Compositions comprising a casein and methods of producing the same

1. A substitute dairy food composition comprising:

(i) a recombinant β-lactoglobulin protein and a κ-casein protein, wherein the recombinant β-lactoglobulin protein comprises an amino acid sequence that is at least 90% identical to an amino acid sequence of a wild-type mammalian β-lactoglobulin protein, and the κ-casein protein comprises an amino acid sequence that is at least 90% identical to an amino acid sequence of a wild-type mammalian κ-casein protein;

(ii) ash; and

(iii) optionally, one or more lipids,

wherein:

the recombinant β-lactoglobulin protein and the k-casein protein confer on the substitute dairy food composition one or more characteristics of a dairy food product selected from the group consisting of: taste, aroma, appearance, handling, mouthfeel, density, structure, texture, elasticity, springiness, coagulation, binding, leavening, aeration, foaming, creaminess and emulsification and

the substitute dairy food composition does not comprise any other milk proteins than the recombinant β-lactoglobulin protein and the κ-casein protein in (i).

2. The substitute dairy food composition of claim 1 , wherein the recombinant β-lactoglobulin protein has been produced by a fungal cell or a bacterial cell.

3. The substitute dairy food composition of claim 2 , wherein the fungal cell is from a genus selected from the group consisting of: Saccharomyces, Kluyveromyces, Pichia, Tetrahymena, Yarrowia, Hansenula, Blastobotrys, Zygosaccharomyces, Debravomyces, Aspergillus, Candida, Trichoderma, Chrysosporium, Fusarium , and Neurospora.

4. The substitute dairy food composition of claim 2 , wherein the recombinant β-lactoglobulin protein has been secreted by the fungal cell or bacterial cell.

5. The substitute dairy food composition of claim 1 , wherein the recombinant β-lactoglobulin protein has a non-mammalian glycosylation pattern and/or a non-mammalian phosphorylation pattern.

6. The substitute dairy food composition of claim 1 , wherein the κ-casein protein is a recombinant κ-casein protein.

7. The substitute dairy food composition of claim 6 , wherein the recombinant κ-casein protein has been produced by a fungal cell or a bacterial cell.

8. The substitute dairy food composition of claim 7 , wherein the fungal cell is from a genus selected from the group consisting of: Saccharomyces, Kluyveromyces, Pichia, Tetrahymena, Yarrowia, Hansenula, Blastobotrys, Zygosaccharomyces, Debravomyces, Aspergillus, Candida, Trichoderma, Chrysosporium, Fusarium , and Neurospora.

9. The substitute dairy food composition of claim 7 , wherein the recombinant κ-casein protein has been secreted by the fungal cell or bacterial cell.

10. The substitute dairy food composition of claim 6 , wherein the recombinant κ-casein protein has a non-mammalian glycosylation pattern and/or a non-mammalian phosphorylation pattern.

11. The substitute dairy food composition of claim 6 , wherein:

(a) the recombinant β-lactoglobulin protein has been produced by a fungal cell or a bacterial cell; and

(b) the recombinant κ-casein protein has been produced by a fungal cell or a bacterial cell.

12. The substitute dairy food composition of claim 11 , wherein the fungal cell in (a) and (b) are each independently from a genus selected from the group consisting of: Saccharomyces, Kluyveromyces, Pichia, Tetrahymena, Yarrowia, Hansenula, Blastobotrys, Zygosaccharomyces, Debravomyces, Aspergillus, Candida Trichoderma, Chrysosporium, Fusarium , and Neurospora.

13. The substitute dairy food composition of claim 11 , wherein the recombinant β-lactoglobulin protein has been secreted by the fungal cell or bacterial cell in (a); and the recombinant κ-casein protein has been secreted by the fungal cell or bacterial cell in (b).

14. The substitute dairy food composition of claim 6 , wherein both the recombinant β-lactoglobulin protein and the recombinant κ-casein protein have a non-mammalian glycosylation pattern and/or a non-mammalian phosphorylation pattern.

15. The substitute dairy food composition of claim 1 , wherein the substitute dairy food composition does not include an animal-derived compound.

16. The substitute dairy food composition of claim 1 , wherein the substitute dairy food composition does not include any compound isolated from a milk produced by a mammal.

17. The substitute dairy food composition of claim 1 , wherein the substitute dairy food composition comprises a plurality of micelles.

18. The substitute dairy food composition of claim 17 , wherein the plurality of micelles comprises the recombinant β-lactoglobulin protein.

19. The substitute dairy food composition of claim 17 , wherein the plurality of micelles comprises the κ-casein protein.

