Beverage products including brazzein sweet protein

Abstract

Described herein are beverage product formulations, including lemonade, soda, sparkling juice, fruit punch, mocktail mixer, cocktail mixer, and concentrates thereof, that include at least one recombinant sweet protein, and methods of producing the same.

Description

BEVERAGE PRODUCTS INCLUDING BRAZZEIN SWEET PROTEINCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application Serial No. 63 / 639,315, filed April 26, 2024 for which the entire contents are herein incorporated by reference.SEQUENCE LISTING

[0002] This application contains a Sequence Listing that has been submitted electronically as an XML file named 54282-0013W01_SL_ST26.xml. The XML file, created on April 25, 2025, is 6,012 bytes in size. The material in the XML file is hereby incorporated by reference in its entirety.TECHNICAL FIELD

[0003] This invention relates to compositions including sweet proteins. In particular, this invention relates to beverage formulations, compositions, and products including sweet proteins (e.g., brazzein and monellin).BACKGROUND

[0004] Various sweeteners are available, including naturally occurring carbohydrates, such as those obtainable from plants, and artificial sweeteners. However, high levels of sugar consumption have been linked to obesity and associated health problems, including an increased risk of high blood pressure, high cholesterol, Type 2 diabetes, and cardiovascular disease. Yet, sweet beverages, such as sodas and fruit juices, are favored by a large proportion of the population. Therefore, there is a need to provide healthy sweetener alternatives that deliver similar or improved qualities to currently available sugar and sugar alternatives, such as artificial sweeteners.SUMMARY

[0005] There is a need for alternative sweeteners and sweetened formulations that are healthier alternatives to available sweeteners such as sugar and sugar alternatives, particularly to provide sweeteners that don’t affect blood sugar levels and / or modulate thegut microbiome. In addition, there is a need to deliver not only an acceptable level of sweetness, but also the timing and longevity of sweetness that make sweet beverages desirable, and even craveable.

[0006] Accordingly, in one aspect, the present disclosure provides a sweetened beverage comprising one or more sweet proteins, wherein at least one of the one or more sweet proteins is present in an amount between 5 ppm and 584 ppm.

[0007] In some embodiments, the sweetened beverage comprises brazzein-53. In some embodiments, the brazzein-53 is present in an amount between 5 ppm and 300 ppm. In some embodiments, the brazzein-53 is the only sweet protein in the sweetened beverage and the brazzein-53 is present in an amount between 28 ppm and 300 ppm.

[0008] In some embodiments, the sweetened beverage comprises brazzein-54. In some embodiments, the brazzein-54 is present in an amount between 11 ppm and 584 ppm. In some embodiments, the brazzein-54 is the only sweet protein in the sweetened beverage and the brazzein-54 is present in an amount between 65 ppm and 584 ppm.

[0009] In some embodiments, the sweetened beverage comprises monellin. In some embodiments, the monellin is present in an amount between 8 ppm and 440 ppm. In some embodiments, the monellin is the only sweet protein in the sweetened beverage and the monellin is present in an amount between 46 ppm and 440 ppm.

[0010] In some embodiments, the sweetened beverage comprises only two sweet proteins.

[0011] In some embodiments, the two sweet proteins are brazzein-53 and brazzein-54. In some embodiments, the brazzein-53 is present in an amount between 7 ppm and 131 ppm and the brazzein-54 is present in an amount between 16 ppm and 256 ppm.

[0012] In some embodiments, the two sweet proteins are brazzein-53 and monellin. In some embodiments, the brazzein-53 is present in an amount between 7 ppm and 131 ppm and the monellin is present in an amount between 11 ppm and 193 ppm. In some embodiments, the brazzein-53 is present in an amount between 10 ppm and 265 ppm and the monellin is present in an amount between 20 ppm and 140 ppm.

[0013] In some embodiments, the two sweet proteins are brazzein-54 and monellin. In some embodiments, the brazzein-54 is present in an amount between 16 ppm and 256 ppm and the monellin is present in an amount between 11 ppm and 193 ppm.

[0014] In some embodiments, the sweetened beverage comprises brazzein-53, brazzein- 54 and monellin. In some embodiments, the brazzein-53 is present in an amount between 5 ppm and 87 ppm, the brazzein-54 is present in an amount between 11 ppm and 169 ppm, and the monellin is present in an amount between 8 ppm and 127 ppm.

[0015] In some embodiments, the sweetened beverage is carbonated. In some embodiments, pH of the sweetened beverage is between about 2.7 and about 3.5. In some embodiments, the sweetened beverage comprises between 0% and 3% sugar. In some embodiments, the sweetened beverage comprises a fruit juice, a fruit concentrate or a combination thereof.

[0016] In some embodiments, the sweetened beverage is a lemonade. In some embodiments, the sweetened beverage is a sparkling juice. In some embodiments, the sweetened beverage is a fruit punch. In some embodiments, the sweetened beverage is a soda. In some embodiments, the sweetened beverage is a mocktail mixer or a cocktail mixer. In certain embodiments, the mocktail mixer or the cocktail mixer is savory or spicy.

[0017] In some embodiments, the sweetened beverage does not contain added sugar. In some embodiments, the sweetened beverage does not contain any high intensity sweeteners selected from the group consisting of rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, rebaudioside I; Rebaudioside J, rebaudioside K, rebaudioside M, rebaudioside N, rebaudioside O, rebaudioside Q, rebaudioside R, dulcoside A, dulcoside C; Rubusoside, steviol monoside, steviol bioside, stevioside, mogroside V, and saccharin.

[0018] In some embodiments, at least one of the one or more sweet proteins is a recombinant protein.

[0019] In some embodiments, the recombinant protein is produced by fermentation of a microbial organism.

[0020] In some embodiments, the microbial organism is a yeast, optionally Komagataella phaffii.

[0021] In some embodiments, the recombinant protein is selected from the group consisting of brazzein-53, brazzein-54, monellin and any combination thereof.DETAILED DESCRIPTION

[0022] Described herein are sweetened beverage products that include a sweet protein, such as brazzein-53, brazzein-54, monellin, or any combinations thereof.I. Taste and Taste Receptors

[0023] Taste is a form of chemoreception which occurs in taste receptors in the mouth and is one of a variety of mechanisms used by mammals to sense stimuli and the external environment.

[0024] A taste can change over the process of consuming a beverage product. A tasting profile can be described as having a starting taste / flavor, for example an initial sensation of sweetness, a curve or shape (such as a Gaussian curve or roundness) of sweetness from start to finish (or a portion thereof) and a finishing taste / flavor. A starting taste and finishing taste can have a temporal element, i.e., how fast the starting taste, such as sweetness, is detected by a subject or how long the finishing flavor is detectable by a subject. A finishing taste / flavor can be described as lingering, meaning the finishing taste / flavor such as sweetness, is discernable by a subject for a long time (e.g., 5 second, 10 seconds, 30 seconds, or 1 minute) after swallowing the beverage product responsible for causing the taste. A curve or shape of the sweetness provides a profile of the timing and magnitude of sweetness of the course when the sweetness is discernible. The presence, amounts, and ratios of one or more sweet proteins in a beverage product can affect the taste profile (e.g., sweetness characteristics) of the beverage product.II. Sweet Proteins

[0025] Sweet proteins (also referred to herein as sweet polypeptides), such as brazzein- 53, brazzein-54 and monellin, can be formulated into a variety of beverages to achieve a desired level and timing of sweetness. In some cases, the sweet polypeptide can yield a sweet taste when consumed by a subject (e.g., a human). The present disclosure describes compositions, formulations, and products with a sweet polypeptide, such as brazzein-53 brazzein-54, monellin, each alone or in various combinations. In some implementations of the formulations described herein, the sweet polypeptide is a recombinant sweetprotein that, for example, can impart, elicit, and / or enhance a sweet taste when formulated into products such as the beverages described herein.

[0026] In some cases, the present disclosure describes the sweet protein, such as brazzein-53, brazzein-54 and monellin, that can generate a sweet taste and / or enhance sweetness when tasted or consumed by humans. In some cases, the sweet protein can be sweet-tasting and can taste sweet, for example, to a human subject. The disclosed sweet protein can generate a sweet taste and / or enhance a sweet taste of a beverage product, for example, as an ingredient or component of a composition, formulation, or product.

[0027] A sweet polypeptide described herein can have a sweeter taste than naturally occurring sugars, sugar alternatives and / or artificial sweeteners. In some cases, the sweetness of a sweet protein, such as brazzein-53, brazzein-54, monellin, alone or in various combinations, as determined by a human subject, for example, in a side-by-side gustatory comparison, can be greater than that of sugars, sugar alternatives and / or artificial sweeteners. In some embodiments, a sweet polypeptide, such as brazzein-53, brazzein-54, monellin, alone or in various combinations described herein can have an equivalent or greater level of sweetness than a naturally occurring sugar, a sugar alternative and / or an artificial sweetener, and additionally do so without affecting glycemic index (e.g., blood sugar levels) and without modifying the gut microbiome.

[0028] The terms “polypeptide,” “peptide,” and “protein” are used interchangeably herein to refer to polymers of amino acids of any length. The polymer can be linear or branched, it can include modified amino acids (e.g., pyroglutamate (pGlu)), and / or it can be interrupted by non-amino acids. The terms also encompass an amino acid polymer that has been modified, for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation, such as conjugation with a labeling component. As used herein, the term “amino acid” refers to either natural and / or unnatural or synthetic amino acids, including pyroglutamate (pGlu) and both the D or L optical isomers, amino acid analogs, and / or peptidomimetics.Organoleptic Properties of Sweet Proteins

[0029] The amount of sweet protein (brazzein-53, brazzein-54, monellin, or combinations thereof) present in any of the beverage products disclosed herein is anyamount that confers one or more improved organoleptic properties of the beverage product to which the sweet protein combination is added (e.g., taste, sight, smell, and texture (e.g., mouthfeel)) and / or health-related properties (e.g., less change or no change in glycemic index, or less modification or no modification of the gut microbiome), as compared to a similar beverage made with sugar, sugar alternatives and / or other artificial sweeteners. In a particular case, the improved organoleptic property is associated with taste. In one case, improving one or more organoleptic property results in the improvement of the taste profile. The overall taste profile of a beverage product is an interplay of several different tastes and senses, such as sweetness, sourness, bitterness, and the like. Examples of improved organoleptic properties can include, for example, a reduction in bitterness, a reduction in astringent and / or licorice notes, faster onset of sweetness, slower onset of sweetness, a reduction in lingering sweetness, a reduction in lingering bitterness, a reduction in bitter aftertaste, a reduction in metallic aftertaste, a reduction in chemical and synthetic aftertaste, a roundness to the sweetness profile, and a combination thereof.

[0030] In a particular case, the term “improved organoleptic properties” means that the beverage product will have one or more improved organoleptic properties for the majority of users, i.e., between about 50% to about 100% of a group of humans (e.g., between about 60% to about 100%, between about 70% to about 100%, between about 80% to about 100%, or between about 90% to about 100% of a group of humans). The improvement can be expressed qualitatively, e.g., as a gustatory comparison or a comparison taste test, or quantitatively, e.g., as a percentage improvement or as a value on a comparative scale. For example, a human or a group of humans can compare any of the beverage products described herein to a similar commercially available product, to any of the other beverage products described herein, or to a similar beverage product made with sugar (e.g., cane sugar) or with an artificial sweetener. Any of the beverage products described herein can have any of the improved organoleptic properties, as determined by any of the methods described herein. For example, any of the beverage products described herein can have an increase of sweetness, a reduction of sweetness, a reduction in bitterness, a faster onset of sweetness, or a reduction in lingering sweetness.

[0031] In some embodiments, the amount of sweet protein present in any of the beverage products disclosed herein is any amount that provides a desired level, curve and / or timing of sweetness and additionally provides a health-related property such as less change or no change in the glycemic index and / or less modification or no modification of the gut microbiome, to the individual consuming the product as compared to the same properties imparted by a beverage product made with sugar, sugar alternatives and / or other artificial sweeteners.Types of Sweet Proteins lira-- ein and Recombinant lira-- ein

[0032] In some cases, the sweet protein is brazzein-53. In some cases, the sweet protein is brazzein-54. Brazzein is a small, sweet-tasting protein originally isolated from the fruit of West African plant known as the Oubli plant (Pentadiplandra brazzeana Baillon).(See, for example, Ming D et al., FEES Lett. (1994) 20 355: 106-108, incorporated by reference in its entirety). It is a monomer protein with a molecular weight of 6.5 kDa. As a member of the CsPa fold family, it contains four disulfide bonds that lend a high degree of thermal and pH stability to its structure. Specifically, the sweet taste of brazzein remains after incubation at 98°C for 2 hours and at 80°C for 4.5 hours in the pH range of 2.5-8. It is also water soluble (>50mg / mL). It is a highly soluble protein (more than 50 g / L) with an isoelectric point of 5.4. Residues 29-33, 36, and 39-43, as well as the C- terminus of the protein, can be involved in the sweet taste of the protein. The charge of the protein can also play a role in its interaction with the sweet taste receptor.

[0033] At least three forms of the protein are known, although only two forms are present in ripe fruit. The major form is 54 amino acids long (pGlu-brazzein, referred to as “brazzein-54”), is about 80% of the total brazzein protein content in ripe fruit, and contains a pyroglutamate (pGlu) residue at its N-terminus (SEQ ID NO: 2, Table 1). The minor form is 53 amino acids long (des-pGlu-brazzein), referred to as “brazzein-53” herein, is about 20% of the total brazzein protein in ripe fruit, and lacks the final pyroglutamate residue (SEQ ID NO: 1, Table 1).

[0034] On a weight basis, brazzein can taste 500 to 2000 times sweeter than sucrose, as compared to a 10% sucrose and a 2% sucrose solution, respectively, or for example about700 times sweeter when compared to a 5% sucrose solution. Sucrose, a commonly used sweetener, is a disaccharide molecule composed of glucose and fructose.

[0035] In the beverages described herein, brazzein can be the minor form (i.e., brazzein- 53) or the major form (i.e., brazzein-54). In addition, in the beverages described herein, brazzein can be naturally produced by the fruit of the Oubli plant or produced recombinantly by, for example, microbial fermentation of a bacterium, a yeast, or a filamentous fungus that is genetically engineered to produce recombinant brazzein-53 or recombinant brazzein-54.Monellin and Recombinant Monellin

[0036] In some cases, the sweet protein used herein is a monellin. Monellin refers to a sweet protein originally discovered in the fruit of the West African shrub known as serendipity berry (Dioscoreophyllum cumminsii). Monellin consists of two non- covalently associated polypeptide chains, an A chain of 44 amino acid residues (SEQ ID NO: 3, Table 1) and a B chain of 50 amino acid residues (SEQ ID NO: 4, Table 1). As used herein, “monellin” is a fusion of chain A and chain B with a single glycine as a linker, referred to as a single-chain monellin (SEQ ID NO:5, Table 1). Monellin has a secondary structure of five beta-strands that forms an antiparallel beta-sheet and a 17- residue alpha-helix. On a molar basis, monellin can taste up to 100,000 times sweeter than commonly used sugars. On a weight basis, monellin can taste several thousand times sweeter that commonly used sugars. For example, monellin can taste between 800-2000 times sweeter than a 5-7% (w / w) sucrose solution.

[0037] In the beverages described below, monellin can be monellin or single-chain monellin. In addition, in the beverages described herein, monellin can be naturally produced by the fruit or produced recombinantly by, for example, microbial fermentation of a bacterium, a yeast, or a filamentous fungi that is genetically engineered to produce recombinant monellin.TABLE 1. Exemplary Sweet Polypeptide SequencesIII. Methods of Producing Sweet ProteinRecombinant Sweet Protein Expression

[0038] Sweet proteins, such as brazzein-53, brazzein-54 and monellin, described herein can be isolated from naturally occurring sources and / or produced recombinantly by a variety of protein expression systems, including, but not limited to, cell-based expression systems, and cell-free expression systems. Sweet proteins produced recombinantly are recombinant proteins.

[0039] As used herein, “recombinant protein” is a protein produced from a recombinant nucleic acid. A “recombinant nucleic acid” is a sequence that is not naturally occurring or has a sequence that is made by an artificial combination of two or more otherwise separated segments of nucleic acid sequence. This artificial combination is often accomplished by chemical synthesis or by the artificial manipulation of isolated segments of nucleic acids (e.g., by genetic engineering techniques). The term recombinant nucleic acids includes nucleic acids that have been altered solely by addition, substitution, or deletion of a portion of the nucleic acid. Frequently, a recombinant nucleic acid can include a nucleic acid sequence operably linked to a promoter sequence. Such a recombinant nucleic acid can be part of a vector that is used, for example, to transform a cell.

[0040] Non-limiting examples of protein expression systems useful in producing sweet polypeptides disclosed herein (e.g., brazzein-53, brazzein-54, and / or monellin (such as a single chain monellin)) include prokaryotic cell-based expression systems (e.g., archaeal systems, bacterial systems) and eukaryotic cell-based expression systems (e.g., filamentous or non-filamentous fungal cells, yeast cells, insect cells, and mammalian cells). In some cases, recombinant sweet protein is produced by a filamentous fungus. In some cases, recombinant sweet protein is produced by a yeast. In some cases, recombinant sweet protein is produced by a Pichia species, such as Pichia pastoris (also known as Komagataella phaffii).

[0041] To express any of the sweet polypeptides disclosed herein in a cell-based expression system, a gene encoding for the polypeptide can be introduced into a host cell of the expression system by, for example, an expression vector. A polynucleotide gene encoding for the sweet protein can be made by various methods, including molecular cloning and synthesis. Molecular cloning methods can involve mutagenesis (e.g., site- directed mutagenesis), restriction enzyme-mediated cloning (e.g., restriction enzyme digestion and ligation), polymerase chain reaction (PCR), and overlap extension.Synthesis can include chemical synthesis (e.g., gene synthesis). The gene sequence can be codon optimized for any desired expression system.

[0042] In some cases, a vector including a nucleic acid sequence encoding a sweet polypeptide disclosed herein is provided. In some cases, a vector including a nucleic acid sequence encoding for the sweet polypeptide brazzein (e.g., a nucleic acid encoding a recombinant brazzein-53, a recombinant brazzein-54) and / or a monellin protein (e.g., a nucleic acid encoding a recombinant monellin (such as a single chain monellin)) is provided.

