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WO2025173761A1 - Nouveau produit laitier et son procédé de production - Google Patents

Nouveau produit laitier et son procédé de production

Info

Publication number
WO2025173761A1
WO2025173761A1 PCT/JP2025/004905 JP2025004905W WO2025173761A1 WO 2025173761 A1 WO2025173761 A1 WO 2025173761A1 JP 2025004905 W JP2025004905 W JP 2025004905W WO 2025173761 A1 WO2025173761 A1 WO 2025173761A1
Authority
WO
WIPO (PCT)
Prior art keywords
coagulum
milk
composition
cheese
protein
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/JP2025/004905
Other languages
English (en)
Inventor
Abdul-Hamid KLANDAR
Martin HERBINIERE
Teppei Ogawa
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Ajinomoto Co Inc
Original Assignee
Ajinomoto Co Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Ajinomoto Co Inc filed Critical Ajinomoto Co Inc
Publication of WO2025173761A1 publication Critical patent/WO2025173761A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
    • A23C19/00—Cheese; Cheese preparations; Making thereof
    • A23C19/06—Treating cheese curd after whey separation; Products obtained thereby
    • A23C19/063—Addition of, or treatment with, enzymes or cell-free extracts of microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
    • A23C19/00—Cheese; Cheese preparations; Making thereof
    • A23C19/06—Treating cheese curd after whey separation; Products obtained thereby
    • A23C19/064—Salting
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
    • A23C19/00—Cheese; Cheese preparations; Making thereof
    • A23C19/06—Treating cheese curd after whey separation; Products obtained thereby
    • A23C19/068—Particular types of cheese
    • A23C19/0684—Soft uncured Italian cheeses, e.g. Mozarella, Ricotta, Pasta filata cheese; Other similar stretched cheeses
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING OR TREATMENT THEREOF
    • A23C19/00—Cheese; Cheese preparations; Making thereof
    • A23C19/06—Treating cheese curd after whey separation; Products obtained thereby
    • A23C19/068—Particular types of cheese
    • A23C19/08—Process cheese preparations; Making thereof, e.g. melting, emulsifying, sterilizing
    • A23C19/084—Treating the curd, or adding substances thereto, after melting

Definitions

  • Whey obtained from raw milk comprises proteins (e.g., whey protein), saccharides and minerals which may be re-used in food products.
  • proteins e.g., whey protein
  • saccharides and minerals which may be re-used in food products.
  • whey proteins typically have undesirable textures. Therefore, methods to recover proteins from cheese whey require several processing steps.
  • EP 0 966 887 states that the dissolution stability of whey proteins can be affected by aggregation of cheese whey proteins. To dissolve these aggregates, partial denaturation of the whey protein and enzymatic treatments have been attempted.
  • a crosslinking and/or protein deamidating enzyme e.g., TG, laccase, tyrosinase, protein glutaminase
  • TG crosslinking and/or protein deamidating enzyme
  • EP 0 966 887 (Ajinomoto Co., Inc. and Amano Enzyme Inc.) WO 2006/075772 (JP) / EP 1 839 491 (Ajinomoto Co., Inc.) WO 2012/172179 (Valio LTD) WO 2015/150637 (Valio LTD) WO 2006/026811 (Smith 2006) JP-A-64-27471 JP-B-1-50382 JP-A-1-300889 JP-A-6-225775 JP-A-7-23737 JP-A-64-27471 JP-A-2000-50887 JP-A-2001-218590 US20220046888A1
  • the present invention solves the problem of providing a dairy product, in particular cheese, like pasta filata cheese, which has excellent organoleptic properties, whilst avoiding by-products (e.g., waste products) like whey or other liquid compositions contaminated with protein or other soluble components derived from the dairy product, like contaminated water derived from the traditional combined cooking-stretching step and contaminated brine resulting from the traditional brining step.
  • by-products e.g., waste products
  • whey or other liquid compositions contaminated with protein or other soluble components derived from the dairy product like contaminated water derived from the traditional combined cooking-stretching step and contaminated brine resulting from the traditional brining step.
  • the invention provides several steps that each individually achieves an improvement over the art.
  • the present invention provides a method for producing a dairy product comprising a conversion of a coagulum to the dairy product comprising the steps of: (a) milling the coagulum to obtain a composition comprising coagulum pieces; (b) subjecting the composition obtained by step (a) to an enzymatic treatment wherein a transglutaminase and/or a protein-glutaminase act on the coagulum pieces; and (c) subjecting the composition obtained by step (b) to a steam stretching step.
  • an enzymatic treatment step prior to the stretching step for dairy product processing e.g., an enzymatic treatment step in the second phase of cheesemaking including the conversion of a coagulum to a dairy product
  • the physical and chemical properties of milk proteins such as solubility, hydration capacity, thermal stability, emulsifying and foaming properties, viscosity, elasticity, and gelation can be improved.
  • fermented coagulum pieces also referred to as “grains” are subjected to an enzyme preparation comprising a crosslinking and/or protein deamidating enzyme (a transglutaminase and/or a protein-glutaminase).
  • the present inventors also found that the traditional combined cooking-stretching step for pasta filata processing, results in protein hydration of the composition comprising coagulum pieces.
  • Protein hydration also referred to as “protein swelling” leads to a dairy product characterized by an undesirable microstructure, product texture and rheological/viscoelastic properties.
  • coagulum components such as oils, proteins (in the form of fines) and salt, may be lost during the traditional combined cooking-stretching step. Due to these diluted components, the water cannot be re-used or directly disposed (see, e.g., WO 2006/026811).
  • the traditional combined cooking-stretching step can be substituted with a steam stretching step (“dry stretching”).
  • a steam stretching step after the enzymatic treatment step, the microstructure of the composition is rearranged without loss of by-products such as whey, oil/fat or proteins (in the form of fines) and the crosslinking and/or protein deamidating enzyme is inactivated.
  • the obtained dairy product has the desired textural properties (e.g., a plastic, pliable, homogeneous, stringy structure including fines) and chemical composition (e.g., no whey expulsion during processing).
  • dairy product yield e.g., higher recovery of proteins and fats
  • traditional methods e.g., no loss of coagulum pieces, proteins or fats during processing
  • milk components e.g., whey and milk proteins
  • the invention provides a step of converting a milk composition to a coagulum from a reconstituted milk composition.
  • a dairy product such as the pasta filata cheese product
  • the flavour and texture of the obtained dairy product can be adjusted and improved, e.g., when the quality and/or quantity of available raw milk is poor or limited.
  • the composition of milk components such as the protein composition, amount of fat and liquids can be adjusted. For instance, the liquid content and the amount of whey protein may be reduced, so that a whey drainage step may be omitted.
  • a traditional brining step (see Figure 1A, column “traditional”, processing step No. 15) may be omitted.
  • the salt content of a coagulum and/or a dairy product may be adjusted by one or more salting step(s) (also referred to as “partial salting”) during the first phase (conversion of a milk composition to a coagulum), and/or the second phase (conversion of the coagulum to a dairy product).
  • the one or more salting step(s) allow, e.g., to modulate the coagulation and/or fermentation reaction(s) in the LAB starter culture, to modulate the protein-crosslinking reaction during the enzymatic treatment step and/or enhance flavour of the obtained dairy product.
  • brining and the resulting by-products (brines) can be avoided. This may be beneficial, as brines are a problematic waste product due to their high salinity, pH, and high organic content.
  • these steps are combined to result in a process of making a dairy product with excellent organoleptic properties and an improved dairy product yield, whilst avoiding or minimizing the generation of by-products (e.g., waste products).
  • Elements that interact in achieving these aims are: ⁇
  • the use of reconstituted milk as a starting material, as reconstituted milk has a defined composition and may have a reduced solvent and whey protein content. This obviates the loss of whey in the generation of a coagulum, as the whey present in the reconstituted milk can be recovered into the coagulum due to the enzymatic treatment step. It also provides a means for controlling the characteristics of the generated coagulum, such as texture.
  • reconstituted milk in combination with the subsequent processing steps may therefore allow the omission of a whey drainage step.
  • This enzymatic treatment step improves the emulsifying and hydrating properties of the proteins.
  • TG further cross-links the protein. This reduces the amount of low molecular weight protein complexes, such as monomeric and dimeric caseins.
  • a steam stretching step in combination with enzymatic treatment step allows for the retention of liquids such as water and oils, as well as the retention of milk proteins, as steam stretching may be performed until all liquids and solids are absorbed. As a result, there is no loss of soluble substances (that would result in a loss of yield) during the stretching step and no by-products are generated. In addition, unlike traditional methods, no wastewater is generated during this step.
  • One or more salting step(s) allow for direct salting, e.g., in the form of one or more step(s) distributed over various method steps to avoid the brining step. Again, this avoids the generation of a waste solution contaminated with soluble components from the dairy product, including oils, sugars and proteins which are usually associated with traditional brining.
  • the process can be such that liquid waste products are either reduced or even eliminated.
  • each of the steps contributes to avoiding loss of soluble material on the way from the starting material to the finished dairy product. Avoiding this loss increases the yield of final product that is obtained from a given amount of starting material as compared to a conventional process.
  • the combined use of the enzyme treatment step and steam stretching results in dairy product with excellent properties.
  • a pasta filata cheese can be obtained that is characterized by a plastic, pliable, homogeneous, stringy structure (e.g., an equal or an improved product texture as compared with the product texture of a pasta filata cheese obtained from raw milk, e.g., natural whole milk) and a moisture content of more than 40 and less than 80 MFFB%, wherein MFFB% is a percentage moisture on a fat free basis.
  • MFFB% is a percentage moisture on a fat free basis.
  • a method for producing a dairy product comprising a conversion of a coagulum to the dairy product comprising the steps of: (a) milling the coagulum to obtain a composition comprising coagulum pieces; (b) subjecting the composition obtained by step (a) to an enzymatic treatment wherein a transglutaminase and/or a protein-glutaminase act on the coagulum pieces; and (c) subjecting the composition obtained by step (b) to a steam stretching step.
  • step (a) the coagulum is subjected to a device configured to mill a block of said coagulum into coagulum pieces having an average diameter of 20 to 60 mm, wherein the average diameter is defined as the average longest distance between two opposing points along the coagulum piece boundary in a 2-dimensional image of the coagulum pieces.
  • step (b) an enzyme preparation is added to the coagulum pieces, and wherein the enzyme preparation comprises the transglutaminase and/or the protein-glutaminase; optionally wherein the enzyme preparation further comprises a caseinate.
  • step (c) the steam stretching step comprises a thermomechanical treatment, wherein steam heating is performed simultaneously to mechanically working, stirring, kneading, and/or stretching the composition.
  • any one of items 1-7 comprising a preceding conversion of a milk composition to a coagulum, wherein the preceding conversion comprises the steps of: (a) preparing the milk composition to obtain a composition having a pH in the range of 6.6 to 6.8; and (b) adding an acidifier and a coagulant to the milk composition obtained by step (a), optionally, wherein the acidifier and the coagulant are added simultaneously.
