of a dairy protein composition containing sodium caseinate and its use in the production of yogurt.

[0349] Sodium caseinate was prepared as described in Example 1 above.

[0350] Sodium caseinate contains 91.9% protein and 40mg calcium per 100g sodium caseinate.

[0351] Caseinate was used to prepare set yoghurt using the method described in Example 4.

[0352] The composition and properties of the caseinates are given in Table 8 below. The composition and properties of the set yoghurt are summarised in Table 9. [Table 11] [Table 12]

[0353] 8. Example 8

[0354] This example describes the preparation of a food product containing a dairy protein composition of the present invention.

[0355] MPC with reduced calcium was prepared according to the method described in US Pat. No. 7,157,108.

[0356] The following MPC and SMP powders, as shown in Table 10, were reconstituted in water at 25-55°C using an overhead stirrer for 1 hour to produce an aqueous composition containing 10% protein by weight, and 7% protein from MPC B. MPC B (70% protein, 326mg calcium per 100g powder) Skim milk powder (protein 33%, calcium 1240mg 100g)

[0357] The composition was cooled to 5° C. and the appropriate amount of DuPont Food Pro PNL enzyme (as per the manufacturer's recommendations) was added.

[0358] The composition was incubated at 5° C. and neutral pH for 30 minutes to allow protein hydrolysis to occur.

[0359] The composition was then heated to 90°C and held for 5 minutes to inactivate the enzymes, then cooled to 43°C.

[0360] The mixture was inoculated with a starter (Chr Hansen YF-L702).

[0361] The mixture was placed in a pot.

[0362] The mixture was incubated at 43°C for approximately 9-16 hours until the final pH was approximately 4.6 to form yogurt.

[0363] The yogurt was stored chilled at 4°C.

[0364] Using the method of the present invention, the pH, breaking force and firmness of the yogurt were evaluated. The sensory properties of the yogurt were also evaluated informally.

[0365] A summary of the MPC including ingredients, composition, hydrolysis conditions, and properties of the partial hydrolysis products is given in Table 10 below.

[0366] These values ​​can be compared with samples A or C in Table 5 as controls. [Table 13]

[0367] Example 8A

[0368] This example describes the preparation of a food product containing a dairy protein composition of the present invention.

[0369] As detailed in Table 10A, the MPC and SMP powders listed below were reconstituted in water at 50-55°C with an overhead stirrer for 1 hour to produce aqueous compositions containing 6% or 10% protein by weight. MPC B (70% protein, 326mg calcium per 100g powder) MPC D (protein 82.4%, calcium 270mg per 100g powder) MPC F (protein 81.1%, calcium 2160mg per 100g powder) Skim milk powder (protein 33%, calcium 1240mg 100g)

[0370] The composition was cooled to 5° C. and enzyme treated as in Example 8.

[0371] Yogurt was prepared as described in Example 8. A summary of the composition, including ingredient composition, hydrolysis conditions, and description of the properties of the yogurt, is provided below in Table 10A. [Table 14-1] [Table 14-2]

[0372] 9. Example 9

[0373] In this example, the preparation of an acidic dairy beverage using a dairy protein composition of the invention comprising MPC is described.

[0374] The sour milk beverage was produced in the following manner.

[0375] Slowly add the modified MPC21 powder to the 55°C water, stirring the water at a speed sufficient to create a vortex but without creating excessive foaming. Reduce the speed and allow the powder to hydrate for 15 minutes.

[0376] Gellan gum and saccharide 1 were mixed and added to the MPC mixture and allowed to hydrate for 1 hour.

[0377] Pectin and sugar (2 parts) are mixed and added to water (2 parts), and the mixture is heated at high shear for 80 o Heated to C.

[0378] The MPC mixture was added to the pectin solution.

[0379] The mixing speed was increased and the citric acid was added, the pH checked and adjusted if necessary to pH 4.2.

[0380] The mixture was heat treated at 95°C for 12 minutes.

[0381] The mixture was homogenized at 150 / 50 bar (single pass).

[0382] The composition was aseptically packaged at 4°C or hot-filled at 80-85°C and the product was cooled as quickly as possible. [Table 15]

[0383] The viscosity, particle size distribution and pH were measured.

