Compositions comprising a casein and methods of producing the same

Abstract

Disclosed herein are methods and compositions including casein, and methods for making these compositions.

What is claimed is: 
     
         1 . A substitute dairy food composition comprising:
 (i) a recombinant β-lactoglobulin protein and a β-casein protein;   (ii) ash; and   (iii) optionally, one or more lipids;   wherein the recombinant β-lactoglobulin protein comprises an amino acid sequence that is at least 80% identical to an amino acid sequence of a mammalian β-lactoglobulin protein;   wherein the β-casein protein comprises an amino acid sequence that is at least 50% identical to an amino acid sequence of a non-human mammalian β-casein protein;   wherein the recombinant β-lactoglobulin protein and the β-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   wherein the substitute dairy food composition does not comprise any other milk proteins than the recombinant β-lactoglobulin protein and the β-casein protein in (i).   
     
     
         2 . The substitute dairy food composition of  claim 1 , wherein the recombinant β-lactoglobulin protein comprises an amino acid sequence that is at least 90% identical to an amino acid sequence of a mammalian β-lactoglobulin protein. 
     
     
         3 . The substitute dairy food composition of  claim 1 , wherein the recombinant β-lactoglobulin protein comprises an amino acid sequence that is at least 95% identical to an amino acid sequence of a mammalian β-lactoglobulin protein. 
     
     
         4 . The substitute dairy food composition of  claim 1 , wherein the recombinant β-lactoglobulin protein comprises an amino acid sequence that is at least 98% identical to an amino acid sequence of a mammalian β-lactoglobulin protein. 
     
     
         5 . The substitute dairy food composition of  claim 1 , wherein the β-casein protein comprises an amino acid sequence that is at least 70% identical to an amino acid sequence of a non-human mammalian β-casein protein. 
     
     
         6 . The substitute dairy food composition of  claim 1 , wherein the β-casein protein comprises an amino acid sequence that is at least 90% identical to an amino acid sequence of a non-human mammalian β-casein protein. 
     
     
         7 . The substitute dairy food composition of  claim 1 , wherein the β-casein protein comprises an amino acid sequence that is at least 95% identical to an amino acid sequence of a non-human mammalian β-casein protein. 
     
     
         8 . The substitute dairy food composition of  claim 1 , wherein the β-casein protein comprises an amino acid sequence that is at least 98% identical to an amino acid sequence of a non-human mammalian β-casein protein. 
     
     
         9 . The substitute dairy food composition of  claim 1 , wherein the mammalian β-lactoglobulin protein is selected from the group consisting of cow, human, sheep, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, mountain goat, monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboons, gibbons, orangutan, mandrill, pig, wolf, fox, lion, tiger, echidna, and woolly mammoth β-lactoglobulin proteins. 
     
     
         10 . The substitute dairy food composition of  claim 1 , wherein the non-human mammalian β-casein protein is selected from the group consisting of cow, sheep, goat, buffalo, camel, horse, donkey, lemur, panda, guinea pig, squirrel, bear, macaque, gorilla, chimpanzee, mountain goat, monkey, ape, cat, dog, wallaby, rat, mouse, elephant, opossum, rabbit, whale, baboons, gibbons, orangutan, mandrill, pig, wolf, fox, lion, tiger, echidna, and woolly mammoth β-casein proteins. 
     
     
         11 . 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. 
     
     
         12 . The substitute dairy food composition of  claim 1 , wherein the substitute dairy food composition comprises 0.02 weight % to 15 weight % of the β-casein. 
     
     
         13 . The substitute dairy food composition of  claim 1 , wherein the recombinant β-lactoglobulin protein is produced by a fungal cell or a bacterial cell. 
     
     
         14 . The substitute dairy food composition of  claim 13 , 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, Fusarium, Chrysosporium , and  Neurospora.    
     
     
         15 . The substitute dairy food composition of  claim 13 , wherein the recombinant β-lactoglobulin protein is secreted by the fungal cell or the bacterial cell. 
     
     
         16 . 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. 
     
     
         17 . The substitute dairy food composition of  claim 1 , wherein the β-casein protein is a recombinant β-casein protein. 
     
     
         18 . The substitute dairy food composition of  claim 17 , wherein the recombinant β-casein protein is produced by a fungal cell or a bacterial cell. 
     
     
         19 . The substitute dairy food composition of  claim 18 , 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, Fusarium, Chrysosporium , and  Neurospora.    
     
     
         20 . The substitute dairy food composition of  claim 18 , wherein the recombinant β-casein protein is secreted by the fungal cell or the bacterial cell. 
     
     
         21 . The substitute dairy food composition of  claim 17 , wherein the recombinant β-casein protein has a non-mammalian glycosylation pattern and/or a non-mammalian phosphorylation pattern. 
     
     
         22 . The substitute dairy food composition of  claim 17 , wherein (a) the recombinant β-lactoglobulin protein is produced by a fungal cell or a bacterial cell, and/or (b) the recombinant β-casein protein is produced by a fungal cell or a bacterial cell. 
     