[0043] The coding sequence can be inserted into a vector by a variety of procedures, including, but not limited to restriction enzyme digestion, ligation, and homologous recombination. The vector can be capable of replicating and expressing the polynucleotides in prokaryotic and / or eukaryotic host cells of expression systems. A vector can contain various components that can be adjusted and optimized for compatibility with the particular host cell. A cloning vector and / or expression vector can include additional nucleic acid sequences, including but not limited to, a signal sequence,an origin of replication, a marker gene (e.g., a selection marker such as an antibiotic resistance gene), an enhancer element, a promoter, a ribosome binding site, a signal sequence, and a transcription termination sequence.

[0044] In some cases, an expression vector containing a polynucleotide encoding a sweet protein, such as brazzein-53, brazzein-54, and / or monellin as disclosed herein, can further include a promoter. Promoters include, but are not limited to, constitutive promoters, inducible promoters, and hybrid promoters. Promoters can include, but are not limited to, acu-5, adhl+, alcohol dehydrogenase (ADH1, ADH2, ADH4), AHSB4m, AINV, alcA, a- amylase, alternative oxidase (AOD), alcohol oxidase I (AOX1), alcohol oxidase 2 (AOX2), AXDH, B2, CaMV, cellobiohydrolase I (cbhl), ccg-1, cDNAl, cellular filament polypeptide (cfp), cpc-2, ctr4+, CUP1, dihydroxyacetone synthase (DAS), enolase (ENO, ENO1), formaldehyde dehydrogenase (FLD1), FMD, formate dehydrogenase (FMDH), Gl, G6, GAA, GALI, GAL2, GAL3, GAL4, GAL5, GAL6, GAL7, GAL8, GAL9, GAL10, GCW14, gdhA, gla-1, a-glucoamylase (glaA), glyceraldehyde-3- phosphate dehydrogenase (gpdA, GAP, GAPDH), phosphoglycerate mutase (GPM1), glycerol kinase (GUT1), HSP82, invl+, isocitrate lyase (ICL1), acetohydroxy acid isomeroreductase (ILV5), KAR2, KEX2, b-galactosidase (lac4), LEET2, melO, MET3, methanol oxidase (MOX), nmtl, NSP, pcbC, PET9, peroxin 8 (PEX8), phosphoglycerate kinase (PGK, PGK1), phol, PH05, PH089, phosphatidylinositol synthase (PIS1), PYK1, pyruvate kinase (pkil), RPS7, sorbitol dehydrogenase (SDH), 3-phospho serine aminotransferase (SERI), SSA4, SV40, TEF, translation elongation factor 1 alpha (TEF1), THI11, homoserine kinase (THR1), tpi, TPS1, triose phosphate isomerase (TPI1), XRP2, and YPT1.

[0045] In some instances, an expression vector containing a gene encoding a sweet polypeptide or protein disclosed herein (e.g., brazzein-53, brazzein-54, and / or monellin) can include a polynucleotide sequence encoding a signal peptide. A signal peptide, also known as a signal sequence, targeting signal, localization signal, localization sequence, secretion signal, transit peptide, leader sequence, or leader peptide, can support secretion of a protein or polynucleotide. Extracellular secretion of a recombinantly expressed protein from a host cell can facilitate protein purification. For example, recovery of a recombinant protein from a cell culture supernatant can be preferable to lysing host cellsto release a complex mixture of proteins, including intracellular proteins of the host cell. Secretion, in some cases, can reduce deleterious effects that intracellular overexpression of a heterologous protein can have on a host cell such as toxicity or decreased growth rate. Secretion, in some cases, can allow increased protein production compared to intracellular expression in a host cell of limited volume to store the synthesized proteins. Secretory production of a protein, in some cases, can facilitate post-translational modification or processing (e.g., protein folding, formation of disulfide bonds, and glycosylation).

[0046] As different host cells can have characteristics and specific mechanisms for the posttranslational processing and modification of protein products, appropriate cell lines or host systems can be chosen to ensure the desired modification and processing of the expressed protein. In some cases, a host cell can be selected from the group consisting of: bacteria, fungi, plant cells, insect cells, mammalian cells, and any combination thereof. A host cell can be grown, fermented, or cultured in any appropriate medium that allows for host cell growth and sweet protein (e.g., recombinant brazzein-53, recombinant brazzein- 54, and / or recombinant monellin (such as a single chain monellin)) expression.

[0047] In some cases, a host cell selected for expression of the sweet polypeptide (e.g., brazzein-53, brazzein-54, and / or monellin) disclosed herein is an archaeal cell. Nonlimiting examples of archaeal host cells include Pyrococcus furiosus, Metallosphera sedula, Thermococcus litoralis, Methanobacterium thermoautotrophicum, Methanococcus jannaschii, Pyrococcus abyssi, Sulfolobus solfataricus, Pyrococcus woesei, Sulfolobus shibatae, and variants thereof.

[0048] In some cases, a host cell selected for expression a sweet polypeptide (e.g., brazzein-53, brazzein-54, and / or monellin) disclosed herein is a bacterial cell. Suitable bacterial host cells include, but are not limited to, BL21 E. coli, DE3 strain E. coli, E. coli M15, DH5u, DH103, HB101, B. subtilis cells, Pseudomonas jluorescens cells, and cyanobacterial cells such as Synechococcus elongates cells.

[0049] In some cases, a host cell selected for expression of a sweet polypeptide (e.g., brazzein-53, brazzein-54, and / or monellin) disclosed herein can be a fungal cell. The fungal cell can be a yeast cell or a filamentous fungi cell. A yeast or filamentous fungal cell can include, but is not limited to, Arxula spp., Arxula adeninivorans, Kluyveromycesspp., Kluyveromyces lactis, Pichia spp., Pichia angusta, Pichia pastoris (also known as Komagataella phaffii), Saccharomyces spp., Saccharomyces cerevisiae, Schizosaccharomyces spp., Schizosaccharomyces pombe, Tetrahymena sp., Hansenula sp., Blastobotrys sp., Candida sp., Zygosaccharomyces sp., Debaryomyces sp. Yarrowia spp., and Yarrowia lipolytica. Fungi can include, but are not limited to, Agaricus spp., Agaricus bisporus, Aspergillus spp., Aspergillus aw amor i, Aspergillus fumigatus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Colletotrichum spp., Colletotrichum gloeosporiodes, Endothia spp., Endothia parasitica, Fusarium spp., Fusarium graminearum, Fusarium solani, Mucor spp., Mucor miehei, Mucor pusillus, Myceliophthora spp., Myceliophthora thermophila, Neurospora spp., Neurospora crassa, Penicillium spp., Penicillium camemberti, Penicillium canescens, Penicillium chrysogenum, Penicillium (T alaromyces) emersonii, Penicillium funiculo sum, Penicillium purpurogenum, Penicillium roqueforti, Pleurotus spp., Pleurotus ostreatus, Rhizomucor spp., Rhizomucor miehei, Rhizomucor pusillus, Rhizopus spp., Rhizopus arrhizus, Rhizopus oligosporus, Rhizopus oryzae, Trichoderma spp., Trichoderma altroviride, Trichoderma reesei, and Trichoderma vireus cells. In some cases, the host cell for expression of recombinant brazzein-53, recombinant brazzein-54, and / or recombinant monellin is a Pichia spp. cell, such as Pichia pastoris cell.

[0050] In some cases, a host cell selected for expression of a sweet polypeptide (e.g., brazzein-53, brazzein-54, and / or monellin) disclosed herein can be an insect cell. Suitable insect host cells include, but are not limited to, Sf9 cells from Spodoptera frugiperda, Sf21 cells from Spodoptera frugiperda, Hi -Five cells, BTI-TN-5B1-4 Trichophusiani cells, Schneider 2 (S2) cells from Drosophila melanogaster , and Schneider 3 (S3) cells from Drosophila melanogaster .

[0051] In some cases, a host cell selected for expression of a sweet polypeptide (e.g., brazzein-53, brazzein-54, and / or monellin) disclosed herein can be a mammalian cell. Non-limiting examples of mammalian host cells include HEK293 cells, HeLa cells, CHO cells, COS cells, Jurkat cells, NSO hybridoma cells, baby hamster kidney (BHK) cells, MDCK cells, NIH-3T3 fibroblast cells, and any other immortalized cell line derived from a mammalian cell.

[0052] In some cases, a sweet polypeptide described herein (e.g., brazzein-53, brazzein- 54 and / or monellin (such as a single chain monellin)) can be produced in in vitro translation systems. An in vitro translation system generally refers to a translation system which is a cell-free extract containing elements for translation of an RNA molecule into a protein. An in vitro translation system can include ribosomes, tRNAs, initiator methionyl- tRNAMet, proteins or complexes involved in translation, e.g., eIF2, eIF3, the cap-binding (CB) complex, including the cap-binding protein (CBP) and eukaryotic initiation factor 4F (eIF4F). A variety of in vitro translation systems are available. Non-limiting examples of in vitro translation systems include eukaryotic lysates, such as rabbit reticulocyte lysates, rabbit oocyte lysates, human cell lysates, insect cell lysates, and wheat germ extracts.Synthesis of Polypeptides

[0053] A sweet peptide described herein (e.g., brazzein-53, brazzein-54, and / or monellin) can be chemically synthesized. Method of peptide chemical synthesis are known in the art. See, for example, Isidro-Llobet et. al. 2019. Journal of Organic Chemistry .84, 8, 4615-4628 or Stawikowski and Fields. 2002. Curr. Protoc. Protein Sci. doi: 10.1002 / 0471140864. psl801s26.Recovery and Purification of Sweet Polypeptides

[0054] A sweet polypeptide such as recombinant brazzein-53, a recombinant brazzein- 54, and / or a recombinant monellin described herein, can be expressed in a host cell, recovered, isolated, and / or purified from the supernatant of a cell culture, for example, if the expressed protein is secreted by the host cell into the cell culture medium. A sweet polypeptide described herein can, in some cases, be recovered from the host cell, for example, if the protein is expressed intracellularly. Proteins can be recovered from the intracellular space by disrupting the host cell, for example, by osmotic shock, sonication, or lysis. Once the cells are disrupted, cell debris can be removed by centrifugation or filtration, and the expressed proteins can be removed or recovered from the cell lysate.

[0055] In some cases, the sweet polypeptide (e.g., brazzein-53, brazzein-54, and / or monellin) can be produced using in vitro or cell-free protein synthesis systems, for example, using a cell-free translation system including a cell extract such as Escherichiacoli cell extract, rabbit reticulocyte cell extract, wheat germ cell extract, or insect cell extract. The expressed protein can be recovered, isolated, and / or optionally purified from the cell extract by any suitable method.

[0056] A sweet polypeptide (e.g., brazzein-53, brazzein-54, and / or monellin) described herein can be recovered and / or isolated from supernatants and lysates by any of a variety of methods, including, but not limited to, chemical extraction, column chromatography, and filtration. The polypeptides can be purified using any of a variety of methods including liquid chromatography such as normal or reversed phase, high-performance liquid chromatography (HPLC), fast protein liquid chromatography (FPLC), and the like; affinity chromatography such as with inorganic ligands, monoclonal antibodies (e.g., immunoaffinity), and ion exchange (e.g., anion exchange, cation exchange); hydrophobic interaction chromatography; size exclusion chromatography; immobilized metal chelate chromatography; gel electrophoresis; ethanol precipitation; and any combination thereof. In some cases, the polypeptides can be purified by centrifugation and / or filtration, including sterile filtration, depth filtration, tangential flow filtration, ultrafiltration (UF), diafiltration (DF), and ultrafiltration / diafiltration (UF ZDF).

[0057] A recovered and / or isolated sweet polypeptide described herein (e.g., brazzein-53, brazzein-54, and / or monellin) can be concentrated, dehydrated, dewatered, or dried by freeze drying, vacuum tray drying, spray drying, rotary drum drying, or any combination thereof. Drying can be done in a desiccator, vacuum dryer, conical dryer, spray dryer, fluid bed or any method known in the art. Preferably, methods are chosen that yield a dried product (e.g., a powder) with the greatest sweetness. The dried sweet protein can be optionally blended, pestled, milled, conched, pulverized, or other methods of processing known in the art. The resulting concentrated liquid or solid powder can be stored, diluted, rehydrated, or used in the formulation of food and beverage products, such as the lemonade, soda, sparkling juice, fruit punch, mocktail mixer, and / or cocktail mixer described herein. In some cases, the concentrated liquid or solid powder can be stored at - 2o°C, 4°C, or room temperature without significant loss in sweetness. In some cases, the concentrated liquid can be diluted using water. In some cases, the solid powder can be rehydrated as a paste or a liquid using water or another liquid in which the solid powder is soluble.

[0058] Where the term “substantially purified” is used, this will refer to a composition in which the specific protein, peptide, or peptide forms the major component of the composition, such as constituting about 50% of the peptides in the composition or more. In preferred cases, a substantially purified peptide will constitute more than 60%, 70%, 80%, 90%, 95%, 99% or even more of the peptides in the composition.

[0059] A peptide, polypeptide or protein that is “purified to homogeneity,” as used herein, means that the peptide, polypeptide or protein has a level of purity where the peptide, polypeptide or protein is substantially purified or free from other proteins / peptides and biological components. For example, a purified peptide, polypeptide, or protein will often be sufficiently free of other protein / peptide components so that degradative sequencing can be performed successfully.

[0060] Although preferred for use in some cases, there is no general requirement that the protein, polypeptide, or peptide always be provided in their most purified state. Indeed, it is contemplated that less substantially purified protein, polypeptide or peptide, which are nonetheless enriched in the desired peptide compositions, relative to the natural state, will have utility in some cases. As used herein, “enriched” refers to a component, such as a peptide or protein that is found at a high concentration in a solution or composition than in nature when produced the same way. Methods of enrichment of a component, such as enrichment of a peptide, are known in the art and are often similar to those of purification of components or peptides.

[0061] In other cases, a preparation enriched with any of the sweet peptides described herein can be used instead of a purified preparation. In this document, whenever purified is used, enriched can be used also. A preparation can be enriched not only by methods of purification, but also by the over-expression or over-production of the peptide by the host cell when compared to a wild-type host cell. This can be accomplished using recombinant methods, or by selecting conditions which will induce the expression of the peptide from the wild-type cells.Additives

[0062] The sweet proteins (e.g., brazzein-53, brazzein-54, and / or monellin) disclosed herein can be added to or mixed with one or more additives. In some cases, an additivecan increase the shelf-life of the beverage product, improve the stability and consistency of sweetness and / or preserve flavor and taste over time.IV. Beverage Products with Sweet Protein

[0063] Described herein are sweetened beverage products that include one or more sweet proteins, such as, brazzein-53, brazzein-54, monellin, and any combinations thereof. Non-limiting examples of a beverage product (e.g., sweetened beverage product) of the present disclosure include lemonade, soda, sparkling juice, fruit punch, mocktail mixer and cocktail mixer. In some embodiments, a beverage product (e.g., a sweetened beverage product) described herein is savory or spicy. For example, a mocktail mixer and / or a cocktail mixer of the present disclosure can be savory or spicy. In some embodiments, a beverage product (e.g., a sweetened beverage product) described herein is not savory or spicy. In some embodiments, a beverage product described herein is carbonated. In some embodiments, a beverage product described herein is not carbonated.

[0064] Any of the beverage products described herein can further include an additional ingredient such as one or more of a plant-based sugar composition, an acid, a bulking agent, an emulsifier, and / or an additional flavoring agent. In some cases, the beverage products described herein, while further including an additional ingredient, specifically do not include a high intensity sweetener. In some cases, the beverage products, such as lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer, described herein do not include sugar alternatives, stevia-derived sweetener, a rebaudioside or an artificial sweetener such as aspartame or saccharin.

[0065] In some cases, a beverage described herein is a flavored beverage, such as, a fruit flavored beverage. A fruit flavored beverage can include fruit flavor, fruit pulp, fruit puree, fruit peel, fruit seed, fruit oil, fruit juice, fruit concentrate, whole fruit, or any combination thereof as a flavoring agent. Non-limiting examples of flavoring agents that can impart a fruit flavor include flavor, pulp, puree, peel, seed, oil, juice, concentrate, whole and / or pieces of banana, blood orange, cherries, citron, coconut, cucumber, grape, grapefruit, key lime, kiwi, lemon, lime, mango, mulberries, nectarine, orange, orange peel, papaya, passionfruit, peach, pineapple, plums, pomegranate, raisins, raspberry, strawberry, watermelon, yuzu, or combinations thereof.

[0066] The beverage products described herein can include additional flavoring agents. Additional flavoring agents can impart additional flavors or tastes. Non-limiting exemplary additional flavoring agents include fruit flavors (e.g., fruit juice, fruit puree, pieces of fruit, dried fruit, whole fruit, fruit flavor, fruit concentrate, fruit pulp, fruit peel, fruit seed, or an oil derived from a fruit), herbs such as mint, flavor-imparting oils, alcoholic beverages (e.g., vodka, malt liquor, whiskey, bourbon, or red wine), and flavor extracts. In some cases, a beverage described here can include one or more flavoring agents. Non-limiting flavoring agents include anise seed, ashwagandha, barley, bergamot, butterfly pea, calendula, cardamom seed, chamomile, chicory, chrysanthemum, cinnamon, cloves, dandelion, Echinacea, elderberry flower, ginger, gingko, gotu kola, guava leaf, hibiscus, honeybush, iceland moss, jasmine, jiaogulan, kava, kuding, lapacho bark, lavender, lemon balm, lemongrass, licorice root, linden flower, marshmallow root, moringa, olive leaf, osmanthus, passionflower, peppermint, pine needle, raspberry leaf, rooibos, rose, rosehip, rosemary, sage, stinging nettle, thyme, tulsi, turmeric, valerian root, yarrow, yerba mate, or combinations thereof. In some cases, a beverage described herein does not include any tea.

[0067] Any of the flavoring agents described herein can be present in an amount between about 0.1% wt / wt and about 25% wt / wt (e.g., between about 0.4% wt / wt and about 0.6% wt / wt, or between about 3% wt / wt and about 4% wt / wt).