[0384] Acidic dairy beverages containing the compositions of the present invention have acceptable viscosity, particle size distribution and pH, and are free of objectionable flavors, as compared to acidic dairy beverages containing the corresponding acid compositions.

[0385] Example 9A

[0386] This example describes the preparation of an acidic dairy beverage using the dairy protein composition of the present invention.

[0387] With reference to the compositions in Table 11A, sour milk beverages were prepared using the following method.

[0388] Slowly add the MPC powder to 1 part of the water at 55°C and stir the water at a speed sufficient to create a vortex but not excessive foaming. Reduce the speed and allow the powder to hydrate for 1 hour.

[0389] Part 1 sugar was added to the MPC mixture.

[0390] The gellan gum, pectin and part 2 sugar were mixed and then added to water 2 under high shear at 80°C using an Ultratrax. Reconstituted at 10,000 rpm and mixed at 15,000 rpm. Once all the powder mixture was added, it was mixed for 1 minute.

[0391] The dissolution stabilizer / sugar solution was added to the MPC mixture and mixed for 5 minutes.

[0392] The mixture was cooled to 25° C. and shear adjusted to pH 4.2 with 10% citric acid solution.

[0393] The solution was made up to a final volume of 1000 g with water.

[0394] The mixture was heated to 60°C.

[0395] The mixture was homogenized at 200 / 50 bar (single pass).

[0396] The mixture was heated to 90° C. for 1 minute.

[0397] The composition was packaged sterile at 4°C. [Table 16-1] [Table 16-2]

[0398] The viscosity, particle size distribution, and pH were measured using the following methods.

[0399] Viscosity and particle size were measured as described in Section 5.

[0400] 10. Example 10

[0401] This example describes the preparation of an acidic dairy beverage using a dairy protein composition of the invention comprising MPC.

[0402] With reference to the composition in Table 12, a sour milk beverage was prepared using the following method.

[0403] The modified MPC powder (from Sample 21 in Example 3) was slowly added to 1 water at 55°C, which was stirred fast enough to create a vortex but not excessive foaming. After mixing for 15 minutes, skim milk powder (33% protein, 1240 mg calcium per 100 g flour) and whole milk powder (25% protein, 980 mg calcium per 100 g flour) were slowly added. The speed was reduced and the powders were allowed to hydrate for 1 hour.

[0404] To this mixture was added part 1 sugar.

[0405] Gellan gum and pectin were mixed with the second sugar and then mixed using the Ultratrax. ℃ The powder mixture was added to water 2 under high shear at 10,000 rpm. Reconstituted at 10,000 rpm and mixed at 15,000 rpm. Once all of the powder mixture was added, it was mixed for 1 minute.

[0406] The dissolved stabilizer / sugar solution was added to the MPC-milk powder-sugar 1 mixture, mixed for 5 minutes, adjusted to pH 4.2 with 10% citric acid solution cooled to 25°C and heated to 60°C.

[0407] The mixture was homogenized at 200 / 50 bar (single pass). It was then heated to 90° C. for 1 min. The milk drink was aseptically packaged at 4° C. [Table 17]

[0408] 11. Example 11

[0409] This example describes the preparation of a protein bar using a dairy protein composition of the present invention.

[0410] The MPCs used are: MPC D (protein 82.4%, calcium 270mg per 100g MPC) MPC F (protein 81.1%, calcium 2160mg per 100g powder) Modified MPC D Sample 21 from Example 3

[0411] Protein bars were prepared in the following manner.

[0412] Maltodextrin and MPC were mixed. Glucose syrup and glycerin were mixed and heated to 50-55°C. Hydrogenated coconut oil and lecithin were heated until the fat melted.

[0413] The grape syrup and glycerin mixture was added to the maltodextrin and MPC mixture, followed by the oil and lecithin mixture. The mixture was mixed for 90 seconds on speed 1 using a Hobart mixer (Model N-50) and the bowl was scraped down. Mixing (speed 2) was continued until a uniform mass was obtained.