  • the milk composition comprises a raw milk and optionally one or more components selected from a protein component, a fat component and a solvent.
  • the milk composition comprises a reconstituted milk, wherein the reconstituted milk comprises a protein component, a fat component and a solvent, and/or wherein the milk composition comprises a moisture content of between 58-65 MFFB%, wherein MFFB% is a percentage moisture on a fat free basis.
  • the fat component is a non-dairy fat or a dairy fat, optionally a fat component selected from a butteroil and an anhydrous milk fat.
  • the milk composition, the coagulum and/or the dairy product further comprise(s) one or more additional ingredients selected from the group of a buffering agent, a colouring agent, a flavoring agent, and a preservative.
  • the pasta filata cheese is a Mozzarella cheese selected from whole milk Mozzarella cheese, low moisture Mozzarella cheese, part-skim Mozzarella cheese, and low moisture part-skim Mozzarella cheese based on the moisture and fat-on-dry basis contents.
  • the dairy product comprises one or more dairy-like properties selected from the group consisting of stretch or stringiness, melt, elasticity of melted dairy product, and oiling after heating the dairy product.
  • a method for producing a coagulum consisting of the steps of: (a) preparing a reconstituted milk composition to obtain a composition having a pH in the range of 6.6 to 6.8, wherein the reconstituted milk composition comprises a protein component, a fat component and a solvent, and wherein the reconstituted milk comprises a moisture content of between 58 to 65 MFFB%, wherein MFFB% is a percentage moisture on a fat free basis; (b) pasteurizing the composition obtained by step (a); (c) adding an acidifier and a coagulant simultaneously to the composition obtained by step (b) for less than 10 min, wherein the acidifier is a lactic acid bacteria (LAB) starter culture and the coagulant is rennet; and (d) fermenting the coagulum obtained by step (c).
  • LAB lactic acid bacteria
  • a coagulum obtainable by the method according to item 29.
  • Figure 1A is a comparison between a traditional cheesemaking process (see column “traditional”) and an exemplary cheesemaking process according to the present invention (see column “invention”).
  • Figure 1B is an exemplary cheesemaking process according to the present invention (e.g., for pasta filata cheese).
  • the method comprises one or more salting step(s) (see column “salting step(s)”) added simultaneously to the processing step indicated in the table (any one of or a combination of the process step(s) of coagulant addition, demoulding, milling, enzymatic treatment and/or stretching).
  • Figure 1C is an exemplary cheesemaking process according to the present invention (e.g., for pasta filata cheese) comprising (i) a conversion of a milk composition to a coagulum according to the traditional method, and (ii) a conversion of the coagulum to the dairy product according to the present invention.
  • the combination of processing steps has the advantage that loss of fines in the obtained cheese product is minimized or avoided.
  • Figure 1D is an exemplary cheesemaking process according to the present invention for producing a low-moisture pasta filata cheese (e.g., a pizza cheese).
  • Figure 2A shows photographs of an exemplary manufacturing process for pasta filata cheese starting from a reassembled milk composition; coagulum (pre-cheese) preparation by steam stretching in a cooker (e.g., a Stephan cooker).
  • Figure 2B shows photographs of an exemplary manufacturing process for pasta filata cheese starting from a reassembled milk composition; coagulum after fermentation.
  • Figure 2C shows photographs of an exemplary manufacturing process for pasta filata cheese starting from a reassembled milk composition; coagulum after fermentation; the obtained cheese was manually moulded under 5lbs loaves shapes.
  • Figure 3 is an exemplary analysis of cheese shreddability of a control cheese (A), a TG-treated cheese (B) and a TG-and-PG (“TG/PG”) treated cheese (C).
  • Figure 4 is an exemplary analysis of a cheese bake-off property (e.g., melting) of a control cheese (A), a TG-treated cheese (B) and a TG-and-PG (“TG/PG”) treated cheese (C).
  • the photograph shows cheese bake-off property (e.g., melting) at 2 weeks of storage at 5°C.
  • Figure 5 is an exemplary analysis of a cheese bake-off property (e.g., stretching) of a control cheese (A), a TG-treated cheese (B) and a TG-and-PG (“TG/PG”) treated cheese (C).
  • the photograph shows cheese bake-off property (e.g., stretching) at 2 weeks of storage at 5°C. Cheese stretching was evaluated after baking at 63°C.
  • the term “comprising” and variations of the word typically is not limiting and thus does not exclude other features, which may be for example technical features, additives, components, or steps.
  • this also includes a special embodiment in which this word is understood as limiting; in this particular embodiment the word “comprise” has the meaning of the term “consist of”.
  • the term “similar”, as used herein, is interchangeable for alike, analogous, comparable, corresponding, and -like, and is meant to have the same or common characteristics, and/or in a quantifiable manner to show comparable results i.e., with a variation of maximum 20 %, 10 %, more preferably 5 %, or even more preferably 1 %, or less.
  • “Dairy product”, as used herein, refers to a milk product obtained by coagulating all or part of the protein of milk, skimmed milk, partly skimmed milk, cream, whey cream, buttermilk, or reconstituted milk, or any combination of these materials, through the action of suitable coagulating agents (e.g., rennet).
  • suitable coagulating agents e.g., rennet
  • the dairy product may be characterized by firmness (e.g., according to MFFB% or a designation selected from extra hard, hard, firm/semi-hard, and soft), ripening (e.g., ripened, mould ripened, unripened/fresh, in brine), milk fat content (e.g., as a percentage by mass; as a percentage of fat in dry matter; in grams per serving as quantified in the label provided that the number of servings is stated).
  • the dairy product may be a cheese (e.g., a pasta filata cheese).
  • the chemical composition of an (intermediate) product or composition may be analysed by a spectroscopy method such as visible (Vis) and near-infrared (NIR) spectroscopy wherein the concentration of biologically important bonds (e.g., aliphatic C-H, aromatic or alkene C-H, amine N-H and O-H) that absorb in the VIS or NIR range is measured.
  • a spectroscopy method such as visible (Vis) and near-infrared (NIR) spectroscopy wherein the concentration of biologically important bonds (e.g., aliphatic C-H, aromatic or alkene C-H, amine N-H and O-H) that absorb in the VIS or NIR range is measured.
  • biologically important bonds e.g., aliphatic C-H, aromatic or alkene C-H, amine N-H and O-H
  • Studies have assigned specific wavelengths to predictions of the fat, protein, and moisture contents of dairy products such as cheese (see,
  • Coagulum refers to a “curd”, “protein gel”, “grain” or “pre-cheese”.
  • the coagulum consists of a milk product obtained by coagulating wholly or partly the protein of milk, e.g., according to phase (i) of a traditional method or phase (i) of the present invention
  • a coagulum may be obtained according to the processing steps shown in Figure 1A.
  • “Cheese”, as used herein, refers to the fresh or matured dairy product obtained after coagulation of milk, cream, skimmed, or partly skimmed milk, or a combination of some or all of these products, and including any cheese that conforms to the requirements of the Food and Drug Administration for cheeses and related cheese products (21 CFR part 133), and any cheese that conforms to the global standards for the definition of cheese according to the Codex Alimentarius (https://www.fao.org/3/i2085e/i2085e00.pdf [current as of January 2024]).
  • “Milling” or “cutting”, as used herein, refers to reducing the size of a coagulum, e.g., to coagulum pieces, particles or grains necessary for facilitating subsequent processing steps (e.g., the enzymatic treatment step, and for providing even heating during stretching).
  • a coagulum is subjected to a device configured to mill a block of said coagulum into coagulum pieces wherein the term “configured to” means adapted to carry out the respective steps/functions (rather than merely means suitable for carrying them out).
  • Coagulum pieces or “coagulum particles”, as used herein, refers to the product obtained from milling.
  • the obtained coagulum pieces have a structure that is optimized for facilitating subsequent processing steps (e.g., the enzymatic treatment step, and for providing even heating during stretching).
  • the coagulum pieces are characterized in - an average surface area (mm 2 ); - an average size (e.g., an average diameter of 20 to 60 mm; and/or - an average weight (e.g., 2 to 8 g) and/or average density.
  • Particle size (mm), particle shape (simple or complex) and/or particle surface area (mm 2 ) may be measured by, e.g., sieving methods or 2-dimensional (2D) image analysis (e.g., according to Aldalur et al. 2019, doi: 10.3168/jds.2018-15177).
  • Particle size refers to a diameter of a particle.
  • Diameter refers to a distance (mm) between two opposing points passing through the centre of a particle (i.e., the centre of a 3-dimensional or a 2-dimensional particle).
  • Average diameter refers to the average longest distance between two opposing points along the coagulum piece boundary in a 2-dimensional image of the coagulum pieces.
  • the coagulum pieces are characterized in an average diameter of 20 to 60 mm.
  • Density is the mass of a material per unit volume.
  • density may be defined as true or apparent density.
  • average density may be measured by, e.g., a water displacement method (see, e.g., Iezzi et al. 2013, doi:10.1111/jfpe.12008, page 464, section “Cheese Density Measurements”).
  • Enzymatic treatment refers to a step wherein a protein crosslinking enzyme (a transglutaminase and/or a protein-glutaminase) is used.
  • Enzyme preparation refers to a composition comprising crosslinking and/or protein deamidating enzyme(s), e.g., a transglutaminase (TG) and/or a protein-glutaminase (PG).
  • the composition may comprise a further crosslinking and/or protein deamidating enzyme selected from a group consisting of transglutaminase (TG), laccase, tyrosinase, peroxidase, sulfhydryl oxidase, glucose oxidase, and protein-glutaminase (PG).
  • the composition further may comprise a caseinate.
  • Transglutaminase or “TG”, as used herein, refers to a protein-glutamine gamma-glutamyltransferase (EC 2.3.2.13) which catalyses the formation of covalent bonds between the amino acid residues glutamine and lysine in proteins.
  • the milk protein caseins particularly K-casein and to a lesser extent ⁇ -casein, are substrates for transglutaminase because they are rich in glutamine and lysine.
  • Transglutaminases are divided into calcium independent and calcium dependent types. Both can be used in the present invention.
  • Examples of the former include those derived from microorganisms such as Actinomycetes, Bacillus subtilis and the like (see, for example, JP-A-64-27471).
  • Examples of the latter include those derived from guinea pig liver (see, for example, JP-B-1-50382), those derived from microorganisms such as oomycetes, those derived from animals such as bovine blood, pig blood and the like, those derived from fish such as salmon, red sea bream and the like (see, for example, Seki Nobuo et al., Nippon Suisan Gakkaishi, vol.56, NO.1, pp.125-132 (1990)), those derived from oyster etc.
  • transglutaminase can be produced by genetic engineering (see, for example, JP-A-1-300889, JP-A-6-225775, JP-A-7-23737).
  • any of these transglutaminases may be used without any specific limitation as to origin and preparation, insofar as they can catalyse the acyl exchange reaction between the gamma carboxy-amide group of a glutamine residue and the epsilon amino group of a lysine residue, which are present in the protein.
  • the calcium-independent transglutaminases are preferable.