[0068] In some cases, a beverage described herein is a lemonade. Any of the lemonades described herein can include a flavored lemonade, such as, a fruit-flavored lemonade. For example, a lemonade described herein can include flavor, pulp, puree, peel, seed, oil, juice, concentrate, whole and / or pieces of one or more of lemon, lime, other citrus fruits (e.g., yuzu, orange, blood orange, citron, grapefruit, nectarine, clementine, mandarin (or mandarin orange), key lime), strawberry, raspberry, apple, avocado, banana, cherry, coconut, cucumber, grape, kiwi, mango, mulberry, papaya, passionfruit, peach, pineapple, plum, pomegranate, raisin, tomato, jalapeno (or other spicy pepper), and / or watermelon. Additionally, or in the alternative, a lemonade of the present disclosure can include one or more flavoring agents described herein. In some cases, a lemonade described herein does not include any tea. In some embodiments, a lemonade of the present disclosure is a low tartness lemonade. In some embodiments, a lemonade of thepresent disclosure has lower sweetness (e.g., as compared to other lemonades or other sweetened beverages). For example, a lemonade of the present disclosure can have low tartness and lower sweetness, as compared to other sweetened beverages. In some embodiments, a lemonade with low tartness and lower sweetness is a fruit-flavored lemonade, such as a fruit-flavored lemonade described herein (e.g., a strawberry lemonade).

[0069] In some cases, a beverage described herein is a soda (e.g., a carbonated drink). Any of the sodas described herein can include a flavored soda, such as, a fruit-flavored soda. For example, a soda described herein can include the flavor, pulp, puree, peel, seed, oil, juice, concentrate, whole and / or pieces of one or more of lemon, lime, other citrus fruits (e.g., yuzu, orange, blood orange, citron, grapefruit, nectarine, clementine, mandarin (or mandarin orange), key lime), strawberry, raspberry, apple, avocado, banana, cherry, coconut, cucumber, grape, kiwi, mango, mulberry, papaya, passionfruit, peach, pineapple, plum, pomegranate, raisin, tomato, jalapeno (or other spicy pepper), and / or watermelon. Additionally, or in the alternative, a soda of the present disclosure can include one or more flavoring agents described herein. In some cases, a soda described herein does not include any tea.

[0070] In some cases, a beverage described herein is a sparkling juice (e.g., a juice that has been carbonated). Any of the sparkling juices described herein can include flavor of one or more fruits. For example, a sparkling juice described herein can include the flavor, pulp, puree, peel, seed, oil, juice, concentrate, whole and / or pieces of one or more of lemon, lime, other citrus fruits (e.g., yuzu, orange, blood orange, citron, grapefruit, nectarine, clementine, mandarin (or mandarin orange), key lime), strawberry, raspberry, apple, avocado, banana, cherry, coconut, cucumber, grape, kiwi, mango, mulberry, papaya, passionfruit, peach, pineapple, plum, pomegranate, raisin, tomato, jalapeno (or other spicy pepper), and / or watermelon. Additionally, or in the alternative, a sparkling juice of the present disclosure can include one or more flavoring agents described herein. In some cases, a sparkling juice described herein does not include any tea.

[0071] In some cases, a beverage described herein is a fruit punch. Any of the fruit punches described herein can include a fruit punch flavoring. For example, a fruit punch can include the flavor, pulp, puree, peel, seed, oil, juice concentrate, or flavoring pre-made flavoring powder or drink mix, such as a fruit punch flavoring packet. In some examples the pre-made flavoring powder, drink mix or packet is unsweetened (i.e., doesn’t contain a sugar, artificial sugar, or sugar substitute).

[0072] In some cases, a beverage described herein is a mocktail mixer (e.g., a margarita mocktail mixer). Any of the mocktail mixers described herein can include a flavored mocktail mixer, such as, a fruit-flavored mocktail mixer. For example, a mocktail mixer described herein can include the flavor, pulp, puree, peel, seed, oil, juice, concentrate, whole and / or pieces of one or more of lemon, lime, other citrus fruits (e.g., yuzu, orange, blood orange, citron, grapefruit, nectarine, clementine, mandarin (or mandarin orange), key lime), strawberry, raspberry, apple, avocado, banana, cherry, coconut, cucumber, grape, kiwi, mango, mulberry, papaya, passionfruit, peach, pineapple, plum, pomegranate, raisin, tomato, jalapeno (or other spicy pepper), and / or watermelon. Additionally, or in the alternative, a mocktail mixer of the present disclosure can include one or more flavoring agents described herein. In some cases, a mocktail mixer described herein does not include any tea.

[0073] In some cases, a beverage described herein is a cocktail mixer. Any of the cocktail mixers described herein can include a flavored cocktail mixer, such as, a fruit-flavored cocktail mixer. For example, a cocktail mixer described herein can include the flavor, pulp, puree, peel, seed, oil, juice, concentrate, whole and / or pieces of one or more of lemon, lime, other citrus fruits (e.g., yuzu, orange, blood orange, citron, grapefruit, nectarine, clementine, mandarin (or mandarin orange), key lime), strawberry, raspberry, apple, avocado, banana, cherry, coconut, cucumber, grape, kiwi, mango, mulberry, papaya, passionfruit, peach, pineapple, plum, pomegranate, raisin, tomato, jalapeno (or other spicy pepper), and / or watermelon. Additionally, or in the alternative, a cocktail mixer of the present disclosure can include one or more flavoring agents described herein. In some cases, a cocktail mixer described herein does not include any tea.Sweet Proteins and Recombinant Sweet Proteins in Beverages

[0074] The beverage products of the present disclosure (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) contain sweet proteins, such as brazzein-53 and / or recombinant brazzein-53, brazzein-54 and / or recombinant brazzein-54, monellin and / or recombinant monellin, and combinations thereof as described herein. In some embodiments, a beverage of the present disclosure contains only one sweet protein. For example, in some instances, brazzein-53 protein and / or recombinant brazzein-53 protein is the only sweet protein in a beverage described herein. In some instances, brazzein-54 protein and / or recombinant brazzein-54 protein is the only sweet protein in a beverage described herein. In some instances, monellin protein and / or recombinant monellin protein is the only sweet protein in a beverage described herein. In some embodiments, a beverage of the present disclosure contains two sweet proteins. For example, in some instances, brazzein-53 protein (and / or recombinant brazzein-53 protein) and brazzein-54 protein (and / or recombinant brazzein-54 protein) are the two sweet proteins in a beverage described herein. In some instances, brazzein-54 protein (and / or recombinant brazzein-54 protein) and monellin protein (and / or recombinant monellin protein) are the two sweet proteins in a beverage described herein. In some instances, brazzein-53 protein (and / or recombinant brazzein-53 protein) and monellin protein (and / or recombinant monellin protein) are the two sweet proteins in a beverage described herein. In some embodiments, a beverage of the present disclosure contains three sweet proteins (e.g., all three sweet proteins described herein). For example, in some instances, brazzein-53 protein (and / or recombinant brazzein-53 protein), brazzein- 54 protein (and / or recombinant brazzein-54 protein), and monellin protein (and / or recombinant monellin protein) are the three sweet proteins in a beverage described herein.

[0075] In some cases, at least one sweet protein is added to any of the beverage products described herein as a powder, slurry, solution or other preparation, and in some cases, the preparation contains other components in addition to the sweet protein. Within such preparations, the amount of sweet protein in the preparation can be from about 20% to about 99% (wt / total preparation wt, also referred to herein as “purity” or “% pure”). For example, within such preparations, the amount of sweet protein in the preparation can be from about 50% purity to about 75% purity. In some cases, the preparation of sweet protein is at least 35% pure and / or no more than 85% pure. For example, the purity of sweet protein and / or recombinant sweet protein powder, before it is added to any of thebeverage products described herein can be about 50%, about 55%, about 60%, about 65%, about 70%, or about 75% pure.

[0076] A beverage product of the present disclosure (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) can include at least one (e.g., one, two, or three) of the sweet proteins described here. For example, a beverage product described here can include at least one (e.g., one, two, or three) of brazzein-53 and / or recombinant brazzein-53, brazzein-54 and / or recombinant brazzein-54, and monellin and / or recombinant monellin. In some embodiments, the sweet protein (e.g., the at least one sweet protein) is present in the beverage in an amount between 2 ppm and 300 ppm, between 2 ppm and 275 ppm, between 2 ppm and 250 ppm, between 2 ppm and 225 ppm, between 2 ppm and 200 ppm, between 4 ppm and 175 ppm, between 4 ppm and 170 ppm, between 4 ppm and 164 ppm, between 4 ppm and 160 ppm, between 4 ppm and 150 ppm, between 4 ppm and 140 ppm, between 4 ppm and 50 ppm, between 4 ppm and 40 ppm, between 4 ppm and 30 ppm, between 6 ppm and 125 ppm, between 6 ppm and 120 ppm, between 6 ppm and 115 ppm, between 8 ppm and 110 ppm, between 8 ppm and 100 ppm, between 10 ppm and 100 ppm, between 10 ppm and 75 ppm, between 15 ppm and 115 ppm, between 15 ppm and 110 ppm, between 15 ppm and 100 ppm, between 20 ppm and 115 ppm, between 20 ppm and 110 ppm, between 20 ppm and 100 ppm, or between 20 ppm and 80 ppm.

[0077] In some embodiments, the sweet protein (e.g., the at least one sweet protein) is present in the beverage in an amount between 5 ppm and 584 ppm, between 5 ppm and 583 ppm, between 5 ppm and 582 ppm, between 5 ppm and 581 ppm, between 5 ppm and 580 ppm, between 5 ppm and 575 ppm, between 5 ppm and 550 ppm, between 5 ppm and 525 ppm, between 5 ppm and 500 ppm, between 5 ppm and 475 ppm, between 5 ppm and 450 ppm, between 5 ppm and 425 ppm, between 5 ppm and 400 ppm, between 5 ppm and 375 ppm, between 5 ppm and 350 ppm, between 5 ppm and 325 ppm, between 5 ppm and 300 ppm, between 5 ppm and 275 ppm, between 5 ppm and 250 ppm, between 5 ppm and 225 ppm, between 5 ppm and 200 ppm, between 5 ppm and 175 ppm, between 5 ppm and 150 ppm, between 5 ppm and 125 ppm, between 5 ppm and 100 ppm, between 5 ppm and 75 ppm, between 5 ppm and 50 ppm, between 5 ppm and 25 ppm, between 5 ppm and 15 ppm, or between 5 ppm and 10 ppm.

[0078] In some embodiments, the sweet protein (e.g., the at least one sweet protein) is present in the beverage in an amount between 25 ppm and 550 ppm, between 25 ppm and 525 ppm, between 25 ppm and 500 ppm, between 25 ppm and 475 ppm, between 25 ppm and 450 ppm, between 25 ppm and 425 ppm, between 25 ppm and 400 ppm, between 25 ppm and 375 ppm, between 25 ppm and 350 ppm, between 25 ppm and 325 ppm, between 25 ppm and 300 ppm, between 25 ppm and 275 ppm, between 25 ppm and 250 ppm, between 25 ppm and 225 ppm, between 25 ppm and 200 ppm, between 25 ppm and 175 ppm, between 25 ppm and 150 ppm, between 25 ppm and 125 ppm, between 25 ppm and 100 ppm, between 25 ppm and 75 ppm, or between 25 ppm and 50 ppm.

[0079] In some embodiments, the sweet protein (e.g., the at least one sweet protein) is present in the beverage in an amount between 28 ppm and 500 ppm, between 28 ppm and 475 ppm, between 28 ppm and 450 ppm, between 28 ppm and 425 ppm, between 28 ppm and 400 ppm, between 28 ppm and 375 ppm, between 28 ppm and 350 ppm, between 28 ppm and 325 ppm, between 28 ppm and 300 ppm, between 28 ppm and 275 ppm, between 28 ppm and 250 ppm, between 28 ppm and 225 ppm, between 28 ppm and 200 ppm, between 28 ppm and 175 ppm, between 28 ppm and 150 ppm, between 28 ppm and 125 ppm, between 28 ppm and 120 ppm, between 28 ppm and 115 ppm, between 28 ppm and 110 ppm, between 28 ppm and 100 ppm, between 28 ppm and 75 ppm, between 28 ppm and 50 ppm, between 28 ppm and 40 ppm, or between 28 ppm and 30 ppm.

[0080] In some embodiments, the sweet protein (e.g., the at least one sweet protein) is present in the beverage in an amount between 50 ppm and 450 ppm, between 50 ppm and 425 ppm, between 50 ppm and 400 ppm, between 50 ppm and 375 ppm, between 50 ppm and 350 ppm, between 50 ppm and 325 ppm, between 50 ppm and 300 ppm, between 50 ppm and 275 ppm, between 50 ppm and 250 ppm, between 50 ppm and 225 ppm, between 50 ppm and 200 ppm, between 50 ppm and 175 ppm, between 50 ppm and 150 ppm, between 50 ppm and 125 ppm, between 50 ppm and 120 ppm, between 50 ppm and 115 ppm, between 50 ppm and 110 ppm, between 50 ppm and 100 ppm, or between 50 ppm and 75 ppm.

[0081] In some embodiments, the sweet protein (e.g., the at least one sweet protein) is present in the beverage in an amount between 60 ppm and 400 ppm, between 60 ppm and 375 ppm, between 60 ppm and 350 ppm, between 60 ppm and 325 ppm, between 60 ppmand 300 ppm, between 60 ppm and 290 ppm, between 60 ppm and 280 ppm, between 60 ppm and 270 ppm, between 60 ppm and 260 ppm, between 60 ppm and 250 ppm, between 60 ppm and 240 ppm, between 60 ppm and 230 ppm, between 60 ppm and 225 ppm, between 60 ppm and 200 ppm, between 60 ppm and 175 ppm, between 60 ppm and 150 ppm, between 60 ppm and 125 ppm, between 60 ppm and 100 ppm, or between 60 ppm and 75 ppm.

[0082] In some embodiments, the sweet protein (e.g., the at least one sweet protein) is present in the beverage in an amount between 75 ppm and 400 ppm, between 75 ppm and 375 ppm, between 75 ppm and 350 ppm, between 75 ppm and 325 ppm, between 75 ppm and 300 ppm, between 75 ppm and 275 ppm, between 75 ppm and 250 ppm, between 75 ppm and 225 ppm, between 75 ppm and 200 ppm, between 75 ppm and 175 ppm, between 75 ppm and 150 ppm, between 75 ppm and 125 ppm, or between 75 ppm and 100 ppm.

[0083] In some embodiments, the sweet protein (e.g., the at least one sweet protein) is present in the beverage in an amount between 100 ppm and 350 ppm, between 100 ppm and 325 ppm, between 100 ppm and 300 ppm, between 100 ppm and 275 ppm, between 100 ppm and 250 ppm, between 100 ppm and 225 ppm, between 100 ppm and 200 ppm, between 100 ppm and 175 ppm, between 100 ppm and 150 ppm, or between 100 ppm and 125 ppm.

[0084] In some embodiments, the sweet protein (e.g., the at least one sweet protein) is present in the beverage in an amount between 200 ppm and 300 ppm, between 200 ppm and 275 ppm, between 200 ppm and 250 ppm, or between 200 ppm and 225 ppm.

[0085] In some embodiments, the sweet protein (e.g., the at least one sweet protein) is present in the beverage in an amount between 300 ppm and 500 ppm, between 300 ppm and 475 ppm, between 300 ppm and 450 ppm, between 300 ppm and 425 ppm, between 300 ppm and 400 ppm, between 300 ppm and 375 ppm, between 300 ppm and 350 ppm, or between 300 ppm and 325 ppm.

[0086] In some embodiments, a sweet protein (e.g., at least one sweet protein) can be present in a beverage of the present disclosure in an amount of about 584 ppm, about 583 ppm, about 582 ppm, about 581 ppm, about 580 ppm, about 579 ppm, about 578 ppm, about 577 ppm, about 576 ppm, about 575 ppm, about 550 ppm, about 525 ppm, about500 ppm, about 475 ppm, about 450 ppm, about 425 ppm, about 400 ppm, about 375 ppm, about 350 ppm, about 325 ppm, about 300 ppm, about 275 ppm, about 250 ppm, about 225 ppm, about 200 ppm, about 175 ppm, about 150 ppm, about 125 ppm, about 100 ppm, about 75 ppm, about 50 ppm, about 45 ppm, about 40 ppm, about 35 ppm, about 30 ppm, about 29 ppm, about 28 ppm, about 27 ppm, about 26 ppm, about 25 ppm, about 24 ppm, about 23 ppm, about 22 ppm, about 21 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 17 ppm, about 16 ppm, about 15 ppm, about 14 ppm, about 13 ppm, about 12 ppm, about 11 ppm, about 10 ppm, about 9 ppm, about 8 ppm, about 7 ppm, about 6 ppm, about 5 ppm, about 4 ppm, about 3 ppm, or about 2 ppm.Beverage Products with Brazzein-53

[0087] Any of the beverage products (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) described herein can include, between 2 ppm and 584 ppm of brazzein-53 and / or recombinant brazzein-53 protein; such as, between 2 ppm and 300 ppm, between 5 ppm and 584 ppm, between 5 ppm and 300 ppm, or between 28 ppm and 300 ppm of brazzein-53 and / or recombinant brazzein-53 protein. In some embodiments, the amount of brazzein-53 protein and / or recombinant brazzein-53 protein in a beverage of the present disclosure is between 5 ppm and 500 ppm, between 5 ppm and 475 ppm, between 5 ppm and 450 ppm, between 5 ppm and 425 ppm, between 5 ppm and 400 ppm, between 5 ppm and 375 ppm, between 5 ppm and 350 ppm, between 5 ppm and 325 ppm, between 5 ppm and 300 ppm, between 5 ppm and 275 ppm, between 5 ppm and 250 ppm, between 5 ppm and 225 ppm, between 5 ppm and 200 ppm, between 5 ppm and 175 ppm, between 5 ppm and 150 ppm, between 5 ppm and 125 ppm, between 5 ppm and 100 ppm, between 5 ppm and 75 ppm, between 5 ppm and 50 ppm, between 5 ppm and 25 ppm, between 5 ppm and 15 ppm, between 5 ppm and 10 ppm, between 6 ppm and 125 ppm, between 6 ppm and 120 ppm, between 6 ppm and 115 ppm, between 7 ppm and 55 ppm, between 7 ppm and 50 ppm, between 7 ppm and 48 ppm, between 7 ppm and 47 ppm, between 7 ppm and 45 ppm, between 8 ppm and 110 ppm, between 8 ppm and 100 ppm, between 10 ppm and 300 ppm, between 10 ppm and 275 ppm, between 10 ppm and 265 ppm, between 10 ppm and 200 ppm, between 10 ppm and 150 ppm, between 10 ppm and 100 ppm, between 10 ppm and 75 ppm, between 10 and 72 ppm,between 10 ppm and 69 ppm, between 10 ppm and 50 ppm, between 10 ppm and 30 ppm, between 10 ppm and 100 ppm, between 10 ppm and 75 ppm, between 10 and 72 ppm, between 10 ppm and 69 ppm, between 10 ppm and 105 ppm, between 12 ppm to 30 ppm, between 12 ppm to 120 ppm, between 12 ppm to 18 ppm, between 12 ppm to 15 ppm, between 15 ppm and 115 ppm, between 15 ppm and 110 ppm, between 15 ppm and 105 ppm, between 15 ppm and 100 ppm, between 19 ppm and 131 ppm, between 20 ppm and 160 ppm, between 20 ppm and 150 ppm, between 20 ppm and 140 ppm, between 20 ppm and 115 ppm, between 20 ppm and 110 ppm, between 20 ppm and 100 ppm, or between 20 ppm and 80 ppm.