[0414] The mixture was poured into a rectangular frame (approximately 16mm deep) placed on oiled baking paper and spread evenly. A piece of Gradwrap was sprayed with oil and the wrap was placed oiled side down on top of the mixture, completely covering the mixture. The mixture was spread flush with the frame and the excess was cut off. The mixture was left overnight to set.

[0415] The mixture was loosened from the mold and cut into strips of 30 mm x 100 mm. These chocolate bars were packed in aluminium foil pouches for storage prior to use. The composition and properties of the protein bars are given in Table 13.

[0416] The breaking force (g), water activity, color and sensory properties of the bars can be measured as described in Section 6. [Table 18-1] [Table 18-2] [Table 18-3]

[0417] 12. Example 12

[0418] This example describes the preparation of a dairy protein composition of the present invention and an ambient temperature yogurt comprising the dairy protein composition. Production of Milk Protein Compositions

[0419] The MPCD is prepared according to the methods described in US Pat. No. 7,157,108 to form a composition containing 6.9% protein by weight.

[0420] The composition was heated to 45° C. and the appropriate amount of DuPont Food Pro PNL (as per the manufacturer's recommendations) was added.

[0421] The compositions were heated and inactivated under the conditions outlined in Table 14 below.

[0422] A summary of the MPC including components, composition, hydrolysis conditions, and molecular weight distribution of the partial hydrolysis products is shown in Table 14. [Table 19]

[0423] Ambient yoghurt was prepared as follows. 1. The modified MPC was mixed with all powdered ingredients and added to water to reconstitute the powder and mixed thoroughly. The solution was mixed at 60°C for 20 minutes. 2. Cream was added and mixed for 10 minutes to produce the yogurt mixture. 3. The yoghurt mixture was homogenised at 150 / 50 bar and subjected to traditional yoghurt milk heat treatment and then cooled to 43°C. 4. Starter (Chr Hansen YF-L702) was added and stirred until well mixed. The inoculated mixture was incubated at 43° C. to a pH of approximately 4.2 (approximately 14 hours). 5. The yogurt was cooled to 20°C, flattened to break down the gel, heated at 75°C for 30 seconds and packaged.

[0424] The ingredients, composition, production conditions, and properties of stirred yogurt are summarized in Table 15. [Table 20]

[0425] 13. Example 13

[0426] This example describes preparing process cheese lollipops containing a dairy protein composition of the present invention.

[0427] The process cheese lollipops were prepared according to the following process. 1. Heat the water to 50℃. 2. Slowly add the dry mixed powder at 50℃, 3. Add cream cheese and butter. 4. Mix the composition for about 4 minutes. 5. Dilute lactic acid solution was used to adjust the pH of the mixture to pH 5.5. 6. The composition was heated to 90° C. with stirring for approximately 5 minutes. 7. The composition was packaged at a temperature above 65°C and cooled with 4°C airflow. ●Sample evaluation: ●The thickness is visualized, making mold processing and filling easier. Informal sensory analysis of flavor and texture (using blind codes to compare test differences and including blind replicates of controls) Yield stress

[0428] As described in Section 7, the yield stress of the processed cheese lollipops was measured at 13 °C. [Table 21-1] [Table 21-2]

[0429] 14. Example 14

[0430] This example describes an individually wrapped slice (IWS) for preparing a process cheese containing a dairy protein composition of the present invention.