  • the microorganism-derived transglutaminases fulfil all conditions and are preferred enzymes.
  • Protein-glutaminase or “PG”, as used herein, refers to a protein-glutaminase (EC 3.5.1.44) which catalyses the deamidation of glutamine residues within proteins, and glutamine is converted to glutamic acid.
  • the type of protein glutaminase in the present invention is not particularly limited as long as it acts directly on an amide group of a protein to deamidate it without cleaving a peptide bond or cross-linking the protein.
  • Powder composition refers to a composition obtained by drying (e.g., consisting of; or comprising a protein component). Drying may be carried out by any method generally used in the field, such as spray drying.
  • the powder composition can be recombined into a solvent to provide the composition in liquid form.
  • a powder composition comprising the enzyme preparation can be recombined into the composition comprising coagulum pieces.
  • a powder composition of a protein component e.g., casein or whey protein
  • water e.g., casein or whey protein
  • “Slurry”, as used herein, refers to a composition comprising the enzyme preparation in liquid form.
  • the enzyme preparations including transglutaminase (TG) and/or protein-glutaminase (PG) is dissolved in a solvent, preferably an aqueous solution (e.g., 20% w/w).
  • the slurry may also include further components, such as caseinate powders; particularly preferred are sodium caseinate and/or reduced calcium or demineralized casein micelles concentrate. Preferred concentrations of caseinate powder in the slurry may be between 5-15 % w/w.
  • the slurry may comprise lactose, acidity regulators (lactic acid and calcium lactates or sodium phosphate), and salt (e.g., NaCl). The concentrations of lactose and salt may be adjusted to correspond to the concentrations of the coagulum to avoid dilution of the coagulum.
  • Milk composition refers to the starting material for the dairy product-making process, e.g., a composition comprising or consisting of raw milk or a reconstituted milk composition.
  • the properties of the starting milk composition may be adjusted including total protein content (e.g., milk proteins such as casein and whey protein), the protein-to-fat ratio, pH value, temperature, calcium (Ca) content, casein interactions and others (Kindstedt, P.S., Hillier, A.J. and Mayes, J.J. (2010). Technology, Biochemistry and Functionality of Pasta Filata/Pizza Cheese. In Technology of Cheesemaking (eds B.A. Law and A.Y. Tamime); Goncalves MC, Cardarelli HR. (2021). Food Technol Biotechnol.).
  • “Raw milk“ or “fresh milk”, as used herein, denotes an edible milk such as cow milk, buffalo milk, goat milk, sheep milk, horse milk or any other animal producing milk suitable for animal or human nourishment.
  • the raw milk may be supplemented with ingredients generally used in the production of milk products, such as fat, protein, mineral and/or sugar fractions or the like.
  • the raw milk may thus be, for instance, whole milk.
  • Protein component refers to proteins obtainable from a raw milk (natural milk proteins, e.g., a casein, and/or a whey protein), synthetic variants of natural milk proteins and other proteins. Casein can be roughly classified into ⁇ -, ⁇ -, and ⁇ -caseins. In milk, caseins are generally present as casein micelles. ⁇ -casein is thought to be predominantly localized on the micellar surface where is stabilises the micelle structure through its interaction with calcium.
  • ⁇ -lactoglobulin ⁇ -LG, for short
  • ⁇ -LA ⁇ -lactalbumin
  • IG immunoglobulins
  • BSA bovine serum albumin
  • BLF bovine lactoferrin
  • LP lactoperoxidase
  • Demineralized refers to a composition with reduced mineral salts (e.g., calcium, phosphorus, and/or magnesium) comprising a protein component (e.g., a milk protein such as casein or whey protein).
  • a protein component e.g., a milk protein such as casein or whey protein.
  • demineralized casein powder or concentrate refers a reduced-Ca Micellar Casein Concentrated (MCC) powder, i.e., at least 40% of colloidal calcium have been removed from the native MCC powder.
  • Demineralization is carried out during liquid MCC manufacturing by adding a Ca-chelating agents such as CO 2 and/or citric acid.
  • the preferred concentration of calcium in demineralized powder is preferably close to the final calcium in cheese (e.g., calculated as mg Ca / g casein in pasta filata cheese).
  • Acidifier refers to a compound or composition for adjusting the pH of the milk composition to a pH suitable for coagulation, for example, a starter culture. Acidification can be achieved by adding a starter culture (traditional method) or by direct acidification of milk (industrial method) with organic acids.
  • An acidifier according to the present invention can be either of a starter culture comprising acid-producing bacteria (e.g., lactic acid bacteria, LAB) or an organic acid.
  • LAB starter cultures may be selected from “thermophilic” and “mesophilic” LAB species.
  • Coagulant refers to coagulants in cheese manufacture for coagulating a milk.
  • a coagulant may be selected from a calf rennet, a microbial rennet (e.g., derived from Rhizomucor miehei, Rhizomucor pusillus and Cryophenectria parasitica), purified versions thereof or pure chymosin.
  • the coagulant comprises a protease which acts on ⁇ -casein as a substrate.
  • the protease cleaves the bond between phenylalanine, the 105th amino acid, and methionine, the 106th amino acid, both from the N-terminus of ⁇ -casein.
  • This cleavage point is the boundary between the hydrophilic and hydrophobic parts of the casein.
  • the protease separates the hydrophilic part from the ⁇ -casein and exposes the hydrophobic part outside the casein micelle.
  • the individual hydrophobic parts gradually aggregate by interaction, become more unstable in the presence of calcium ions and precipitate, e.g., when the temperature is raised.
  • This precipitate is called coagulum (or curd) and the water-soluble part that has not curdled is separated as whey.
  • the coagulum may be characterised by a defined ratio of clotting-to-proteolysis, e.g., the higher said ratio value, the lower is the proteolysis after clotting.
  • Salting step refers to the addition of salt to affect microbial growth and activity, control of the various enzyme activities used in the processing steps, physicochemical, functional and flavour characteristics of the obtained dairy product (e.g., moisture content, texture, protein solubility, protein conformation).
  • the salt is absorbed by (an intermediate product of) the dairy product, and moisture may simultaneously be expelled. Salting may be performed by brining (also referred to as “brine salting”) or direct addition to intermediate products of the dairy product (e.g., to the coagulum).
  • a salt may be selected from sodium chloride (NaCl) and other salts typically used in processed cheese manufacture.
  • NaCl sodium chloride
  • a dairy product such as a pasta filata cheese obtained from raw/fresh milk (a natural pasta filata cheese)
  • sodium chloride (NaCl) is used for a salting step according to the present invention.
  • a salt may be selected from sodium chloride (NaCl) and other salts typically used in processed cheese manufacture (e.g., emulsifying/melting/chelating salts which are Na, K or Ca-salts of citrate or phosphates such as trisodium citrate or sodium aluminium phosphate).
  • NaCl sodium chloride
  • other salts typically used in processed cheese manufacture e.g., emulsifying/melting/chelating salts which are Na, K or Ca-salts of citrate or phosphates such as trisodium citrate or sodium aluminium phosphate.
  • “Pasta filata cheese” or “stretched curd cheese”, as used herein, refers to a finished cheese with several distinct functionalities compared to other cheese, such as - melt characteristics (e.g., as measured by the Arnott and Schreiber tests where discs of cheese are melted under standard conditions in a convection oven and changes in the diameter of the discs after heating are determined); - stretch characteristics (e.g., determined by measuring how far the melted cheese will stretch when a constant force is applied either vertically or horizontally); - the potential to release free-oil when heated (e.g. as measured using a modified Babcock test). These feature result from the unique thermising and texturising process that occurs during stretching of pasta filata cheese.
  • - melt characteristics e.g., as measured by the Arnott and Schreiber tests where discs of cheese are melted under standard conditions in a convection oven and changes in the diameter of the discs after heating are determined
  • - stretch characteristics e.g.,
  • the chemical composition of the (intermediate) products and compositions of the present invention may measured by standard methods known to the skilled person such as a spectroscopy method.
  • a spectroscopy method for example, dry matter, content of fat and/or protein may be measured by a spectroscopy method such as visible (Vis) and near-infrared (NIR) spectroscopy (e.g., using a FoodScan device).
  • Vis visible
  • NIR near-infrared
  • Total weight refers to the weight of all components comprised in a composition at the respective production steps, unless otherwise defined. For instance, at and after the final production step, total weight refers to the total weight of the dairy product such as the cheese.
  • Dry matter refers to that portion of the dairy product or composition that remains after removal of the water contained. Dry matter may be measured by a spectroscopy method such as visible (Vis) and near-infrared (NIR) spectroscopy (e.g., using a FoodScan device) in grams per 100 grams total weight of a starting material (e.g., a milk composition), any intermediate product (e.g., a coagulum) or a final dairy product (e.g., a cheese).
  • a starting material e.g., a milk composition
  • any intermediate product e.g., a coagulum
  • a final dairy product e.g., a cheese
  • Moisture content refers to a percentage of moisture, i.e., the portion of the dairy product or composition that is not dry matter. Moisture may thereby be calculated by measuring the amount of dry matter, DM in grams per 100 grams total weight of a starting material (e.g., a milk composition), any intermediate product (e.g., a coagulum) or a final dairy product (e.g., a cheese), i.e.,
  • Weight of moisture 100 - DM (g/100 g)
  • FDM Far-on-Dry-Matter
  • FDS Far-dry solid basis
  • the respective fat content and the moisture content may be determined by a spectroscopy method such as visible (Vis) and near-infrared (NIR) spectroscopy (e.g., using a FoodScan device) in grams per 100 grams total weight of a starting material (e.g., a milk composition), any intermediate product (e.g., a coagulum) or a final dairy product (e.g., a cheese).
  • a starting material e.g., a milk composition
  • any intermediate product e.g., a coagulum
  • final dairy product e.g., a cheese
  • MNFS Melten-in-Non-Fat-Substances
  • MFFB Melten-free basis
  • the fat and the moisture content may be determined by a spectroscopy method such as visible (Vis) and near-infrared (NIR) spectroscopy (e.g., using a FoodScan device) in grams per 100 grams total weight of a starting material (e.g., a milk composition), any intermediate product (e.g., a coagulum) or a final dairy product (e.g., a cheese).
  • a starting material e.g., a milk composition
  • any intermediate product e.g., a coagulum
  • a final dairy product e.g., a cheese
  • soft or “firm/semi-hard” as used herein includes cheeses meeting the definition of a soft or firm/semi-hard cheese according to the CODEX ALIMENTARIUS by FAO and WHO (see CXS 283-1978, 2022 Amendment; see table below).
  • the term also includes soft or firm/semi-hard cheeses as defined by other local, regional, national, or international agencies or organizations.
  • pasta filata cheese as used herein, has a moisture content of 40 to 80 MFFB%. Depending on the moisture content, pasta filata cheese can be in the “soft cheese” or “firm/semi-hard cheese” category (see table above), or in between the two, depending upon their moisture content.
  • “Mozzarella” cheese has a minimum milkfat content of 45% by weight of the solids and a moisture content of more than 52% but not more than 60% by weight.