20. The substitute dairy food composition of claim 1 , wherein the substitute dairy food composition comprises one or more lipids selected from the group consisting of: oils, monoglycerides, diglycerides, triglycerides, free fatty acids, and phospholipids.

21. The substitute dairy food composition of claim 1 , wherein the substitute dairy food composition further comprises one or more sweetening agents.

22. The substitute dairy food composition of claim 21 , wherein the one or more sweetening agents are selected from the group consisting of: glucose, mannose, maltose, fructose, galactose, lactose, sucrose, monatin, tagatose, stevia , aspartame, cyclamate, saccharin, sucralose, mogrosides, brazzein, curculin, erythritol, glycyrrhizin, inulin isomalt, lacititol, mabinlin, malititol, mannitol, miraculin, monatin, monelin, osladin, pentadin sorbitol thaumatin, xylitol, acesulfame potassium advantame, alitame, aspartame-acesulfame sodium cyclamate, dulcin, glucin, neohesperidin dihydrochalcone, neotame, and P-4000.

23. The substitute dairy food composition of claim 1 , wherein the substitute dairy food composition further comprises one or more saccharides.

24. The substitute dairy food composition of claim 1 , wherein the ash comprises one or more of: calcium, phosphorous, phosphate, potassium, sodium, citrate, sulfate, carbonate, chloride, magnesium, iron, copper, zinc, manganese, selenium, iodine, retinol, carotene, vitamins, vitamin D, vitamin E, vitamin B 12 (cobalamin), vitamin B1 (thiamin), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B6 (pyridoxine), vitamin C, folate, vitamin A, and vitamin K, or a salt(s) thereof.

25. The substitute dairy food composition of claim 1 , wherein the ash has a final concentration in the substitute dairy food composition of about 0.1 weight % to about 2 weight %, or about 5% w/w to about 7% w/w.

26. The substitute dairy food composition of claim 1 , wherein the substitute dairy food composition is selected from the group consisting of a substitute buttermilk composition, a substitute yogurt composition, a substitute caseinate, a substitute cream, a substitute ice cream, a substitute frozen custard, a substitute cottage cheese, a substitute cream cheese, a substitute crème fraiche, a substitute curd, a substitute milk, and a substitute cheese.

27. The substitute dairy food composition of claim 1 , wherein the substitute dairy food composition is a powder composition.

28. The substitute dairy food composition of claim 1 , wherein the substitute dairy food composition comprises 0.5 weight % to 15 weight % of the recombinant β-lactoglobulin.

29. The substitute dairy food composition of claim 1 , wherein the substitute dairy food composition comprises 0.02 weight % to 15 weight % of the k-casein.

30. The substitute dairy food composition of claim 17 , wherein the plurality of micelles comprises the recombinant β-lactoglobulin and the k-casein protein.

31. The substitute dairy food composition of claim 1 , wherein the recombinant β-lactoglobulin forms a polymer matrix gel.

32. The substitute dairy food composition of claim 1 , wherein the one or more lipids comprise or consist of one or more lipids derived from a plant or a microbe.

33. The substitute dairy food composition of claim 32 , wherein the plant is selected from the group consisting of: soy, sunflower, coconut, peanut, cottonseed, olive, palm, rapeseed, safflower, sesame seed, soybean, almond, beech nut, brazil nut, cashew, hazelnut, macadamia nut, mongongo nut, pecans, pine nut, pistachio, walnut, avocado, corn, canola, safflower, flax seed, palm, palm kernel, palm fruit, babassu, shea butter, mango, cocoa, wheat, and rice.

34. The substitute dairy food composition of claim 1 , wherein the one or more lipids comprise one or more monoglycerides, diglycerides, triglycerides, and/or phospholipids comprising acyl chains selected from the group consisting of butyric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, α-linolenic acid, vaccenic acid, arachidic acid, behenic acid, lignoceric acid, cerotic acid, myristoleic acid, sapienic acid, elaidic acid, linoelaidic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, octanoic acid, hexanoic acid, and decanoic acid.

35. The substitute dairy food composition of claim 1 , wherein the one or more lipids comprise one or more trans-esterified monoglycerides, diglycerides, triglycerides, and/or phospholipids.

36. The substitute dairy food composition of claim 1 , wherein the one or more lipids comprise one or more monoglycerides, diglycerides, triglycerides, and/or phospholipids comprising acyl chains in a percentage that resembles a percentage of the fatty acyl chains found in a mammal-derived milk or dairy product.

