DAIRY PRODUCT AND PROCESS

Abstract

The invention relates to cheese and cheese-like products comprising about 0.2 to about 6% by weight ?.sub.s-casein, ?-casein, or a combination thereof, at least about 5% by weight of one or more hydrocolloids comprising one or more starches, and wherein the cheese or cheese-like product has one or more characteristics of a dairy product, such as flavour, taste, aroma, body, appearance, texture, firmness, handling, density, structure, coagulation, binding, leavening, aeration, foaming, emulsification, elasticity, viscoelasticity, melt, creaminess and mouthfeel.

Claims

1. A cheese or cheese-like product comprising a) about 0.2 to about 6% by weight of ?.sub.s-casein, ?-casein, or a combination thereof, b) at least about 5% by weight of one or more hydrocolloids, wherein the one or more hydrocolloids comprises one or more starches, and c) about 0.2 to about 20% by weight total dairy protein, wherein total casein in the cheese or cheese-like product comprises i) at least about 55% by weight ?.sub.s-casein, and/or ii) at least about 40% by weight ?-casein.

2. The cheese or cheese-like product according to claim 1, comprising 0.2 to about 6% by weight ?.sub.s-casein, wherein the total casein in the cheese or cheese-like product comprises at least about 55% by weight ?.sub.s-casein, and wherein the cheese or cheese-like product optionally comprises ?-casein, ?-casein, p-?-casein, or any combination of any two or more thereof.

3. (canceled)

4. The cheese or cheese-like product according to claim 1, comprising 0.2 to about 6% by weight ?-casein, wherein the total casein in the cheese or cheese-like product comprises at least about 40% by weight ?-casein, and wherein the cheese or cheese-like product optionally comprises ?.sub.s-casein, ?-casein, p-?-casein, or any combination of any two or more thereof.

5. (canceled)

6. The cheese or cheese-like product according to claim 1, wherein the ?.sub.s-casein comprises ?.sub.s1-casein, ?.sub.s2-casein, or a combination thereof.

7. The cheese or cheese-like product according to claim 1, comprising a) from about 0.2 to about 15% or about 0.2 to about 10% by weight total dairy protein, b) from about 0.2 to about 20%, about 0.2 to about 15% or about 0.2 to about 10% by weight total casein, c) at least about 10%, 15% or 20% by weight of one or more hydrocolloids, d) from about 0.2 to about 10%, about 0.2 to about 6%, about 0.2 to about 5%, or e) about 0.2 to about 3% by weight total protein, or e) any combination of any two or more of (a) to (d).

8. (canceled)

9. The cheese or cheese-like product according to claim 1, wherein the ?.sub.s-casein and/or ?-casein is provided by a protein ingredient comprising an ?.sub.s-casein enriched fraction, a ?-casein enriched fraction, recombinant ?.sub.s1-casein, recombinant ?.sub.s2-casein, recombinant ?-casein, or any combination of any two or more thereof.

10. (canceled)

11. The cheese or cheese-like product of claim 1, wherein the cheese or cheese-like product comprises a) at least about 10% by weight starch, b) at least about 15% by weight starch, or c) about 5% to about 30% by weight starch.

12. The cheese or cheese-like product according to claim 1, wherein the one or more starches are selected from the group comprising potato starch, maize starch, rice starch, tapioca (cassava) starch, wheat starch, pea starch, sweet potato starch, banana starch, barley starch, oat starch, sago starch, amaranth starch, arrowroot starch, canna starch, sorghum starch, chia seed starch, fava bean starch, jackfruit starch, bamboo starch, chickpea starch, cassava starch, carrot starch, and any combination of any two or more thereof.

13. The cheese or cheese-like product according to claim 12, wherein the one or more starches comprises one or more modified starches, optionally selected from the group comprising pre-gelatinised starch, thin-boiling starch, oxidised starch, octenyl succinic anhydride (OSA) modified starch, or a combination of any two or more thereof.

14. (canceled)

15. The cheese or cheese-like product of claim 1, wherein the one or more hydrocolloids further comprises one or more gums, optionally selected from the group comprising xanthan gum, guar gum, locust bean gum, tragacanth gum, acacia gum, karaya gum, tara gum, gellan gum, konjac gum (konjac mannan), carboxymethylcellulose (CMC), tragacanth, agar, pectin, alginate, carrageenan (?-carrageenan, .Math.-carrageenan, ?-carrageenan), Arabic gum, pectin, alginate, flaxseed gum, guar gum, or any combination of any two or more thereof.

16. (canceled)

17. (canceled)

18. The cheese or cheese-like product of claim 1, wherein the cheese or cheese-like product comprises one or more non-dairy proteins, the one or more non-dairy proteins comprising or consisting of one or more plant proteins or hydrolysed plant proteins.

19. (canceled)

20. The cheese or cheese-like product according to claim 1, wherein the cheese or cheese-like product comprises a) one or more emulsifying salts, b) about 15 to about 65% by weight moisture, c) about 0.05 to about 35% by weight lipid, or d) any combination of any two or more of a)-c).

21. A method for preparing a cheese or cheese-like product, the method comprising a) providing a cheese composition comprising i) ?.sub.s-casein, ?-casein, or a combination thereof, ii) one or more hydrocolloids, comprising one or more starches, iii) optionally one or more lipids, and iv) optionally one or more additional ingredients, b) heating the composition at a temperature and for a time sufficient to produce a heated cheese mass or cheese-like mass, and c) cooling the heated cheese mass or cheese-like mass to produce the cheese or cheese-like product, wherein the cheese or cheese-like product comprises i) about 0.2 to about 6% by weight of ?.sub.s-casein, ?-casein, or a combination thereof, and ii) at least about 5% by weight of one or more hydrocolloids, and iii) about 0.2 to about 20% by weight total dairy protein, wherein total casein in the cheese or cheese-like product comprises 1. at least about 55% by weight ?.sub.s-casein, and/or 2. at least about 40% by weight ?-casein.

22. The method according to claim 21, comprising a) providing an aqueous composition comprising ?.sub.s-casein, ?-casein, or a combination thereof, b) adding the one or more hydrocolloids, lipid and, optionally, the one or more additional ingredients, to the aqueous composition to produce a cheese composition, c) heating the cheese composition at a temperature and for a time sufficient to produce a heated cheese mass or cheese-like mass, and d) cooling the heated cheese mass or cheese-like mass to produce the cheese or cheese-like product; or a) providing an aqueous composition comprising ?.sub.s-casein, ?-casein, or a combination thereof, b) adding lipid to the aqueous composition and mixing to form an emulsion, c) adding the emulsion to a second aqueous composition comprising the one or more hydrocolloids and, optionally, the one or more additional ingredients, to produce a cheese, d) heating the cheese composition at a temperature and for a time sufficient to produce a heated cheese mass or cheese-like mass, and e) cooling the heated cheese mass or cheese-like mass to produce the cheese or cheese-like product.

23. (canceled)

24. The method of claim 22, comprising a) dispersing the ?.sub.s-casein and/or ?-casein in water at a temperature of from about 20 to about 50? C. to substantially completely dissolve the ?.sub.s-casein and/or ?-casein to produce the aqueous composition b) dispersing ?.sub.s-casein in water at a temperature of from about 40 to about 55? C. to substantially completely dissolve the ?.sub.s-casein to produce the aqueous composition, or c) dispersing ?-casein in water at a temperature of from about 15 to about 30? C., or at ambient temperature, to substantially completely dissolve the ?-casein to produce the aqueous composition.

25. (canceled)

26. (canceled)

27. The method according to claim 21, comprising providing an aqueous composition comprising a protein ingredient comprising an ?.sub.s-casein enriched fraction, a ?-casein enriched fraction, recombinant ?.sub.s1-casein, recombinant ?.sub.s2-casein, recombinant ?-casein, or any combination of any two or more thereof.

28. The cheese or cheese-like product of claim 9, wherein the protein ingredient comprises an ?.sub.s-casein enriched fraction, and wherein: a) total casein in the protein ingredient comprises at least about 50% or at least about 75% by weight ?.sub.s-casein, and/or b) the protein ingredient comprises ?.sub.s-casein and ?-casein in a weight ratio of at least about 50:50, preferably at least about 80:20, and/or the protein ingredient comprises a ?-casein enriched fraction, and wherein: c) total casein in the ?-casein enriched fraction comprises at least about 50% by weight ?-casein, and/or d) the ?-casein enriched fraction comprises ?-casein and ?.sub.s-casein in a weight ratio of at least about 50:50.

29. The cheese or cheese-like product of claim 1 wherein the cheese or cheese-like product is a processed cheese or processed cheese-like product.

30. A cheese or cheese-like product comprising a) about 0.2 to about 6% by weight ?.sub.s-casein, b) at least about 5% by weight of one or more hydrocolloids, comprising at least one starch, c) about 0.5 to about 20% by weight total protein; wherein the ?.sub.s-casein comprises i) at least about 60% of the casein in the cheese or cheese-like product, and/or ii) at least about 55% by weight of the total protein in the cheese or cheese-like product.

31. (canceled)

Description

DETAILED DESCRIPTION OF THE INVENTION

[0243] The present invention relates to the use of protein compositions comprising one or more caseins as functional ingredients in the preparation of cheeses, cheese-like products and methods for manufacture of such products, particularly processed cheeses and processed cheese-like products. Preferably, the cheese and cheese-like products of the invention are produced with minimal amounts of milk proteins, or with recombinant milk proteins that are not derived from milk, to produce cheese-like products that are acceptable to those who wish to lower their dairy intake, or who identify as vegetarians and vegans. The invention particularly makes use of ?.sub.s-casein, ?-casein, and/or blends of ?.sub.s-caseins, ?-caseins, and, optionally, specific ?.sub.s-casein and/or ?-casein fractions that, when combined with starches and/or gums, produce processed cheese-like structure, flavour and mouthfeel, using surprisingly low amounts of protein.

[0244] The present invention utilises casein proteins that interact with other proteins, simple carbohydrates, complex carbohydrates, fats, oils, gums, emulsifying agents, salt (sodium and/or potassium chloride), ions (both cations and anions), starches and/or other GRAS ingredients to produce functional ingredients and foods.

Caseins and Casein Compositions

[0245] The present invention utilises casein proteins or blends of casein proteins, in particular, ?.sub.s-casein and/or ?-casein, to produce cheese, such as processed cheese, and cheese-like products.

[0246] Following casein synthesis in mammary gland tissue, posttranslational phosphorylation occurs at selected serine residues in the primary structures of each of these caseins. The caseins in milk then form ionic bonds with calcium phosphate in combination with hydrophobic interactions between other caseins to create aggregates identified as casein micelles. (Farrell, J. M., Jr., R. Jimenez-Flores, G. T. Bleck, E. M. Brown, J. E. Butler, L. K. Creamer, C. L. Hicks, C. M. Hollar, K. F. Ng-Kwai-Hang, and H. E. Swaisgood. 2004. Nomenclature of the proteins of cow's milk, 6th revision. Dairy Sci. 87:1641-1674).

[0247] Casein proteins for use herein may include ?.sub.s1-casein, ?.sub.s2-casein, ?-casein, ?-casein, ?-casein, p-?-casein, and/or variants thereof. In various embodiments, the casein may be ?.sub.s-casein or ?-casein.

[0248] Cow's milk typically comprises about 48% ?.sub.s-casein as a proportion of total casein, and about 36% ?-casein as a proportion of total casein, as shown in Table 1 below (Davies and Law. 1980. J. Dairy Res. 47:83-90).

[0249] An average ratio of casein proteins in cow's milk is about 3:0.8:3:1 of ?.sub.s1-casein:?.sub.s2-casein:?-casein:?-casein (Brown and Ernstrom. 1988. Milk clotting enzymes and cheese chemistry Part I: Milk clotting enzymes in Fundamentals of dairy chemistry 3.sup.rd ed.; Schmidt, 1982. Association of caseins and casein micelle structure. In Dairy Chemistry. Col. 1. P. F. Fox, et. c Appl. Sci. Pub., Ltd., Essex, England).

TABLE-US-00001 TABLE 1 Theoretical milk casein content Casein type Casein (% by weight) ?.sub.s1-casein 38 ?.sub.s2-casein 10 ?-casein 36 ?-casein 13 ?-casein 3 Total 100.00

[0250] In various embodiments the ?.sub.s-casein may comprise ?.sub.s1-casein, ?.sub.s2-casein, or a combination thereof.

[0251] In various embodiments, the ?.sub.s-casein may be encoded by a protein sequence selected from SEQ ID NO. 1-26 or a variant thereof with at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 98%, or 99% sequence identity.

[0252] In various embodiments the ?.sub.s-casein comprises, or is provided by an ingredient that comprises or is enriched in, ?.sub.s1-casein and/or ?.sub.s2-casein, for example, ?.sub.s-casein-enriched fractions.

[0253] The term ?.sub.s-casein-enriched fraction as used herein refers to compositions fractionated from milk having a ratio of ?.sub.s-casein to ?-casein that is greater than that of skim milk (1:0.94, as measured by polyacrylamide gel electrophoresis followed by staining with Coomassie Blue and densitometry. Other suitable analytical methods are also known to the skilled addressee). Preferably, the ratio is higher than 1.3:1, more preferably higher than 1.6:1, more preferably higher than 2:1, most preferably higher than 3:1. ?.sub.s-Casein-enriched fractions are enriched in at least one of ?.sub.s1 and ?.sub.s2-casein, generally both, relative to casein in the casein source from which it was prepared (generally cows' milk).