  • "Low-moisture mozzarella” cheeses have a minimum milkfat content of 45% by weight of the solids and the moisture content is more than 45% but not more than 52% by weight.
  • Part-skim mozzarella has a moisture content of more than 52% but not more than 60% by weight, and a milk fat content that is less than 45% but not less than 30% calculated on the solid’s basis.
  • “Low-moisture part-skim” mozzarella has a moisture content of more than 45% but not more than 52% by weight and a milkfat content, calculated on the solid’s basis, of less than 45% but not less than 30%. Further details regarding these various mozzarella cheeses are provided by 21 CFR. ⁇ 133.155-133.158 (https://www.ecfr.gov/current/title-21/chapter-I/subchapter-B/part-133 [current as of January 2024]).
  • a manufacturing method according to the present invention comprises (i) a conversion of a milk composition to a coagulum; and/or (ii) a conversion of a coagulum to a dairy product.
  • a novel method for producing a coagulum from a milk composition comprising (i) a conversion of a milk composition to a coagulum.
  • a novel method for producing a dairy product e.g., pasta filata cheese
  • a novel method for producing a dairy product comprising the consecutive steps of (i) a conversion of a milk composition to a coagulum; and (ii) a conversion of the coagulum to the dairy product is provided.
  • any edible milk such as cow milk, buffalo milk, goat milk, sheep milk, horse milk or any other animal producing milk suitable for animal or human nourishment may be used, preferably bovine milk is used.
  • milk protein concentrate MPC
  • cream may be added to the raw milk and may be adjusted to fall within the above-mentioned ranges according to conventional methods of adjusting the concentration of milk protein and fat.
  • Commercially available milk protein concentrates are available at protein concentrations ranging from 42% to 85%.
  • a milk protein concentrate may be added to the raw milk which consists of 80% casein and 20% whey proteins.
  • the cream to be added to the raw milk may comprise 30 - 45 % milk fat.
  • the casein concentrate may additionally contain lactose at 0 to about 2 weight-% and calcium at about 2500 - about 12500 mg/kg.
  • the milk composition according to the present application may be obtained from reconstituted milk.
  • Reconstituted milk refers to a milk composition comprising a protein component, a fat component and a solvent.
  • protein concentrates are available as a protein component with varying range in protein content such as a milk protein concentrate (MPC), a micellar casein concentrate (MCC) or whey protein concentrate (WPC),
  • MPC milk protein concentrate
  • WPC micellar casein concentrate
  • the protein component comprised in the reconstituted milk is a casein-rich concentrate or powder, preferably a micellar casein concentrate (MCC).
  • the protein composition may comprise more than 92% casein and/or less than 8% whey protein.
  • the protein composition comprises between 92% to 100% casein and 0 to 8% whey.
  • the protein powder may be demineralized.
  • the demineralised protein powder may a demineralized casein powder such as a reduced-Ca Micellar Casein Concentrated (MCC) powder.
  • MCC reduced-Ca Micellar Casein Concentrated
  • at least 40% of colloidal calcium may have been removed from the native MCC powder.
  • the concentration of calcium may be adjusted to the final calcium concentration in the dairy product.
  • the final calcium is calculated as mg Ca per g casein in the dairy product.
  • Calcium chloride should not be added in an amount more than 0.02 percent (calculated as anhydrous calcium chloride) of the weight of the dairy ingredients, i.e., it may be in the range of 5-20 g per 100 kg milk composition.
  • a fat component may be any fat suitable for use in dairy products, preferably a dairy fat, even more preferably a butteroil or anhydrous milk fat. For instances, where non-dairy fat is used, a colouring agent may be added.
  • a solvent may be any food grade solvent capable of dissolving the milk powder components, preferably the solvent is water.
  • the reconstituted milk may also contain lactose.
  • the lactose may be added as a lactose concentrate powder or skim milk.
  • lactose may be added in an amount of 0 to 4.7 g/100 g total weight of reconstituted milk, such as 4.2-4.6 g lactose/100 g reconstituted milk (corresponding to the amount of lactose in raw bovine milk).
  • Lactose may be preferably added at a concentration of 2.0 - 6.0% w/w.
  • the amounts of protein and fat in the reconstituted milk can be suitably adjusted to fall within the desired ranges necessary to produce the desired dairy product.
  • the reconstituted milk may comprise 18-22 g of protein per 100g total weight of the milk composition, and/or 16 - 26 g of fat per 100g total weight of the milk composition.
  • the reconstituted milk may comprise a fat-to-protein ratio in the range of 0.5 - 1.5, preferably between 0.73 - 1.44.
  • the solvent may be adjusted so that the moisture content of the milk composition is between 58-65 MFFB% (percentage moisture on a fat free basis).
  • the production of the reassembled milk may comprise the steps of: 1. Melting the dairy and/or non-dairy concentrated fat; 2. Adding warm water to melted concentrated fat and mixing under vacuum; 3. Adding one part of demineralized milk protein powder; 4. Gradually adding the remaining milk protein powder and blending by slowly mixing to avoid clumping; 5. Adjusting the mixing speed until a smooth and uniformly thick liquid is produced.
  • the reconstituted milk may be prepared in multipurpose cooker, such as a Stephan cooker (STEPHAN Machinery GmbH), RobotQbo cooker (CADIXPRO), or SPX cooker. Temperatures for melting depend on the type of fat used, such at 50°C. Stirring speeds may be adjusted in the range from 10 - 40 rpm.
  • multipurpose cooker such as a Stephan cooker (STEPHAN Machinery GmbH), RobotQbo cooker (CADIXPRO), or SPX cooker.
  • Temperatures for melting depend on the type of fat used, such at 50°C.
  • Stirring speeds may be adjusted in the range from 10 - 40 rpm.
  • the pH of the reconstituted milk may be adjusted to be in the pH range of raw milk, such as a pH of approximately 6.7.
  • the milk composition has a pH in the range of 6.6 to 6.8.
  • Pasteurisation The reconstituted milk may be pasteurised.
  • pasteurization is attained by maintaining a specified temperature for a specified period of time. The specified temperature is usually attained by heating. The temperature and duration may be selected in order to kill or inactivate certain bacteria, such as harmful bacteria.
  • Pasteurization may be performed at 65 - 78°C for 14 - 120 seconds. For instance, pasteurization of recombined milk may be performed at 65 - 76°C for 15 - 30 seconds.
  • Pasteurization of raw milk may be performed at 72 - 75°C for 14 -18 seconds. A rapid cooling step may follow.
  • the acidifier may be an organic acid or a starter culture.
  • the organic acid used in the direct acidification method may be any food-grade organic acid, such as citric acid, citric, lactic, acetic, etc. The skilled person is aware of methods for acidifying the coagulum by direct acidification.
  • the acidifier is a LAB starter culture.
  • starter culture is a culture or composition which comprises a mixture of bacterial strains (such as lactic acid bacteria strains).
  • lactic acid bacteria or “lactic acid bacterium” (“LAB”) designates food-grade bacteria producing lactic acid as the major metabolic end-product of carbohydrate fermentation. These bacteria are related by their common metabolic and physiological characteristics and are Gram positive, low-GC, acid tolerant, non-sporulating, rod-shaped bacilli or cocci. During the fermentation stage, the consumption of carbohydrate by these bacteria causes the formation of lactic acid, reducing the pH and leading to the formation of a protein coagulum.
  • the lactic acid starter culture may comprise Streptococcus thermophilus.
  • the amount of the starter to be added can be determined by a conventional method according to the kind of the milk and starter.
  • the amount of the lactobacillus starter culture may be 0.1 to 0.4 U/Kg of the milk composition. Reaction times and temperatures depend on the lactic acid bacterium used and can by adjusted by the skilled person. If the starter lactic acid culture comprises mesophilic cultures, the temperatures may be adjusted between 25°C and 40°C. In instances, where the starter lactic acid culture comprises thermophilic cultures, the temperatures may between 25 and 50°C. For example, if the starter culture contains Streptococcus thermophilus, the temperature can be adjusted to 38-45°C. In some cases, it is preferable to use mixed strain cultures, which contain two or more bacterial strains. Mixed strain cultures may consist of a blend of either mesophilic or thermophilic bacteria or a combination of both.
  • the acidifier may be added prior to addition of the coagulum.
  • the acidifier may be added 1-60 min prior to adding the coagulant.
  • the pH decreases, this is called pre-acidification or pre-ripening.
  • the pH prior to renneting depends on reaction times, temperatures and the type of acidifier added. In instances, where a LAB starter culture was added to the milk composition 60 min prior to renneting, the pH may be 6.50 - 6.20. In such instances, the pH at the time of coagulant addition may be 6.50 - 6.20.
  • the method of the present invention may not comprise a pre-ripening or pre-acidification step prior to addition of the microbial rennet. Therefore, the pH at the time of coagulant addition may be 6.60 - 6.80.
  • a coagulant may be added after the acidifier has been added or may be added at the same time as the acidifier.
  • the coagulant may be any suitable coagulant that may be used in the production of the dairy product of the present application.
  • coagulants include those derived from animals such as the most commonly used calf rennet, but also those such as bovine rennet, pig pepsin, poultry rennet, sheep rennet and goat rennet.
  • the rennet (coagulant) may also be chymosin (EC 3.4.23.4), for instance a chymosin derived from microorganisms or plants which can be recombinantly produced.
  • the coagulant is a microbial rennet.
  • the coagulant may be added in sufficient amounts.
  • the amount may be suitably adjusted based on the activity of the coagulant (e.g., units of chymosin) to 1 gram of protein (e.g., casein) in a milk composition (w/w), and/or the ratio of clotting-to-proteolysis.
  • the coagulant may be added in sufficient amounts.
  • chymosin enzyme may be used in an amount from about 0.0001% w/w to about 0.05% w/w, preferably about 0.002% w/w.
  • Coagulum formation Physical characteristics of the coagulum vary depending on coagulation time. In general, longer coagulation times may be associated with a softer curd and a more lactic-yoghurt-like structure than rennet rigid (or rennet-strengthened) curds.
  • the coagulation time may be between 3-60 min, such as between 3-8 min. In absence of a whey draining step, short coagulation times may be preferred. In contrast, in presence of a whey draining step, longer coagulation times may be preferred.
  • the coagulation time may be less than 10 min, e.g., between 3-8 min.
  • the method comprises the steps of coagulum cutting and whey drainage step
  • the coagulation time may be between 15-60 min, such as 30 min.
  • Coagulum cutting The formed coaglum may be chopped and whey may be released from the coagulum.
  • a coagulum cutting step is preferably comprised in the method for producing a coagulum, in instances, where the coagulation time may be between 15-60 min. Suitable equipment is available to determine the progress of coagulation and the optimal time for cutting.
  • the coagulum may be cut with 10 - 20 mm knives. After cutting, the coagulum may heal for 5-10 min.
  • the method of the present invention may not include a coagulum cutting step.
  • a coagulum cutting step is comprised in the method for producing a coagulum.