[0088] In some embodiments, the amount of brazzein-53 protein and / or recombinant brazzein-53 protein in a beverage of the present disclosure is between 25 ppm and 300 ppm, between 25 ppm and 275 ppm, between 25 ppm and 250 ppm, between 25 ppm and 225 ppm, between 25 ppm and 200 ppm, between 25 ppm and 175 ppm, between 25 ppm and 150 ppm, between 25 ppm and 125 ppm, between 25 ppm and 100 ppm, between 25 ppm and 75 ppm, or between 25 ppm and 50 ppm.

[0089] In some embodiments, the amount of brazzein-53 protein and / or recombinant brazzein-53 protein in a beverage of the present disclosure is between 40 ppm and 300 ppm, between 40 ppm and 250 ppm, between 40 ppm and 200 ppm, between 41 ppm and 250 ppm, between 41 ppm and 200 ppm, between 41 ppm and 175 ppm, between 41 ppm and 164 ppm, between 41 ppm and 160 ppm, between 41 ppm and 150 ppm, between 45 ppm and 164 ppm, between 50 ppm and 300 ppm, between 50 ppm and 275 ppm, between 50 ppm and 250 ppm, between 50 ppm and 225 ppm, between 50 ppm and 200 ppm, between 50 ppm and 175 ppm, between 50 ppm and 150 ppm, between 50 ppm and 125 ppm, between 50 ppm and 100 ppm, between 50 ppm and 75 ppm, between 60 ppm and 300 ppm, between 60 ppm and 290 ppm, between 60 ppm and 280 ppm, between 60 ppm and 270 ppm, between 60 ppm and 260 ppm, between 60 ppm and 250 ppm, between 60 ppm and 240 ppm, between 60 ppm and 230 ppm, between 60 ppm and 225 ppm, between 60 ppm and 200 ppm, between 60 ppm and 175 ppm, between 60 ppm and 150 ppm, between 60 ppm and 125 ppm, between 60 ppm and 100 ppm, or between 60 ppm and 75 ppm.

[0090] In some embodiments, the amount of brazzein-53 protein and / or recombinant brazzein-53 protein in a beverage of the present disclosure is between 75 ppm and 300 ppm, between 75 ppm and 275 ppm, between 75 ppm and 250 ppm, between 75 ppm and 225 ppm, between 75 ppm and 200 ppm, between 75 ppm and 175 ppm, between 75 ppm and 150 ppm, between 75 ppm and 125 ppm, or between 75 ppm and 100 ppm.

[0091] In some embodiments, the amount of brazzein-53 protein and / or recombinant brazzein-53 protein in a beverage of the present disclosure is between 100 ppm and 200 ppm, between 100 ppm and 175 ppm, between 100 ppm and 150 ppm, or between 100 ppm and 125 ppm.

[0092] In some embodiments, the amount of brazzein-53 protein and / or recombinant brazzein-53 protein in a beverage of the present disclosure is about 500 ppm, about 475 ppm, about 450 ppm, about 425 ppm, about 400 ppm, about 375 ppm, about 350 ppm, about 325 ppm, about 300 ppm, about 275 ppm, about 250 ppm, about 240 ppm, about 230 ppm, about 225 ppm, about 200 ppm, about 175 ppm, about 164 ppm, about 160 ppm, about 150 ppm, about 130 ppm, about 131 ppm, about 125 ppm, about 105 ppm, about 100 ppm, about 95 ppm, about 90 ppm, about 85 ppm, about 82 ppm, about 80 ppm, about 75 ppm, about 72 ppm, about 70 ppm, about 69 ppm, about 65 ppm, about 60 ppm, about 55 ppm, about 50 ppm, about 48 ppm, about 47 ppm, about 45 ppm, about 40 ppm, about 41 ppm, about 40 ppm, about 35 ppm, about 30 ppm, about 28 ppm, about 27 ppm, about 25 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 15 ppm, about 12 ppm, about 10 ppm, about 7 ppm, about 5 ppm, about 4 ppm, about 3 ppm, about 2 ppm, or about 1 ppm.

[0093] In some embodiments, brazzein-53 protein and / or recombinant brazzein-53 protein is the only sweet protein in a beverage (e.g., a lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) described herein. In some instances, brazzein-53 protein and / or recombinant brazzein-53 is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 6 ppm and 125 ppm, between 6 ppm and 120 ppm, between 6 ppm and 115 ppm, between 8 ppm and 110 ppm, between 8 ppm and 100 ppm, between 10 ppm and 100 ppm, between 10 ppm and 75 ppm, between 15 ppm and 115 ppm, between 15 ppm and 110 ppm, between15 ppm and 100 ppm, between 20 ppm and 115 ppm, between 20 ppm and 110 ppm, between 20 ppm and 100 ppm, or between 20 ppm and 80 ppm.

[0094] In some instances, brazzein-53 protein and / or recombinant brazzein-53 is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 28 ppm and 300 ppm, between 28 ppm and 275 ppm, between 28 ppm and 250 ppm, between 28 ppm and 225 ppm, between 28 ppm and 200 ppm, between 28 ppm and 175 ppm, between 28 ppm and 150 ppm, between 28 ppm and 125 ppm, between 28 ppm and 120 ppm, between 28 ppm and 115 ppm, between 28 ppm and 110 ppm, between 28 ppm and 100 ppm, between 28 ppm and 75 ppm, between 28 ppm and 50 ppm, between 28 ppm and 40 ppm, or between 28 ppm and 30 ppm.

[0095] In some instances, brazzein-53 protein and / or recombinant brazzein-53 is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 60 ppm and 300 ppm, between 60 ppm and 290 ppm, between 60 ppm and 280 ppm, between 60 ppm and 270 ppm, between 60 ppm and 260 ppm, between 60 ppm and 250 ppm, between 60 ppm and 240 ppm, between 60 ppm and 230 ppm, between 60 ppm and 225 ppm, between 60 ppm and 200 ppm, between 60 ppm and 175 ppm, between 60 ppm and 150 ppm, between 60 ppm and 125 ppm, between 60 ppm and 100 ppm, or between 60 ppm and 75 ppm.

[0096] In some instances, brazzein-53 protein and / or recombinant brazzein-53 is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 50 ppm and 300 ppm, between 50 ppm and 275 ppm, between 50 ppm and 250 ppm, between 50 ppm and 225 ppm, between 50 ppm and 200 ppm, between 50 ppm and 175 ppm, between 50 ppm and 150 ppm, between 50 ppm and 125 ppm, between 50 ppm and 100 ppm, or between 50 ppm and 75 ppm.

[0097] In some instances, brazzein-53 protein and / or recombinant brazzein-53 is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 75 ppm and 300 ppm, between 75 ppm and 275 ppm, between 75 ppm and 250 ppm, between 75 ppm and 225 ppm, between 75 ppm and 200 ppm, between 75 ppm and 175 ppm, between 75 ppm and 150 ppm, between 75 ppm and 125 ppm, or between 75 ppm and 100 ppm.

[0098] In some instances, brazzein-53 protein and / or recombinant brazzein-53 is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 100 ppm and 300 ppm, between 100 ppm and 275 ppm, between 100 ppm and 250 ppm, between 100 ppm and 225 ppm, between 100 ppm and 200 ppm, between 100 ppm and 175 ppm, between 100 ppm and 150 ppm, or between 100 ppm and 125 ppm.

[0099] In some instances, brazzein-53 protein and / or recombinant brazzein-53 is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 150 ppm and 250 ppm, between 150 ppm and 225 ppm, between 150 ppm and 200 ppm, or between 150 ppm and 175 ppm.

[0100] In some instances, brazzein-53 protein and / or recombinant brazzein-53 protein is the only sweet protein in a beverage, and is present in the beverage in an amount of about 300 ppm, about 275 ppm, about 250 ppm, about 245 ppm, about 240 ppm, about 235 ppm, about 230 ppm, about 225 ppm, about 200 ppm, about 175 ppm, about 170 ppm, about 165 ppm, about 160 ppm, about 155 ppm, about 150 ppm, about 125 ppm, about 100 ppm, about 90 ppm, about 89 ppm, about 88 ppm, about 87 ppm, about 86 ppm, about 85 ppm, about 84 ppm, about 83 ppm, about 82 ppm, about 81ppm, about 80 ppm, about 79 ppm, about 78 ppm, about 77 ppm, about 76 ppm, about 75 ppm, about 50 ppm, about 49 ppm, about 48 ppm, about 47 ppm, about 46 ppm, about 45 ppm, about 44 ppm, about 43 ppm, about 42 ppm, about 41 ppm, about 40 ppm, about 39 ppm, about 38 ppm, about 37 ppm, about 36 ppm, about 35 ppm, about 34 ppm, about 33 ppm, about 32 ppm, about 31 ppm, about 30 ppm, about 29 ppm, about 28 ppm, about 27 ppm, about 26 ppm, about 25 ppm, about 24 ppm, about 23 ppm, about 22 ppm, about 21 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 17 ppm, about 16 ppm, about 15 ppm, about 14 ppm, about 13 ppm, about 12 ppm, about 11 ppm, about 10 ppm, about 9 ppm, about 8 ppm, about 7 ppm, about 6 ppm, about 5 ppm, about 4 ppm, about 3 ppm, or about 2 ppm.Beverage Products with Brazzein-54

[0101] Any of the beverage products (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) described herein can include, between 5 ppm and 584 ppm of brazzein-54 and / or recombinant brazzein-54 protein. In someembodiments, the amount of brazzein-54 protein and / or recombinant brazzein-54 protein in a beverage of the present disclosure is between 5 ppm and 584 ppm, between 5 ppm and 580 ppm, between 5 ppm and 575 ppm, between 5 ppm and 550 ppm, between 5 ppm and 525 ppm, between 5 ppm and 500 ppm, between 5 ppm and 475 ppm, between 5 ppm and 450 ppm, between 5 ppm and 425 ppm, between 5 ppm and 400 ppm, between 5 ppm and 375 ppm, between 5 ppm and 350 ppm, between 5 ppm and 325 ppm, between 5 ppm and 300 ppm, between 5 ppm and 275 ppm, between 5 ppm and 250 ppm, between 5 ppm and 225 ppm, between 5 ppm and 200 ppm, between 5 ppm and 175 ppm, between 5 ppm and 150 ppm, between 5 ppm and 125 ppm, between 5 ppm and 100 ppm, between 5 ppm and 75 ppm, between 5 ppm and 50 ppm, between 5 ppm and 25 ppm, between 5 ppm and 20 ppm, between 5 ppm and 15 ppm, or between 5 ppm and 10 ppm. In some embodiments, the amount of brazzein-54 protein and / or recombinant brazzein-54 protein in a beverage of the present disclosure is between 10 ppm and 300 ppm, between 10 ppm and 275 ppm, between 10 ppm and 265 ppm, between 10 ppm and 200 ppm, between 10 ppm and 150 ppm, between 10 ppm and 100 ppm, between 10 ppm and 75 ppm, between 10 and 72 ppm, between 10 ppm and 69 ppm, between 10 ppm and 50 ppm, between 10 ppm and 30 ppm. In some embodiments, the amount of brazzein-54 protein and / or recombinant brazzein-54 protein in a beverage of the present disclosure is between 11 ppm and 584 ppm, between 11 ppm and 580 ppm, between 11 ppm and 575 ppm, between 11 ppm and 550 ppm, between 11 ppm and 525 ppm, between 11 ppm and 500 ppm, between 11 ppm and 475 ppm, between 11 ppm and 450 ppm, between 11 ppm and 425 ppm, between 11 ppm and 400 ppm, between 11 ppm and 375 ppm, between 11 ppm and 350 ppm, between 11 ppm and 325 ppm, between 11 ppm and 300 ppm, between 11 ppm and 275 ppm, between 11 ppm and 250 ppm, between 11 ppm and 225 ppm, between 11 ppm and 200 ppm, between 11 ppm and 175 ppm, between 11 ppm and 150 ppm, between 11 ppm and 125 ppm, between 11 ppm and 100 ppm, between 11 ppm and 75 ppm, between 11 ppm and 50 ppm, between 11 ppm and 25 ppm, between 11 ppm and 20 ppm, or between 11 ppm and 15 ppm.

[0102] In some embodiments, the amount of brazzein-54 protein and / or recombinant brazzein-54 protein in a beverage of the present disclosure is between 15 ppm and 200 ppm, between 15 ppm and 150 ppm, between 15 ppm and 100 ppm,between 16 ppm and 120 ppm, between 16 ppm and 114 ppm, between 16 ppm and 100 ppm, between 17 ppm and 200 ppm, between 17 ppm and 150 ppm, between 17 ppm and 117 ppm, between 17 ppm and 110 ppm, between 18 ppm and 114 ppm, between 20 ppm and 200 ppm, between 20 ppm and 160 ppm, between 20 ppm and 150 ppm, between 20 ppm and 140 ppm, between 20 ppm and 114 ppm, between 24 ppm and 200 ppm, between 24 ppm and 169 ppm, between 24 ppm and 150 ppm, between 24 ppm and 100 ppm, between 25 ppm and 584 ppm, between 25 ppm and 580 ppm, between 25 ppm and 575 ppm, between 25 ppm and 550 ppm, between 25 ppm and 525 ppm, between 25 ppm and 500 ppm, between 25 ppm and 475 ppm, between 25 ppm and 450 ppm, between 25 ppm and 425 ppm, between 25 ppm and 400 ppm, between 25 ppm and 375 ppm, between 25 ppm and 350 ppm, between 25 ppm and 325 ppm, between 25 ppm and 300 ppm, between 25 ppm and 275 ppm, between 25 ppm and 250 ppm, between 25 ppm and 225 ppm, between 25 ppm and 200 ppm, between 25 ppm and 175 ppm, between 25 ppm and 150 ppm, between 25 ppm and 125 ppm, between 25 ppm and 100 ppm, between 25 ppm and 75 ppm, between 25 ppm and 50 ppm, between 28 ppm and 300 ppm, between 28 ppm and 200 ppm, between 28 ppm and 194 ppm, between 30 ppm and 190 ppm, between 35 ppm and 300 ppm, between 35 ppm and 260 ppm, between 37 ppm and 256 ppm. between 37 ppm and 200 ppm, or between 40 ppm and 200 ppm.

[0103] In some embodiments, the amount of brazzein-54 protein and / or recombinant brazzein-54 protein in a beverage of the present disclosure is between 50 ppm and 500 ppm, between 50 ppm and 475 ppm, between 50 ppm and 450 ppm, between 50 ppm and 425 ppm, between 50 ppm and 400 ppm, between 50 ppm and 375 ppm, between 50 ppm and 350 ppm, between 50 ppm and 325 ppm, between 50 ppm and 300 ppm, between 50 ppm and 275 ppm, between 50 ppm and 250 ppm, between 50 ppm and 225 ppm, between 50 ppm and 200 ppm, between 50 ppm and 175 ppm, between 50 ppm and 150 ppm, between 50 ppm and 125 ppm, between 50 ppm and 100 ppm, between 50 ppm and 75 ppm, between 60 ppm and 300 ppm, between 65 ppm and 300 ppm, between 65 ppm and 260 ppm, between 65 ppm and 250 ppm, between 65 ppm and 200 ppm, between 67 ppm and 300 ppm, between 67 ppm and 268 ppm, between 67 ppm and 250 ppm, between 67 ppm and 200 ppm, between 70 ppm and 250 ppm, or between 70 ppm and 200 ppm.

[0104] In some embodiments, the amount of brazzein-54 protein and / or recombinant brazzein-54 protein in a beverage of the present disclosure is between 100 ppm and 400 ppm, between 100 ppm and 375 ppm, between 100 ppm and 350 ppm, between 100 ppm and 325 ppm, between 100 ppm and 300 ppm, between 100 ppm and 275 ppm, between 100 ppm and 250 ppm, between 100 ppm and 225 ppm, between 100 ppm and 200 ppm, between 100 ppm and 175 ppm, between 100 ppm and 150 ppm, between 100 ppm and 125 ppm, between 111 ppm and 444 ppm, between 111 ppm and 440 ppm, between 111 ppm and 420 ppm, between 111 ppm and 400 ppm, between 111 ppm and 350 ppm, between 111 ppm and 300 ppm, between 111 ppm and 250 ppm, between 111 ppm and 200 ppm, between 125 ppm and 400 ppm, between 125 ppm and 375 ppm, between 125 ppm and 350 ppm, between 125 ppm and 300 ppm, between 125 ppm and 275 ppm, between 125 ppm and 250 ppm, or between 125 ppm and 225 ppm.