[0431] IWS slices were prepared in a heated mixing pot by the following method. 1. Dry ingredients (weight, butter, grated cheese (e.g. TSC, DSP, SMP, gelling enzymes casein, citric acid, potassium sorbate, salt) were weighed out according to Table 17a. The grated cheese was refrigerated. 2. Place the dry ingredients (by weight), butter, and cheese ingredients (by weight) in the bowl of a Magimix food processor and process with the metal blade until the cheese becomes a roughly uniform paste (approximately 30-60 seconds). 3. Measure out the amount of water needed for this mixture and add it to a food processor and process until it lightens in color and becomes a uniform paste (about 30-60 seconds). 4. The cheese mixture was transferred from the Maggimix bowl to the Thermomix bowl. 5. 9 g of water (per 1000 g lot) was added to account for moisture loss during cooking. 6. The Thermomix temperature target was set to 95°C and the ingredients were mixed until melted, glossy and smooth (all cheese was fully melted). During this time the mixing speed (between speed 1-4) was manually adjusted at various times to ensure the product was moving well around the drum. 7. Once the mixture was melted, shiny and smooth, the pH was checked and adjusted to pH 5.70-5.75 with dilute citric acid. 8. Heating continued and when the temperature reached 87°C (manual measurement), the temperature setting on the Thermomix was reduced to 90°C and the product was mixed at speed 3-4 for 6 minutes. 9. During cooking, the slice forming device and other sample containers (plastic IWS film, 3 mm thick metal rod, marble rolling pin, and sample container) were prepared on the table next to the hot mixing machine. 10. Pour or scoop some of the hot mixture onto a polypropylene film and cover with another film, then press into a 3mm thick sheet using a rolling pin and metal rod to control the thickness. Repeat this to get 2-3 slices. 11. The film-coated slices were placed on pre-chilled metal trays and transferred to a cooler for further cooling. After 45 minutes, the chilled slices were batched together and stored in ziplock bags at 4°C. [Table 22]

[0432] The samples were evaluated as follows: Observation during the cooking process Hardness at 13°C 7 days after manufacture using a TA-HD Texture Analyzer Melting test for 7 days after production Informal analysis by processed cheese experts 7-10 days after manufacture Oven drying moisture

[0433] IWS hardness was measured by stacking 10 slices as described in Section 7 .

[0434] The melting test measured the melting and flow of cheese. Two 40 mm circles, approximately 3 mm thick, were cut from the stacked hardness test cheese samples. The circles were placed one on top of the other in the center of a glass petri dish and covered with a glass cover. • Samples were conditioned at 4°C for 10 minutes and then placed in the centre of a conventional preheated fan oven at 232°C for 5 minutes. After cooling, an uncovered sample was placed on the chart and the cheese's flow in six directions was measured. An average score of 0 indicates no flow in all six directions. The highest score, 12, indicates the sample was flowing in all directions toward the edge of the dish. Six flow measurements were averaged and recorded as the melt score.

[0435] IWS Humidity Measurements: ● Wet dishes were conditioned by heating in an oven and then cooling in a hair dryer. Approximately 10–10.5 g of sample was placed into a metal disk. This was heated in a controlled oven / incubator at 105°C for 16 hours. The dried sample was placed in a dryer to cool. The samples were weighed and the difference between the initial and final weight was considered as the moisture content of the cheese sample. [Table 23]

[0436] It is not intended to limit the scope of the invention to the above examples: as one skilled in the art will appreciate, many variations are possible without departing from the scope of the invention as defined in the claims.

[0437] Industrial Applicability

[0438] The dairy protein compositions of the present invention are useful in the preparation of low viscosity protein-containing foods, including acidified and fermented foods such as yogurt and acidified dairy drinks.

Claims

1. A milk protein concentrate (MPC), milk protein isolate (MPI), or a combination thereof, wherein a) the MPC, MPI, or combination thereof contains at least about 40% by weight of total protein based on dry matter; b) the total protein contains less than about 79% by weight of peptides having a molecular weight greater than about 20 kDa; c) the MPC, MPI, or combination thereof i. contains less than about 2 g of calcium per 100 g of the total protein, and / or ii. contains less than about 1.4 g of calcium per 100 g of dry matter, a composition.

2. The MPC, MPI, or combination thereof according to claim 1, wherein the total protein contains about 20% to about 79% by weight of peptides having a molecular weight greater than about 20 kDa.

3. The MPC, MPI, or combination thereof according to claim 1, wherein the total protein contains about 15% to about 55% by weight of peptides having a molecular weight of about 5 to about 20 kDa.

4. The MPC, MPI, or combination thereof according to claim 1, wherein the total protein contains about 5% to about 30% by weight, or about 5% to about 22% by weight of peptides having a molecular weight less than about 5 kDa.

5. The MPC, MPI, or combination thereof according to claim 1, wherein the total protein contains about 2% to about 20% by weight, or about 2% to about 12% by weight of peptides having a molecular weight of about 1 to about 5 kDa.