  • Whey drainage The method of the present application may comprise a whey drainage step.
  • the whey released from the coagulum may be drained.
  • Whey drainage may be performed in 2 steps or more than 2 steps, i.e., whey drainage may be performed before and after the stirring/scalding/cooking step.
  • up to 90% of the whey may be drained.
  • fermentation may be carried out until a pH of 4.9-5.5 is reached.
  • Reaction times and temperatures depend on the lactic acid bacterium used and can be adjusted by the skilled person. Fermentation may be carried out at temperatures between 38 - 45 °C.
  • the fermented composition may be demoulded into slabs or blocks.
  • the coagulum is subjected to a milling step to obtain a composition comprising coagulum pieces.
  • the size/weight/surface area of the resulting coagulum pieces may be suitably adjusted to facilitate subsequent processing steps (e.g., the enzymatic treatment step, and for providing even heating during stretching).
  • Said coagulum may be subjected to a device configured to mill a block of said coagulum into coagulum pieces (e.g., a miller).
  • the coagulum may be subjected to a miller comprising at least one or more rotor(s), e.g., one rotor or two rotors including a bottom rotor and/or a top rotor (e.g., stainless-steel cyclones).
  • said rotor(s) may be a blade or knife.
  • the milling speed of the rotor(s) may be adjusted, e.g., to reduce the size of the coagulum to a size suitable for facilitating subsequent processing steps (e.g., the obtained coagulum pieces have an average diameter of 20 to 60 mm, and/or an average weight of 2 to 8 g).
  • the coagulum is subjected to a miller comprising a bottom rotor and a top rotor for a milling time of at least 10 min.
  • the milling speed of the bottom rotor may be adjusted to obtain coagulum pieces having an average diameter of 20 to 60 mm, wherein the average diameter is defined as the average longest distance between two opposing points along the coagulum piece boundary in a 2-dimensional image of the coagulum pieces.
  • the coagulum is subjected to a miller comprising a bottom rotor and a top rotor for a milling time of 10 to 40 min, optionally wherein the rotation speed of the bottom rotor and/or of the top rotor operate(s) between 50 to 300 rpm. In some instances, the coagulum is subjected to a miller comprising a bottom rotor and a top rotor for a milling time of 10 to 20 min, optionally wherein the coagulum is subjected to said miller at a rotation speed of the bottom rotor of 5 rpm and a rotation speed of the top rotor of to 3 rpm.
  • the defined final size of the coagulum pieces can be adjusted by adjusting an aperture size of the bottom sieves or by adjusting a hole size or position of the rotor(s) (e.g., blade), e.g., the aperture size of the bottom sieves or the hole size in the blade may be between 20-60 mm.
  • the feed hopper and machine frame may be made of stainless steel.
  • the coagulum miller may be fixed over a table or can feed directly onto a stainless-steel tray.
  • the coagulum may be milled to obtain a composition comprising coagulum pieces having an average diameter of 20 to 60 mm.
  • a coagulum piece may be obtained in a shape of a cuboid (cube-like), and/or the cuboid may be characterized by dimensions in the range of 20-40 mm x 15-40 mm x 20-60 mm.
  • the weight of the coagulum piece generated by milling the coagulum may be between 2 - 8 g.
  • the composition comprising coagulum pieces may be subjected to an enzymatic treatment wherein a transglutaminase and/or a protein-glutaminase act on the coagulum pieces.
  • a transglutaminases or protein glutaminase of the present invention a commercially available one or one produced from a culture medium of a microorganism producing protein glutaminase can be used, as disclosed herein.
  • Suitable enzyme preparations include transglutaminase (TG) and/or protein-glutaminase (PG).
  • the enzyme preparations include transglutaminase (TG) and protein-glutaminase (PG).
  • the protein-glutaminase is a protein-glutaminase derived from Chryseobacterium.
  • the transglutaminase is a transglutaminase (EC 2.3.2.13).
  • Commercially available preparations including transglutaminase and protein-glutaminase are available (ACTIVA(Registered Trademark) SYG (Ajinomoto, Japan).
  • Transglutaminases and protein-glutaminase that will be developed and found in the future can also be used in the invention as long as they have the transglutaminase activity.
  • the use of such transglutaminases and protein-glutaminase is, as a matter of course, included in the present invention.
  • Enzyme activity may be measured in U ( ⁇ mol/min) which is defined as the amount of enzyme that catalyses the conversion of one micromole of substrate per minute.
  • the amount(s) of transglutaminase and/or protein-glutaminase may be adjusted depending on the enzyme activity.
  • the enzyme dosages may be adjusted based on the total weight (g) of a composition comprising coagulum pieces and optionally further components (e.g., chopped coagulum including the drained whey and protein level). The total weight of said composition may be measured by methods known to the skilled person.
  • transglutaminases or protein-glutaminase to be added may be less than 2 U, preferably less than 1 U, more preferably less than 0.9 U, per 1 g of protein present in the coagulum, as measured by a spectroscopy method such as visible (Vis) and near-infrared (NIR) spectroscopy (e.g., using a FoodScan device).
  • a spectroscopy method such as visible (Vis) and near-infrared (NIR) spectroscopy (e.g., using a FoodScan device).
  • transglutaminase and/or protein-glutaminase may be used in an amount of 0.1 U/g to 0.9 U/g (U enzyme/g protein in the coagulum), preferably the enzyme is adjusted to 0.3 U/g protein in the coagulum.
  • the amount of enzyme preparation in grams (g) to be added to the coagulum may be calculated in an exemplary embodiment as follows:
  • [protein (g/100 g)] refers to the protein content of the composition comprising coagulum pieces after milling
  • [batch volume] refers to the total weight (in kg) of the composition to be treated (comprising, e.g., a blend of milled coagulum and whey)
  • [enzyme activity (target)] in U/g protein refers to the final dosage of PG or TG in the reaction, which is between 0.1 - 0.9 U/g protein
  • [enzyme activity (initial)] refers to the PG or TG activity in U/g in the enzymatic preparation/ enzyme stock.
  • the enzyme preparation comprising the transglutaminase and/or the protein-glutaminase may further comprise additional components.
  • the enzyme preparation comprises caseinate powders; preferably sodium caseinate and/or reduced calcium or demineralized casein micelles concentrate.
  • the enzyme preparation may comprise lactose and salt. The concentrations of lactose and salt may be adjusted to correspond to the concentrations of the coagulum to avoid dilution of the coagulum.
  • the enzyme preparation may be added as a slurry or as a powder.
  • the expression "slurry” means that the enzyme preparations include transglutaminase (TG) and/or protein-glutaminase (PG) is dissolved in a solvent, preferably an aqueous solution.
  • TG transglutaminase
  • PG protein-glutaminase
  • the enzyme preparation comprising transglutaminase and/or protein-glutaminase is prepared as a slurry in aqueous solution.
  • the enzyme concentration in the slurry may be adjusted depending on the activity (U) of TG and/or PG, preferably the enzyme concentration is between 10-50 % w/w, e.g., about 10% w/w or about 20% w/w.
  • the slurry comprising transglutaminase and/or protein-glutaminase may further comprise additional components.
  • the slurry may comprise the enzyme preparation, caseinate and salt. Additionally, the slurry may comprise lactose.
  • the concentrations of the slurry, caseinate and salt may be adjusted to correspond to the concentrations of the coagulum to avoid dilution of the coagulum.
  • the casein amount in the slurry may be 2-15% w/w.
  • the slurry may further comprise a buffering agent or an acid regulator. Any food-grade buffering agent or acid regulator suitable to prevent a pH change in the coagulum due to the addition of the enzyme preparation may be added. For instance, lactic acid may be added, so that the pH remains in the range of 5-5.4, preferably around pH 5.2.
  • the enzyme preparation may be added gradually, and the reaction times depend on the temperature of the coagulum.
  • an enzyme preparation may be added onto the coagulum at an addition rate of 0.5-10% depending on the concentration of the enzyme(s) in the enzyme preparation (e.g., comprising an enzyme concentration of 10 to 50% w/w).
  • An addition rate of 0.5 - 10% refers to the addition rate of the enzyme preparation to the total weight of the milled coagulum.
  • a slurry comprising 10 - 50 % w/w (preferably at 10% w/w or 20% w/w) is prepared and added to the milled coagulum at a higher addition rate, preferably 3 - 5%.
  • the enzyme preparation is added to the milled coagulum at lower addition rates (e.g., at 0.5-3%).
  • the enzyme preparation may be added at a temperature range between 5 to 35°C.
  • the incubation/reaction time with the enzyme preparation depends on the amount of enzyme added and the temperature. For instance, the reaction time ranges from between 20 - 60 min at temperature between 35 - 58° C and between 1 - 14 h at temperatures between 4-35°C.
  • the enzymatic treatment step is performed to achieve a complete reabsorption of water and oil at the final stretching step in a steam cooker stretcher.
  • Preferred combinations may be: an amount of enzyme preparation of 600 ppm (mg/kg w/w), e.g., for an enzyme preparation containing 20 U of PG and 35 U of TG; a temperature ramping of 45 - 55°C; and a reaction time of 25 - 35 min.
  • PG enhances the protein emulsification properties and thus allows for an increased reabsorption of dairy product components/ingredients (e.g., fats/oils and water) into the protein matrix.
  • dairy product components/ingredients e.g., fats/oils and water
  • PG may impact casein functionality. For instance, if a low-fat pasta filata cheese is produced by this inventive method, then PG may enhance the solubilization of caseins in the coagulum (e.g., by rehydration and/or emulsification) upon steam stretching in a multipurpose cooker, such as a Stephan cooker (STEPHAN Machinery GmbH), RobotQbo cooker (CADIXPRO), or SPX cooker.
  • a multipurpose cooker such as a Stephan cooker (STEPHAN Machinery GmbH), RobotQbo cooker (CADIXPRO), or SPX cooker.
  • the structural features of the obtained dairy product may be different, e.g., a gel may be formed instead of a viscous mass (e.g., depending on the milk composition; see also Table 1).
  • PG may impact smoothing the milled coagulum (and, the resulting dairy product) together with a starter culture at pH 5.2 due to lowering the isoelectric point of paracasein micelles at 38 - 45°C.
  • PG may impact sensory aspects of the dairy product (e.g., dryness and/or crispyness) and/or enhance stretchability of the dairy product (e.g., pasta filata cheese).
  • TG enhances (or further enhances, e.g., if used in combination with PG) the protein hydration properties and crosslinks caseins and thus allows the capture of fines.
  • the enzyme(s) may react with the milk proteins (e.g., casein and whey protein).
  • TG and PG may also modify and crosslink the whey proteins.
  • the enzyme reaction prior to stretching is particularly associated with the ability of curd to plasticize and form strings / sheets when heated and extended. This impact is also evidenced by the modulating effect of casein mineralization level on the above properties.
  • the enzymatic treatment of coagulum has a major impact on the subsequent steam stretching step, e.g., in terms of product yield (e.g., recovery of fines and/or dairy product components/ingredients such as liquid), and the textural, rheological, viscoelastic, cooking sensory properties of the obtained dairy product.