37. The substitute dairy food composition of claim 1 , wherein the one or more lipids comprise one or more free fatty acid selected from the group consisting of butyric acid, caproic acid, caprylic acid, and capric acid.

38. The substitute dairy food composition of claim 1 , wherein the substitute dairy food composition further comprises a flavor compound.

39. The substitute dairy food composition of claim 38 , wherein the flavor compound is selected from the group consisting of: δ-decalactone, ethyl butyrate, 2-furyl methyl ketone, 2,3-pentanedione, γ-undecalactone, δ-undecalactone, acetoin, furfuryl alcohol, furfural, 2-methylfurfural, 2-methylpyrazine, chocolate flavoring, coffee flavoring, strawberry flavoring, almond flavoring, hazelnut flavoring, vanilla flavoring, green tea flavoring, Irish cream flavoring, and coconut flavoring.

40. A powder food composition comprising:

(i) a recombinant β-lactoglobulin protein and a κ-casein protein, wherein the recombinant β-lactoglobulin protein comprises an amino acid sequence that is at least 90% identical to an amino acid sequence of a wild-type mammalian β-lactoglobulin protein, and wherein the κ-casein protein comprises an amino acid sequence that is at least 90% identical to an amino acid sequence of a wild-type mammalian κ-casein protein;

(ii) ash; and

(iii) optionally, one or more lipids, wherein:

the powder food composition does not comprise any other milk proteins other than the recombinant β-lactoglobulin protein lactoglobulin and the κ-casein protein in (i); and

reconstitution of the powder food composition results in the production of a substitute dairy food composition, wherein the recombinant β-lactoglobulin protein and the k-casein protein confer on the substitute dairy food composition one or more characteristics of a dairy food product selected from the group consisting of: taste, aroma, appearance, handling, mouthfeel, density, structure, texture, elasticity, springiness, coagulation, binding, leavening, aeration, foaming, creaminess, and emulsification.

41. The powder food composition of claim 40 , wherein the recombinant β-lactoglobulin protein has been produced by a fungal cell or a bacterial cell.

42. The powder food composition of claim 41 , wherein the fungal cell is selected from a genus selected from the group consisting of: Saccharomyces, Kluyveromyces, Pichia, Tetrahymena, Yarrowia, Hansenula, Blastobotrys, Zygosaccharomyces, Debrayomyces, Aspergillus, Candida, Trichoderma, Chrysosporium, Fusarium , and Neurospora.

43. The powder food composition of claim 41 , wherein the recombinant β-lactoglobulin protein has been secreted by the fungal cell or bacterial cell.

44. The powder food composition of claim 40 , wherein the recombinant β-lactoglobulin protein has a non-mammalian glycosylation pattern and/or a non-mammalian phosphorylation pattern.

45. The powder food composition of claim 40 , wherein the κ-casein protein is a recombinant κ-casein protein.

46. The powder food composition of claim 45 , wherein the recombinant κ-casein protein has been produced by a fungal cell or a bacterial cell.

47. The powder food composition of claim 46 , wherein the fungal cell is selected from a genus selected from the group consisting of: Saccharomyces, Kluyveromyces, Pichia, Tetrahymena, Yarrowia, Hansenula, Blastobotrys, Zygosaccharomyces, Debrayomyces, Aspergillus, Candida, Trichoderma, Chrysosporium, Fusarium , and Neurospora.

48. The powder food composition of claim 46 , wherein the recombinant κ-casein protein has been secreted by the fungal cell or bacterial cell.

49. The powder food composition of claim 45 , wherein the recombinant κ-casein protein has a non-mammalian glycosylation pattern and/or a non-mammalian phosphorylation pattern.

50. The powder food composition of claim 45 , wherein:

(a) the recombinant β-lactoglobulin protein has been produced by a fungal cell or bacterial cell; and

(b) the recombinant κ-casein protein has been produced by a fungal cell or bacterial cell.

51. The powder food composition of claim 50 , wherein the fungal cell is selected from a genus selected from the group consisting of: Saccharomyces, Kluyveromyces, Pichia, Tetrahymena, Yarrowia, Hansenula, Blastobotrys, Zygosaccharomyces, Debrayomyces, Aspergillus, Candida, Trichoderma, Chrysosporium , and Neurospora.

52. The powder food composition of claim 50 , wherein the recombinant β-lactoglobulin protein has been secreted by the fungal cell or bacterial cell in (a); and the recombinant κ-casein protein has been secreted by the fungal cell or bacterial cell in (b).

53. The powder food composition of claim 45 , wherein both the recombinant β-lactoglobulin protein and the recombinant κ-casein protein have a non-mammalian glycosylation pattern and/or a non-mammalian phosphorylation pattern.