6. The MPC, MPI, or combination thereof according to claim 1, wherein the total protein contains about 2% to about 20% by weight, or about 2% to about 9% by weight of peptides having a molecular weight less than about 1 kDa.

7. The total protein is a) about 20% to about 79% by weight of peptides having a molecular weight greater than about 20 kDa, and b) about 15% to about 54% by weight of peptides having a molecular weight of about 5 to about 20 kDa, and c) about 2% to about 17% by weight of peptides having a molecular weight of about 1 to about 5 kDa, and d) about 2% to about 20% by weight of peptides having a molecular weight less than about 1 kDa, the MPC, MPI, or combination thereof according to claim 1.

8. The MPC, MPI, or combination thereof according to claim 1, a dry composition, preferably a powder.

9. The total protein is a) about 35% to about 65% by weight of peptides having a molecular weight greater than about 20 kDa, b) about 25 to about 50% by weight of a peptide having a molecular weight of about 5 to about 20 kDa, c) about 4 to about 12% by weight of a peptide having a molecular weight of about 1 to about 5 kDa, d) about 2 to about 6% by weight of a peptide having a molecular weight of less than about 1 kDa, or e) the MPC, MPI, or a combination thereof according to claim 1, comprising any combination of two or more of a) to d). **Claim 10**: A method for preparing MPC, MPI, or a combination thereof, a) providing an aqueous composition comprising a milk protein concentrate, a milk protein isolate, or a combination thereof, and containing about 0.5 to about 20% by weight of total protein; b) exposing the aqueous composition to the action of one or more proteolytic enzymes; c) inactivating the one or more proteolytic enzymes to produce MPC, MPI, or a combination thereof, wherein 1. the MPC, MPI, or a combination thereof contains at least about 40% by weight of total protein based on dry matter; 2. the total protein in the MPC, MPI, or a combination thereof contains less than about 79% of peptides having a molecular weight greater than about 20 kDa; 3. the MPC, MPI, or a combination thereof i. contains less than about 2 g of calcium per 100 g of total protein, and / or ii. contains less than about 1.4 g of calcium per 100 g of dry matter. **Claim 11** The method according to claim 10, comprising subjecting the milk protein composition to ion exchange chromatography, calcium chelation, and / or ultrafiltration under acidic conditions to produce an aqueous composition. **Claim 12** The method according to claim 11, wherein the calcium in the MPC, MPI, or a combination thereof is reduced by at least about 40% by weight. **Claim 13** The method according to claim 10, wherein the proteolytic enzyme is an endopeptidase. **Claim 14** The method according to claim 10, wherein the proteolytic enzyme is derived from the genus Bacillus, Aspergillus, or Fusarium. **Claim 15** The method according to claim 10, wherein the proteolytic enzyme has optimal activity at a pH of about pH 6 to about pH 11. **Claim 16** a) a temperature of about 0°C to about 85°C, b) about 30 seconds to about 48 hours, and / or c) exposing the aqueous composition to the action of one or more proteolytic enzymes at a pH of from about pH 6 to about pH 8, the method according to claim 10.

17. The method according to claim 10, further comprising the step of drying the MPC, MPI, or a combination thereof to form a powdered (dry) MPC, MPI, or a combination thereof.

18. A method for producing a protein-containing food, comprising: a) providing an aqueous composition comprising MPC, MPI, or a combination thereof according to any one of claims 1 to 9; and b) mixing with one or more additional ingredients to produce a protein-containing food.

19. The method according to claim 18, wherein the food is a liquid nutritional composition, a beverage, ice cream, an acidified product, an acidified milk beverage, yogurt, cheese, processed cheese, cheese analog, buttermilk, a fermented product, drinking yogurt, set yogurt, Greek yogurt, stirred yogurt, milk, kefir, skyr, puto svejs, ambient yogurt, a fermented milk beverage, a smoothie, sour cream, quark, pudding, a frozen dessert, a coffee whitener, a gel, a bar, or a baked good.

20. The method according to claim 18, wherein the one or more additional ingredients comprise one or more lipids, carbohydrates, proteins, flavors, vitamins, minerals, dairy products, water, food additives, dyes, fruit preparations, or any combination of two or more of these ingredients.

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