[0105] In some embodiments, the amount of brazzein-54 protein and / or recombinant brazzein-54 protein in a beverage of the present disclosure is between 140 ppm and 584 ppm, between 146 ppm and 584 ppm, between 150 ppm and 580 ppm, between 150 ppm and 500 ppm, between 150 ppm and 400 ppm, between 150 ppm and 350 ppm, between 150 ppm and 300 ppm, between 200 ppm and 300 ppm, between 200 ppm and 275 ppm, between 200 ppm and 250 ppm, or between 200 ppm and 225 ppm.

[0106] In some embodiments, the amount of brazzein-54 protein and / or recombinant brazzein-54 protein in a beverage of the present disclosure is about 584 ppm, about 580 ppm, about 575 ppm, about 550 ppm, about 525 ppm, about 500 ppm, about 475 ppm, about 450 ppm, about 445 ppm, about 444 ppm, about 440 ppm, about 430 ppm, about 425 ppm, about 400 ppm, about 375 ppm, about 350 ppm, about 325 ppm, about 300 ppm, about 275 ppm, about 270 ppm, about 269 ppm, about 268 ppm, about 267 ppm, about 266 ppm, about 265 ppm, about 264 ppm, about 263 ppm, about 262 ppm, about 261 ppm, about 260 ppm, about 259 ppm, about 258 ppm, about 257 ppm, about 256 ppm, about 255 ppm, about 254 ppm, about 253 ppm, about 252 ppm, about 251 ppm, about 250 ppm, about 225 ppm, about 200 ppm, about 199 ppm, about 198 ppm, about 197 ppm, about 196 ppm, about 195 ppm, about 194 ppm, about 193 ppm, about 192 ppm, about 191 ppm, about 190 ppm, about 189 ppm, about 188 ppm, about 175 ppm, about 170 ppm, about 169 ppm, about 168 ppm, about 167 ppm, about166 ppm, about 165 ppm, about 160 ppm, about 155 ppm, about 150 ppm, about 149 ppm, about 148 ppm, about 147 ppm, about 146 ppm, about 145 ppm, about 144 ppm, about 143 ppm, about 142 ppm, about 141 ppm, about 140 ppm, about 139 ppm, about 138 ppm, about 137 ppm, about 136 ppm, about 135 ppm, about 134 ppm, about 133 ppm, about 132 ppm, about 131 ppm, about 130 ppm, about 129 ppm, about 128 ppm, about 127 ppm, about 126 ppm, about 125 ppm, about 120 ppm, about 119 ppm, about 118 ppm, about 117 ppm, about 116 ppm, about 115 ppm, about 114 ppm, about 113 ppm, about 112 ppm, about 111 ppm, about 110 ppm, about 105 ppm, about 100 ppm, about 95 ppm, about 90 ppm, about 85 ppm, about 80 ppm, about 79 ppm, about 78 ppm, about 77 ppm, about 76 ppm, about 75 ppm, about 74 ppm, about 73 ppm, about 72 ppm, about 71 ppm, about 70 ppm, about 69 ppm, about 68 ppm, about 67 ppm, about 66 ppm, about 65 ppm, about 64 ppm, about 63 ppm, about 62 ppm, about 61 ppm, about 60 ppm, about 55 ppm, about 50 ppm, about 45 ppm, about 40 ppm, about 39 ppm, about 38 ppm, about 37 ppm, about 36 ppm, about 35 ppm, about 30 ppm, about 29 ppm, about 28 ppm, about 27 ppm, about 26 ppm, about 25 ppm, about 24 ppm, about 23 ppm, about 22 ppm, about 21 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 17 ppm, about 16 ppm, about 15 ppm, about 14 ppm, about 13 ppm, about 12 ppm, about 11 ppm, about 10 ppm, or about 5 ppm.

[0107] Brazzein-54 protein and / or recombinant brazzein-54 protein can be the only sweet protein in a beverage (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) described herein. In some instances, brazzein-54 protein and / or recombinant brazzein-54 is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 65 ppm and 584 ppm, between 65 ppm and 580 ppm, between 65 ppm and 575 ppm, between 65 ppm and 550 ppm, between 65 ppm and 525 ppm, between 65 ppm and 500 ppm, between 65 ppm and 475 ppm, between 65 ppm and 450 ppm, between 65 ppm and 425 ppm, between 65 ppm and 400 ppm, between 65 ppm and 375 ppm, between 65 ppm and 350 ppm, between 65 ppm and 325 ppm, between 65 ppm and 300 ppm, between 65 ppm and 275 ppm, between 65 ppm and 250 ppm, between 65 ppm and 225 ppm, between 65 ppm and 200 ppm, between 65 ppm and 175 ppm, between 65 ppm and 150 ppm, between 65 ppm and 125 ppm, between 65 ppm and 100 ppm, between 65 ppm and 75 ppm, between 65 ppmand 70 ppm, between 67 ppm and 300 ppm, between 67 ppm and 268 ppm, between 67 ppm and 250 ppm, between 67 ppm and 200 ppm, between 70 ppm and 250 ppm, or between 70 ppm and 200 ppm.

[0108] In some instances, brazzein-54 protein and / or recombinant brazzein-54 is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 100 ppm and 584 ppm, between 100 ppm and 580 ppm, between 100 ppm and 575 ppm, between 100 ppm and 550 ppm, between 100 ppm and 525 ppm, between 100 ppm and 500 ppm, between 100 ppm and 475 ppm, between 100 ppm and 450 ppm, between 100 ppm and 425 ppm, between 100 ppm and 400 ppm, between 100 ppm and 375 ppm, between 100 ppm and 350 ppm, between 100 ppm and 325 ppm, between 100 ppm and 300 ppm, between 100 ppm and 275 ppm, between 100 ppm and 250 ppm, between 100 ppm and 225 ppm, between 100 ppm and 200 ppm, between 100 ppm and 175 ppm, between 100 ppm and 150 ppm, between 100 ppm and 125 ppm, between 111 ppm and 444 ppm, between 111 ppm and 440 ppm, between 111 ppm and 420 ppm, between 111 ppm and 400 ppm, between 111 ppm and 350 ppm, between 111 ppm and 300 ppm, between 111 ppm and 250 ppm, between 111 ppm and 200 ppm, between 125 ppm and 400 ppm, between 125 ppm and 375 ppm, between 125 ppm and 350 ppm, between 125 ppm and 300 ppm, between 125 ppm and 275 ppm, between 125 ppm and 250 ppm, or between 125 ppm and 225 ppm.

[0109] In some instances, brazzein-54 protein and / or recombinant brazzein-54 is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 146 ppm and 584 ppm, between 150 ppm and 580 ppm, between 150 ppm and 500 ppm, between 150 ppm and 400 ppm, between 150 ppm and 350 ppm, between 150 ppm and 300 ppm, between 200 ppm and 500 ppm, between 200 ppm and 475 ppm, between 200 ppm and 450 ppm, between 200 ppm and 425 ppm, between 200 ppm and 400 ppm, between 200 ppm and 375 ppm, between 200 ppm and 350 ppm, between 200 ppm and 325 ppm, between 200 ppm and 300 ppm, between 200 ppm and 275 ppm, between 200 ppm and 250 ppm, or between 200 ppm and 225 ppm.

[0110] In some instances, brazzein-54 protein and / or recombinant brazzein-54 is the only sweet protein in a beverage described herein, and is present in the beverage in anamount between 300 ppm and 400 ppm, between 300 ppm and 375 ppm, between 300 ppm and 350 ppm, or between 300 ppm and 325 ppm.

[0111] In some instances, brazzein-54 protein and / or recombinant brazzein-54 is the only sweet protein in a beverage described herein, and is present in the beverage in an amount of about 584 ppm, about 580 ppm, about 575 ppm, about 550 ppm, about 525 ppm, about 500 ppm, about 475 ppm, about 450 ppm, about 445 ppm, about 444 ppm, about 440 ppm, about 430 ppm, about 425 ppm, about 400 ppm, about 375 ppm, about 350 ppm, about 325 ppm, about 300 ppm, about 275 ppm, about 270 ppm, about 269 ppm, about 268 ppm, about 267 ppm, about 266 ppm, about 265 ppm, about 264 ppm, about 263 ppm, about 262 ppm, about 261 ppm, about 260 ppm, about 259 ppm, about 258 ppm, about 257 ppm, about 256 ppm, about 255 ppm, about 254 ppm, about 253 ppm, about 252 ppm, about 251 ppm, about 250 ppm, about 225 ppm, about 200 ppm, about 199 ppm, about 198 ppm, about 197 ppm, about 196 ppm, about 195 ppm, about 194 ppm, about 193 ppm, about 192 ppm, about 191 ppm, about 190 ppm, about 189 ppm, about 188 ppm, about 175 ppm, about 150 ppm, about 149 ppm, about 148 ppm, about 147 ppm, about 146 ppm, about 145 ppm, about 144 ppm, about 143 ppm, about 142 ppm, about 141 ppm, about 140 ppm, about 139 ppm, about 138 ppm, about 137 ppm, about 136 ppm, about 135 ppm, about 134 ppm, about 133 ppm, about 132 ppm, about 131 ppm, about 130 ppm, about 129 ppm, about 128 ppm, about 127 ppm, about 126 ppm, about 125 ppm, about 120 ppm, about 119 ppm, about 118 ppm, about 117 ppm, about 116 ppm, about 115 ppm, about 114 ppm, about 113 ppm, about 112 ppm, about 111 ppm, about 110 ppm, about 105 ppm, about 100 ppm, about 95 ppm, about 90 ppm, about 85 ppm, about 80 ppm, about 75 ppm, about 74 ppm, about 73 ppm, about 72 ppm, about 71 ppm, about 70 ppm, about 69 ppm, about 68 ppm, about 67 ppm, about 66 ppm, about 65 ppm, about 64 ppm, about 63 ppm, about 62 ppm, about 61 ppm, about 60 ppm, or about 55 ppm.Beverage Products with Monellin

[0112] Any of the beverage products (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) described herein can include between 5 ppm and 584 ppm of monellin protein and / or recombinant monellin protein. In someembodiments, the amount of monellin protein and / or recombinant monellin protein in a beverage of the present disclosure is between 5 ppm and 500 ppm, between 5 ppm and 475 ppm, between 5 ppm and 450 ppm, between 5 ppm and 425 ppm, between 5 ppm and400 ppm, between 5 ppm and 375 ppm, between 5 ppm and 350 ppm, between 5 ppm and325 ppm, between 5 ppm and 300 ppm, between 5 ppm and 275 ppm, between 5 ppm and250 ppm, between 5 ppm and 225 ppm, between 5 ppm and 200 ppm, between 5 ppm and175 ppm, between 5 ppm and 150 ppm, between 5 ppm and 125 ppm, between 5 ppm and100 ppm, between 5 ppm and 75 ppm, between 5 ppm and 50 ppm, between 5 ppm and 25 ppm, between 5 ppm and 15 ppm, or between 5 ppm and 10 ppm.

[0113] In some embodiments, the amount of monellin protein and / or recombinant monellin protein in a beverage of the present disclosure is between 8 ppm and 440 ppm, between 8 ppm and 430 ppm, between 8 ppm and 425 ppm, between 8 ppm and 400 ppm, between 8 ppm and 375 ppm, between 8 ppm and 350 ppm, between 8 ppm and 325 ppm, between 8 ppm and 300 ppm, between 8 ppm and 275 ppm, between 8 ppm and 250 ppm, between 8 ppm and 225 ppm, between 8 ppm and 200 ppm, between 8 ppm and 175 ppm, between 8 ppm and 150 ppm, between 8 ppm and 125 ppm, between 8 ppm and 100 ppm, between 8 ppm and 75 ppm, between 8 ppm and 60 ppm, between 8 ppm and 55 ppm, between 8 ppm and 54 ppm, between 8 ppm and 53 ppm, between 8 ppm and 52 ppm, between 8 ppm and 51 ppm, between 8 ppm and 50 ppm, between 8 ppm and 49 ppm, between 8 ppm and 48 ppm, between 8 ppm and 45 ppm, between 8 ppm and 40 ppm, between 8 ppm and 30 ppm, between 8 ppm and 25 ppm, between 8 ppm and 20 ppm, between 8 ppm and 15 ppm, or between 8 ppm and 10 ppm.

[0114] In some embodiments, the amount of monellin protein and / or recombinant monellin protein in a beverage of the present disclosure is between 10 ppm and 400 ppm, between 10 ppm and 350 ppm, between 10 ppm and 300 ppm, between 10 ppm and 275 ppm, between 10 ppm and 265 ppm, between 10 ppm and 200 ppm, between 10 ppm and 100 ppm, between 10 ppm and 75 ppm, between 10 and 72 ppm, between 10 ppm and 69 ppm, between 10 ppm and 105 ppm, between 10 ppm and 50 ppm, between 10 ppm and 30 ppm, between 11 ppm and 275 ppm, between 11 ppm and 250 ppm, between 11 ppm and 225 ppm, between 11 ppm and 200 ppm, between 11 ppm and 175 ppm, between 11 ppm and 150 ppm, between 11 ppm and 125 ppm, between 11 ppm and 100ppm, between 11 ppm and 90 ppm, between 11 ppm and 85 ppm, between 11 ppm and 80 ppm, between 11 ppm and 75 ppm, between 11 ppm and 74 ppm, between 11 ppm and 73 ppm, between 11 ppm and 72 ppm, between 11 ppm and 71 ppm, between 11 ppm and 70 ppm, between 11 ppm and 65 ppm, between 11 ppm and 60 ppm, between 11 ppm and 50 ppm, between 13 ppm and 150 ppm, between 13 ppm and 125 ppm, between 13 ppm and 100 ppm, between 13 ppm and 90 ppm, between 13 ppm and 89 ppm, between 13 ppm and 88 ppm, between 13 ppm and 87 ppm, between 13 ppm and 86 ppm, between 13 ppm and 85 ppm, between 13 ppm and 80 ppm, between 13 ppm and 75 ppm, between 13 ppm and 70 ppm, between 13 ppm and 50 ppm, between 13 ppm and 25 ppm, between 13 ppm and 20 ppm, between 15 ppm and 200 ppm, between 15 ppm and 150 ppm, between 15 ppm and 140 ppm, between 15 ppm and 130 ppm, between 15 ppm and 120 ppm, between 16 ppm and 115 ppm, between 16 ppm and 113 ppm, between 16 ppm and 112 ppm, between 16 ppm and 110 ppm, between 16 ppm and 105 ppm, between 16 ppm and 80 ppm, between 16 ppm and 70 ppm, between 18 ppm and 200 ppm, between 18 ppm and 150 ppm, between 18 ppm and 140 ppm, between 18 ppm and 130 ppm, between 18 ppm and 127 ppm, between 18 ppm and 125 ppm, between 18 ppm and 120 ppm, between 18 ppm and 115 ppm, between 18 ppm and 105 ppm, between 18 ppm and 80 ppm, or between 18 ppm and 70 ppm.

[0115] In some embodiments, the amount of monellin protein and / or recombinant monellin protein in a beverage of the present disclosure is between 20 ppm and 440 ppm, between 20 ppm and 430 ppm, between 20 ppm and 425 ppm, between 20 ppm and 400 ppm, between 20 ppm and 375 ppm, between 20 ppm and 350 ppm, between 20 ppm and 325 ppm, between 20 ppm and 300 ppm, between 20 ppm and 275 ppm, between 20 ppm and 250 ppm, between 20 ppm and 225 ppm, between 20 ppm and 200 ppm, between 25 ppm and 175 ppm, between 20 ppm and 150 ppm, between 20 ppm and 140 ppm, between 20 ppm and 125 ppm, between 20 ppm and 100 ppm, between 20 ppm and 75 ppm, between 20 ppm and 50 ppm, between 28 ppm and 250 ppm, between 28 ppm and 220 ppm, between 28 ppm and 200 ppm, between 28 ppm and 195 ppm, between 28 ppm and 193 ppm, between 28 ppm and 190 ppm, between 28 ppm and 185 ppm, between 28 ppm and 180 ppm, between 28 ppm and 175 ppm, between 28 ppm and 150 ppm,between 28 ppm and 125 ppm, between 28 ppm and 100 ppm, between 28 ppm and 75 ppm, or between 28 ppm and 50 ppm.

[0116] In some embodiments, the amount of monellin protein and / or recombinant monellin protein in a beverage of the present disclosure is between 25 ppm and 440 ppm, between 25 ppm and 430 ppm, between 25 ppm and 425 ppm, between 25 ppm and 400 ppm, between 25 ppm and 375 ppm, between 25 ppm and 350 ppm, between 25 ppm and 325 ppm, between 25 ppm and 300 ppm, between 25 ppm and 275 ppm, between 25 ppm and 250 ppm, between 25 ppm and 225 ppm, between 25 ppm and 200 ppm, between 25 ppm and 175 ppm, between 25 ppm and 150 ppm, between 25 ppm and 125 ppm, between 25 ppm and 100 ppm, between 25 ppm and 75 ppm, between 25 ppm and 50 ppm, between 28 ppm and 250 ppm, between 28 ppm and 225 ppm, between 28 ppm and 200 ppm, between 28 ppm and 195 ppm, between 28 ppm and 193 ppm, between 28 ppm and 190 ppm, between 28 ppm and 185 ppm, between 28 ppm and 180 ppm, between 28 ppm and 175 ppm, between 28 ppm and 150 ppm, between 28 ppm and 125 ppm, between 28 ppm and 100 ppm, between 28 ppm and 75 ppm, or between 28 ppm and 50 ppm.

[0117] In some embodiments, the amount of monellin protein and / or recombinant monellin protein in a beverage of the present disclosure is between 46 ppm and 300 ppm, between 46 ppm and 275 ppm, between 46 ppm and 250 ppm, between 46 ppm and 225 ppm, between 46 ppm and 200 ppm, between 46 ppm and 190 ppm, between 46 ppm and 185 ppm, between 46 ppm and 182 ppm, between 46 ppm and 181 ppm, between 46 ppm and 180 ppm, between 46 ppm and 175 ppm, between 46 ppm and 150 ppm, between 46 ppm and 125 ppm, between 50 ppm and 400 ppm, between 50 ppm and 375 ppm, between 50 ppm and 350 ppm, between 50 ppm and 325 ppm, between 50 ppm and 300 ppm, between 50 ppm and 275 ppm, between 50 ppm and 250 ppm, between 50 ppm and 225 ppm, between 50 ppm and 200 ppm, between 50 ppm and 175 ppm, between 50 ppm and 150 ppm, between 50 ppm and 125 ppm, between 50 ppm and 100 ppm, between 50 ppm and 75 ppm, between 60 ppm and 300 ppm, between 60 ppm and 275 ppm, between 60 ppm and 250 ppm, between 60 ppm and 225 ppm, between 60 ppm and 200 ppm, between 64 ppm and 300 ppm, between 64 ppm and 275 ppm, between 64 ppm and 260 ppm, between 64 ppm and 255 ppm, between 64 ppm and 250 ppm, between 64 ppm and225 ppm, between 64 ppm and 200 ppm, between 65 ppm and 200 ppm, between 65 ppm and 150 ppm, between 65 ppm and 175 ppm, between 65 ppm and 150 ppm, or between 65 ppm and 125 ppm.