[0254] In some embodiments, the ?.sub.s-casein-enriched fractions comprise at least about 55% by weight ?.sub.s-casein relative to total casein, preferably about 60%, more preferably about 65%, more preferably about 70%, more preferably about 75%, most preferably about 80% by weight ?.sub.s-casein relative to total casein. In a preferred embodiment, the ?.sub.s-casein-enriched fractions comprise at least about 55% by weight ?.sub.s-casein relative to total casein.

[0255] In some embodiments, the ?.sub.s-casein-enriched fractions may be prepared as described in published PCT application WO2007/100264 or WO 2009/108074 (hereby incorporated by reference in their entirety). Other methods that may be used will be apparent to those skilled in the art. For example, cold microfiltration may be used for at least part of the enrichment or depletion, for example, as described in U.S. Pat. No. 5,169,666.

[0256] In various embodiments, ?.sub.s-casein enriched fractions are prepared by fractionating lactic casein as described herein in the examples. Briefly, pasteurised skim milk is incubated with dairy starter cultures under acid conditions to produce lactic casein.

[0257] The lactic casein is dispersed in water to produce a slurry comprising solubilised casein. The temperature is adjusted to less than about 7? C. and the pH of the slurry adjusted to about pH 10. Calcium chloride (dihydrate) is added to the slurry to produce a coagulum. The curd is obtained, washed and dried to provide an ?.sub.s-casein enriched fraction.

[0258] In various embodiments the ?.sub.s-casein comprises one or more recombinant ?.sub.s-casein proteins. The use of recombinant proteins provides for the manufacture of cheeses or cheese-like products that comprise no ingredients derived from animals.

[0259] In various embodiments the recombinant ?.sub.s-casein protein comprises a variant of an ?.sub.s-casein protein.

[0260] In various embodiments the recombinant ?.sub.s-casein comprises a native ?.sub.s1-casein protein, a native ?.sub.s2-casein protein, a variant of ?.sub.s1-casein or ?.sub.s2-casein, or a combination thereof. In various embodiments the recombinant ?.sub.s-casein protein has an amino acid sequence of any one of SEQ ID Nos: 1-26. In various embodiments the variant of ?.sub.s1-casein or ?.sub.s2-casein has an amino acid sequence having at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 98%, or 99% sequence identity to a sequence of SEQ ID Nos: 1-26.

[0261] In various embodiments, the ?-casein may be encoded by a protein sequence selected from SEQ ID NO. 27-40 or a variant thereof with at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 98%, or 99% sequence identity.

[0262] In various embodiments the ?-casein comprises, or is provided by an ingredient that comprises or is enriched in, ?-casein, for example, ?-casein-enriched fractions.

[0263] The term ?-casein-enriched fraction as used herein refers to compositions fractionated from milk having a ratio of ?-casein to ?.sub.s-casein that is greater than that of skim milk (0.94:1, as measured by polyacrylamide gel electrophoresis followed by staining with Coomassie Blue and densitometry. Other suitable analytical methods are also known to the skilled addressee). Preferably, the ratio is higher than 1:1, more preferably higher than 1.2:1, more preferably higher than 1.4:1, more preferably higher than 1.5:1, more preferably higher than 1.6:1, most preferably higher than 1.7:1. ?-casein enriched fractions are enriched in ?-casein relative to casein in the casein source from which it was prepared (generally cows' milk).

[0264] In some embodiments, the ?-casein-enriched fractions comprise at least about 40% by weight ?-casein relative to total casein, preferably at least about 45%, more preferably at least about 50%, more preferably at least about 52%, most preferably at least about 54%, by weight ?-casein relative to total casein. In a preferred embodiment, the ?-casein-enriched fractions comprise at least about 40% by weight ?-casein relative to total casein.

[0265] In some embodiments, the ?-casein-enriched fractions may be prepared as described in published PCT application WO2007/100264 or WO2009/108074 (hereby incorporated by reference in their entirety). Other methods that may be used will be apparent to those skilled in the art. For example, cold microfiltration may be used for at least part of the enrichment or depletion, for example, as described in U.S. Pat. No. 5,169,666.

[0266] In various embodiments, ?-casein enriched fractions are prepared by fractionating lactic casein as described herein in the examples. Briefly, pasteurised skim milk is incubated with dairy starter cultures under acid conditions to produce lactic casein.

[0267] The lactic casein is dispersed in water to produce a slurry comprising solubilised casein. The temperature is adjusted to less than about 7? C. and the pH of the slurry adjusted to about pH 10. Calcium chloride (dihydrate) is added to the slurry to produce a coagulum. The liquid is obtained, and pH adjusted to 4.3-4.4 to produce a second coagulum. This second coagulum is washed and dried to provide a ?-casein enriched fraction.

[0268] In various embodiments the ?-casein comprises one or more recombinant ?-casein proteins.

[0269] In various embodiments the recombinant ?-casein comprises a native recombinant ?-casein, a variant of ?-casein, or a combination thereof. In various embodiments the recombinant ?-casein protein has an amino acid sequence of any one of SEQ ID Nos: 27-40. In various embodiments the variant of ?.sub.s1-casein or ?.sub.s2-casein has an amino acid sequence having at least about 70%, 75%, 80%, 85%, 90%, 95%, 96%, 98%, or 99% sequence identity to a sequence of SEQ ID Nos: 27-40.

Recombinant Expression

[0270] One or more proteins used in the formation of the cheese or cheese-like products may be produced recombinantly. In some cases, the ?.sub.s-casein and/or ?-casein is produced recombinantly.

[0271] ?.sub.s1-casein, ?.sub.s2-casein, and/or ?-casein can have an amino acid sequence from any species. For example, recombinant ?.sub.s-casein and/or ?-casein may have an amino acid sequence of cow, human, sheep, goat, buffalo, bison, pig, horse or camel ?.sub.s-casein and/or ?-casein. The casein nucleotide sequence may be codon-optimized for increased efficiency of production. Exemplary ?.sub.s-casein protein sequences are provided as SEQ ID NOs: 1-26 and exemplary ?-casein protein sequences are provided as SEQ ID NOs: 27-40 in Table 2 below. Recombinant ?.sub.s-casein and/or ?-casein can be a non-naturally occurring variant of an ?.sub.s-casein and/or ?-casein. Such a variant can comprise one or more amino acid insertions, deletions, or substitutions relative to a native casein sequence. For example, such variants may include an amino acid sequence or peptide truncated at the N-terminus and/or C-terminus, preferably by from 1 to 10 amino acids, for example by 1, 2, 3, or about 5 amino acids, relative to a native casein sequence. Such variants may include an amino acid sequence or peptide elongated at the N-terminus and/or C-terminus, preferably by from 1 to 10 amino acids, for example by 1, 2, 3, or about 5 amino acids, relative to a native casein sequence. In some embodiments, variants may contain from 1 to 10 amino acid insertions, deletions, and/or substitutions (collectively) with respect to the native sequence. For example, such variants may have from 1 to about 5 amino acid insertions, deletions, and/or substitutions (collectively) with respect to the native sequence. The degree and location of glycosylation or other post-translation modifications may vary depending on the chosen host cells and the nature of the host cellular environment. Partial hydrolytic products of ?.sub.s-casein and/or ?-casein are also encompassed by the term variant.

[0272] Such a variant can have at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NOs: 1-40. The term sequence identity as used herein in the context of amino acid sequences is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in a selected sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2 or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full-length of the sequences being compared.

[0273] A recombinant ?.sub.s-casein and/or ?-casein is recombinantly expressed in a host cell. As used herein, a host or host cell denotes any protein production host selected or genetically modified to produce a desired product. Exemplary hosts include fungi, such as filamentous fungi, as well as bacteria, yeast, algae, plant, insect, and mammalian cells.

[0274] Host cells comprising genetic constructs, such as expression constructs, as disclosed herein are useful in methods well known in the art (e.g. Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Press, 1987; Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing, 1987) for recombinant production of polypeptides disclosed herein. Such methods may involve the culture of host cells in an appropriate medium in conditions suitable for or conducive to expression of a polynucleotide or polypeptide disclosed herein. The expressed recombinant polypeptide, which may optionally be secreted into the culture, may then be separated from the medium, host cells or culture medium by methods well known in the art (e.g. Deutscher, Ed, 1990, Methods in Enzymology, Vol 182, Guide to Protein Purification).

[0275] In some cases, a bacterial host cell such as Lactococcus lactis, Bacillus subtilis or Escherichia coli may be used to produce ?-, ?-and/or ?-casein proteins. Other host cells include bacterial host such as, but not limited to, Lactococci sp., Lactococcus lactis, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus licheniformis and Bacillus megaterium, Brevibacillus choshinensis, Mycobacterium smegmatis, Rhodococcus erythropolis and Corynebacterium glutamicum, Lactobacilli sp., Lactobacillus fermentum, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus plantarum and Synechocystis sp. 6803.

[0276] In some embodiments, the host cell is a yeast cell selected from the list consisting of Saccharomyces cerevisiae, Schizosaccharomyces pombe, Yarrowia lipolytica, Candida glabrata, Ashbya gossypii, Cyberlindnera jadinii, Pichia pastoris, Pichia methanolica, Kluyveromyces lactis, Hansenula polymorpha, Candida boidinii, Arxula adeninivorans, Xanthophyllomyces dendrorhous, and Candida albicans species. In some embodiments, the yeast cell is a Saccharomycete.

[0277] In some embodiments, the host cell is a fungal cell selected from the list consisting of Aspergillus spp. and Trichoderma spp.

[0278] Expression of a target protein can be provided by an expression vector, a plasmid, a nucleic acid integrated into the host genome or other means. For example, a vector for expression can include: (a) a promoter element, (b) a signal peptide, (c) a heterologous casein sequence, and (d) a terminator element.

[0279] Methods for producing and using genetic constructs and expression vectors are well known in the art and are described generally in Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Press, 1987; Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing, 1987).

[0280] Expression vectors that can be used for expression of ?.sub.s-casein and/or ?-casein include those containing an expression cassette with elements (a), (b), (c) and (d). In some embodiments, the signal peptide (c) need not be included in the vector. In some cases, a signal peptide may be part of the native signal sequence of the casein protein, for instance, the protein may comprise a native signal sequence as bolded in SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, or 39. In some cases, the vector comprises a polynucleotide encoding a protein sequence as exemplified in SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, or 39. In some cases, the vector may comprise a polynucleotide encoding a mature protein sequence, as exemplified in SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, or 40 with a heterologous signal sequence. In general, the expression cassette is designed to mediate the transcription of the transgene when integrated into the genome of a cognate host microorganism or when present on a plasmid or other replicating vector maintained in a host cell.

[0281] To aid in the amplification of the vector prior to transformation into the host microorganism, a replication origin (e) may be contained in the vector. To aid in the selection of microorganism stably transformed with the expression vector, the vector may also include a selection marker (f). The expression vector may also contain a restriction enzyme site (g) that allows for linearization of the expression vector prior to transformation into the host microorganism to facilitate the expression vectors stable integration into the host genome. In some embodiments the expression vector may contain any subset of the elements (b), (e), (f), and (g), including none of elements (b), (e), (f), and (g). Other expression elements and vector element known to one of skill in the art can be used in combination or substituted for the elements described herein. For example, because many microorganisms are capable of expressing multiple gene products from a polycistronic mRNA, multiple polypeptides can be expressed under the control of a single regulatory region for those microorganisms, if desired. These might include one or more ?.sub.s-casein proteins, one or more ?-casein proteins, one or more different dairy proteins (e.g. one or more caseins), and/or one or more non-dairy proteins.