  • product yield e.g., recovery of fines and/or dairy product components/ingredients such as liquid
  • the textural, rheological, viscoelastic, cooking sensory properties of the obtained dairy product e.g., the mechanical fluidity properties of a pasta filata cheese are a key factor contributing to cheese performance as an ingredient (e.g., in a pizza application).
  • the molar ratios of TG to PG can be varied to modulate the fat, protein and moisture content of the resulting dairy product.
  • the molar ratios of TG to PG may be suitably adjusted based on the dairy product to be produced with the method of the present application. As shown in the Table 1 below, for instance, when a dairy product is a low-moisture part-skim pasta filata cheese, PG may be added in excess to TG. In instances, where the dairy product is a low-moisture full fat pasta filata cheese, TG may be added in excess to PG. In instances, where the dairy product is a high moisture mozzarella cheese, only PG may be added, or PG and a small amount of TG may be added.
  • TG may be added, or TG and a small amount of PG may be added.
  • the molar ratio of transglutaminase to protein-glutaminase may be adjusted between 5:1 to 1:5 (TG:PG).
  • TG may enhance moisture (e.g., water) binding properties
  • PG may enhance emulsifying properties.
  • a higher fat and/or protein content of the coagulum requires a lower dosage of PG as compared to TG, and vice versa.
  • Particularly high fat and/or protein content may be treated with TG only, or in combination with very little PG.
  • low fat and/or protein content of the coagulum requires a higher dosage of PG as compared to TG.
  • a coagulum with a low-fat content may be treated with PG only, or in combination with very little TG.
  • the term “higher”, as used herein for the dosage of PG and/or TG enzyme(s), refers to 0.6 - 1.0 U enzyme(s) /1 gram protein.
  • the ratio of TG:PG within the enzyme preparation can be in the range of 1.5:1 to 5:1, preferably in the range of 2:1 to 3:1.
  • transglutaminase (TG) and protein-glutaminase (PG) may be used at molar ratios of 2.3:1.
  • the heating and stretching of the acidified curd is the defining operation in the manufacture of pasta filata pizza cheese.
  • the coagulum is molten/plasticized during heating and form strings / sheets when mechanically treated in the stretching step.
  • stretching may be carried out in a combined cooking-stretching step in a liquid heating medium, also called hot water stretching.
  • hot water stretching the composition may be subjected to a liquid at a temperature between 60 to 95°C ( Figure 1A).
  • the method of the present application uses steam stretching (“dry” stretching) instead of hot water stretching. Hence, in instances where the coagulum is subjected to a steam stretching step, the method does not comprise a hot water stretching step.
  • the steam stretching step may be performed in a steam cooker.
  • the steam cooker uses steam infusion to heat the coagulum and melt it, and thus transform it into the plastic and workable consistency.
  • Steam heating may be performed at a steam temperature above 100 °C and below 150 °C, preferably between 110-120 °C, even more preferably 115 °C.
  • the steam injection may occur in 2 - 4 steps.
  • the temperature of the molten composition during steam stretching may be above 60 °C and below 80 °C, preferably between 64 - 76°C.
  • the steam injection might be gradual.
  • Steam heating may be performed in more than 2 steps.
  • the temperature of the molten mass in the steam cooker stretcher may be measured by a probe integrated in the steam cooker stretcher and further verified at the end of stretching by means of a standard calibrated temperature probe.
  • a temperature probe may thereby be inserted into the heart of the molten mass at the end of stretching.
  • the plastic curd may also be treated mechanically.
  • the steam stretching step comprises a thermomechanical treatment, wherein steam heating is performed simultaneously to mechanically working, stirring, kneading, and/or stretching the composition.
  • Mechanical treatment may be performed by a dull rotary mixer.
  • a mechanical treatment may be performed at a speed of about 250 rpm.
  • Any commercially available steam cooker can be used for the steam stretching step.
  • Examples of commercially available steam cookers are Stephan Cheese Cooking Machine (STEPHAN Machinery GmbH), and a RoboQbo (CADIXPRO).
  • the maximum and minimum batch weight depend on the volume of the product and the cooker used e.g., a maximum batch weight between 6 and 7 KG and minimum batch weight between 2-3 KG may be used in RoboQbo (CADIXPRO).
  • the steam cooker may comprise a dull rotary mixer, a scraper to remove products form the inner walls and prevent burning and may perform gradual direct-steam injection. Rotation speed (rpm) and rotation time (seconds) of the blades can be set up according to the manufacturer’s recommendations.
  • the time for steam stretching may be adjusted to achieve full plasticization and formation of strings / sheets.
  • the total residence time in the steam cooker may range from 5 - 30 min, preferably 5-20 min.
  • the stream stretching step may be performed until all liquids, such as oil and water, are absorbed. Hence, during the steam stretching step, there may be no loss of liquids and solids. Therefore, the net sum of physical and chemical ingredients of the output may not change compared to the input.
  • the hot plastic curd may be moulded.
  • the temperature may be between 60 to 72°C.
  • the curd may be forced under pressure into a mould. Moulding may be performed using a twin-screw extruder.
  • the temperature of the curd may also be decreased.
  • the curd may be cooled by using cold water. The water may have a temperature of 4°C and the cooling time may be between 10 - 30 min.
  • Brining Brining may be carried out in a solution saturated with salt.
  • the salt concentration in the brine may be 18 - 25%.
  • the pH and calcium content may be adjusted to prevent changes in calcium distribution and loss of calcium from the cheese during brining.
  • the pH of the brining solution may be adjusted to a pH of 5.2 with an acidifier, such as lactic acid.
  • the brine may further comprise additional components, such as calcium.
  • the calcium content of the brine may be 0.06 g 100 g -1 .
  • Brining may be carried out at low temperatures such as 1-4 °C. Residence time in the brine and brine concentration are key parameters that affect total salt uptake and moisture loss during brining. Hence, the duration of brining may be suitably adjusted based on to desired salt content. For instance, brining may be carried out for 60 min. This may be followed by a drying step.
  • the brining step may be omitted.
  • the method of the present application may not comprise a brining step.
  • the resulting dairy product may be packed under vacuum.
  • the dairy product may be store at a temperature between 4 to 5°C for 3 - 9 months, such for 6 months at a temperature of 4°C.
  • the chemical composition and functionality of the dairy product may be analyzed.
  • the chemical composition that may be analyzed are for example the percentages of Fat-on-Dry-Base (FDB), Moisture-in-Non-Fat-Substances (MNFS) and protein level, as well as the pH value of the dairy product.
  • FDB Fat-on-Dry-Base
  • MNFS Moisture-in-Non-Fat-Substances
  • protein level as well as the pH value of the dairy product.
  • Functional properties may be selected from the stretch or stringiness, melt, elasticity of melted cheese, and oiling after heating the cheese.
  • shreddability/sliceability and bake-off properties may be analysed.
  • Partial Salting In contrast to traditional methods, the method of the present application does not comprise a brining step.
  • the method further comprises at least one (“dry”) salting step.
  • the salt may be food-grade NaCl.
  • partial salting is carried out by directly adding salt during various production step.
  • the salt can be added in combination with the rennet; in combination with the enzyme preparation, and/or during the stretching step.
  • salt is added during the stretching step before complete absorption of liquids.
  • the salting rate may be between 0.1 to 0.5% (w/w).
  • the (w/w) as used herein, is the weight of the dry salt in g per total weight.
  • the salt content in the final composition of the dairy product may range from 1.0 to 1.5 g/100g dairy product.
  • a person skilled in the art is able to adjust the amount of salt added during the at least one salting step to arrive at from 1.0 - 1.5 g of salt per 100 g of final dairy product.
  • the salt concentrations may thereby be suitably adjusted to avoid the inhibition of the enzymes and bacteria, such as the enzymes in the enzymatic preparation, the coagulation enzymes or the lactic acid bacteria.
  • the one or more salting step(s) may father allow, e.g., to modulate the coagulation and/or fermentation reaction(s) in the LAB starter culture, to modulate the protein-crosslinking reaction during the enzymatic treatment step and/or enhance flavour of the obtained dairy product.
  • Additional ingredients selected from the group of a buffering agent, a colouring agent, a flavouring agent, and a preservative can be added at any of production steps.
  • a buffering agent may be added when preparing the slurry
  • a colouring agent is added when a non-dairy fat is used
  • a flavouring agent can be added when bacterial cultures are not performing the buttery flavour.
  • a preservative may be added when the cheese is shredded or sliced, such as a food grade anti-fungicide.
  • Dairy products obtained by the methods of the present invention may be produced with 70 to 100% yield, such as between 80 to 100%, preferably 90 to 100%, more preferably 99 to 100% yield.
  • the economic yield may be determined as weight percentage (% w/w) and is defined as the weight of coagulum or dairy product (e.g., cheese) in kg produced per kg of milk composition.
  • the yield of the stretching step (% w/w) may be determined as the weight of the final dairy product after stretching per weight of the coagulum before stretching. In a preferred embodiment, the yield of the stretching step is 100%.
  • the improved yield relates to protein and fat recoveries reflecting on the composition of final product (Fat-on-Dry-Basis Matter FDB and Protein content).
  • FDB and Protein content For example, higher protein content and higher FDM (23 - 26% w/w Protein; 43 - 48% FDB for enzymatic treated cheese versus 20 - 23 w/w % and 39 - 43 % in control).
  • the dairy product obtained by the methods described herein is a pasta filata cheese.
  • Types of pasta filata cheese include “low moisture” pasta filata cheese (e.g., firm/semi-hard such as pizza cheese, Kashkaval cheese), and “high moisture” pasta filata cheese (e.g., soft cheese such fresh Mozzarella for use as a table cheese).
  • the pasta filata cheese may be a low-moisture pasta filata cheese and may have has a moisture level in the range of 40 - 56 wt. %.
  • the cheese is preferably used as an ingredient in the preparation of other foods.
  • the consumable composition is a food product selected from the group consisting of pizza, pasta, lasagna, gratin, fondue, and cheese sauce.
  • the dairy product comprises one or more dairy-like properties selected from the group consisting of viscoelasticity, fluidity, elasticity, adhesiveness, stretch, firmness, viscosity, tackiness, chewiness, resilience, springiness, mouthfeel, melt, hardness, creaminess, and flexibility.
  • the filata cheese of the present application has a stretch or stringiness, melt, elasticity of melted cheese, oiling after heating the cheese comparable or better than a pasta filata cheese produced with traditional methods.
  • the "cheese texture” refers to the softness and smoothness on the tongue when eaten.
  • chee stretchability refers to the property of being stretched without breaking when physically separated.
  • elasticity refers to the property of trying to return to its original shape when physically separated. Since cheese preferably has elasticity in addition to high stretchability to achieve good texture, both stretchability and elasticity were evaluated in the present invention.
  • the "meltability" refers to the property of the cheese analogue to liquefy and spread.
  • flavour is an important quality factor for dairy products, and consumer acceptance is often closely linked to their flavour profiles.