     
         23 . The substitute dairy food composition of  claim 22 , wherein the fungal cell in (a) and the fungal cell in (b) are each independently selected from a genus selected from the group consisting of  Saccharomyces, Kluyveromyces, Pichia, Tetrahymena, Yarrowia, Hansenula, Blastobotrys, Zygosaccharomyces, Debrayomyces, Aspergillus, Candida, Trichoderma, Fusarium, Chrysosporium , and  Neurospora.    
     
     
         24 . The substitute dairy food composition of  claim 22 , wherein the recombinant β-casein protein is secreted by the fungal cell or bacterial cell in (a), and/or the recombinant β-casein protein is secreted by the fungal cell or bacterial cell in (b). 
     
     
         25 . The substitute dairy food composition of  claim 17 , wherein one or both of the recombinant β-lactoglobulin protein and the recombinant β-casein protein have a non-mammalian glycosylation pattern and/or a non-mammalian phosphorylation pattern. 
     
     
         26 . The substitute dairy food composition of  claim 1 , wherein the substitute dairy food composition comprises a plurality of micelles. 
     
     
         27 . The substitute dairy food composition of  claim 26 , wherein the plurality of micelles comprises the recombinant β-lactoglobulin protein. 
     
     
         28 . The substitute dairy food composition of  claim 26 , wherein the plurality of micelles comprises the β-casein protein. 
     
     
         29 . The substitute dairy food composition of  claim 26 , wherein the plurality of micelles comprises the recombinant β-lactoglobulin protein and the β-casein protein. 
     
     
         30 . The substitute dairy food composition of  claim 1 , wherein the recombinant β-lactoglobulin forms a polymer matrix gel. 
     
     
         31 . 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. 
     
     
         32 . The substitute dairy food composition of  claim 31 , 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. 
     
     
         33 . 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, pamitoleic 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. 
     
     
         34 . 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. 
     
     
         35 . 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. 
     
     
         36 . The substitute dairy food composition of  claim 1 , wherein the one or more lipids comprise one or more free fatty acids selected from the group consisting of butyric acid, caproic acid, caprylic acid, and capric acid. 
     
     
         37 . The substitute dairy food composition of  claim 1 , wherein the substitute dairy food composition further comprises one or more sweetening agents. 
     
     
         38 . The substitute dairy food composition of  claim 37 , 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, monelin, osladin, pentadin, sorbitol, thaumatin, xylitol, acesulfame potassium, advantame, alitame, aspartame-acesulfame, sodium cyclamate, dulcin, glucin, neohesperidin dihydrochalcone, neotame, and P-4000. 
     
     
         39 . The substitute dairy food composition of  claim 1 , wherein the substitute dairy food composition further comprises one or more saccharides. 
     
     
         40 . 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, molybdenum, iron, copper, zinc, manganese, selenium, iodine, retinol, carotene, vitamins, vitamin D, vitamin E, vitamin B12 (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. 
     
     
         41 . 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. 
     
     
         42 . The substitute dairy food composition of  claim 1 , wherein the substitute dairy food composition further comprises at least one flavor compound. 
     
     
         43 . The substitute dairy food composition of  claim 42 , wherein the at least one 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, artificial flavors, chocolate flavoring, coffee flavoring, strawberry flavoring, almond flavoring, hazelnut flavoring, vanilla flavoring, green tea flavoring, Irish cream flavoring, and coconut flavoring. 
     
     
         44 . The substitute dairy food composition of  claim 1 , wherein the substitute dairy food composition is selected from the group consisting of: a substitute ice cream composition, a substitute milk composition, a substitute yoghurt composition, a substitute cream composition, a substitute cheese composition, a substitute frozen custard composition, a substitute crème fraiche composition, a substitute curd composition, a substitute cottage cheese composition, a substitute cream cheese composition, a substitute buttermilk composition, a substitute butter composition, and a substitute caseinate composition. 
     
     
         45 . The substitute dairy food composition of  claim 1 , wherein the substitute dairy food composition is a powder composition. 
     
     
         46 . A powder food composition comprising:
 (i) a recombinant β-lactoglobulin protein and a β-casein protein;   (ii) ash; and   (iii) optionally, one or more lipids;   wherein the recombinant β-lactoglobulin protein comprises an amino acid sequence that is at least 80% identical to an amino acid sequence of a mammalian β-lactoglobulin protein;   wherein the β-casein protein comprises an amino acid sequence that is at least 50% identical to an amino acid sequence of a non-human mammalian β-casein protein;   wherein the powder food composition does not comprise any other milk proteins other than the recombinant β-lactoglobulin protein and the β-casein protein in (i); and   wherein reconstitution of the powder food composition results in production of a substitute dairy food composition wherein the recombinant β-lactoglobulin protein and the β-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.   
     
     
         47 . 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 non-human β-casein protein;   wherein the recombinant β-lactoglobulin protein comprises an amino acid sequence that is at least 80% identical to an amino acid sequence of a mammalian β-lactoglobulin protein;   wherein the recombinant non-human β-casein protein comprises an amino acid sequence that is at least 50% identical to an amino acid sequence of a non-human mammalian β-casein protein;   wherein the recombinant β-lactoglobulin protein and the recombinant non-human β-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   wherein the supplemented dairy food composition does not comprise any recombinant milk proteins other than the recombinant milk proteins in (i).