TABLE-US-00002 TABLE2 Sequences SEQIDNo. Name Sequence 1 Bovine?.sub.s1-casein MKLLILTCLVAVALARPKHPIKHQGLPQEVLNENLLRFFV APFPEVFGKEKVNELSKDIGSESTEDQAMEDIKQMEAESI SSSEEIVPNSVEQKHIQKEDVPSERYLGYLEQLLRLKKYKV PQLEIVPNSAEERLHSMKEGIHAQQKEPMIGVNQELAYFY PELFRQFYQLDAYPSGAWYYVPLGTQYTDAPSFSDIPNPIG SENSEKTTMPLW 2 Bovine?.sub.s1-casein RPKHPIKHQGLPQEVLNENLLRFFVAPFPEVFGKEKVNELS matureprotein KDIGSESTEDQAMEDIKQMEAESISSSEEIVPNSVEQKHI QKEDVPSERYLGYLEQLLRLKKYKVPQLEIVPNSAEERLHS MKEGIHAQQKEPMIGVNQELAYFYPELFRQFYQLDAYPSG AWYYVPLGTQYTDAPSFSDIPNPIGSENSEKTTMPLW 3 Ovine?.sub.s1-casein MKLLILTCLVAVALARPKHPIKHQGLSSEVLNENLLRFV VAPFPEVFRKENINELSKDIGSESIEDQAMEDAKQMKAGS SSSSEEIVPNSAEQKYIQKEDVPSERYLGYLEQLLRLKKYN VPQLEIVPKSAEEQLHSMKEGNPAHQKQPMIAVNQELAYF YPQLFRQFYQLDAYPSGAWYYLPLGTQYTDAPSFSDIPNPI GSENSGKITMPLW 4 Ovine?.sub.s1-casein RPKHPIKHQGLSSEVLNENLLRFVVAPFPEVFRKENINELS matureprotein KDIGSESIEDQAMEDAKQMKAGSSSSSEEIVPNSAEQKYI QKEDVPSERYLGYLEQLLRLKKYNVPQLEIVPKSAEEQLHS MKEGNPAHQKQPMIAVNQELAYFYPQLFRQFYQLDAYPS GAWYYLPLGTQYTDAPSFSDIPNPIGSENSGKITMPLW 5 Caprine?.sub.s1- MKLLILTCLVAVALARPKHPINHRGLSPEVPNENLLRFVV casein APFPEVFRKENINELSKDIGSESTEDQAMEDAKQMKAGSS SSSEEIVPNSAEQKYIQKEDVPSERYLGYLEQLLRLKKYNV PQLEIVPKSAEEQLHSMKEGNPAHQKQPMIAVNQELAYFY PQLFRQFYQLDAYPSGAWYYLPLGTQYTDAPSFSDIPNPIG SENSGKTTMPLW 6 Caprine?.sub.s1- RPKHPINHRGLSPEVPNENLLRFVVAPFPEVFRKENINELS caseinmature KDIGSESTEDQAMEDAKQMKAGSSSSSEEIVPNSAEQKY protein IQKEDVPSERYLGYLEQLLRLKKYNVPQLEIVPKSAEEQLH SMKEGNPAHQKQPMIAVNQELAYFYPQLFRQFYQLDAYPS GAWYYLPLGTQYTDAPSFSDIPNPIGSENSGKTTMPLW 7 Buffalo?.sub.s1-casein MKLLILTCLVAVALARPKQPIKHQGLPQGVLNENLLRFF VAPFPEVFGKEKVNELSTDIGSESTEDQAMEDIKQMEAES ISSSEEIVPISVEQKHIQKEDVPSERYLGYLEQLLRLKKYNV PQLEIVPNLAEEQLHSMKEGIHAQQKEPMIGVNQELAYFY PQLFRQFYQLDAYPSGAWYYVPLGTQYPDAPSFSDIPNPI GSENSEKTTMPLW 8 Buffalo?.sub.s1-casein RPKQPIKHQGLPQGVLNENLLRFFVAPFPEVFGKEKVNELS matureprotein TDIGSESTEDQAMEDIKQMEAESISSSEEIVPISVEQKHIQ KEDVPSERYLGYLEQLLRLKKYNVPQLEIVPNLAEEQLHSM KEGIHAQQKEPMIGVNQELAYFYPQLFRQFYQLDAYPSGA WYYVPLGTQYPDAPSFSDIPNPIGSENSEKTTMPLW 9 Equine?.sub.s1-casein MKLLILTCLVAVALARPKLPHRQPEIIQNEQDSREKVLK ERKFPSFALEYINELNRQRELLKEKQKDEHKEYLIEDPEQQ ESSSTSSSEEVVPINTEQKRIPREDMLYQHTLEQLRRLSKY NQLQLQAIHAQEQLIRMKENSQRKPMRVVNQEQAYFYLE PFQPSYQLDVYPYAAWFHPAQIMQHVAYSPFHDTAKLIAS ENSEKTDIIPEW 10 Equine?.sub.s1-casein RPKLPHRQPEIIQNEQDSREKVLKERKFPSFALEYINELNR matureprotein QRELLKEKQKDEHKEYLIEDPEQQESSSTSSSEEVVPINTE QKRIPREDMLYQHTLEQLRRLSKYNQLQLQAIHAQEQLIR MKENSQRKPMRVVNQEQAYFYLEPFQPSYQLDVYPYAAW FHPAQIMQHVAYSPFHDTAKLIASENSEKTDIIPEW 11 Camel?.sub.s1-casein MKLLILTCLVAVALARPKYPLRYPEVFQNEPDSIEEVLNK RKILELAVVSPIQFRQENIDELKDTRNEPTEDHIMEDTERK ESGSSSSEEVVSSTTEQKDILKEDMPSQRYLEELHRLNKY KLLQLEAIRDQKLIPRVKLSSHPYLEQLYRINEDNHPQLGE PVKVVTQEQAYFHLEPFPQFFQLGASPYVAWYYPPQVMQY IAHPSSYDTPEGIASEDGGKTDVMPQWW 12 Camel?.sub.s1-casein RPKYPLRYPEVFQNEPDSIEEVLNKRKILELAVVSPIQFRQE matureprotein NIDELKDTRNEPTEDHIMEDTERKESGSSSSEEVVSSTTE QKDILKEDMPSQRYLEELHRLNKYKLLQLEAIRDQKLIPRV KLSSHPYLEQLYRINEDNHPQLGEPVKVVTQEQAYFHLEPF PQFFQLGASPYVAWYYPPQVMQYIAHPSSYDTPEGIASED GGKTDVMPQWW 13 Human?.sub.s1- MRLLILTCLVAVALARPKLPLRYPERLQNPSESSEPIPLE casein SREEYMNGMNRQRNILREKQTDEIKDTRNESTQNCVVAE PEKMESSISSSSEEMSLSKCAEQFCRLNEYNQLQLQAAHA QEQIRRMNENSHVQVPFQQLNQLAAYPYAVWYYPQIMQY VPFPPFSDISNPTAHENYEKNNVMLQW 14 Human?.sub.s1- RPKLPLRYPERLQNPSESSEPIPLESREEYMNGMNRQRNIL caseinmature REKQTDEIKDTRNESTQNCVVAEPEKMESSISSSSEEMSL protein SKCAEQFCRLNEYNQLQLQAAHAQEQIRRMNENSHVQVP FQQLNQLAAYPYAVWYYPQIMQYVPFPPFSDISNPTAHEN YEKNNVMLQW 15 Bovine?.sub.s2-casein MKFFIFTCLLAVALAKNTMEHVSSSEESIISQETYKQEK NMAINPSKENLCSTFCKEVVRNANEEEYSIGSSSEESAEV ATEEVKITVDDKHYQKALNEINQFYQKFPQYLQYLYQGPIV LNPWDQVKRNAVPITPTLNREQLSTSEENSKKTVDMESTE VFTKKTKLTEEEKNRLNFLKKISQRYQKFALPQYLKTVYQH QKAMKPWIQPKTKVIPYVRYL 16 Bovine?.sub.s2-casein KNTMEHVSSSEESIISQETYKQEKNMAINPSKENLCSTFC matureprotein KEVVRNANEEEYSIGSSSEESAEVATEEVKITVDDKHYQK ALNEINQFYQKFPQYLQYLYQGPIVLNPWDQVKRNAVPIT PTLNREQLSTSEENSKKTVDMESTEVFTKKTKLTEEEKNRL NFLKKISQRYQKFALPQYLKTVYQHQKAMKPWIQPKTKVI PYVRYL 17 Ovine?.sub.s2-casein MKFFIFTCLLAVALAKHKMEHVSSSEEPINISQEIYKQE KNMAIHPRKEKLCTTSCEEVVRNADEEEYSIRSSSEESAE VAPEEVKITVDDKHYQKALNEINQFYQKFPQYLQYLYQGPI VLNPWDQVKRNAGPFTPTVNREQLSTSEENSKKTIDMES TEVFTKKTKLTEEEKNRLNFLKKISQYYQKFAWPQYLKTVD QHQKAMKPWTQPKTNAIPYVRYL 18 Ovine?.sub.s2-casein KHKMEHVSSSEEPINISQEIYKQEKNMAIHPRKEKLCTTS matureprotein CEEVVRNADEEEYSIRSSSEESAEVAPEEVKITVDDKHYQ KALNEINQFYQKFPQYLQYLYQGPIVLNPWDQVKRNAGPF TPTVNREQLSTSEENSKKTIDMESTEVFTKKTKLTEEEKNR LNFLKKISQYYQKFAWPQYLKTVDQHQKAMKPWTQPKTN AIPYVRYL 19 Caprine?.sub.s2- MKFFIFTCLLAVALAKHKMEHVSSSEEPINIFQEIYKQEK casein NMAIHPRKEKLCTTSCEEVVRNANEEEYSIRSSSEESAEV APEEIKITVDDKHYQKALNEINQFYQKFPQYLQYPYQGPIV LNPWDQVKRNAGPFTPTVNREQLSTSEENSKKTIDMEST EVFTKKTKLTEEEKNRLNFLKKISQYYQKFAWPQYLKTVD QHQKAMKPWTQPKTNAIPYVRYL 20 Caprine?.sub.s2- KHKMEHVSSSEEPINIFQEIYKQEKNMAIHPRKEKLCTTSC caseinmature EEVVRNANEEEYSIRSSSEESAEVAPEEIKITVDDKHYQKA protein LNEINQFYQKFPQYLQYPYQGPIVLNPWDQVKRNAGPFTP TVNREQLSTSEENSKKTIDMESTEVFTKKTKLTEEEKNRLN FLKKISQYYQKFAWPQYLKTVDQHQKAMKPWTQPKTNAI PYVRYL 21 Buffalo?.sub.s2-casein MKFFIFTCLLAVALAKHTMEHVSSSEESIISQETYKQEK NMAIHPSKENLCSTFCKEVIRNANEEEYSIGSSSEESAEVA TEEVKITVDDKHYQKALNEINQFYQKFPQYLQYLYQGPIVL NPWDQVKRNAVPITPTLNREQLSTSEENSKKTVDMESTE VFTKKTKLTEEDKNRLNFLKKISQHYQKFAWPQYLKTVYQ YQKAMKPWTQPKTNVIPYVRYL 22 Buffalo?.sub.s2-casein KHTMEHVSSSEESIISQETYKQEKNMAIHPSKENLCSTFC matureprotein KEVIRNANEEEYSIGSSSEESAEVATEEVKITVDDKHYQK ALNEINQFYQKFPQYLQYLYQGPIVLNPWDQVKRNAVPIT PTLNREQLSTSEENSKKTVDMESTEVFTKKTKLTEEDKNR LNFLKKISQHYQKFAWPQYLKTVYQYQKAMKPWTQPKTN VIPYVRYL 23 Equine?.sub.s2-casein MKFFIFTCLLAVALAKHNMEHRSSSEDSVNISQEKFKQ EKYVVIPTSKESICSTSCEEATRNINEMESAKFPTEREEKE VEEKHHLKQLNKINQFYEKLNFLQYLQALRQPRIVLTPWD QTKTGDSPFIPIVNTEQLFTSEEIPKKTVDMESTEVVTEKT ELTEEEKNYLKLLYYEKFTLPQYFKIVRQHQTTMDPRSHRK TNSYQIIPVLRYF 24 Equine?.sub.s2-casein KHNMEHRSSSEDSVNISQEKFKQEKYVVIPTSKESICSTS matureprotein CEEATRNINEMESAKFPTEREEKEVEEKHHLKQLNKINQFY EKLNFLQYLQALRQPRIVLTPWDQTKTGDSPFIPIVNTEQL FTSEEIPKKTVDMESTEVVTEKTELTEEEKNYLKLLYYEKFT LPQYFKIVRQHQTTMDPRSHRKTNSYQIIPVLRYF 25 Camel?.sub.s2-casein MKFFIFTCLLAVVLAKHEMDQGSSSEESINVSQQKFKQ VKKVAIHPSKEDICSTFCEEAVRNIKEVESAEVPTENKISQ FYQKWKFLQYLQALHQGQIVMNPWDQGKTRAYPFIPTVN TEQLSISEESTEVPTEESTEVFTKKTELTEEEKDHQKFLNKI YQYYQTFLWPEYLKTVYQYQKTMTPWNHIKRYF 26 Camel?.sub.s2-casein KHEMDQGSSSEESINVSQQKFKQVKKVAIHPSKEDICSTF matureprotein CEEAVRNIKEVESAEVPTENKISQFYQKWKFLQYLQALHQ GQIVMNPWDQGKTRAYPFIPTVNTEQLSISEESTEVPTEE STEVFTKKTELTEEEKDHQKFLNKIYQYYQTFLWPEYLKTV YQYQKTMTPWNHIKRYF 27 Bovine?-casein MKVLILACLVALALARELEELNVPGEIVESLSSSEESITRI NKKIEKFQSEEQQQTEDELQDKIHPFAQTQSLVYPFPGPIP NSLPQNIPPLTQTPVVVPPFLQPEVMGVSKVKEAMAPKHK EMPFPKYPVEPFTESQSLTLTDVENLHLPLPLLQSWMHQP HQPLPPTVMFPPQSVLSLSQSKVLPVPQKAVPYPQRDMPI QAFLLYQEPVLGPVRGPFPIIV 28 Bovine?-Casein RELEELNVPGEIVESLSSSEESITRINKKIEKFQSEEQQQTE Matureprotein DELQDKIHPFAQTQSLVYPFPGPIPNSLPQNIPPLTQTPVV VPPFLQPEVMGVSKVKEAMAPKHKEMPFPKYPVEPFTESQ SLTLTDVENLHLPLPLLQSWMHQPHQPLPPTVMFPPQSVL SLSQSKVLPVPQKAVPYPQRDMPIQAFLLYQEPVLGPVRG PFPIIV 29 Ovine?-Casein MKVLILACLVALALAREQEELNVVGETVESLSSSEESIT HINKKIEKFQSEEQQQTEDELQDKIHPFAQAQSLVYPFTG PIPNSLPQNILPLTQTPVVVPPFLQPEIMGVPKVKETMVPK HKEMPFPKYPVEPFTESQSLTLTDVEKLHLPLPLVQSWMH QPPQPLPPTVMFPPQSVLSLSQPKVLPVPQKAVPQRDMPI QAFLLYQEPVLGPVRGPFPILV 30 Ovine?-casein REQEELNVVGETVESLSSSEESITHINKKIEKFQSEEQQQT matureprotein EDELQDKIHPFAQAQSLVYPFTGPIPNSLPQNILPLTQTPV VVPPFLQPEIMGVPKVKETMVPKHKEMPFPKYPVEPFTESQ SLTLTDVEKLHLPLPLVQSWMHQPPQPLPPTVMFPPQSVL SLSQPKVLPVPQKAVPQRDMPIQAFLLYQEPVLGPVRGPFP ILV 31 Caprine?-casein MKVLILACLVALAIAREQEELNVVGETVESLSSSEESIT HINKKIEKFQSEEQQQTEDELQDKIHPFAQAQSLVYPFTG PIPNSLPQNILPLTQTPVVVPPFLQPEIMGVPKVKETMVPK HKEMPFPKYPVEPFTESQSLTLTDVEKLHLPLPLVQSWMH QPPQPLSPTVMFPPQSVLSLSQPKVLPVPQKAVPQRDMPI QAFLLYQEPVLGPVRGPFPILV 32 Caprine?-casein REQEELNVVGETVESLSSSEESITHINKKIEKFQSEEQQQT matureprotein EDELQDKIHPFAQAQSLVYPFTGPIPNSLPQNILPLTQTPV VVPPFLQPEIMGVPKVKETMVPKHKEMPFPKYPVEPFTESQ SLTLTDVEKLHLPLPLVQSWMHQPPQPLSPTVMFPPQSVL SLSQPKVLPVPQKAVPQRDMPIQAFLLYQEPVLGPVRGPFP ILV 33 Buffalo?-casein MKVLILACLVALALARELEELNVPGEIVESLSSSEESITH INKKIEKFQSEEQQQMEDELQDKIHPFAQTQSLVYPFPGPI PKSLPQNIPPLTQTPVVVPPFLQPEIMGVSKVKEAMAPKHK EMPFPKYPVEPFTESQSLTLTDVENLHLPLPLLQSWMHQPP QPLPPTVMFPPQSVLSLSQSKVLPVPQKAVPYPQRDMPIQ AFLLYQEPVLGPVRGPFPIIV 34 Buffalo?-casein RELEELNVPGEIVESLSSSEESITHINKKIEKFQSEEQQQM matureprotein EDELQDKIHPFAQTQSLVYPFPGPIPKSLPQNIPPLTQTPVV VPPFLQPEIMGVSKVKEAMAPKHKEMPFPKYPVEPFTESQ SLTLTDVENLHLPLPLLQSWMHQPPQPLPPTVMFPPQSVL SLSQSKVLPVPQKAVPYPQRDMPIQAFLLYQEPVLGPVRG PFPIIV 35 Equine?-casein MKILILACLVALALAREKEELNVSSETVESLSSNEPDSS SEESITHINKEKLQKFKHEGQQQREVERQDKISRFVQPQP VVYPYAEPVPYAVVPQSILPLAQPPILPFLQPEIMEVSQAKE TILPKRKVMPFLKSPIVPFSERQILNPTNGENLRLPVHLIQP FMHQVPQSLLQTLMLPSQPVLSPPQSKVAPFPQPVVPYPQ RDTPVQAFLLYQDPRLGPTGELDPATQPIVAVHNPVIV 36 Equine?-casein REKEELNVSSETVESLSSNEPDSSSEESITHINKEKLQKFK matureprotein HEGQQQREVERQDKISRFVQPQPVVYPYAEPVPYAVVPQS ILPLAQPPILPFLQPEIMEVSQAKETILPKRKVMPFLKSPIVP FSERQILNPTNGENLRLPVHLIQPFMHQVPQSLLQTLMLPS QPVLSPPQSKVAPFPQPVVPYPQRDTPVQAFLLYQDPRLG PTGELDPATQPIVAVHNPVIV 37 Camel?-casein MKVLILACRVALALAREKEEFKTAGEALESISSSEESITH INKQKIEKFKIEEQQQTEDEQQDKIYTFPQPQSLVYSHTEP IPYPILPQNFLPPLQPAVMVPFLQPKVMDVPKTKETIIPKRK EMPLLQSPVVPFTESQSLTLTDLENLHLPLPLLQSLMYQIPQ PVPQTPMIPPQSLLSLSQFKVLPVPQQMVPYPQRAMPVQA VLPFQEPVPDPVRGLHPVPQPLVPVIA 38 Camel?-casein REKEEFKTAGEALESISSSEESITHINKQKIEKFKIEEQQQT matureprotein EDEQQDKIYTFPQPQSLVYSHTEPIPYPILPQNFLPPLQPAV MVPFLQPKVMDVPKTKETIIPKRKEMPLLQSPVVPFTESQS LTLTDLENLHLPLPLLQSLMYQIPQPVPQTPMIPPQSLLSLS QFKVLPVPQQMVPYPQRAMPVQAVLPFQEPVPDPVRGLHP VPQPLVPVIA 39 Human?-casein MKVLILACLVALALARETIESLSSSEESITEYKQKVEKVK HEDQQQGEDEHQDKIYPSFQPQPLIYPFVEPIPYGFLPQNI LPLAQPAVVLPVPQPEIMEVPKAKDTVYTKGRVMPVLKSPT IPFFDPQIPKLTDLENLHLPLPLLQPLMQQVPQPIPQTLALP PQPLWSVPQPKVLPIPQQVVPYPQRAVPVQALLLNQELLLN PTHQIYPVTQPLAPVHNPISV 40 Human?-casein RETIESLSSSEESITEYKQKVEKVKHEDQQQGEDEHQDKI matureprotein YPSFQPQPLIYPFVEPIPYGFLPQNILPLAQPAVVLPVPQPEI MEVPKAKDTVYTKGRVMPVLKSPTIPFFDPQIPKLTDLENL HLPLPLLQPLMQQVPQPIPQTLALPPQPLWSVPQPKVLPIP QQVVPYPQRAVPVQALLLNQELLLNPTHQIYPVTQPLAPVH NPISV