  • the flavour of dairy products is dependent on various aspects of taste, including, but not limited to, e.g., the acidity (pH of the cheese and/or packing water), the freshness, the bitterness, the creaminess (assessed by expert panellists).
  • the method for producing a dairy product comprises or consist of the following consecutive steps (1) to (8): (1) preparing a milk composition to obtain a composition having a pH in the range of 6.6 to 6.8, wherein the milk composition is obtained from a reconstituted milk; (2) pasteurizing the milk composition obtained in step (1); (3) adding an acidifier and a coagulant to the milk composition obtained by step (2) to form a rennet-induced gel or coagulum ; (4) fermenting the coagulum obtained by step (3); (5) milling the coagulum obtained by step (4) to obtain a composition comprising coagulum pieces; wherein said milling reduces the size of the coagulum to a size suitable for facilitating subsequent processing steps; (6) subjecting the composition obtained by step (5) to an enzymatic treatment wherein a transglutaminase and/or a protein-glutaminase act on the coagulum pieces; (7) subjecting the composition
  • the reconstituted milk comprises a protein component, a fat component and a solvent (e.g., water).
  • the fat component of the milk composition of step (1) of item (A) is a non-dairy fat or a dairy fat, optionally the fat component is selected from a butteroil and an anhydrous milk fat.
  • the solvent of the milk composition of step (1) of item (A) is water.
  • the reconstituted milk of in step (1) of item (A) is reconstituted with the solvent from a powder comprising the protein component; preferably wherein the protein powder is demineralized.
  • the protein component of the reconstructed milk in step (1) of item (A) comprises a casein, and/or a whey protein, preferably wherein the protein component comprises at least 90% (w/w) (e.g., 92% (w/w)) casein.
  • the milk composition comprises a fat-to-protein ratio in the range of 0.5 to 1.5; preferably wherein a milk composition comprising a fat-to-protein ratio of 0.5 is treated with protein-glutaminase.
  • the milk composition comprises a moisture content of between 58-65 MFFB%, wherein MFFB% is a percentage moisture on a fat free basis.
  • the obtained milk composition may constitute a viscous masshaving a moisture content of 50-56% w/w.
  • the obtained milk composition constitutes a mass having a moisture content of 83-86% w/w.
  • step (1) of item (A) wherein the milk composition is obtained from a reconstituted milk without the formation of a gel (gelation), to avoid said gelation, in particularly when using reconstituted milk for manufacturing a low fat dairy product (low fat pasta filata cheese).
  • a protein glutaminase (PG) may be added to fluidise a viscous mass.
  • step (3) of item (A) the acidifier and the coagulant are added simultaneously.
  • the acidifier is a lactic acid bacteria (LAB) starter culture, preferably a thermophilic LAB, even more preferably Streptococcus thermophilus or a blend of Streptococcus thermophilus and Lactobacillus bulgaricus or Lactobacillus helveticus.
  • the coagulant is a rennet, preferably a microbial rennet.
  • coagulation is carried out for less than 10 min, preferably for 3-8 min; and/or the temperature is 38 to 45°C.
  • step (4) of item (A) the coagulum is fermented to obtain a composition having a pH in the range of 4.9-5.
  • the coagulum is subjected to a device configured to mill a block of said coagulum into coagulum pieces (e.g., a miller).
  • a device configured to mill a block of said coagulum into coagulum pieces
  • the coagulum may be subjected to a miller comprising at least one or more rotor(s), e.g., one rotor or two rotors including a bottom rotor and/or a top rotor (e.g., stainless-steel cyclones).
  • said rotor(s) may be a blade or knife.
  • the milling speed of the rotor(s) may be adjusted, e.g., to reduce the size of the coagulum to a size suitable for facilitating subsequent processing steps (e.g., the obtained coagulum pieces have an average diameter of 20 to 60 mm, and/or an average weight of 2 to 8 g).
  • the milling speed of the bottom rotor may be adjusted to 5 rpm and the milling speed of the top rotor to 3 rpm to obtain coagulum pieces having an average diameter of 20 to 60 mm, wherein the average diameter is defined as the average longest distance between two opposing points along the coagulum piece boundary in a 2-dimensional image of the coagulum pieces.
  • an enzyme preparation comprising the transglutaminase and/or the protein-glutaminase is added to the coagulum pieces.
  • the enzyme preparation further comprises a caseinate, preferably sodium caseinate, reduced calcium or demineralized casein micelles concentrate.
  • the enzyme preparation is provided as a powder composition or a slurry, preferably wherein the enzyme preparation is provided as a slurry.
  • the protein-glutaminase is a protein-glutaminase derived from Chryseobacterium; and/or wherein the transglutaminase is a protein-glutamine gamma-glutamyltransferase.
  • the transglutaminase and/or protein-glutaminase are added in an amount of 0.1 U/g to 0.9 U/g (U of enzyme per grams, g of protein of the composition as measured by, e.g., near-infrared spectroscopy (NIR)), such as 0.3 U enzyme/g protein.
  • NIR near-infrared spectroscopy
  • the molar ratio of transglutaminase to protein-glutaminase may be between 5:1 to 1:5 (TG:PG), more preferably between 3:1 to 1:3 (TG:PG).
  • step (7) of item (A) the composition is heated to above 60 °C and below 80 °C (preferably, between 64 °C and 76 °C) by using steam heating, preferably by direct steam injections.
  • the steam heating is performed at a steam temperature above 100 °C and below 150 °C.
  • the steam stretching step comprises a thermomechanical treatment, wherein steam heating is performed simultaneously to mechanically working, stirring, kneading, and/or stretching the composition, optionally mechanical treatment is performed by a dull rotary mixer.
  • step (7) of item (A) the steam heating is performed in more than 2 steps (e.g., between 2 to 6 steps) wherein each step comprises a direct steam injection (e.g., at 55-62 °C for 1 min; or at 62-66 °C for 0.5 min).
  • step (7) of item (A) steam stretching is performed for 1-30 min, preferably 5-20 min.
  • step (7) of item (A) steam stretching is performed to obtain a composition wherein all liquids, such as oil and water, are absorbed.
  • step (8) of item (A) the composition obtained in step (7) is moulded and cooled.
  • the at least one salting step of item (A) is performed simultaneously to - adding an acidifier and/or to adding a coagulant according to step (3); - milling according to step (5); - enzymatic treatment according to step (6); and/or - steam stretching according to step (7).
  • the at least one salting step of item (A) is performed simultaneously to the steam stretching according to step (7).
  • the at least one salting step of item (A) is performed at a salting rate between 1.0 to 1.5 g/100g total weight.
  • the salt used in the at least one salting step is NaCl.
  • FIG. 1B An exemplary method for producing a dairy product according to item (A) is illustrated in Figure 1B, wherein the method further comprises a step of demoulding the coagulum obtained by step (4) prior to step (5) (e.g., into slabs or blocks), and wherein the method further does not comprise a whey drainage step and/or a brining step.
  • the method for producing a dairy product comprises or consist of the following consecutive steps (1) to (12): (1) preparing a milk composition to obtain a composition having a pH in the range of 6.6 to 6.8; wherein the milk composition is obtained from a raw milk; (2) pasteurizing the milk composition obtained in step (1); (3) adding an acidifier to the milk composition obtained by step (2) to obtain a milk composition with a pH in the range of 6.50-6.20; (4) adding a coagulant to the milk composition obtained in step (3); (5) cutting to coagulum obtained by step (4); (6) stirring/scalding/cooking the coagulum obtained in step (5); (7) draining the whey obtained by step (6); (8) fermenting the coagulum obtained by step (7); (9) milling the coagulum obtained by step (8) to obtain a composition comprising coagulum pieces; (10) subjecting the composition obtained by step (9) to an enzymatic treatment wherein a transglutaminase and/or a protein-glutamina
  • the milk composition comprises a fat-to-protein ratio in the range of 0.5 to 1.5.
  • the milk composition comprises a raw milk and optionally one or more components selected from a protein component, a fat component and a solvent.
  • the fat component may be a non-dairy fat or a dairy fat, optionally a fat component selected from a butteroil and an anhydrous milk fat.
  • the protein component may comprise a casein, and/or a whey protein, preferably the protein component comprises at least 90% casein.
  • the acidifier is a lactic acid bacteria (LAB) starter culture, preferably a thermophilic LAB starter culture or a blend of a thermophilic LAB and mesophilic LAB starter culture.
  • LAB lactic acid bacteria
  • the coagulant is a rennet, preferably a microbial rennet.
  • the coagulum of step (5) is obtained 15 to 60 min after addition of the coagulant in step (4).
  • step (5) of item (B) the coagulum is cut with 10 to 20 mm knives.
  • step (6) of item (B) scalding is carried out at temperatures between 42 to 45 °C with continuous stirring for 20 - 50 min.
  • a step of cooking may be carried out for less than one hour (e.g., 20 to 50 min) with stirring at temperature between 42 to 45 °C.
  • step (8) of item (B) the coagulum is fermented to obtain a composition having a pH in the range of 4.9 to 5.5.
  • the coagulum is subjected to a device configured to mill a block of said coagulum into coagulum pieces (e.g., a miller).
  • a device configured to mill a block of said coagulum into coagulum pieces
  • the coagulum may be subjected to a miller comprising at least one rotor, e.g., one rotor or two rotors including a bottom rotor and/or a top rotor (e.g., stainless-steel cyclones).
  • said rotor may be a blade or knife.
  • the milling speed of the rotor(s) may be adjusted, e.g., to reduce the size of the coagulum to a size suitable for facilitating subsequent processing steps (e.g., the obtained coagulum pieces have an average diameter of 20 to 60 mm, and/or an average weight of 2 to 8 g).
  • the milling speed of the bottom rotor may be adjusted to 5 rpm and the milling speed of the top rotor to 3 rpm to obtain coagulum pieces having an average diameter of 20 to 60 mm, wherein the average diameter is defined as the average longest distance between two opposing points along the coagulum piece boundary in a 2-dimensional image of the coagulum pieces.
  • an enzyme preparation comprising the transglutaminase and/or the protein-glutaminase is added to the composition comprising coagulum pieces.
  • the enzyme preparation further comprises a caseinate, preferably sodium caseinate; reduced calcium or demineralized casein micelles concentrate.
  • the enzyme preparation is provided as a powder composition or a slurry, preferably wherein the enzyme preparation is provided as a slurry.
  • the protein-glutaminase is a protein-glutaminase derived from Chryseobacterium; and/or wherein the transglutaminase is a protein-glutamine gamma-glutamyltransferase.
  • the transglutaminase and/or protein-glutaminase are added in an amount of 0.1 U/g to 0.9 U/g (U enzyme/g protein of the composition as measured by near-infrared spectroscopy (NIR)), such as 0.3 U enzyme/g protein.
  • the molar ratio of transglutaminase to protein-glutaminase may be between 5:1 to 1:5 (TG:PG), preferably between 3:1 to 1:3 (TG:PG).
  • step (11) of item (B) the composition is heated to above 60 °C and below 80 °C (preferably, between 64 °C and 76 °C) by using steam heating.