[0118] In some embodiments, the amount of monellin protein and / or recombinant monellin protein in a beverage of the present disclosure is between 75 ppm and 300 ppm, between 75 ppm and 275 ppm, between 75 ppm and 250 ppm, between 75 ppm and 225 ppm, between 75 ppm and 200 ppm, between 75 ppm and 175 ppm, between 75 ppm and 150 ppm, between 75 ppm and 125 ppm, or between 75 ppm and 100 ppm.

[0119] In some embodiments, the amount of monellin protein and / or recombinant monellin protein in a beverage of the present disclosure is between 100 ppm and 440 ppm, between 100 ppm and 400 ppm, between 100 ppm and 300 ppm, between 100 ppm and 200 ppm, between 100 ppm and 175 ppm, between 100 ppm and 150 ppm, between 100 ppm and 125 ppm, between 110 ppm and 440 ppm, between 110 ppm and 430 ppm, between 110 ppm and 420 ppm, between 110 ppm and 400 ppm, between 110 ppm and 350 ppm, between 110 ppm and 300 ppm, between 110 ppm and 250 ppm, between 110 ppm and 200 ppm, between 110 ppm and 175 ppm, between 110 ppm and 150 ppm, or between 110 ppm and 125 ppm.

[0120] In some embodiments, the amount of monellin protein and / or recombinant monellin protein in a beverage of the present disclosure is about 500 ppm, about 450 ppm, about 440 ppm, about 430 ppm, about 425 ppm, about 400 ppm, about 375 ppm, about 350 ppm, about 325 ppm, about 300 ppm, about 275 ppm, about 265 ppm, about 260 ppm, about 255 pm, about 250 ppm, about 225 ppm, about 210 ppm, about 200 ppm, about 193 ppm, about 190 ppm, about 185, about 182 ppm, about 180 ppm, about 175 ppm, about 150 ppm, about 140 ppm, about 135, about 130 ppm, about 129, about 128 ppm, about 127 ppm, about 126 ppm, about 125 ppm, about 120, about 115 ppm, about 112 ppm, about 110 ppm, about 105 ppm, about 100 ppm, about 95 ppm, about 90 ppm, about 88 ppm, about 85 ppm, about 80 ppm, about 75 ppm, about 74 ppm, about 73 ppm, about 72 ppm, about 71 ppm, about 70 ppm, about 65 ppm, about 64 ppm, about 60 ppm, about 55 ppm, about 54 ppm, about 53 ppm, about 52 ppm, about 51 ppm, about 50 ppm, about 46 ppm, about 45 ppm, about 40 ppm, about 35 ppm, about 30 ppm, about 28 ppm, about 25 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 17 ppm, about 16 ppm,about 15 ppm, about 14 ppm, about 13 ppm, about 12 ppm, about 11 ppm, about 10 ppm, about 9 ppm, about 8 ppm, about 7 ppm, about 6 ppm, or about 5 ppm.

[0121] Monellin protein and / or recombinant monellin protein can be the only sweet protein in a beverage (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) described herein. In some instances, monellin protein and / or recombinant monellin protein is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 46 ppm and 440 ppm, between 46 ppm and 430 ppm, between 46 ppm and 425 ppm, between 46 ppm and 400 ppm, between 46 ppm and 375 ppm, between 46 ppm and 350 ppm, between 46 ppm and 325 ppm, between 46 ppm and 300 ppm, between 46 ppm and 275 ppm, between 46 ppm and 250 ppm, between 46 ppm and 225 ppm, between 46 ppm and 200 ppm, between 46 ppm and 190 ppm, between 46 ppm and 185 ppm, between 46 ppm and 182 ppm, between 46 ppm and 181 ppm, between 46 ppm and 180 ppm, between 46 ppm and 175 ppm, between 46 ppm and 150 ppm, between 46 ppm and 125 ppm, between 46 ppm and 100 ppm, between 46 ppm and 75 ppm, or between 46 ppm and 50 ppm.

[0122] In some instances, monellin protein and / or recombinant monellin protein is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 50 ppm and 400 ppm, between 50 ppm and 375 ppm, between 50 ppm and 350 ppm, between 50 ppm and 325 ppm, between 50 ppm and 300 ppm, between 50 ppm and 275 ppm, between 50 ppm and 250 ppm, between 50 ppm and 225 ppm, between 50 ppm and 200 ppm, between 50 ppm and 175 ppm, between 50 ppm and 150 ppm, between 50 ppm and 125 ppm, between 50 ppm and 100 ppm, between 50 ppm and 75 ppm, between 60 ppm and 300 ppm, between 60 ppm and 275 ppm, between 60 ppm and 250 ppm, between 60 ppm and 225 ppm, between 60 ppm and 200 ppm, between 64 ppm and 300 ppm, between 64 ppm and 275 ppm, between 64 ppm and 260 ppm, between 64 ppm and 255 ppm, between 64 ppm and 250 ppm, between 64 ppm and 225 ppm, between 64 ppm and 200 ppm, between 65 ppm and 200 ppm, between 65 ppm and 150 ppm, between 65 ppm and 175 ppm, between 65 ppm and 150 ppm, or between 65 ppm and 125 ppm.

[0123] In some instances, monellin protein and / or recombinant monellin protein is the only sweet protein in a beverage described herein, and is present in the beverage inan amount between 75 ppm and 300 ppm, between 75 ppm and 275 ppm, between 75 ppm and 250 ppm, between 75 ppm and 225 ppm, between 75 ppm and 200 ppm, between 75 ppm and 175 ppm, between 75 ppm and 150 ppm, between 75 ppm and 125 ppm, or between 75 ppm and 100 ppm.

[0124] In some instances, monellin protein and / or recombinant monellin protein is the only sweet protein in a beverage described herein, and is present in the beverage in an amount between 100 ppm and 440 ppm, between 100 ppm and 400 ppm, between 100 ppm and 300 ppm, between 100 ppm and 200 ppm, between 100 ppm and 175 ppm, between 100 ppm and 150 ppm, between 100 ppm and 125 ppm, between 110 ppm and 440 ppm, between 110 ppm and 430 ppm, between 110 ppm and 420 ppm, between 110 ppm and 400 ppm, between 110 ppm and 350 ppm, between 110 ppm and 300 ppm, between 110 ppm and 250 ppm, between 110 ppm and 200 ppm, between 110 ppm and 175 ppm, between 110 ppm and 150 ppm, or between 110 ppm and 125 ppm.

[0125] In some instances, monellin protein and / or recombinant monellin protein is the only sweet protein in a beverage described herein, and is present in the beverage in an amount of about 440 ppm, about 430 ppm, about 425 ppm, about 400 ppm, about 375 ppm, about 350 ppm, about 325 ppm, about 300 ppm, about 275 ppm, about 265 ppm, about 260 ppm, about 255 pm, about 250 ppm, about 225 ppm, about 210 ppm, about 200 ppm, about 193 ppm, about 190 ppm, about 182 ppm, about 180 ppm about 175 ppm, about 150 ppm, about 140 ppm, about 130 ppm, about 128 ppm, about 127 ppm, about 126 ppm, about 125 ppm, about 115 ppm, about 112 ppm, about 110 ppm, about 100 ppm, about 90 ppm, about 88 ppm, about 85 ppm, about 80 ppm, about 75 ppm, about 74 ppm, about 73 ppm, about 72 ppm, about 71 ppm, about 70 ppm, about 65 ppm, about 64 ppm, about 60 ppm, about 55 ppm, about 54 ppm, about 53 ppm, about 52 ppm, about 51 ppm, about 50 ppm, about 49 ppm, about 48 ppm, about 47 ppm, about 46 ppm, about 45 ppm, about 40 ppm, about 35 ppm, or about 30 ppm.Beverage Products with Brazzein-53 and Brazzein-54

[0126] A beverage product (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) of the present disclosure can include two sweet proteins, such as, brazzein-53 (and / or recombinant brazzein-53) protein and brazzein-54(and / or recombinant brazzein-54) protein. In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and brazzein-54 (and / or recombinant brazzein-54) protein, and brazzein-53 protein is present in that beverage in an amount between 7 ppm and 131 ppm, between 7 ppm and 130 ppm, between 7 ppm and 120 ppm, between 7 ppm and 110 ppm, between 7 ppm and 100 ppm, between 7 ppm and 90 ppm, between 7 ppm and 80 ppm, between 7 ppm and 70 ppm, between 7 ppm and 60 ppm, between 7 ppm and 50 ppm, between 7 ppm and 48 ppm, between 7 ppm and 45 ppm, between 7 ppm and 40 ppm, between 7 ppm and 35 ppm, between 7 ppm and 30 ppm, between 7 ppm and 25 ppm, between 7 ppm and 20 ppm, between 7 ppm and 15 ppm, or between 7 ppm and 10 ppm.

[0127] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and brazzein-54 (and / or recombinant brazzein-54) protein, and brazzein-53 protein is present in that beverage in an amount between 10 ppm and 131 ppm, between 10 ppm and 130 ppm, between 10 ppm and 120 ppm, between 10 ppm and 110 ppm, between 10 ppm and 100 ppm, between 10 ppm and 90 ppm, between 10 ppm and 80 ppm, between 10 ppm and 72 ppm, between 10 ppm and 70 ppm, between 10 ppm and 60 ppm, between 10 ppm and 50 ppm, between 10 ppm and 45 ppm, between 10 ppm and 40 ppm, between 10 ppm and 35 ppm, between 10 ppm and 30 ppm, between 10 ppm and 25 ppm, between 10 ppm and 20 ppm, or between 10 ppm and 15 ppm.

[0128] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and brazzein-54 (and / or recombinant brazzein-54) protein, and brazzein-53 protein is present in that beverage in an amount between 15 ppm and 131 ppm, between 15 ppm and 125 ppm, between 15 ppm and 120 ppm, between 15 ppm and 115 ppm, between 15 ppm and 110 ppm, between 15 ppm and 105 ppm, between 15 ppm and 100 ppm, between 19 ppm and 131 ppm, between 20 ppm and 125 ppm, between 20 ppm and 120 ppm, between 20 ppm and 115 ppm, between 20 ppm and 110 ppm, between 20 ppm and 100 ppm, between 20 ppm and 80 ppm, between 25 ppm and 131 ppm, between 25 ppm and 130 ppm, between 25 ppm and 120 ppm, between 25 ppm and 110 ppm, between 25 ppm and100 ppm, between 25 ppm and 90 ppm, between 25 ppm and 80 ppm, between 25 ppm and 70 ppm, between 25 ppm and 60 ppm, between 25 ppm and 50 ppm, between 25 ppm and 45 ppm, between 25 ppm and 40 ppm, between 25 ppm and 35 ppm, or between 25 ppm and 30 ppm.

[0129] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and brazzein-54 (and / or recombinant brazzein-54) protein, and brazzein-53 protein is present in that beverage in an amount between 50 ppm and 100 ppm, between 50 ppm and 90 ppm, between 50 ppm and 80 ppm, between 50 ppm and 70 ppm, or between 50 ppm and 60 ppm.

[0130] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and brazzein-54 (and / or recombinant brazzein-54) protein, and brazzein-53 protein is present in that beverage in an amount of about 131 ppm, about 130 ppm, about 125 ppm, about 120 ppm, about 115 ppm, about 110 ppm, about 105 ppm, about 100 ppm, about 95 ppm, about 90 ppm, about 85 ppm, about 80 ppm, about 75 ppm, about 72 ppm, about 70 ppm, about 69 ppm, about 65 ppm, about 60 ppm, about 55 ppm, about 50 ppm, about 48 ppm, about 47 ppm, about 45 ppm, about 40 ppm, about 41 ppm, about 40 ppm, about 35 ppm, about 30 ppm, about 28 ppm, about 27 ppm, about 25 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 15 ppm, about 12 ppm, about 10 ppm, about 7 ppm, or about 5 ppm.

[0131] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and brazzein-54 (and / or recombinant brazzein-54) protein, and brazzein-54 protein is present in that beverage in an amount between 16 ppm and 256 ppm, between 16 ppm and 250 ppm, between 16 ppm and 225 ppm, between 16 ppm and 200 ppm, between 16 ppm and 175 ppm, between 16 ppm and 150 ppm, between 16 ppm and 125 ppm, between 16 ppm and 120 ppm, between 16 ppm and 114 ppm, between 16 ppm and 100 ppm, between 16 ppm and 90 ppm, between 16 ppm and 80 ppm, between 16 ppm and 70 ppm, between 16 ppm and 60 ppm, between 16 ppm and 50 ppm, between 16 ppm and 45 ppm, between 16 ppm and 40 ppm, between 16 ppm and 35 ppm, between 16 ppm and 30 ppm, between 16ppm and 25 ppm, between 16 ppm and 20 ppm, between 17 ppm and 150 ppm, between 17 ppm and 117 ppm, or between 17 ppm and 110 ppm,.

[0132] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and brazzein-54 (and / or recombinant brazzein-54) protein, and brazzein-54 protein is present in that beverage in an amount between 25 ppm and 256 ppm, between 25 ppm and 250 ppm, between 25 ppm and 225 ppm, between 25 ppm and 200 ppm, between 25 ppm and 175 ppm, between 25 ppm and 150 ppm, between 25 ppm and 125 ppm, between 25 ppm and 100 ppm, between 25 ppm and 90 ppm, between 25 ppm and 80 ppm, between 25 ppm and 70 ppm, between 25 ppm and 60 ppm, between 25 ppm and 50 ppm, between 25 ppm and 45 ppm, between 25 ppm and 40 ppm, between 25 ppm and 35 ppm, between 25 ppm and 30 ppm, between 28 ppm and 256 ppm, between 28 ppm and 200 ppm, between 28 ppm and 194 ppm, between 30 ppm and 256 ppm, between 30 ppm and 190 ppm, between 35 ppm and 256 ppm, between 35 ppm and 200 ppm, between 37 ppm and 256 ppm, between 37 ppm and 200 ppm, or between 40 ppm and 200 ppm.

[0133] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and brazzein-54 (and / or recombinant brazzein-54) protein, and brazzein-54 protein is present in that beverage in an amount between 50 ppm and 256 ppm, between 50 ppm and 250 ppm, between 50 ppm and 225 ppm, between 50 ppm and 200 ppm, between 50 ppm and 175 ppm, between 50 ppm and 150 ppm, between 50 ppm and 125 ppm, between 50 ppm and 100 ppm, between 50 ppm and 90 ppm, between 50 ppm and 80 ppm, between 50 ppm and 70 ppm, or between 50 ppm and 60 ppm.

[0134] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and brazzein-54 (and / or recombinant brazzein-54) protein, and brazzein-54 protein is present in that beverage in an amount between 100 ppm and 200 ppm, between 100 pm and 175 ppm, between 100 ppm and 150 ppm, or between 100 ppm and 125 ppm.

[0135] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and brazzein-54 (and / or recombinant brazzein-54) protein, and brazzein-54 protein is presentin that beverage in an amount of about 256 ppm, about 255 ppm, about 254 ppm, about 253 ppm, about 252 ppm, about 251 ppm, about 250 ppm, about 225 ppm, about 200 ppm, about 199 ppm, about 198 ppm, about 197 ppm, about 196 ppm, about 195 ppm, about 194 ppm, about 193 ppm, about 192 ppm, about 191 ppm, about 190 ppm, about 189 ppm, about 188 ppm, about 175 ppm, about 150 ppm, about 125 ppm, about 120 ppm, about 119 ppm, about 118 ppm, about 117 ppm, about 116 ppm, about 115 ppm, about 114 ppm, about 113 ppm, about 112 ppm, about 111 ppm, about 110 ppm, about 105 ppm, about 100 ppm, about 95 ppm, about 90 ppm, about 85 ppm, about 80 ppm, about 75 ppm, about 74 ppm, about 73 ppm, about 72 ppm, about 71 ppm, about 70 ppm, about 69 ppm, about 68 ppm, about 67 ppm, about 66 ppm, about 65 ppm, about 64 ppm, about 63 ppm, about 62 ppm, about 61 ppm, about 60 ppm, about 55 ppm, about 50 ppm, about 45 ppm, about 40 ppm, about 39 ppm, about 38 ppm, about 37 ppm, about 36 ppm, about 35 ppm, about 30 ppm, about 29 ppm, about 28 ppm, about 27 ppm, about 26 ppm, about 25 ppm, about 24 ppm, about 23 ppm, about 22 ppm, about 21 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 17 ppm, about 16 ppm, about 15 ppm, about 14 ppm, about 13 ppm, about 12 ppm, about 11 ppm, or about 10 ppm.Beverage Products with Brazzein-53 and Monellin

[0136] A beverage product (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) of the present disclosure can include two sweet proteins, such as, brazzein-53 (and / or recombinant brazzein-53) protein and monellin (and / or recombinant monellin) protein. In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and monellin (and / or recombinant monellin) protein, and brazzein- 53 protein is present in that beverage in an amount between 7 ppm and 300 ppm, between7 ppm and 280 ppm, between 7 ppm and 265 ppm, between 7 ppm and 250 ppm, between7 ppm and 200 ppm, between 7 ppm and 150 ppm, between 7 ppm and 131 ppm, between7 ppm and 130 ppm, between 7 ppm and 120 ppm, between 7 ppm and 110 ppm, between7 ppm and 100 ppm, between 7 ppm and 90 ppm, between 7 ppm and 80 ppm, between 7 ppm and 70 ppm, between 7 ppm and 60 ppm, between 7 ppm and 50 ppm, between 7 ppm and 48 ppm, between 7 ppm and 45 ppm, between 7 ppm and 40 ppm, between 7ppm and 35 ppm, between 7 ppm and 30 ppm, between 7 ppm and 25 ppm, between 7 ppm and 20 ppm, between 7 ppm and 15 ppm, or between 7 ppm and 10 ppm.