[0282] Gram positive bacteria (such as Lactococcus lactis and Bacillus subtilis) may be used to secrete target proteins into the media, and gram-negative bacteria (such as Escherichia coli) may be used to secrete target proteins into periplasm or into the media. In some embodiments, the bacterially-expressed proteins expressed may not have any post-translational modifications (PTMs), which means they are not glycosylated and/or may not be phosphorylated.

[0283] Target casein proteins may be expressed and produced in L. lactis both in a nisin-inducible expression system (regulated by PnisA promoter), lactate-inducible expression system (regulated by P170 promoter) or other similar inducible systems, as well as a constitutively expressed system (regulated by P secA promoter), wherein both are in a food-grade selection strain, such as NZ3900 using vector pNZ8149 (lacF gene supplementation/rescue principle). The secretion of functional proteins may be enabled by the signal peptide of Usp45 (SP(usp45)), the major Sec-dependent protein secreted by L. lactis. For example, ?.sub.s1-casein and ?.sub.s2-casein may be co-expressed or individually expressed in L. lactis using a synthetic operon.

Cheese and Cheese-Like Products

[0284] The present invention generally relates to cheese or cheese-like products comprising [0285] a) about 0.2 to about 6% by weight of ?.sub.s-casein, ?-casein, or a combination thereof, [0286] b) at least about 5% by weight of one or more hydrocolloids, wherein the one or more hydrocolloids comprise one or more starches, and [0287] c) from about 0.2 to about 20% by weight total dairy protein, wherein total casein in the cheese or cheese-like product comprises [0288] i) at least about 55% by weight ?.sub.s-casein, and/or [0289] ii) at least about 40% by weight ?-casein.

[0290] Also described herein are cheese and cheese-like products comprising [0291] a) about 0.2 to about 35% by weight ?.sub.s-casein, [0292] b) one or more hydrocolloids, comprising at least one of a starch or a gum.

[0293] The invention particularly reduces the total casein content required to form cheese and cheese-like products, particularly processed cheese and processed cheese-like products, while still achieving the desirable properties of such cheeses and cheese-like products.

[0294] The invention also may reduce or eliminate the amount of emulsifying salts needed to produce processed cheese and/or analogue type products. Processed cheese manufacture requires the use of emulsifying salts to modify casein conformation into forms that simultaneously emulsify milk fat, while creating a gel upon cooling that produces the desired body and texture. However, the use of ?.sub.s-casein and/or ?-casein according to the invention may both emulsify fat and form the desired body and texture upon cooling without the use of emulsifying salts.

[0295] The term cheese-like product as used herein, refers to products having one or more characteristics of a particular cheese, for example, products that, when consumed, impart the sensation of consuming cheese. In some embodiments, the cheese-like products may not meet the standard for a particular cheese, for example the Codex Alimentarius standard (found at http://www.fao.org/fao-who-codexalimentarius/codex-texts/list-standards/en/). In some embodiments the cheese-like product comprises no animal-derived ingredients. Such products are frequently referred to as vegan cheeses. Cheese-like products may comprise proteins, fats and/or lipids made by genetic engineering and/or fermentation to replicate the protein and/or fatty acid composition of milk fat.

[0296] In an exemplary embodiment the cheese-like product is a processed cheese-like product.

[0297] As used herein, the term processed cheese-like product refers to products having one or more of the characteristics of a processed cheese, and includes products such as processed cheese spread or an analogue cheese. As used herein, the term processed cheese refers to cheeses prepared by cooking and melting cheese or a blend of cheeses, and includes processed cheeses prepared as described herein in the Examples. When the hot processed cheese is formed, it is a homogeneous pumpable, fluid cheese material that may be cast or formed into sheets to produce blocks, loaves, slices or other desired forms using techniques such as rolling and/or cutting. Processed cheese is sometimes referred to as process cheese, and the terms are synonymous.

[0298] In various embodiments the cheese-like product is a vegetarian or vegan cheese.

[0299] The invention also relates to the use of unusually low amounts of ?.sub.s-casein and/or ?-casein to produce cheese and cheese-like products. Typical processed cheeses, such as those described herein in the Examples, may comprise from about 12 to 25% by weight total protein. In contrast, cheeses and cheese-like products produced according to the invention may comprise 0.2 to about 10% by weight total protein, 0.2 to about 9% by weight total protein or about 0.2 to 5% by weight total protein.

[0300] An exemplary cheese or cheese-like product of the invention may comprise: [0301] a) from about 0.05 to about 35% lipid, [0302] b) from about 15 to about 65% by weight moisture, [0303] c) from about 0.25 to about 7.5% by weight of simple carbohydrates, [0304] d) from about 0 to about 45% by weight of complex carbohydrates, [0305] e) from about 0 to about 0.75% by weight of one or more gums, [0306] f) from about 0.25 to about 3.5% salt, and [0307] g) from about 0.15 to about 5.0% by weight of minerals, cations, and/or anions.

[0308] In some embodiments the cheese or cheese-like product may comprise at least about 0.05, 1, 2, 2.5, 3, 4, 5, 6, 7, 7.5, 8, 9, 10, 11, 12, 12.5, 13, 14, 15, 16, 17, 17.5, 18, 19, 20, 21, 22, 22.5, 23, 24, 25, 26, 27, 27.5, 28, 29, 30, 31, 32, 33, 34 or 35% by weight lipid, and useful ranges may be selected between any of these values (for example, from about 0.05 to about 35, 1% to about 30%, about 5% to about 30%, about 10% to about 30%, about 17% to about 20%, about 20% to about 30%, about 25% to about 30%, about 27% to about 30%, about 27.5% to about 30%, about 1% to about 27.5% by weight fat, about 5% to about 27.5%, about 10% to about 27.5%, about 17% to about 27.5%, about 17% to about 27%, about 20% to about 25%, about 1% to about 22% by weight fat, about 5% to about 22%, about 10% to about 22%, about 17% to about 22%, about 1% to about 20%, about 5% to about 20%, about 10% to about 20%, about 1% to about 10% or about 1% to about 5% by weight lipid).

[0309] In one embodiment the cheese or cheese-like product is a reduced fat cheese or cheese-like product. In one embodiment the reduced fat cheese or cheese-like product comprises from about 0.5% to about 10% by weight lipid.

[0310] In some embodiments cheeses and cheese-like products of the invention may have the desired flavour, body, appearance, structure, texture, firmness, fibrosity, density, elasticity, springiness, melt, stretch, water binding capacity, creaminess and/or mouthfeel of equivalent cheeses produced by standard methods. Additionally, the proteins may perform specific physicochemical tasks such as forming desired gels, coagula, emulsions, foams, etc. The elasticity and functional properties of the gels produced by the protein blends of the invention may allow processing on processed cheese manufacturing and packaging equipment, including chill rolls, casting belts, free slice casting equipment, etc. Preferably all ingredients meet vegan/vegetarian specifications and expectations.

[0311] In one embodiment, the processed cheese or processed cheese-like product has a melt value of from about 3 to about 10, preferably from about 6 to about 7 on the Schreiber melt test. The Schreiber melt test was designed for use on processed cheese products, and especially for slice processed cheese products.

[0312] These techniques are described in Park, J., J. R. Rosenau, and M. Peleg. 1984. Comparison of four procedures of cheese meltability evaluation. J. Food Sci. 49:1158-1162, & 1170 and Zehren, V, L., and D. D. Nusbaum, 1962. Process Cheese. Cheese Reporter Publishing Co., Inc. Madison, WI.

[0313] Briefly, the Schreiber melt test is conducted by performing the following steps: [0314] cut a circular disk of cheese to a 39.5 mm diameter and thickness of about 4.7 mm, that is set into a glass (100?20 mm thin walled) petri dish bottom, [0315] place the petri dish cover on the sample and place in the refrigerator for 10 minutes at 4? C., [0316] place prepared sample into a preheated 232? C. forced draft oven and bake for 5 minutes, [0317] cool for 30 minutes, and [0318] remove petri dish lid and place dish upon Schreiber melt test chart (a chart containing a series of concentric circles starting at 39.5 mm in diameter, with the diameter of each succeeding circle 2 mm greater. [0319] report the Schreiber melt test score as listed on the concentric circles (i.e. sequentially 0 to 12).