  • the steam heating is performed at a steam temperature above 100 °C and below 150 °C.
  • the steam stretching step comprises a thermomechanical treatment, wherein steam heating is performed simultaneously to mechanically working, stirring, kneading, and/or stretching the composition.
  • the steam heating is performed in more than 2 steps.
  • steam stretching is performed to obtain a composition wherein all liquids, such as oil and water, are absorbed.
  • the at least one salting step is performed simultaneously to step (4); simultaneously to step (9); simultaneously to step (10); and/or simultaneously to step (11); preferably the at least one salting step is performed simultaneously to step (11).
  • the at least one salting step is performed at a salting rate between 1.0 to 1.5 g/100g total weight.
  • the salt used in the at least one salting step is NaCl.
  • FIG. 1C An exemplary method for producing a dairy product according to item (B) is illustrated in Figure 1C, wherein the method further comprises a step of demoulding the coagulum obtained by step (8) prior to step (9), and wherein the method does not comprise a brining step.
  • the method for producing a dairy product comprises or consist of the following consecutive steps: (1) preparing a reconstituted milk composition comprising a demineralized milk protein powder, a dairy or non-dairy fat, a solvent (e.g., water), a moisture content of 58-65 MFFB% by using a cooker (e.g., Stephane cooker, RobotQbo cooker, SPX cooker); (2) pasteurizing the milk composition obtained in step (1) by a direct-steam injection; and preferably, cooling down the milk composition to a temperature between 38 to 45°C; (3) adding an acidifier (e.g., a LAB starter culture) and a coagulant (e.g., a microbial rennet) to the milk composition obtained by step (2); optionally, partial salting (e.g., at a salting rate of 0.1 - 0.5% (w/w)); (4) fermenting the coagulum obtained by step (3) to reduce pH to 4.9 to 5.5; (5) demoulding the coagulum
  • FIG. 1D An exemplary method for producing a dairy product according to item (C) is illustrated in Figure 1D, wherein the method further does not comprise a whey drainage step and/or a brining step.
  • composition analysis The level of moisture, fat, and protein in the curd and cheese were determined by a FoodScan device (NIR FT-IR Foss inc.). The salt concentration was assessed by titration method. The total calcium content was measured by inductively coupled plasma atomic emission spectroscopy.
  • Shreddability/Sliceability and bake-off properties Shredding/cutting was carried out on 1 week old pasta filata cheese.
  • Shreddability and sliceability include physical attributes, such as ease of machinability/processing, propensity/tendency to stick to equipment, excessive production of fines, and shape and integrity of shreds.
  • the bake-off on pizza crust was performed by using an oven at 270°C for 6 min. Meltability, stretchability and blistering were evaluated at an application laboratory by informal tests and compared with a traditional pizza Mozzarella cheese obtained from raw/fresh milk (natural Low-Moisture Whole-Milk (LMWM) pizza Mozzarella cheese).
  • LMWM Low-Moisture Whole-Milk
  • stringiness i.e., the degree to which a product hangs on the probe when the probe is withdrawn.
  • a vertical traction test was carried out using a mobile harpoon with 6 branches, which stretches the cheese contained in a stainless-steel bowl in a water bath at a constant speed, and the ability of the cheese to form threads under the effect of vertical stretching was measured using a filometer.
  • the stringiness corresponds to the length (expressed in mm) of the last cheese strand when it breaks.
  • the resulting exemplary process of making pasta filata cheese from reassembled milk was as follows: 1. The dairy and/or non-dairy concentrated fat was completely melted at 50°C in a multipurpose cooker such as Stephan cooker, RobotQbo cooker, SPX cooker. 2. Warm water, ca. 50°C, and melted concentrated fat were added and heated to 50°C by double heating jacket with mixing at 10 - 40% rpm under vacuum at -500 mbar. 3. The lid of the cooker was opened and one part of demineralized milk protein powder was added and mixed for 3 - 15 minutes at 10 - 40% rpm and the vacuum valve was set up at -500 mbar. 4. The milk protein powder was added gradually into the blend with slowly mixing to avoid clumping. 5.
  • the mixing speed was increased until a smooth and a consistent thick liquid (viscous mass) was generated.
  • the milk composition (liquid pre-cheese) was pasteurized by a direct-steam injection at 66 - 78°C for 30 - 120 seconds. 7.
  • the liquid pre-cheese was cooled down to 38 - 45°C then lactic acid bacteria (LAB) starter cultures and microbial rennet were added for coagulation and fermented until a of pH 4.9 - 5.5 was reached. 8.
  • LAB lactic acid bacteria
  • the fermented pre-cheese matrix was milled, then the chopped curd was treated with an enzymatic preparation comprising transglutaminase and/or protein-glutaminase which was prepared as a slurry and further comprises caseinates and salt; the enzymatic preparation was added at a rate between 0.5 - 10%; the reaction time depended on the temperature of curd. 9.
  • the curd was heated, kneaded, plasticised, and stretched by using a steam cooker (dry stretching) at 64 - 64°C with a total residence time ranging from 5 - 20 min until a complete absorption of liquid of water/oil pools and an inactivation of the residual activity of enzymes transglutaminase and/or protein-glutaminase was achieved.
  • the cheese was rapidly moulded by using a twin-screw extruder; the cheese loaves were cooled in chilled water for 10 - 30 min without a traditional brining; and the cheese was dried and packaged under vacuum and stored over a period of 3 - 9 months at 5°C.
  • FIG. 2 shows pictures of Cheese Without Whey Separation (CWWS) Mozzarella cheese generated by the above-described methods by preforming steam (“dry”) stretching in a Stephan cooker.
  • Successful dry stretching of the fermented pre-cheese (CWWS) in a Stephan cooker was achieved by using a dull rotary mixer and gradual direct-steam injections for melting and pre-stretching then stretching.
  • a complete re-absorption of water and oil was observed when the temperature of the molten mass reached around 63°C ( Figure 2C).
  • the total residence time was short at around 6 min. It was noticeable that the net sum of physical and chemical ingredients of the resulting cheese did not change compared to the input, i.e., no water and oil was lost during the dry stretching step.
  • the obtained cheese was manually moulded under 5lbs loaf shapes.
  • the cheese made by this novel process has 100% yield and has a quality comparable to low-moisture Mozzarella made by traditional method in terms of composition and functionality. No significant difference (p > 0.05) could be found in moisture-on-non-fat-substances (% MNFS) and fat-on-dry-base (% FDB) as well as shreddability/sliceability and meltability/stretchability when baked on pizza crust.
  • the sensory properties of unmelted pasta filata cheese blocks made by this novel method reveal no significant difference (p ⁇ 0.05) compared to pasta filata cheese made by traditional method.
  • the advantages of curd treatment by enzymatic preparation prior to stretching enhance a higher mouthfeel and shing appearance compared to traditional one.
  • Three types of pasta filata cheese were produced from reassembled milk via a stretching step in a steam cooker stretcher (dry stretching) and without whey separation as described above.
  • the production method of the three types of cheese differed in the enzymatic treatment step carried out on the milled curd/coagulum prior to stretching.
  • the enzymatic treatment step was omitted, one cheese sample was only treated with TG, and one cheese was treated with TG and PG.
  • Table 3 shows the means of compositional and microbiological results (replicates of 3 trials) of three types of 2-week-old pasta filata cheeses.
  • Figure 3 shows pictures of the cheese shreds generated from the control cheese ( Figure 3A), the TG-treated cheese ( Figure 3B) and the TG-and-PG (“TG/PG”) treated cheese ( Figure 3C).
  • the shreds were consistently uniform and smooth in contrast with the TG-treated cheese, and the control cheese samples, which showed a poor integrity of the shreds.
  • Figure 4 shows the melting properties of the control cheese ( Figure 4A), the TG-treated cheese ( Figure 4B) and the TG-and-PG (“TG/PG”) treated cheese ( Figure 4C).
  • the TG or TG/PG treated cheeses had a less crispy appearance than the control.
  • Figure 5 shows the stretchability of the control cheese ( Figure 5A), a TG-treated cheese ( Figure 5B) and a TG-and-PG (“TG/PG”) treated cheese ( Figure 5C).
  • the melting on pizza crust was completed for all cheeses with a release of an oil sheen in cheese sample treated by a blend of TG/PG.
  • a comparison study (difference from control) based on some numerical pre-results measuring free oil after bake-off showed higher values of TG/PG treated sample than control.
  • flavour and texture of these melted cheeses was further evaluated by sensory panellists.
  • 6 panellists evaluated the stretchability after cheese bake-off on pizza crust at 63 - 65 C by using a fork, a digital Camera was used to estimate the dryness aspect of the surface, and the following aspects were tested: cutting the crust by pizza roller, tasting the molten cheese at 55 - 58 C to evaluate the chewiness and pliability (blended study). While the TG-treated and control were found to have a slightly dry flavour and chewy texture, the cheese sample treated by blend of TG/PG were found to be less dry and chewy.
  • pasta filata cheese was successfully made from reassembling protein powder (e.g., casein concentrate powder), anhydrous fat, and water to form a homogeneous pre-cheese matrix followed by a fermentation without whey separation and treatment of enzyme slurry comprising TG or a blend of TG and PG, then a stretching by direct-steam cooker stretcher (steam stretching).
  • the composition and functionality were evaluated, and it was noticeable that coagulum (cheese curd) treated by a blend of TG and PG showed better functionality at lower pH value than a control and TG-treated cheese curd.
  • the sensory results of unmelted cheese blocks on shreds revealed a difference between 3 cheese samples: the cheese sample treated by a blend of TG and PG presented a better mouthfeel compared to control and TG-cheese samples.
  • the present invention is useful in a wide range of fields, including the field of processed dairy products and cheesemaking, in particular pasta filata cheese production.
  • the dairy products obtained by the methods of the present invention support nutrition and are useful as food products.
  • the dairy products are useful as an ingredient for food products (such as pizza, pasta, lasagna, gratin, fondue, and cheese sauce) for animal consumption.
  • the present invention is industrially applicable.

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Microbiology (AREA)
  • Dairy Products (AREA)

Abstract

La présente invention concerne de produits laitiers (tels que du fromage, de préférence du fromage à pâte filée), des compositions consommables contenant ledit produit laitier (par exemple, des produits alimentaires) et leurs procédés de production. La présente invention concerne en particulier un procédé de production d'un produit laitier consistant à convertir un coagulum en produit laitier et comprenant les étapes consistant à : (a) broyer le coagulum pour obtenir une composition comprenant des morceaux de coagulum ; (b) soumettre la composition obtenue à l'étape (a) à un traitement enzymatique dans lequel une transglutaminase et/ou une protéine-glutaminase agissent sur les morceaux de coagulum ; et (c) soumettre la composition obtenue à l'étape (b) à une étape d'étirage à la vapeur.
PCT/JP2025/004905 2024-02-16 2025-02-14 Nouveau produit laitier et son procédé de production Pending WO2025173761A1 (fr)

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