[0137] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and monellin (and / or recombinant monellin) protein, and brazzein-53 protein is present in that beverage in an amount between 10 ppm and 300 ppm, between 10 ppm and 280 ppm, between 10 ppm and 265 ppm, between 10 ppm and 250 ppm, between 10 ppm and 200 ppm, between 10 ppm and 150 ppm, between 10 ppm and 131 ppm, between 10 ppm and 130 ppm, between 10 ppm and 120 ppm, between 10 ppm and 110 ppm, between 10 ppm and 100 ppm, between 10 ppm and 90 ppm, between 10 ppm and 80 ppm, between 10 ppm and 72 ppm, between 10 ppm and 70 ppm, between 10 ppm and 60 ppm, between 10 ppm and 50 ppm, between 10 ppm and 45 ppm, between 10 ppm and 40 ppm, between 10 ppm and 35 ppm, between 10 ppm and 30 ppm, between 10 ppm and 25 ppm, between 10 ppm and 20 ppm, or between 10 ppm and 15 ppm.

[0138] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and monellin (and / or recombinant monellin) protein, and brazzein-53 protein is present in that beverage in an amount between 15 ppm and 300 ppm, between 15 ppm and 280 ppm, between 15 ppm and 265 ppm, between 15 ppm and 250 ppm, between 15 ppm and 200 ppm, between 15 ppm and 150 ppm, between 15 ppm and 131 ppm, between 15 ppm and 130 ppm, between 15 ppm and 125 ppm, between 15 ppm and 120 ppm, between 15 ppm and 115 ppm, between 15 ppm and 110 ppm, between 15 ppm and 105 ppm, between 15 ppm and 100 ppm, between 19 ppm and 131 ppm, between 20 ppm and 115 ppm, between 20 ppm and 110 ppm, between 20 ppm and 100 ppm, between 20 ppm and 80 ppm, between 25 ppm and 131 ppm, between 25 ppm and 130 ppm, between 25 ppm and 120 ppm, between 25 ppm and 110 ppm, between 25 ppm and 100 ppm, between 25 ppm and 90 ppm, between 25 ppm and 80 ppm, between 25 ppm and 70 ppm, between 25 ppm and 60 ppm, between 25 ppm and 50 ppm, between 25 ppm and 45 ppm, between 25 ppm and 40 ppm, between 25 ppm and 35 ppm, or between 25 ppm and 30 ppm.

[0139] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein andmonellin (and / or recombinant monellin) protein, and brazzein-53 protein is present in that beverage in an amount between 50 ppm and 400 ppm, between 50 ppm and 350 ppm, between 50 ppm and 300 ppm, between 50 ppm and 280 ppm, between 50 ppm and 265 ppm, between 50 ppm and 250 ppm, between 50 ppm and 200 ppm, between 50 ppm and 150 ppm, between 50 ppm and 100 ppm, between 50 ppm and 90 ppm, between 50 ppm and 80 ppm, between 50 ppm and 70 ppm, or between 50 ppm and 60 ppm.

[0140] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and monellin (and / or recombinant monellin) protein, and brazzein-53 protein is present in that beverage in an amount of about 300 ppm, about 280 ppm, about 265 ppm, about 263 ppm, about 260 ppm, about 250 ppm, about 230 ppm, about 215 ppm, about 200 ppm, about 185 ppm, about 150 ppm , about 138 ppm, about 131 ppm, about 130 ppm, about 125 ppm, about 120 ppm, about 115 ppm, about 110 ppm, about 105 ppm, about 100 ppm, about 95 ppm, about 90 ppm, about 85 ppm, about 80 ppm, about 75 ppm, about 72 ppm, about 70 ppm, about 69 ppm, about 65 ppm, about 64 ppm, about 60 ppm, about 55 ppm, about 50 ppm, about 48 ppm, about 47 ppm, about 45 ppm, about 40 ppm, about 41 ppm, about 40 ppm, about 35 ppm, about 30 ppm, about 29 ppm, about 28 ppm, about 27 ppm, about 25 ppm, about 24 ppm, about 23 ppm, about 22 ppm, about 21 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 15 ppm, about 12 ppm, about 11 ppm, about 10 ppm, about 7 ppm, or about 5 ppm.

[0141] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and monellin (and / or recombinant monellin) protein, and monellin protein is present in that beverage in an amount between 11 ppm and 193 ppm, between 11 ppm and 190 ppm, between 11 ppm and 180 ppm, between 11 ppm and 170 ppm, between 11 ppm and 160 ppm, between 11 ppm and 150 ppm, between 11 ppm and 140 ppm, between 11 ppm and 130 ppm, between 11 ppm and 120 ppm, between 11 ppm and 110 ppm, between 11 ppm and 100 ppm, between 11 ppm and 90 ppm, between 11 ppm and 85 ppm, between 11 ppm and 80 ppm, between 11 ppm and 75 ppm, between 11 ppm and 70 ppm, between 11 ppm and 60 ppm, between 11 ppm and 50 ppm, between 11 ppm and 45 ppm, between 11 ppm and 40 ppm, between 11 ppm and 35 ppm, between 11 ppm and 30 ppm, between 11ppm and 25 ppm, between 11 ppm and 20 ppm, between 11 ppm and 15 ppm, between 13 ppm and 193 ppm, between 13 ppm and 150 ppm, between 13 ppm and 125 ppm, between 13 ppm and 100 ppm, between 13 ppm and 90 ppm, between 13 ppm and 89 ppm, between 13 ppm and 88 ppm, between 13 ppm and 87 ppm, between 13 ppm and 86 ppm, between 13 ppm and 85 ppm, between 13 ppm and 80 ppm, between 13 ppm and 75 ppm, between 13 ppm and 70 ppm, between 13 ppm and 50 ppm, between 13 ppm and 25 ppm, between 13 ppm and 20 ppm, between 15 ppm and 193 ppm, between 15 ppm and 150 ppm, between 15 ppm and 140 ppm, between 15 ppm and 130 ppm, between 15 ppm and 120 ppm, between 16 ppm and 115 ppm, between 16 ppm and 113 ppm, between 16 ppm and 112 ppm, between 16 ppm and 110 ppm, between 16 ppm and 105 ppm, between 16 ppm and 80 ppm, or between 16 ppm and 70 ppm.

[0142] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and monellin (and / or recombinant monellin) protein, and monellin protein is present in that beverage in an amount between 20 ppm and 193 ppm, between 20 ppm and 190 ppm, between 20 ppm and 180 ppm, between 20 ppm and 170 ppm, between 20 ppm and 160 ppm, between 20 ppm and 150 ppm, between 20 ppm and 140 ppm, between 20 ppm and 130 ppm, between 20 ppm and 120 ppm, between 20 ppm and 110 ppm, between 20 ppm and 100 ppm, between 20 ppm and 90 ppm, between 20 ppm and 80 ppm, between 20 ppm and 70 ppm, between 20 ppm and 60 ppm, between 20 ppm and 50 ppm, between 20 ppm and 45 ppm, between 20 ppm and 40 ppm, between 20 ppm and 35 ppm, and between 20 ppm and 30 ppm.

[0143] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and monellin (and / or recombinant monellin) protein, and monellin protein is present in that beverage in an amount between 25 ppm and 193 ppm, between 25 ppm and 190 ppm, between 25 ppm and 180 ppm, between 25 ppm and 170 ppm, between 25 ppm and 160 ppm, between 25 ppm and 150 ppm, between 25 ppm and 140 ppm, between 25 ppm and 130 ppm, between 25 ppm and 120 ppm, between 25 ppm and 110 ppm, between 25 ppm and 100 ppm, between 25 ppm and 90 ppm, between 25 ppm and 80 ppm, between 25 ppm and 70 ppm, between 25 ppm and 60 ppm, between 25 ppm and 50 ppm, between 25ppm and 45 ppm, between 25 ppm and 40 ppm, between 25 ppm and 35 ppm, between 25 ppm and 30 ppm, between 28 ppm and 193 ppm, between 28 ppm and 190 ppm, between 28 ppm and 185 ppm, between 28 ppm and 180 ppm, between 28 ppm and 175 ppm, between 28 ppm and 150 ppm, between 28 ppm and 125 ppm, between 28 ppm and 100 ppm, between 28 ppm and 75 ppm, or between 28 ppm and 50 ppm.

[0144] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and monellin (and / or recombinant monellin) protein, and monellin protein is present in that beverage in an amount between 50 ppm and 193 ppm, between 50 ppm and 190 ppm, between 50 ppm and 180 ppm, between 50 ppm and 170 ppm, between 50 ppm and 160 ppm, between 50 ppm and 150 ppm, between 50 ppm and 140 ppm, between 50 ppm and 130 ppm, between 50 ppm and 120 ppm, between 50 ppm and 110 ppm, between 50 ppm and 100 ppm, between 50 ppm and 90 ppm, between 50 ppm and 80 ppm, between 50 ppm and 70 ppm, between 50 ppm and 60 ppm, or between 50 ppm and 55 ppm.

[0145] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and monellin (and / or recombinant monellin) protein, and monellin protein is present in that beverage in an amount between 100 ppm and 150 ppm, between 100 ppm and 140 ppm, between 100 ppm and 130 ppm, between 100 ppm and 120 ppm, or between 100 ppm and 110 ppm.

[0146] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein and monellin (and / or recombinant monellin) protein, and monellin protein is present in that beverage in an amount of about 193 ppm, about 190 ppm, about 185 ppm, about 182 ppm, about 180 ppm, about 175 ppm, about 170 ppm, about 165 ppm, about 160 ppm, about 155 ppm, about 150 ppm, about 145 ppm, about 140 ppm, about 136 ppm, about 135 ppm, about 130 ppm, about 129, about 128 ppm, about 127 ppm, about 126 ppm, about 125 ppm, about 120 ppm, about 115 ppm, about 112 ppm, about 110 ppm, about 105 ppm, about 100 ppm, about 95 ppm, about 90 ppm, about 88 ppm, about 85 ppm, about 80 ppm, about 75 ppm, about 74 ppm, about 73 ppm, about 72 ppm, about 71 ppm, about 70 ppm, about 65 ppm, about 64 ppm, about 63 ppm, about 61 ppm, about 60 ppm,about 55 ppm, about 54 ppm, about 53 ppm, about 52 ppm, about 51 ppm, about 50 ppm, about 46 ppm, about 45 ppm, about 40 ppm, about 35 ppm, about 30 ppm, about 28 ppm, 25 ppm, about 24 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 17 ppm, about 16 ppm, about 15 ppm, about 14 ppm, about 13 ppm, about 12 ppm, about 11 ppm, about 10 ppm, about 9 ppm, about 8 ppm, about 7 ppm, about 6 ppm, or about 5 ppm.Beverage Products with Brazzein-54 and Monellin

[0147] A beverage product (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) of the present disclosure can include two sweet proteins, such as, brazzein-54 (and / or recombinant brazzein-54) protein and monellin (and / or recombinant monellin) protein. In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-54 (and / or recombinant brazzein-54) protein and monellin (and / or recombinant monellin) protein, and brazzein- 54 protein is present in that beverage in an amount between 16 ppm and 256 ppm, between 16 ppm and 250 ppm, between 16 ppm and 225 ppm, between 16 ppm and 200 ppm, between 16 ppm and 175 ppm, between 16 ppm and 150 ppm, between 16 ppm and 125 ppm, between 16 ppm and 114 ppm, between 16 ppm and 100 ppm, between 16 ppm and 90 ppm, between 16 ppm and 80 ppm, between 16 ppm and 70 ppm, between 16 ppm and 60 ppm, between 16 ppm and 50 ppm, between 16 ppm and 45 ppm, between 16 ppm and 40 ppm, between 16 ppm and 35 ppm, between 16 ppm and 30 ppm, between 16 ppm and 25 ppm, between 16 ppm and 20 ppm, between 17 ppm and 200 ppm, between 17 ppm and 150 ppm, between 17 ppm and 117 ppm, between 17 ppm and 110 ppm, between 18 ppm and 114 ppm, or between 20 ppm and 200 ppm,.

[0148] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-54 (and / or recombinant brazzein-54) protein and monellin (and / or recombinant monellin) protein, and brazzein-54 protein is present in that beverage in an amount between 25 ppm and 256 ppm, between 25 ppm and 250 ppm, between 25 ppm and 225 ppm, between 25 ppm and 200 ppm, between 25 ppm and 175 ppm, between 25 ppm and 150 ppm, between 25 ppm and 125 ppm, between 25 ppm and 100 ppm, between 25 ppm and 90 ppm, between 25 ppm and 80 ppm, between 25 ppm and 70 ppm, between 25 ppm and 60 ppm, between 25 ppm and 50 ppm, between 25 ppmand 45 ppm, between 25 ppm and 40 ppm, between 25 ppm and 35 ppm, between 25 ppm and 30 ppm, between 28 ppm and 256 ppm, between 28 ppm and 200 ppm, between 28 ppm and 194 ppm, between 30 ppm and 190 ppm, between 35 ppm and 256 ppm, between 37 ppm and 256 ppm, between 37 ppm and 200 ppm, or between 40 ppm and 200 ppm.

[0149] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-54 (and / or recombinant brazzein-54) protein and monellin (and / or recombinant monellin) protein, and brazzein-54 protein is present in that beverage in an amount between 50 ppm and 256 ppm, between 50 ppm and 250 ppm, between 50 ppm and 225 ppm, between 50 ppm and 200 ppm, between 50 ppm and 175 ppm, between 50 ppm and 150 ppm, between 50 ppm and 125 ppm, between 50 ppm and 100 ppm, between 50 ppm and 90 ppm, between 50 ppm and 80 ppm, between 50 ppm and 70 ppm, or between 50 ppm and 60 ppm.

[0150] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-54 (and / or recombinant brazzein-54) protein and monellin (and / or recombinant monellin) protein, and brazzein-54 protein is present in that beverage in an amount between 100 ppm and 200 ppm, between 100 pm and 175 ppm, between 100 ppm and 150 ppm, or between 100 ppm and 125 ppm.

[0151] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-54 (and / or recombinant brazzein-54) protein and monellin (and / or recombinant monellin) protein, and brazzein-54 protein is present in that beverage in an amount of about 256 ppm, about 255 ppm, about 254 ppm, about 253 ppm, about 252 ppm, about 251 ppm, about 250 ppm, about 249 ppm, about 248 ppm, about 247 ppm, about 246 ppm, about 245 ppm, about 244 ppm, about 243 ppm, about 242 ppm, about 241 ppm, about 240 ppm, about 239 ppm, about 238 ppm, about 237 ppm, about 236 ppm, about 235 ppm, about 234 ppm, about 233 ppm, about 232 ppm, about 231 ppm, about 230 ppm, about 229 ppm, about 228 ppm, about 227 ppm, about 226 ppm, about 225 ppm, about 224 ppm, about 223 ppm, about 222 ppm, about 221 ppm, about 220 ppm, about 215 ppm, about 214 ppm, about 213 ppm, about 212 ppm, about 211 ppm, about 210 ppm, about 209 ppm, about 208 ppm, about 207 ppm, about 206 ppm, about 205 ppm, about 200 ppm, about 199 ppm, about 198 ppm, about 197ppm, about 196 ppm, about 195 ppm, about 194 ppm, about 193 ppm, about 192 ppm, about 191 ppm, about 190 ppm, about 189 ppm, about 188 ppm, about 175 ppm, about 170 ppm, about 169 ppm, about 168 ppm, about 167 ppm, about 166 ppm, about 165 ppm, about 160 ppm, about 150 ppm, about 149 ppm, about 148 ppm, about 147 ppm, about 146 ppm, about 145 ppm, about 144 ppm, about 143 ppm, about 142 ppm, about 141 ppm, about 140 ppm, about 139 ppm, about 138 ppm, about 137 ppm, about 136 ppm, about 135 ppm, about 134 ppm, about 133 ppm, about 132 ppm, about 131 ppm, about 130 ppm, about 129 ppm, about 128 ppm, about 127 ppm, about 126 ppm, about 125 ppm, about 120 ppm, about 119 ppm, about 118 ppm, about 117 ppm, about 116 ppm, about 115 ppm, about 114 ppm, about 113 ppm, about 112 ppm, about 111 ppm, about 110 ppm, about 105 ppm, about 100 ppm, about 95 ppm, about 90 ppm, about 85 ppm, about 80 ppm, about 79 ppm, about 78 ppm, about 77 ppm, about 76 ppm, about 75 ppm, about 74 ppm, about 73 ppm, about 72 ppm, about 71 ppm, about 70 ppm, about 69 ppm, about 68 ppm, about 67 ppm, about 66 ppm, about 65 ppm, about 64 ppm, about 63 ppm, about 62 ppm, about 61 ppm, about 60 ppm, about 55 ppm, about 50 ppm, about 45 ppm, about 40 ppm, about 39 ppm, about 38 ppm, about 37 ppm, about 36 ppm, about 35 ppm, about 30 ppm, about 29 ppm, about 28 ppm, about 27 ppm, about 26 ppm, about 25 ppm, about 24 ppm, about 23 ppm, about 22 ppm, about 21 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 17 ppm, about 16 ppm, about 15 ppm, about 14 ppm, about 13 ppm, about 12 ppm, about 11 ppm, about 10 ppm, or about 5 ppm.