[0320] A melt value of from about 6 to about 7 on the Schreiber melt test indicates an acceptable processed cheese performance. A melt value of greater than 6 on the Schreiber melt test indicates an unacceptable cheese performance.

[0321] In various embodiments, the cheese or cheese-like product comprises: [0322] a) about 1 to about 3.5%, preferably about 1.5% to about 2.5% w/w ?.sub.s-casein, [0323] b) about 15 to about 30%, preferably about 18 to about 27% w/w starch, [0324] c) about 0.1 to about 0.24%, preferably about 0.12 to about 0.2% w/w gum, [0325] d) optionally one or more non-dairy proteins, [0326] e) optionally one or more emulsifying salts, [0327] f) about 45 to about 65%, preferably about 50 to about 60% w/w moisture, and [0328] g) about 12 to about 26%, preferably about 15 to about 22% w/w lipid.

[0329] An exemplary cheese or cheese-like product comprises: [0330] a) about 2% w/w ?.sub.s-casein, [0331] b) about 22.5% w/w starch, [0332] c) about 0.16% w/w carrageenan, [0333] d) optionally one or more non-dairy proteins, [0334] e) optionally one or more emulsifying salts, [0335] f) about 58% w/w moisture, [0336] g) about 19% w/w vegetable fat, and [0337] h) about 1% w/w salt.

[0338] In various embodiments, the cheese or cheese-like product comprises: [0339] a) about 1.0 to about 5.0% total protein, [0340] b) about 0.90 to about 5.0% dairy protein, [0341] c) about 0.5 to about 4.5%, preferably about 0.8% to about 2.5% w/w ?.sub.s-casein, and/or about 1.0% to about 2.0%, preferably about 1.2% to about 1.7% ?-casein, [0342] d) about 15 to about 30%, preferably about 18 to about 27% w/w starch, preferably wherein the starch comprises about 65% to about 75% oxidised thin-boiling potato starch by weight of total starch, about 16% to about [0343] 22% OSA modified potato starch by weight of total starch, and about 11% to about 16% acetylated distarch phosphate modified potato starch by weight of total starch, [0344] e) about 0.1 to about 0.24%, preferably about 0.12 to about 0.2% w/w gum, [0345] f) optionally one or more non-dairy proteins, [0346] g) optionally one or more emulsifying salts, [0347] h) about 45 to about 65%, preferably about 50 to about 60% w/w moisture, and [0348] i) about 12 to about 26%, preferably about 15 to about 22% w/w lipid.

[0349] An exemplary cheese or cheese-like product comprises: [0350] a) about 1.0% w/w total protein, [0351] b) about 1.0% w/w dairy protein, [0352] c) about 0.8% w/w ?.sub.s-casein, [0353] d) about 18.4% w/w starch, [0354] e) about 0.16% w/w carrageenan, [0355] f) optionally one or more non-dairy proteins, [0356] g) optionally one or more emulsifying salts, [0357] h) about 56% w/w moisture, [0358] i) about 19% w/w vegetable fat, and [0359] j) about 1% w/w salt.

[0360] Another exemplary cheese or cheese-like product comprises: [0361] a) about 3% w/w total protein, [0362] b) about 3% w/w dairy protein, [0363] c) about 2.5% w/w ?.sub.s-casein, [0364] d) about 18.4% w/w starch, [0365] e) about 0.16% w/w carrageenan, [0366] f) optionally one or more non-dairy proteins, [0367] g) optionally one or more emulsifying salts, [0368] h) about 56% w/w moisture, [0369] i) about 19% w/w vegetable fat, and [0370] j) about 1% w/w salt.

[0371] Another exemplary cheese or cheese-like product comprises: [0372] a) about 3.0% w/w total protein, [0373] b) about 3.0% w/w dairy protein, [0374] c) about 1.3% w/w ?.sub.s-casein, [0375] d) about 1.3% w/w ?-casein, [0376] e) about 18.4% w/w starch, [0377] f) about 0.16% w/w carrageenan, [0378] g) optionally one or more non-dairy proteins, [0379] h) optionally one or more emulsifying salts, [0380] i) about 56% w/w moisture, [0381] j) about 19% w/w vegetable fat, and [0382] k) about 1% w/w salt.

[0383] Another exemplary cheese or cheese-like product comprises: [0384] a) about 3.0% w/w total protein, [0385] b) about 3.0% w/w dairy protein, [0386] c) about 1.6% w/w ?-casein, [0387] d) about 18.4% w/w starch, [0388] e) about 0.16% w/w carrageenan, [0389] f) optionally one or more non-dairy proteins, [0390] g) optionally one or more emulsifying salts, [0391] h) about 56% w/w moisture, [0392] i) about 19% w/w vegetable fat, and [0393] j) about 1% w/w salt.

[0394] Another exemplary cheese or cheese-like product comprises: [0395] a) about 5% w/w total protein, [0396] b) about 5% w/w dairy protein, [0397] c) about 4.2% w/w ?.sub.s-casein, [0398] d) about 18.4% w/w starch, [0399] e) about 0.16% w/w carrageenan, [0400] f) optionally one or more non-dairy proteins, [0401] g) optionally one or more emulsifying salts, [0402] h) about 56% w/w moisture, [0403] i) about 19% w/w vegetable fat, and [0404] j) about 1% w/w salt.

[0405] In various embodiments, the cheese or cheese-like product comprises: [0406] a) about 1.0 to about 5.0% total protein, [0407] b) about 0.90 to about 5.0% dairy protein, [0408] c) about 0.5 to about 6.0%, preferably about 0.8% to about 4.5% w/w ?.sub.s-casein, [0409] d) about 15 to about 30%, preferably about 18 to about 27% w/w starch, preferably wherein the starch comprises about 65% to about 75% oxidised thin-boiling potato starch by weight of total starch, about 16% to about [0410] 22% OSA modified maize starch by weight of total starch, and about 10% to about 15% hydroxypropyl distarch phosphate modified tapioca starch by weight of total starch, [0411] e) about 0.1 to about 0.24%, preferably about 0.12 to about 0.2% w/w gum, [0412] f) optionally one or more non-dairy proteins, [0413] g) optionally one or more emulsifying salts, [0414] h) about 45 to about 65%, preferably about 50 to about 60% w/w moisture, and [0415] i) about 12 to about 26%, preferably about 15 to about 22% w/w lipid.

[0416] An exemplary cheese or cheese-like product comprises: [0417] a) about 1.0% w/w total protein, [0418] b) about 1.0% w/w dairy protein, [0419] c) about 0.8% w/w ?.sub.s-casein, [0420] d) about 18.5% w/w starch, [0421] e) about 0.15% w/w locust bean gum, [0422] f) optionally one or more non-dairy proteins, [0423] g) optionally one or more emulsifying salts, [0424] h) about 56% w/w moisture, [0425] i) about 19% w/w vegetable fat, and [0426] j) about 1% w/w salt.

[0427] Another exemplary cheese or cheese-like product comprises: [0428] a) about 5.0% w/w total protein, [0429] b) about 5.0% w/w dairy protein, [0430] c) about 4.1% w/w ?.sub.s-casein, [0431] d) about 18.5% w/w starch, [0432] e) about 0.15% w/w locust bean gum, [0433] f) optionally one or more non-dairy proteins, [0434] g) optionally one or more emulsifying salts, [0435] h) about 56% w/w moisture, [0436] i) about 19% w/w vegetable fat, and [0437] j) about 1% w/w salt.

Hydrocolloids

[0438] Hydrocolloids are useful as thickening or gelling agents in food products.

[0439] The cheese or cheese products described herein comprise one or more hydrocolloids. In various embodiments the one or more hydrocolloids comprise one or more starches, one or more gums, or a combination thereof.

[0440] In various embodiments the one or more hydrocolloids comprise one or more starches.

[0441] In various embodiments the starch comprises potato starch, maize starch, rice starch, tapioca (cassava) starch, wheat starch, pea starch, sweet potato starch, banana starch, barley starch, oat starch, sago starch, amaranth starch, arrowroot starch, canna starch, sorghum starch, chia seed starch, fava bean starch, jackfruit starch, bamboo starch, chickpea starch, cassava starch, carrot starch, or any combination of any two or more thereof. Particularly preferred starches include maize, potato, cassava, tapioca and rice starch.

[0442] In some embodiments the starch may be modified. Starches may be modified by physical, chemical or enzymatic means to produce modified starches with altered and/or enhanced properties, such as resistance to gelatinisation and/or retrogradation, improved stability, low viscosity when heated, enhanced gelling, and/or improved emulsification with fat.

[0443] In various embodiments the starch comprises a modified starch, for example, a pre-gelatinised starch, a thin-boiling starch, an oxidised starch, octenyl succinic anhydride (OSA) modified starch, or an acetylated distarch phosphate.

[0444] In various embodiments the starch comprises a modified potato starch, modified maize starch and/or a modified tapioca starch. In various embodiments the modified starch is selected from an oxidised thin boiling potato starch, an octenyl succinic anhydride (OSA) modified potato starch, an acetylated distarch phosphate modified potato starch, an octenyl succinic anhydride (OSA) modified maize starch, or a hydroxypropyl distarch phosphate modified tapioca starch, or a combination of any two or more thereof.

[0445] In various embodiments the one or more gums comprise xanthan gum, guar gum, locust bean gum, tragacanth gum, acacia gum, karaya gum, tara gum, gellan gum, konjac gum (konjac mannan), carboxymethylcellulose (CMC), tragacanth, agar, pectin, alginate, carrageenan (?-carrageenan, .Math.-carrageenan, ?-carrageenan), Arabic gum, pectin, alginate, flaxseed gum, guar gum, or any combination of any two or more thereof.

[0446] Other useful hydrocolloids include carboxymethylcellulose (CMC), tragacanth and plant-derived hydrocolloids, such as agar, alginate, carrageenan (?-carrageenan, .Math.-carrageenan, ?-carrageenan), Arabic gum, and mixtures thereof. Preferred hydrocolloids include, for example, agar, pectin, xanthan gum, carboxymethylcellulose (CMC), and carrageenan (?-carrageenan, .Math.-carrageenan, ?-carrageenan) and mixtures of such. A particularly preferred hydrocolloid is carrageenan.

Lipid

[0447] In various embodiments the cheese or cheese-like product comprises one or more vegetable or animal lipids.

[0448] In various embodiments the one or more lipids may comprise one or more vegetable or animal lipids.

[0449] Exemplary plant oils include, for example coconut oil, corn oil, cottonseed oil, canola oil, olive oil, palm oil, ground nut oil, safflower oil, sesame oil, soybean oil, sunflower oil, nut oil, hazelnut oil, almond oil, cashew oil, macadamia oil, pecan oil, pistachio oil, walnut oil, oils from melon and gourd seeds, bottle gourd oil, buffalo gourd oil, pumpkin seed oil, watermelon seed oil, acai oil, blackcurrant seed oil, borage seed oil, evening primrose oil, carob seed oil, amaranth oil, apricot oil, argan oil, artichoke oil, avocado oil, babassu oil, ben oil, borneo tallow nut oil, cohune oil, coriander seed oil, flax oil, flax seed oil, coriander seeds oil, grape seed oil, hemp oil, kapok seed oil, kiwi fruit oil, lallemantia oil, linseed oil, mustard oil, okra seed oil, perilla seed oil, pequi oil, pine nut oil, poppyseed oil, prune kernel oil, quinoa oil, ramtil oil, rice bran oil, tea oil, wheat germ oil, hydrogenated vegetable oil, cocoa butter, shea butter any derivative of palm kernel oil, cocoa butter or shea butter, for example a derivative obtained by fractionation, partial or full hydrogenation and/or extraction, or any combination of any two or more thereof.

[0450] In various embodiments the vegetable lipid comprises one or more vegetable oils, coconut oil, one or more lecithins, or any combination thereof.

[0451] In various embodiments the vegetable oil comprises canola (rapeseed) oil, corn oil, sunflower oil, olive oil, soybean oil, or any combination of any two or more thereof.

[0452] In various embodiments the one or more lecithins may comprise soy lecithin.

[0453] In various embodiments the lipid source is or comprises a milk lipid. Preferably the milk lipid is selected from cream, high fat cream, plastic cream, beta serum or anhydrous milk fat.

[0454] In various embodiments the vegetable or animal lipid may be derived from natural sources. In other embodiments, the vegetable or animal lipid may be produced recombinantly.

Dairy Proteins

[0455] The term dairy protein as used herein refers to any protein that is naturally present in the milk of a mammal, or any variant thereof. The term includes unpurified proteins as well as partially or completely purified or isolated proteins, and includes full-length and processed (mature) dairy proteins, as well as protein fragments. The term also includes dairy proteins that have been produced from non-milk sources, such as recombinantly expressed dairy proteins.

[0456] Dairy proteins include caseins, such as ?.sub.s1-casein, ?.sub.s2-casein, ?-casein, ?-casein, and p-?-casein; and whey proteins including ?-lactoglobulin, ?-lactalbumin, serum albumin, and immunoglobulins. Some examples of casein proteins are listed in Table 2.

Non-Dairy Proteins

[0457] The term non-dairy protein as used herein refers to any protein, excluding dairy proteins. The term non-dairy animal protein as used herein refers to any non-dairy protein that is naturally present in an animal. Examples of non-dairy animal proteins include gelatin (such as gelatin produced from pork) and collagen.

[0458] In various embodiments the non-dairy protein comprises algal protein, hydrolysed algal protein, plant protein or hydrolysed plant protein.