54. The powder food composition of claim 40 , wherein the powder food composition is an infant formula.

55. A supplemented dairy food composition comprising:

(i) a mammalian-produced milk or a mammal-produced dairy food, and

(ii) a recombinant β-lactoglobulin protein and a recombinant k-casein protein;

wherein:

the recombinant β-lactoglobulin protein comprises an amino acid sequence that is at least 90% identical to an amino acid sequence of a wild-type mammalian β-lactoglobulin protein, and the recombinant κ-casein protein comprises an amino acid sequence that is at least 90% identical to an amino acid sequence of a wild-type mammalian κ-casein protein;

the recombinant β-lactoglobulin protein and the recombinant k-casein protein enhance in the supplemented dairy food composition one or more characteristics of a dairy food product selected from the group consisting of: taste, aroma, appearance, handling, mouthfeel, density, structure, texture, elasticity, springiness, coagulation, binding, leavening, aeration, foaming, creaminess, and emulsification; and

the supplemented dairy food composition does not comprise any recombinant milk proteins other than the recombinant milk proteins in (ii).

56. The supplemented dairy food composition of claim 55 , wherein the recombinant β-lactoglobulin protein has been produced by a fungal cell or a bacterial cell.

57. The supplemented dairy food composition of claim 56 , wherein the fungal cell is selected from a genus selected from the group consisting of: Saccharomyces, Kluyveromyces, Pichia, Tetrahymena, Yarrowia, Hansenula, Blastobotrys, Zygosaccharomyces, Debrayomyces, Aspergillus, Candida, Trichoderma, Chrysosporium, Fusarium , and Neurospora.

58. The supplemented dairy food composition of claim 56 , wherein the recombinant β-lactoglobulin protein has been secreted by the fungal cell or bacterial cell.

59. The supplemented dairy food composition of claim 55 , wherein the recombinant β-lactoglobulin protein has a non-mammalian glycosylation pattern and/or a non-mammalian phosphorylation pattern.

60. The supplemented dairy food composition of claim 55 , wherein the recombinant κ-casein protein has been produced by a fungal cell or a bacterial cell.

61. The supplemented dairy food composition of claim 60 , wherein the fungal cell is selected from a genus selected from the group consisting of: Saccharomyces, Kluyveromyces, Pichia, Tetrahymena, Yarrowia, Hansenula, Blastobotrys, Zygosaccharomyces, Debrayomyces, Aspergillus, Candida, Trichoderma, Chrysosporium, Fusarium , and Neurospora.

62. The supplemented dairy food composition of claim 55 , wherein the recombinant κ-casein protein has been secreted by the fungal cell or bacterial cell.

63. The supplemented dairy food composition of claim 55 , wherein the recombinant κ-casein protein has a non-mammalian glycosylation pattern and/or a non-mammalian phosphorylation pattern.

64. The supplemented dairy food composition of claim 55 , wherein:

(a) the recombinant β-lactoglobulin protein has been produced by a fungal cell or bacterial cell; and

(b) the recombinant κ-casein protein has been produced by a fungal cell or bacterial cell.

65. The supplemented dairy food composition of claim 64 , wherein the fungal cell is selected from a genus selected from the group consisting of: Saccharomyces, Kluyveromyces, Pichia, Tetrahymena, Yarrowia, Hansenula, Blastobotrys, Zygosaccharomyces, Debrayomyces, Aspergillus, Candida, Trichoderma, Chrysosporium , and Neurospora.

66. The supplemented dairy food composition of claim 55 , wherein the recombinant β-lactoglobulin protein has been secreted by the fungal cell or bacterial cell in (a); and the recombinant κ-casein protein has been secreted by the fungal cell or bacterial cell in (b).

67. The supplemented dairy food composition of claim 55 , wherein both the recombinant β-lactoglobulin protein and the recombinant κ-casein protein have a non-mammalian glycosylation pattern and/or a non-mammalian phosphorylation pattern.

68. The supplemented dairy food composition of claim 55 , wherein the supplemented dairy food composition comprises a plurality of micelles comprising the recombinant β-lactoglobulin protein and the recombinant k-casein protein.

69. The supplemented dairy food composition of claim 55 , wherein the mammalian-produced milk or the mammal-produced dairy food is selected from the group consisting of: an ice cream composition, a milk composition, a yoghurt composition, a cream composition, a cheese composition, a frozen custard composition, a crème fraiche composition, a curd composition, a cottage cheese composition, a cream cheese composition, a buttermilk composition, and a caseinate composition.