[0459] In various embodiments the plant protein may comprise, for example, canola (rapeseed), legume, cereal, nut, or seed protein or any combination of any two or more thereof, optionally wherein any one or more of the plant proteins is a hydrolysed plant protein. In various embodiments the legume protein comprises pea, chickpea, bean, lupin, lentil or soy protein, or any combination of any two or more thereof, optionally wherein any one or more of the plant proteins is a hydrolysed plant protein. In various embodiments the cereal protein comprises rice, wheat, sorghum, maize, corn, or barley protein, or any combination of any two or more thereof, optionally wherein any one or more of the plant proteins is hydrolysed plant protein. In various embodiments the nut protein comprises almond, or cashew protein, or any combination thereof, optionally wherein any one or more of the plant proteins is a hydrolysed plant protein. In various embodiments the seed protein comprises chia, hemp or flax (linseed) protein, or any combination of any two or more thereof, optionally wherein any one or more of the plant proteins is hydrolysed plant protein.

[0460] In various embodiments the non-dairy protein comprises a soy protein material, for example soy protein flour, soy protein concentrate, soy protein isolate, and mixtures thereof.

[0461] In various embodiments the cheese or cheese-like product comprises at least about 0, 0.01, 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 16, 17, 18, 19 or at least about 20% w/w non-dairy protein, and various ranges can be selected from between these values, for example, from about 0 to about 17, about 0 to about 15, about 0 to about 10, about 0 to about 8, about 0 to about 5, about 0.01 to about 17, about 0.01 to about 15, about 0.01 to about 10, about 0.01 to about 8, about 0.01 to about 5, about 0.1 to about 17, about 0.1 to about 15, 0.1 to about 10, 0.1 to about 8, 1 to about 8, 1 to about 5, or 5 to about 8% w/w of the non-dairy protein. Preferably the cheese or cheese-like product comprises about 0, 0.01, 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, or at about 8% w/w non-dairy protein, and suitable ranges may be selected from any of these values. In one embodiment, the cheese or cheese-like product contains essentially no non-dairy protein.

[0462] In various embodiments the non-dairy protein may be derived from natural sources. In other embodiments, the non-dairy protein may be produced recombinantly.

Emulsifying Salts

[0463] In various embodiments the cheese or cheese-like product comprises one or more emulsifying salts. In various embodiments the emulsifying salt comprises one or more sodium emulsifying salts, one or more potassium emulsifying salts, one or more calcium emulsifying salts, or any combination thereof. For example, the emulsifying salt may comprise sodium, potassium and/or calcium salts of the mono, di, and polyphosphoric acids, sodium, potassium and/or calcium salts of citric acid, citric acid and/or phosphoric acid with sodium hydrogen carbonate and/or calcium carbonate.

[0464] In various embodiments the potassium emulsifying salt comprises potassium citrate, tripotassium citrate, potassium dihydrogen orthophosphate, dipotassium phosphate, dipotassium dihydrogen orthophosphate, dipotassium dihydrogen orthophosphate-3-hydrate, dipotassium dihydrogen orthophosphate-6-hydrate, dipotassium dihydrogen diphosphate, tripotassium orthophosphate and tripotassium orthophosphate-3-hydrate and tripotassium orthophosphate-6-hydrate, tetrapotassium diphosphate, tetrapotassium diphosphate-3-hydrate, tetrapotassium pyrophosphate, pentapotassium triphosphate, Kurroll's salt ((KPO3)n), or any combination of any two or more thereof.

[0465] In various embodiments the sodium emulsifying salt comprises monosodium phosphate, disodium phosphate, trisodium phosphate, sodium metaphosphate (sodium hexametaphosphate), sodium acid pyrophosphate, tetrasodium pyrophosphate, sodium aluminium phosphate, sodium citrate, sodium tartrate, sodium potassium tartrate, and any combination of any two or more thereof.

[0466] In various embodiments the cheese or cheese-like product may comprise a calcium salt. The calcium salt may be selected from monocalcium phosphate, dicalcium phosphate, tricalcium phosphate, calcium chloride, calcium carbonate, calcium citrate, calcium glubionate, calcium lactate, calcium gluconate, calcium acetate, equivalents thereof and/or combinations thereof.

[0467] In various embodiments the concentration of calcium in the cheese or cheese-like product may be from about 0 mM to about 260 mM. In some embodiments the concentration of calcium in the cheese or cheese-like product may be from about 10 mM to about 55 mM. In some embodiments the concentration of calcium in the cheese or cheese-like product may be at least 10 mM or at most 50 mM. In some embodiments, the concentration of calcium in the cheese or cheese-like product may be 28 mM or no more than 28 mM or may be 55 mM or no more than 55 mM. The concentration of calcium in the cheese or cheese-like product may be 10 mM to 15 mM, 10 mM to 20 mM, 10 mM to 25 mM, 10 mM to 30 mM, 10 mM to 35 mM, 10 mM to 40 mM, 10 mM to 45 mM, 10 mM to 50 mM, 10 mM to 55 mM, 15 mM to 20 mM, 15 mM to 25 mM, 15 mM to 30 mM, 15 mM to 35 mM, 15 mM to 40 mM, 15 mM to 45 mM, 15 mM to 50 mM, 15 mM to 55 mM, 20 mM to 25 mM, 20 mM to 30 mM, 20 mM to 35 mM, 20 mM to 40 mM, 20 mM to 45 mM, 20 mM to 50 mM, 20 mM to 55 mM, 25 mM to 30 mM, 25 mM to 35 mM, 25 mM to 40 mM, 25 mM to 45 mM, 25 mM to 50 mM, 25 mM to 55 mM, 30 mM to 35 mM, 30 mM to 40 mM, 30 mM to 45 mM, 30 mM to 50 mM, 30 mM to 55 mM, 35 mM to 40 mM, 35 mM to 45 mM, 35 mM to 50 mM, 35 mM to 55 mM, 40 mM to 45 mM, 40 mM to 50 mM, 40 mM to 55 mM, 45 mM to 50 mM, 45 mM to 55 mM, or 50 mM to 55 mM. The concentration of calcium in the cheese or cheese-like product may be 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, or 55 mM. The concentration of calcium in the cheese or cheese-like product may be at least 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM or 50 mM. The concentration of calcium in the cheese or cheese-like product may be at most 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM or 55 mM.

Other Ingredients

[0468] In various embodiments the cheese or cheese-like product may comprise one or more additional ingredients.

[0469] In various embodiments the cheese or cheese-like product may comprise a source of milk protein.

[0470] In various embodiments the source of milk protein may be selected from [0471] a) whole fat milk, whole milk retentate/concentrate, semi skimmed milk, skimmed milk, skimmed retentate/concentrate, butter milk, butter milk retentate/concentrate and whey protein retentate/concentrate; or [0472] b) one or more powders, such as whole milk powder, skimmed milk powder, milk protein concentrate powder, micellar milk protein powder, milk protein isolate powder, whey protein concentrate powder, whey protein isolate powder and buttermilk powder or other powders made from milk, reconstituted or dry, singularly or in combination. In various embodiments the source of milk protein may comprise cheese or curd.

[0473] In alternative embodiments the cheese or cheese-like product does not comprise a source of milk protein. In some embodiments, the cheese or cheese-like product does not comprise caseinate (such as sodium caseinate), a milk protein concentrate (MPC), lactic casein and/or rennet casein.

[0474] In various embodiments the cheese or cheese-like product may comprise [0475] a) one or more complex carbohydrates, such as one or more maltodextrins, corn syrup, and corn syrup solids, [0476] b) one or more GRAS cationic compounds including one or more monovalent, divalent or multivalent cations such as potassium, sodium, calcium, magnesium, manganese and/or iron compounds, [0477] c) one or more GRAS anionic compounds such as citrate, propionate, phosphate, and/or carbonate, [0478] d) one or more mineral salts, [0479] e) one or more GRAS acids, including one of more of lactic acid, (L+) lactic acid, acetic acid, citric acid, malic acid, tartaric acid, propionic acid, phosphoric acid, sulfuric acid, and/or hydrochloric acid, and [0480] f) one or more other GRAS ingredients including preservatives (such as sorbates, for example, potassium sorbate), acidity regulators, colours and/or colouring agents such as pigments, flavours, and flow agents.

[0481] In various embodiments the cheese or cheese-like product comprises from about 0.25 to about 7.5% by weight simple carbohydrate and from about 0 to about 45% by weight complex carbohydrate.

[0482] In various embodiments the cheese or cheese-like product may comprise one or more lipid replacers. Lipid replacers are useful for reduced fat or no fat cheeses or cheese-like products. In some embodiments, the lipid replacers may comprise one or more polyols, for example, glycerine, sorbitol, allulose, erythritol, xylitol, and/or maltitol. In some embodiments, the lipid replacers comprise one or more long chain carbohydrates/fibres, for example inulin, galacto-oligosaccharides (GOS), fructo-oligosaccharides (FOS), oligo-fructose, and/or polydextrose. In some embodiments, the lipid replacers comprise both polyols and long chain carbohydrates/fibres.

Manufacturing Methods

[0483] The invention also relates to a method for preparing a cheese or cheese-like product, the method comprising [0484] a) providing a cheese composition comprising [0485] i. ?.sub.s-casein, ?-casein, or a combination thereof, [0486] ii. one or more hydrocolloids, comprising one or more starches, [0487] iii. optionally one or more lipids, and [0488] iv. optionally one or more additional ingredients, [0489] b) heating the composition at a temperature and for a time sufficient to produce a heated cheese mass or cheese-like mass, and [0490] c) cooling the heated cheese mass or cheese-like mass to produce the cheese or cheese-like product, [0491] wherein the cheese or cheese-like product comprises [0492] i. about 0.2 to about 6% by weight of ?.sub.s-casein, ?-casein, or a combination thereof, and [0493] ii. at least about 5% by weight of one or more hydrocolloids, and [0494] iii. from about 0.2 to about 20% by weight total dairy protein, [0495] wherein total casein in the cheese or cheese-like product comprises [0496] 1. at least about 55% by weight ?.sub.s-casein, and/or [0497] 2. at least about 40% by weight ?-casein.

[0498] Also described herein is a method for preparing a cheese or cheese-like product, the method comprising [0499] a) providing a composition comprising [0500] i. ?.sub.s-casein, the ?.sub.s-casein comprising ?.sub.s1-casein, ?.sub.s2-casein, or a combination thereof, [0501] ii. one or more hydrocolloids, comprising at least one of a starch or a gum, [0502] iii. optionally one or more lipids [0503] iv. optionally one or more non-dairy proteins, [0504] v. optionally one or more emulsifying salts, and [0505] vi. optionally one or more additional ingredients, [0506] b) heating the composition at a temperature and for a time sufficient to produce a heated cheese mass or cheese-like mass, and [0507] c) cooling the heated cheese mass or cheese-like mass to produce the cheese or cheese-like product,
wherein the cheese or cheese-like product comprises [0508] 1. about 0.2 to about 35% by weight ?.sub.s-casein, [0509] 2. optionally about 15 to about 65% by weight moisture, and [0510] 3. optionally about 0.05 to about 35% by weight lipid.

[0511] In one embodiment the invention relates to a method for producing a processed cheese or processed cheese product. Exemplary embodiments of the method for producing the processed cheese or processed cheese according to the invention are described as follows. Other suitable variations or alternatives to the embodiments described herein will be apparent to those skilled in the art.

[0512] In various embodiments the ?.sub.s-casein comprises or is provided by an ingredient or composition comprising an ?.sub.s-casein enriched fraction described herein or lyophilised recombinant ?.sub.s-casein proteins. In various embodiments the ?-casein comprises or is provided by an ingredient or composition comprising an ?-casein enriched fraction described herein or lyophilised recombinant ?-casein proteins.

[0513] In various embodiments the method comprises dispersing the ?.sub.s-casein and/or ?-casein in water, preferably at a temperature of about 15 to about 55? C., more preferably about 50? C., to substantially completely dissolve the ?.sub.s-casein and/or ?-casein. In various embodiments, the method comprises dispersing the ?.sub.s-casein in water at a temperature of about 40 to about 55? C., preferably about 50? C. In various embodiments, the method comprises dispersing the ?-casein in water at a temperature of about 15 to about 30? C., preferably about 20? C.

[0514] In one embodiment the water is adjusted to a pH of about 8.5 before adding the ?.sub.s-casein and/or ?-casein. In another embodiment, the ?.sub.s-casein and/or ?-casein is added to water and the pH subsequently adjusted to a pH of about 8.0. In one embodiment a salt such as trisodium citrate is added with the ?.sub.s-casein and/or ?-casein.

[0515] In one embodiment the ?.sub.s-casein and/or ?-casein may be dispersed in water and then added to a mixture of the remaining ingredients or added with other dry ingredients to lipid. In another embodiment the ?.sub.s-casein and/or ?-casein is combined directly with other dry ingredients or is added directly to the lipid. In some embodiments the lipid is mixed with water to form an emulsion before addition of the dry ingredients.

[0516] The ingredients may be combined to produce a composition using any mixer or cheese cooker typically used for cheese preparation. Suitable equipment will be well known to those in the art. Suitable cookers include a Vorwerk Thermomix, and processed cheese cookers. Preferably the ingredients are combined under agitation to ensure even mixing.

[0517] The composition is then heated (cooked) for a period at a temperature and for a time sufficient to produce a heated cheese mass or cheese-like mass. In a preferred embodiment the composition is heated for at least about 3 to about 15 minutes at a temperature of at least about 70? C., preferably about 80 to about 85? C. to produce a heated cheese or cheese-like product. Preferably the composition is heated under agitation.

[0518] The pH may be adjusted at suitable stages, for example, before, during or after cooking.

[0519] In one embodiment the heated cheese mass or cheese-like mass is immediately cooled and packed. In other embodiments the heated cheese is rolled and cast into slices, for example, to form a slice-on-slice (SOS) processed cheese or processed cheese-like product.

[0520] The above methods should be considered in no way limiting and suitable variations or alternatives will be apparent to those skilled in the art.

EXAMPLES

Example 1

[0521] This example describes the preparation of casein proteins for use in preparing the cheese and cheese-like products of the invention.

[0522] Casein was fractionated as described in WO 2009/108074, which is incorporated herein by reference.

[0523] Raw whole milk was separated to produce fresh raw skim milk that was subsequently pasteurized at 72.5? C. The temperature was adjusted to about 18? C. and the skim milk inoculated with a blend of standard dairy starter cultures including Lactobacillus subsp. lactis and Lactobacillus lactis subsp. cremoris. The skim milk was then incubated until microbial fermentation reduced the pH to about 4.6 to produce an acid casein curd. The curd was cut, cooked, separated from the whey, washed, and dried to produce a bulk supply of lactic casein by standard procedures. The composition of the lactic casein is shown below in Table 3.

[0524] Approximately 4.5 kg of lactic casein was dispersed into deionized water using sufficient shear to produce 100 L of a solubilized lactic casein slurry comprising 4% total solids. The temperature of the solubilized lactic casein was adjusted to <7? C. and then the pH adjusted to 10.2 by the addition of sodium hydroxide (1 Mol NaOH). Calcium chloride (dihydrate) was blended and dissolved into the prepared slurry to produce a uniform concentration of 0.272 kg CaCl.sub.2)/kg lactic casein. The treated casein sat quiescently overnight (?14 hours) to produce a coagulum in which syneresis created a curd suspended within a liquid portion. The curd and liquid were separated using a decanting centrifuge and both the curd and liquid were reserved.

[0525] The curd received an initial wash with a hydrochloric acid solution to remove residual calcium, which was followed by an additional acid wash (H.sub.2SO.sub.4). The curd was then dried to produce an ?.sub.s-casein enriched fraction.

[0526] The pH of the reserved liquid was adjusted to 4.3-4.4 by the addition of 1.0 Mol hydrochloric acid. The acidified mixture sat quiescently until another coagulum formed, which was subsequently separated from the second liquid mixture using a decanter, washed by two acid wash treatments (H.sub.2SO.sub.4), and dried to produce the ?-casein enriched fraction.

[0527] The dried casein fractions were each packaged separately and stored at ambient temperature until use.

[0528] Table 3 below shows the compositions of the dried ?.sub.s-casein enriched and ?-casein enriched fractions.

TABLE-US-00003 TABLE 3 Composition of lactic casein, ?.sub.s-casein enriched fraction, and ?-casein enriched fraction. Lactic casein measured ?.sub.s-casein ?-casein Component (-%-) enriched enriched Moisture (g/100 g) 9.31 9.33 10.1 Total solids 90.69 90.67 89.90 (g/100 g) Fat (g/100g) 1.15 0.49 0.41 Total protein 88.87 89.38 88.24 (g/100 g) Total casein NA 89.38 88.24 (g/100 g) ?.sub.s-Casein 35.59.sup.a 74.55.sup.b 27.58.sup.b (g/100 g) ?-Casein 30.91 7.69 47.89 (g/100 g) ?-Casein 11.28.sup.c 7.14 12.77 (g/100 g) CHO >0.1 0.02 0.02 Ash (g/100 g) 1.87 2.5 1.6 Calcium 0.02 175 34.1 (mg/100 g) (mg/100 g) Sodium >0.1 2.81 4.3 (mg/100 g) (mg/100 g) pH 4.38 5.27 3.73 .sup.a?.sub.s1-casein only .sup.b?.sub.s1- and ?.sub.s2-casein .sup.c?-casein was calculated based on ratio in Table 1

Example 2

[0529] This example describes the preparation of a standard slice-on-slice (SOS) processed cheddar cheese for comparison with the cheese and cheese-like products of the invention.

[0530] SOS processed cheddar cheese was prepared using the following method. [0531] a) Temper cheeses to 4 to 6? C. as required to ensure that all cheese is completely thawed. [0532] b) Grind cheese. [0533] c) Weigh citric acid and water (at ambient temperature), and dissolve citric acid into the water. [0534] d) Weigh cheeses and add weighed cheeses to the processed cheese cooker. [0535] e) Start low speed agitation in the selected processed cheese cooker, (most trials conducted using Thermomix, Vorwerk, Wuppertal, Germany, using speed setting 1). Suitable agitation in a standard twin screw, laydown processed cheese cooker is approximately 10 to 15 rpm. [0536] f) Uniformly disperse water/citric acid mixture into the cheese in the cooker. [0537] g) Weigh and combine all remaining dry ingredients. [0538] h) Slowly add the combined dry ingredients to the cheese/water/citric acid mixture in the cooker, uniformly distributing the dry ingredients into the cheese mixture. [0539] i) Moderately increase cooker agitation speed to produce a uniformly mixed blend, such as to speed setting 2 in a Thermomix or about 40 rpm in a twin screw cooker. [0540] j) Increase agitation and initiate cooking, such as to speed setting 4 in a Thermomix or about 120 rpm in a twin screw cooker. [0541] k) Cook at a constant rate, increasing the cheese/ingredient blend temperature from about 17-22? C. to about 85? C. within ?5 minutes. [0542] l) Adjust the agitation speed profile beginning at a moderate speed and increasing to the final agitation speed within about 2.5 min. [0543] m) Maintain final agitation speed throughout remainder of cooking. [0544] n) Halt heat application when cheese blend temperature reaches 85? C. and hold the cooked cheese blend at final agitation speed for 6 min. [0545] o) Discharge molten cheese into cooling and packaging system, roll molten blend with cooling to produce slices of desired thickness (usually 2 mm), cut slices, and wrap in a suitable packaging material. [0546] p) Cool and package wrapped SOS cheese further as required. [0547] q) Hold at refrigerated temperatures (4-6? C.) until evaluation.

[0548] Table 4 shows the formulation of the comparative SOS processed cheese. Table 5 shows the calculated composition of the final processed cheese.

TABLE-US-00004 TABLE 4 Formulation of comparative SOS processed cheddar cheese. Component % Cheddar cheese blend 76.7 Skim milk powder 2.0 Trisodium citrate (dihydrate) 3.6 Salt 0.2 Citric acid (crystallized) 0.2 Sorbic acid 0.2 Water 17.1 Total 100.0

TABLE-US-00005 TABLE 5 Calculated composition of comparative SOS cheddar cheese. Component Amount Total solids 58.5 w/w Moisture 41.5 w/w Fat 29.65 w/w Fat-in-dry matter 50.65 w/W Total protein 20.0 w/W Casein 19.8 w/w Whey protein/NPN 0.2 w/w Lactose 1.1 w/W Non-lactose carbohydrate 0.5 w/w Ash 2.5 w/w Emulsifying salt 3.2 w/w Salt 1.6 w/W Sorbate 0.2 w/w Calcium (mg/100 g) 625 Sodium (mg/100 g) 1475

Example 3

[0549] This example describes the preparation of processed cheese-like products of the invention comprising potato starch, carrageenan gum and ?.sub.s-casein. The example also investigates the effect of varying pH in the products of the invention and compares the properties of the products of the invention with those of a control cheese lacking ?.sub.s-casein.

[0550] The applicant acknowledges Emsland-St?rke GmbH as a supplier of some of the base formulations that may be used in the invention.

Method of Manufacture

[0551] The processed cheese-like products of the invention were prepared as follows: [0552] a) An ?.sub.s-casein enriched fraction prepared according to Example 1 was dispersed into water by [0553] i) Weighing half the water required by the formulation and adjusting to ambient temperature, [0554] ii) Adding trisodium phosphate (TSP) to the water with constant agitation to increase the pH to 8.5 (continuously measuring the solution pH during TSP addition, which is halted immediately when pH 8.5 is reached), [0555] iii) Increasing the temperature of the high pH water/TSP solution to 50? C. with low shear, low speed mixing, and [0556] iv) Adding the ?.sub.s-casein enriched fraction to the 50? C. high pH water and mixing for 5 minutes to ensure that the powder is dispersed and dissolved; [0557] v) Adjust the pH to 8.0, while maintaining agitation and hold for 5 minutes to ensure the casein dissolves. [0558] b) A fat and powdered ingredient mixture was prepared by [0559] i) Heating coconut fat to 60? C., allowing the fat to melt, [0560] ii) Separately combining modified potato starch, salt, carrageenan and potassium sorbate to produce a dry blend mixture; and [0561] iii) Gently mixing the dry blended ingredients into the molten fat for 1 minute to produce a smooth uniform blend; [0562] c) The ingredients were combined and cooked by [0563] i) Combining the remaining water, dissolved ?.sub.s-casein enriched fraction, and the fat-dry ingredient blend in processed cheese cooker, [0564] ii) Heating the blended ingredients to 85? C. at a low shear rate (speed setting 2 on the Thermomix), [0565] iii) Increasing the mixing speed immediately when the temperature reached 85? C. (speed setting 4.0 on the Thermomix) and mixing for 5 minutes, and [0566] d) The pH of the cooked product was adjusted by [0567] i) Removing a subsample of product and allowing the subsample to cool, while continuing to mix blend in cooker at low shear, [0568] ii) Measuring the pH when the subsample temperature was ?65? C., [0569] iii) Reducing the cooker blend sample to target pH by adding citric acid, or increasing the cooker blend pH by adding trisodium phosphate, [0570] iv) Total cook time including pH adjustment was 10 minutes in total. [0571] e) Pouring hot cooked product (85? C.) into suitable plastic containers, [0572] f) Sealing the containers, and [0573] g) Refrigerating the product until further use in testing, cutting into slices as required.

Formulation and Composition of Processed Cheese-Like Products

[0574] Table 6 shows the formulations of the processed cheese-like products and a standard processed cheese prepared according to the method of Example 2, while Table 7 shows the calculated final product composition.

TABLE-US-00006 TABLE 6 Formulations of processed cheese-like products. Control product Inventive product Component (-%-) (-%-) Water 54.8 54.8 Coconut fat 19.0 19.0 Modified potato starch - 15.1 15.1 oxidized thin boiling (e1404) Modified potato starch - 4.23 4.23 OSA (e1450) Modified potato starch - 3.12 3.12 acetylated distarch phosphate (e1414) Pea protein isolate 2.0 0 ?.sub.s-Casein 0 2.0 enriched fraction Salt 1.0 1.0 Potassium sorbate 0.24 0.24 Carrageenan 0.16 0.16 Lactic acid 0.1 0.1 Citric acid 0.1 As Needed Trisodium phosphate (TSP) 0 As Needed Total 100% by 100% by weight weight

TABLE-US-00007 TABLE 7 Calculated composition of processed cheese-like products. Inventive Processed Control product cheese of Component product (at all pH) Example 2 Total solids 41.2 41.7 58.5 (% by weight) Moisture 58.8 58.3 41.5 (% by weight) Fat 19.2 19.1 29.6 (% by weight) Fat-in-dry matter 46.6 46.1 50.65 (% by weight) Total protein 1.6 2.0 20.0 (% by weight) Total casein 0 1.96 19.8 (% by weight) ?.sub.s-Casein 0 1.67 NA (% by weight) ?-Casein 0 0.17 NA (% by weight) ?-Casein 0 0.16 NA (% by weight) Carbohydrate 18.5 18.8 1.6 (% by weight) Ash 1.23 1.32 2.5 (% by weight) Emulsifying salt 0 0.27 3.2 (% by weight) Salt 1 1 1.6 (% by weight) Sodium (mg/100 g) 390 500 1475

Melt Properties and Firmness

[0575] The meltability of the products was assessed using the Schreiber melt test described herein, upon duplicate samples.

[0576] The pH of each product was measured at the time the melt test was conducted.

[0577] The following penetrometer procedure measures firmness of cheeses and cheese-like products by determining the amount of force (measured as Newtons) required to push a cylindrical probe through a prepared sample at a constant speed. The force is measured by a load cell on a texture analyser. The maximum force is determined.

[0578] Penetrometer firmness was measured using by a TA HDplus Texture Analyzer (Stable Micro Systems, Ltd. Godalming, Surrey, UK) at 13? C. using a 6.3 mm cylindrical probe, with the following settings: [0579] Pre-Test speed 1.0 mm/s [0580] Test speed 1.0 mm/s [0581] Post-Test speed 1.0 mm/s [0582] Distance 10.0 mm [0583] Force 0.1 N [0584] Load cell 50 kg [0585] Trigger: auto

[0586] The product temperature was tested at 13? C. Each product was either packaged as a suitable block or consisted of a stack of 25 individual slices.

[0587] The data was collected on a computer and analysed by a commercial computer program. The results are shown in Table 8.

TABLE-US-00008 TABLE 8 Sample pH and mean Schreiber melt test results. Processed cheese of Control Example Inventive Inventive Inventive product 2 product product product pH measured 4.71 5.65 4.79 5.51 5.92 at time of analysis Melt 11.8 8.0 6.2 11.3 10.5 Penetrometer 6.0 12.3 8.4 8.1 8.8 firmness (N)

Viscoelasticity

[0588] The viscoelastic properties of the products were assessed by rheology according to the following method.

[0589] Oscillatory measurements (mechanical spectra) for each sample were performed in a frequency range of 0.1-50 rad/s, using strain values compromised in the linear viscoelastic region for each sample (0.5%). Data were collected using a Modular Compact Rheometer (MCR 300, Anton Paar). The attachment used was a 20 mm smooth parallel plate geometry. Gap was modified based on the thickness of each sample (ensuring the normal force was less than 0.5 N). Experiments were done at 25? C. The recorded data included the components of shear modulus G (storage modulus) and the viscous component G (loss modulus). The component tan (delta) were calculated at frequency 1.2 rad/s by dividing G to G.

[0590] The results are shown in Table 9.

TABLE-US-00009 TABLE 9 Rheological analysis. Control Dairy SOS Inventive Inventive Inventive product control product product product Measured 4.71 5.85 4.79 5.51 5.62 pH (at time of analysis) Storage 67721 42676 35648 21958 53116 modulus G (Pa at 1.2 rad/s) Loss 5915 13775 5330 4626 10598 modulus G (Pa at 1.2 rad/s) Tan ? 0.087 0.323 0.15 0.211 0.20 (at freq 1.2 rad/s)

Sensory Properties

[0591] The sensory properties of the prepared samples were evaluated by a team of 8 judges experienced in grading pasteurized processed cheese slices. The judges evaluated all samples for flavour including salt, acid, bitter, sweet, intensity of starch flavour, and any noticeable defects, such as oxidized flavours. Additionally, the judges evaluated the cheese samples for firmness and mouthfeel.

[0592] The judges generally found the sensory properties of the inventive products to be similar.

[0593] Other observations included that: [0594] a) the flavour of the inventive samples was less powdery with a reduced starch flavour compared with the control product; [0595] b) the fracturability and mouthfeel of the inventive samples more closely resembled a typical cheddar SOS cheese than the control sample; and [0596] c) the firmness and elasticity of the inventive samples more closely resembled a typical cheddar SOS cheese than the control sample.

Example 4

[0597] This example describes the preparation of processed cheese-like products of the invention.

Protein Sources

[0598] ?.sub.s-Casein enriched and ?-casein enriched fractions were obtained as described in Example 1. The total protein, total casein and casein fraction content for these fractions are provided in Table 3.

Formulations

[0599] Table 10 shows the formulations of the processed cheese-like products prepared according to the method outlined below, while Table 11 shows the calculated final product composition.

TABLE-US-00010 TABLE 10 Formulations of processed cheese-like samples made from ?.sub.s-casein enriched and ?-casein enriched fractions 1% Protein 3% Protein 3% Protein 1:1 3% Protein 5% Protein Component 0% Protein high alpha-Cn high alpha-Cn alpha:beta-Cn high beta-Cn high alpha-Cn Water 51.61 43.05 25.46 26.50 26.80 7.87 Coconut fat 19.13 19.13 19.13 19.13 19.13 19.13 Modified potato starch - 15.10 15.10 15.10 15.10 15.10 15.10 oxidized, thin boiling (e1404) Modified Potato starch - 4.23 4.23 4.23 4.23 4.23 4.23 OSA (e1450) Modified potato starch - 3.12 3.12 3.12 3.12 3.12 3.12 acetylated distarch phosphate (e1414) ?.sub.s-Casein enriched fraction 9.60 29.33 6.90 49.05 solution (10.3% protein) ?-Casein enriched fraction 21.40 28.00 solution (10.8% protein) Salt 1.01 1.01 1.01 1.01 1.01 1.01 Potassium sorbate 0.25 0.25 0.25 0.25 0.25 0.25 Carrageenan 0.16 0.16 0.16 0.16 0.16 0.16 Lactose 5.38 4.34 2.20 2.19 2.19 0.07 Citric acid As required As required As required As required As required Trisodium phosphate (TSP) As required As required As required As required As required Total 100% 100% 100% 100% 100% 100% by weight by weight by weight by weight by weight by weight

TABLE-US-00011 TABLE 11 Compositional analysis of processed cheese-like samples made from ?.sub.s-casein enriched and ?-casein enriched fractions - Base 1 (potato starch) 3% Protein 1:1 1% Protein high 3% Protein high alpha:beta-Cn 3% Protein high 5% Protein high 0% Protein alpha-Cn alpha-Cn ?-Enriched & beta-Cn alpha-Cn Dairy ?-Enriched ?-Enriched ?-enriched ?-Enriched ?-Enriched Component protein free fraction fraction fractions fraction fraction Moisture (g/100 g) 55.59 55.59 55.59 55.59 55.59 55.59 Total solids (g/100 g) 44.41 44.41 44.41 44.41 44.41 44.41 Fat (g/100 g) 19.15 19.16 19.17 19.16 19.15 19.18 Total protein (g/100 g) 0.03 1.00 3.00 3.00 3.00 5.00 ?.sub.s-Casein (g/100 g) 0.00 0.81 2.48 1.29 0.93 4.15 ?-Casein g/100 g) 0.00 0.08 0.26 1.29 1.61 0.43 ?-Casein (g/100 g) 0.00 0.08 0.24 0.38 0.43 0.40 Total casein (g/100 g) 0.00 0.97 2.97 2.97 2.97 4.97 Whey protein (g/100 g) 0.00 0.00 0.00 0.00 0.00 0.00 Carbohydrate (total)(g/100 g) 23.85 22.81 20.67 20.66 20.66 18.54 Starch solids (g/100 g) 18.39 18.39 18.39 18.39 18.39 18.39 Carrageenan (%) 0.16 0.16 0.16 0.16 0.16 0.16 Lactose (g/100 g) 5.11 4.12 2.09 2.08 2.08 0.07 Fibre (g/100 g) 0.07 0.07 0.07 0.07 0.07 0.07 Sugars (g/100 g) 5.39 4.35 2.21 2.20 2.20 0.08 Ash (g/100 g) 0.64 0.64 0.64 0.64 0.64 0.64 Potassium sorbate.sup.a (%) 0.25 0.25 0.25 0.25 0.25 0.25 Salt (%) 1.01 1.01 1.01 1.01 1.01 1.01 pH (Lab) 5.06 5.08 5.12 5.18 5.24 5.19 Ca (mg/100 g) 1.56 3.44 7.29 2.97 1.65 11.14 Na (mg/100 g) 433.00 432.85 432.52 432.52 432.52 432.20 .sup.a Potassium sorbate is also included in the ash content.

Method of Manufacture

[0600] Processed cheese manufacture proceeded as follows: [0601] a) Hydrating each individual protein source by [0602] i) dispersing each protein into water at an appropriate temperature with sufficient agitation to create separate solutions with about 89.5% moisture, and about 10.5% solids: [0603] ii) The ?.sub.s-casein enriched casein source solution was made by heating it to 50? C., and adjusting to pH 8 with TSP. [0604] iii) The ?-casein enriched protein source solution was made by heating it to 20? C., adjusting to pH 8 with TSP. [0605] iv) each solution was held quiescently overnight at refrigeration temperatures from 4 to 5? C. [0606] b) Processed cheese manufacture continued the following day by [0607] i) heating hydrogenated coconut fat (Confat 92) to 60? C. until completely melted, [0608] ii) transferring the required amount of protein solution to a Thermomix cooker/mixer, and heating up to 60? C., [0609] iii) adding the required amount of the molten coconut fat into the protein solution, then uniformly blending the mixture to produce a uniform blend. Add the remaining water and mix until temperature reaches 60? C., [0610] iv) adding all of the remaining ingredients including starches, carrageenan, salt, lactose and potassium sorbate and blend to a uniform paste at 60? C., [0611] v) heating the blended ingredients to 85? C. at a low shear rate (speed setting 2 on the Thermomix), and [0612] vi) increasing the mixing speed immediately when the temperature reached 85? C. (speed setting 4.0 on the Thermomix) and mixing for 5 minutes, [0613] c) The pH of the cooked product was adjusted by [0614] i) removing a subsample of product and allowing the subsample to cool, while continuing to mix blend in cooker at low shear, [0615] ii) measuring the pH when the subsample temperature was ?65? C., [0616] iii) reducing the cooker blend sample to target in process pH of 5.2 by adding citric acid, or increasing the cooker blend pH by adding trisodium phosphate, [0617] iv) total cook time including pH adjustment was 10 minutes in total. [0618] d) Pouring hot cooked product (85? C.) into suitable plastic containers, [0619] e) Sealing the containers, and [0620] f) Refrigerating the product until further use in testing, cutting into slices as required.
Properties of processed cheese-like products of the invention

[0621] Table 12 shows the melt, firmness, and viscosity of the processed cheese-like product.

TABLE-US-00012 TABLE 12 Melt, Firmness and Viscosity of processed cheese-like products of the invention (Base 1) 1% Protein 3% Protein 3% Protein 1:1 3% Protein 5% Protein 0% Protein high alpha-Cn high alpha-Cn alpha:beta-Cn high beta-Cn high alpha-Cn Melt (Sch test units) 11.5 9.3 6.4 6.9 5.9 1.4 Penetrometer firmness (N) 6.3 7.3 9.7 11.4 11.0 14.2 ?.sub.Hot (mPas).sup.a 781 841 1143 1277 1162 1203 .sup.a?.sub.Hot = cheese viscosity measured at 85? C. in a Rapid Visco Analyser RVA4 (Newport Scientific, Warriewood, NSW).

Example 5

[0622] This example describes the preparation of processed cheese-like products of the invention.

Protein Sources

[0623] ?.sub.s-Casein enriched and ?-casein enriched fractions were obtained as described in Example 1. The total protein, total casein, casein fraction content, and lactose content for these fractions are provided in Table 3.

Method of Manufacture

[0624] Processed cheese was manufactured as described in Example 4.

Formulation and Composition of Processed Cheese-Like Products

[0625] Formulations of processed cheese-like products of the invention are presented in Table 13.

TABLE-US-00013 TABLE 13 Formulations of processed cheese-like samples made from ?.sub.s-casein enriched fractions 0% 1% Protein 5% Protein Component Protein high alpha-Cn high alpha-Cn Water 51.95 43.39 8.21 Coconut fat 19.13 19.13 19.13 Modified potato starch - 15.10 15.10 15.10 oxidised, thin boiling (e1404) Modified maize starch - 4.32 4.32 4.32 OSA (e1450) Modified tapioca starch - 2.71 2.71 2.71 hydroxypropyl distarch phosphate (e1442) ?.sub.s-Casein enriched 9.60 49.05 fraction solution 10.3% protein) Salt 1.01 1.01 1.01 Potassium sorbate 0.25 0.25 0.25 Locust bean gum 0.15 0.15 0.15 Lactose 5.38 4.34 0.07 Citric acid As required As required Trisodium phosphate (TSP) As required As required Total 100% by 100% by 100% by weight weight weight

[0626] Table 14 shows the target compositions of processed cheese-like products made by the above method.

TABLE-US-00014 TABLE 14 Composition of processed cheese-like products (Base 2) 1% Protein 5% Protein high high 0% Protein alpha-Cn alpha-Cn Dairy ?-Enriched ?-Enriched Component protein free fraction fraction Moisture (g/100 g) 55.59 55.59 55.59 Total solids (g/100 g) 44.41 44.41 44.41 Fat (g/100 g) 19.16 19.16 19.18 Total protein (g/100 g) 0.03 1.00 5.00 ?.sub.s-Casein 0.000 0.812 4.147 (g/100 g) ?-Casein g/100 g) 0.000 0.084 0.428 ?-Casein (g/100 g) 0.000 0.078 0.397 p-?-Casein (g/100 g) 0.000 0.000 0.000 Total Casein (g/100 g) 0.000 0.973 4.974 Whey protein 0.00 0.00 0.00 (g/100 g) Carbohydrate (total) 23.98 22.94 18.67 (g/100 g) Starch solids (g/100 g) 18.46 18.46 18.46 Locust bean gum (%) 0.15 0.15 0.15 Lactose (g/100 g) 5.11 4.12 0.07 Fibre (g/100 g) 0.14 0.14 0.14 Sugars (g/100 g) 5.38 4.34 0.07 Ash (g/100 g) 0.52 0.52 0.52 Potassium sorbate.sup.a 0.25 0.25 0.25 (%) Salt (%) 1.01 1.01 1.01 pH (target) 5.10 5.10 5.10 pH (lab) 5.22 5.07 5.21 Ca (mg/100 g) 1.56 3.16 9.76 Na (mg/100 g) 424 424 423 .sup.aPotassium sorbate is also included in the ash content.

Properties of Processed Cheese-Like Products of the Invention

[0627] Firmness was measured by penetrometry as outlined in previous examples. Melt was measured using the Schreiber Test method as outlined above.

TABLE-US-00015 TABLE 15 Melt, Firmness and viscosity of processed cheese- like products of the invention (Base 2) 0% 1% Protein high 5% Protein high Protein alpha-Cn alpha-Cn Melt (Sch test units) 9.3 9.3 9.8 Penetrometer 4.8 7.2 10.2 firmness (N) ?.sub.Hot (mPas).sup.a 463 510 836 .sup.a?.sub.Hot = cheese viscosity measured at 85? C. in a Rapid Visco Analyser RVA4 (Newport Scientific, Warriewood, NSW).

Sensory Properties

[0628] The sensory properties of the prepared samples were evaluated by a team of 6 judges experienced in grading pasteurized processed cheese slices. The judges evaluated all samples for flavour including salt, acid, bitter, sweet, intensity of starch flavour, and any noticeable defects, such as oxidized flavours. Additionally, the judges evaluated the cheese samples for firmness and mouthfeel.

[0629] The judges generally found the products containing 1% protein were the most similar to a dairy SOS control. Higher levels of protein provided higher firmness, but these products tended to be slower to melt in the mouth and have a drier mouthfeel.

[0630] Any documents referred to herein including, but not limited to, patents, patent applications, journal articles, books, and the like, are incorporated herein by reference in their entirety. Any section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0631] Although the invention has been described by way of example and with reference to particular embodiments, it is to be understood that modifications and/or improvements may be made without departing from the scope or spirit of the invention.