[0152] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-54 (and / or recombinant brazzein-54) protein and monellin (and / or recombinant monellin) protein, and monellin protein is present in that beverage in an amount between 11 ppm and 193 ppm, between 11 ppm and 190 ppm, between 11 ppm and 180 ppm, between 11 ppm and 170 ppm, between 11 ppm and 160 ppm, between 11 ppm and 150 ppm, between 11 ppm and 140 ppm, between 11 ppm and 130 ppm, between 11 ppm and 120 ppm, between 11 ppm and 110 ppm, between 11 ppm and 100 ppm, between 11 ppm and 90 ppm, between 11 ppm and 85 ppm, between 11 ppm and 80 ppm, between 11 ppm and 75 ppm, between 11 ppm and 70 ppm, between 11 ppm and 60 ppm, between 11 ppm and 50 ppm, between 11 ppm and 45 ppm, between 11 ppm and 40 ppm, between 11 ppm and 35 ppm, between 11 ppm and 30 ppm, between 11ppm and 25 ppm, between 11 ppm and 20 ppm, between 11 ppm and 15 ppm, between 13 ppm and 193 ppm, between 13 ppm and 125 ppm, between 13 ppm and 100 ppm, between 13 ppm and 90 ppm, between 13 ppm and 89 ppm, between 13 ppm and 88 ppm, between 13 ppm and 87 ppm, between 13 ppm and 86 ppm, between 13 ppm and 85 ppm, between 13 ppm and 80 ppm, between 13 ppm and 75 ppm, between 13 ppm and 70 ppm, between 13 ppm and 50 ppm, between 13 ppm and 25 ppm, between 13 ppm and 20 ppm, between 15 ppm and 193 ppm, between 15 ppm and 150 ppm, between 15 ppm and 140 ppm, between 15 ppm and 130 ppm, between 15 ppm and 120 ppm, between 16 ppm and 115 ppm, between 16 ppm and 113 ppm, between 16 ppm and 112 ppm, between 16 ppm and 110 ppm, between 16 ppm and 105 ppm, between 16 ppm and 80 ppm, or between 16 ppm and 70 ppm.

[0153] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-54 (and / or recombinant brazzein-54) protein and monellin (and / or recombinant monellin) protein, and monellin protein is present in that beverage in an amount between 25 ppm and 193 ppm, between 25 ppm and 190 ppm, between 25 ppm and 180 ppm, between 25 ppm and 170 ppm, between 25 ppm and 160 ppm, between 25 ppm and 150 ppm, between 25 ppm and 140 ppm, between 25 ppm and 130 ppm, between 25 ppm and 120 ppm, between 25 ppm and 110 ppm, between 25 ppm and 100 ppm, between 25 ppm and 90 ppm, between 25 ppm and 80 ppm, between 25 ppm and 70 ppm, between 25 ppm and 60 ppm, between 25 ppm and 50 ppm, between 25 ppm and 45 ppm, between 25 ppm and 40 ppm, between 25 ppm and 35 ppm, between 25 ppm and 30 ppm, between 28 ppm and 193 ppm, between 28 ppm and 190 ppm, between 28 ppm and 185 ppm, between 28 ppm and 180 ppm, between 28 ppm and 175 ppm, between 28 ppm and 150 ppm, between 28 ppm and 125 ppm, between 28 ppm and 100 ppm, between 28 ppm and 75 ppm, or between 28 ppm and 50 ppm.

[0154] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-54 (and / or recombinant brazzein-54) protein and monellin (and / or recombinant monellin) protein, and monellin protein is present in that beverage in an amount between 50 ppm and 193 ppm, between 50 ppm and 190 ppm, between 50 ppm and 180 ppm, between 50 ppm and 170 ppm, between 50 ppm and 160 ppm, between 50 ppm and 150 ppm, between 50 ppm and 140 ppm, between 50 ppm and130 ppm, between 50 ppm and 120 ppm, between 50 ppm and 110 ppm, between 50 ppm and 100 ppm, between 50 ppm and 90 ppm, between 50 ppm and 80 ppm, between 50 ppm and 70 ppm, between 50 ppm and 60 ppm, or between 50 ppm and 55 ppm.

[0155] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-54 (and / or recombinant brazzein-54) protein and monellin (and / or recombinant monellin) protein, and monellin protein is present in that beverage in an amount between 100 ppm and 150 ppm, between 100 ppm and 140 ppm, between 100 ppm and 130 ppm, between 100 ppm and 120 ppm, or between 100 ppm and 110 ppm.

[0156] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-54 (and / or recombinant brazzein-54) protein and monellin (and / or recombinant monellin) protein, and monellin protein is present in that beverage in an amount of about 193 ppm, about 190 ppm, about 185 ppm, about 182 ppm, about 180 ppm, about 175 ppm, about 170 ppm, about 165 ppm, about 160 ppm, about 155 ppm, about 150 ppm, about 145 ppm, about 140 ppm, about 135 ppm, about 130 ppm, about 129, about 128 ppm, about 127 ppm, about 126 ppm, about 125 ppm, about 120 ppm, about 115 ppm, about 112 ppm, about 110 ppm, about 105 ppm, about 100 ppm, about 95 ppm, about 90 ppm, about 88 ppm, about 85 ppm, about 80 ppm, about 75 ppm, about 74 ppm, about 73 ppm, about 72 ppm, about 71 ppm, about 70 ppm, about 65 ppm, about 64 ppm, about 60 ppm, about 55 ppm, about 54 ppm, about 53 ppm, about 52 ppm, about 51 ppm, about 50 ppm, about 46 ppm, about 45 ppm, about 40 ppm, about 35 ppm, about 30 ppm, about 28 ppm, 25 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 17 ppm, about 16 ppm, about 15 ppm, about 14 ppm, about 13 ppm, about 12 ppm, about 11 ppm, about 10 ppm, about 9 ppm, about 8 ppm, about 7 ppm, about 6 ppm, or about 5 ppm.Beverage Products with Brazzein-53, Brazzein-54, and Monellin

[0157] A beverage product (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) of the present disclosure can include three sweet proteins, such as, brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin)protein. In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin) protein; and brazzein-53 protein is present in that beverage in an amount between 5 ppm and 87 ppm, between 5 ppm and 85 ppm, between 5 ppm and 80 ppm, between 5 ppm and 75 ppm, between 5 ppm and 70 ppm, between 5 ppm and 65 ppm, between 5 ppm and 60 ppm, between 5 ppm and 55 ppm, between 5 ppm and 50 ppm, between 5 ppm and 45 ppm, between 5 ppm and 40 ppm, between 5 ppm and 35 ppm, between 5 ppm and 30 ppm, between 5 ppm and 25 ppm, between 5 ppm and 20 ppm, between 5 ppm and 15 ppm, or between 5 ppm and 10 ppm.

[0158] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin) protein; and brazzein-53 protein is present in that beverage in an amount between 7 ppm and 87 ppm, between 7 ppm and 55 ppm, between 7 ppm and 50 ppm, between 7 ppm and 48 ppm, between 7 ppm and 47 ppm, between 7 ppm and 45 ppm, between 10 ppm and 87 ppm, between 10 ppm and 86 ppm, between 10 ppm and 85 ppm, between 10 ppm and 80 ppm, between 10 ppm and 75 ppm, between 10 ppm and 70 ppm, between 10 ppm and 69 ppm, between 10 ppm and 65 ppm, between 10 ppm and 60 ppm, between 10 ppm and 55 ppm, between 10 ppm and 50 ppm, between 10 ppm and 45 ppm, between 10 ppm and 40 ppm, between 10 ppm and 35 ppm, between 10 ppm and 30 ppm, between 10 ppm and 25 ppm, between 10 ppm and 20 ppm, between 10 ppm and 15 ppm, between 12 ppm and 30 ppm, between 12 ppm and 20 ppm, between 12 ppm and 18 ppm, or between 12 ppm and 15 ppm.

[0159] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin) protein; and brazzein-53 protein is present in that beverage in an amount between 25 ppm and 50 ppm, between 25 ppm and 45 ppm, between 25 ppm and 40 ppm, between 25 ppm and 35 ppm, or between 25 ppm and 30 ppm.

[0160] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin) protein; and brazzein-53 protein is present in that beverage in an amount of about 87 ppm, about 86 ppm, about 85 ppm, about 80 ppm, about 75 ppm, about 70 ppm, about 69 ppm, about 65 ppm, about 60 ppm, about 55 ppm, about 50 ppm, about 48 ppm, about 47 ppm, about 45 ppm, about 40 ppm, about 41 ppm, about 40 ppm, about 35 ppm, about 30 ppm, about 28 ppm, about 27 ppm, about 25 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 15 ppm, about 12 ppm, about 10 ppm, about 7 ppm, about 5 ppm, about 4 ppm, about 3 ppm, about 2 ppm, or about 1 ppm.

[0161] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin) protein; and brazzein-54 protein is present in that beverage in an amount between 11 ppm and 169 ppm, between 11 ppm and 168 ppm, between 11 ppm and 167 ppm, between 11 ppm and 166 ppm, between 11 ppm and 165 ppm, between 11 ppm and 160 ppm, between 11 ppm and 150 ppm, between 11 ppm and 140 ppm, between 11 ppm and 130 ppm, between 11 ppm and 120 ppm, between 11 ppm and 110 ppm, between 11 ppm and 100 ppm, between 11 ppm and 90 ppm, between 11 ppm and 80 ppm, between 11 ppm and 75 ppm, between 11 ppm and 70 ppm, between 11 ppm and 60 ppm, between 11 ppm and 50 ppm, between 11 ppm and 40 ppm, between 11 ppm and 30 ppm, or between 11 ppm and 20 ppm.

[0162] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin) protein; and brazzein-54 protein is present in that beverage in an amount between 15 ppm and 169 ppm, between 15 ppm and 150 ppm, between 15 ppm and 100 ppm, between 16 ppm and 169 ppm, between 16 ppm and 114 ppm, between 16 ppm and 100 ppm, between 17 ppm and 169 ppm, between 17 ppm and 150 ppm, between 17 ppm and 117 ppm, between 17 ppm and 110 ppm, between 18 ppm and 114 ppm, between 20 ppm and 169 ppm, between 20 ppm and 114 ppm, between 24 ppm and 200 ppm,between 24 ppm and 169 ppm, between 24 ppm and 150 ppm, between 24 ppm and 100 ppm, between 25 ppm and 169 ppm, between 25 ppm and 168 ppm, between 25 ppm and 167 ppm, between 25 ppm and 166 ppm, between 25 ppm and 165 ppm, between 25 ppm and 160 ppm, between 25 ppm and 150 ppm, between 25 ppm and 140 ppm, between 25 ppm and 130 ppm, between 25 ppm and 120 ppm, between 25 ppm and 110 ppm, between 25 ppm and 100 ppm, between 25 ppm and 90 ppm, between 25 ppm and 80 ppm, between 25 ppm and 70 ppm, between 25 ppm and 60 ppm, between 25 ppm and 50 ppm, between 25 ppm and 40 ppm, between 25 ppm and 30 ppm, between 28 ppm and 169 ppm, between 28 ppm and 150 ppm, between 28 ppm and 100 ppm, between 30 ppm and 169 ppm, between 30 ppm and 150 ppm, between 30 ppm and 100 ppm, between 35 ppm and 169 ppm, between 35 ppm and 150 ppm, between 35 ppm and 100 ppm, between 37 ppm and 169 ppm, between 37 ppm and 150 ppm, between 37 ppm and 100 ppm, or between 40 ppm and 169 ppm.

[0163] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin) protein; and brazzein-54 protein is present in that beverage in an amount between 50 ppm and 150 ppm, between 50 ppm and 140 ppm, between 50 ppm and 130 ppm, between 50 ppm and 120 ppm, between 50 ppm and 110 ppm, between 50 ppm and 100 ppm, between 50 ppm and 90 ppm, between 50 ppm and 80 ppm, between 50 ppm and 70 ppm, or between 50 ppm and 60 ppm.

[0164] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin) protein; and brazzein-54 protein is present in that beverage in an amount of about 169 ppm, about 168 ppm, about 167 ppm, about 166 ppm, about 165 ppm, about 160 ppm, about 155 ppm, about 150 ppm, about 149 ppm, about 148 ppm, about 147 ppm, about 146 ppm, about 145 ppm, about 144 ppm, about 143 ppm, about 142 ppm, about 141 ppm, about 140 ppm, about 139 ppm, about 138 ppm, about 137 ppm, about 136 ppm, about 135 ppm, about 134 ppm, about 133 ppm, about 132 ppm, about 131 ppm, about 130 ppm, about 129 ppm, about 128 ppm, about 127 ppm, about 126 ppm,about 125 ppm, about 120 ppm, about 119 ppm, about 118 ppm, about 117 ppm, about 116 ppm, about 115 ppm, about 114 ppm, about 113 ppm, about 112 ppm, about 111 ppm, about 110 ppm, about 105 ppm, about 100 ppm, about 95 ppm, about 90 ppm, about 85 ppm, about 80 ppm, about 79 ppm, about 78 ppm, about 77 ppm, about 76 ppm, about 75 ppm, about 74 ppm, about 73 ppm, about 72 ppm, about 71 ppm, about 70 ppm, about 69 ppm, about 68 ppm, about 67 ppm, about 66 ppm, about 65 ppm, about 64 ppm, about 63 ppm, about 62 ppm, about 61 ppm, about 60 ppm, about 55 ppm, about 50 ppm, about 45 ppm, about 40 ppm, about 39 ppm, about 38 ppm, about 37 ppm, about 36 ppm, about 35 ppm, about 30 ppm, about 29 ppm, about 28 ppm, about 27 ppm, about 26 ppm, about 25 ppm, about 24 ppm, about 23 ppm, about 22 ppm, about 21 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 17 ppm, about 16 ppm, about 15 ppm, about 14 ppm, about 13 ppm, about 12 ppm, about 11 ppm, about 10 ppm, or about 5 ppm.

[0165] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin) protein; and monellin protein is present in that beverage in an amount between 8 ppm and 127 ppm, between 8 ppm and 126 ppm, between 8 ppm and 125 ppm, between 8 ppm and 120 ppm, between 8 ppm and 110 ppm, between 8 ppm and 100 ppm, between 8 ppm and 90 ppm, between 8 ppm and 80 ppm, between 8 ppm and 75 ppm, between 8 ppm and 70 ppm, between 8 ppm and 60 ppm, between 8 ppm and 55 ppm, between 8 ppm and 54 ppm, between 8 ppm and 53 ppm, between 8 ppm and 52 ppm, between 8 ppm and 51 ppm, between 8 ppm and 50 ppm, between 8 ppm and 49 ppm, between 8 ppm and 48 ppm, between 8 ppm and 45 ppm, between 8 ppm and 40 ppm, between 8 ppm and 30 ppm, between 8 ppm and 25 ppm, between 8 ppm and 20 ppm, between 8 ppm and 15 ppm, between 8 ppm and 10 ppm, between 11 ppm and 127 ppm, between 11 ppm and 125 ppm, between 11 ppm and 100 ppm, between 11 ppm and 90 ppm, between 11 ppm and 85 ppm, between 11 ppm and 80 ppm, between 11 ppm and 75 ppm, between 11 ppm and 74 ppm, between 11 ppm and 73 ppm, between 11 ppm and 72 ppm, between 11 ppm and 71 ppm, between 11 ppm and 70 ppm, between 11 ppm and 65 ppm, between 11 ppm and 60 ppm, between 11 ppm and 50 ppm, between 13 ppm and 127 ppm, between 13 ppm and 125 ppm, between 13 ppm and 100 ppm, between 13 ppm and 90ppm, between 13 ppm and 89 ppm, between 13 ppm and 88 ppm, between 13 ppm and 87 ppm, between 13 ppm and 86 ppm, between 13 ppm and 85 ppm, between 13 ppm and 80 ppm, between 13 ppm and 75 ppm, between 13 ppm and 70 ppm, between 13 ppm and 50 ppm, between 13 ppm and 25 ppm, between 13 ppm and 20 ppm, between 15 ppm and127 ppm, between 15 ppm and 120 ppm, between 16 ppm and 115 ppm, between 16 ppm and 113 ppm, between 16 ppm and 112 ppm, between 16 ppm and 110 ppm, between 16 ppm and 105 ppm, between 16 ppm and 80 ppm, between 16 ppm and 70 ppm, between 18 ppm and 127 ppm, between 18 ppm and 125 ppm, between 18 ppm and 120 ppm, between 18 ppm and 115 ppm, between 18 ppm and 105 ppm, between 18 ppm and 80 ppm, or between 18 ppm and 70 ppm.

[0166] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin) protein; and monellin protein is present in that beverage in an amount between 10 ppm and 127 ppm, between 10 ppm and 126 ppm, between 10 ppm and 125 ppm, between 10 ppm and 120 ppm, between 10 ppm and 110 ppm, between 10 ppm and 100 ppm, between 10 ppm and 90 ppm, between 10 ppm and 80 ppm, between 10 ppm and 70 ppm, between 10 ppm and 60 ppm, between 10 ppm and 50 ppm, between 10 ppm and 40 ppm, between 10 ppm and 30 ppm, or between 10 ppm and 20 ppm.

[0167] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin) protein; and monellin protein is present in that beverage in an amount between 25 ppm and 100 ppm, between 25 ppm and 90 ppm, between 25 ppm and 80 ppm, between 25 ppm and 70 ppm, between 25 ppm and 60 ppm, between 25 ppm and 50 ppm, between 25 ppm and 40 ppm, or between 25 ppm and 30 ppm.

[0168] In some instances, a beverage product (e.g., sweetened beverage product) described herein includes brazzein-53 (and / or recombinant brazzein-53) protein, brazzein-54 (and / or recombinant brazzein-54) protein, and monellin (and / or recombinant monellin) protein; and monellin protein is present in that beverage in an amount of about 127 ppm, about 126 ppm, about 125 ppm, about 120 ppm, about 115 ppm, about 112ppm, about 110 ppm, about 105 ppm, about 100 ppm, about 95 ppm, about 90 ppm, about 88 ppm, about 85 ppm, about 80 ppm, about 75 ppm, about 74 ppm, about 73 ppm, about72 ppm, about 71 ppm, about 70 ppm, about 65 ppm, about 64 ppm, about 60 ppm, about55 ppm, about 54 ppm, about 53 ppm, about 52 ppm, about 51 ppm, about 50 ppm, about46 ppm, about 45 ppm, about 40 ppm, about 35 ppm, about 30 ppm, about 28 ppm, about25 ppm, about 20 ppm, about 19 ppm, about 18 ppm, about 17 ppm, about 16 ppm, about15 ppm, about 14 ppm, about 13 ppm, about 12 ppm, about 11 ppm, about 10 ppm, about9 ppm, about 8 ppm, about 7 ppm, about 6 ppm, or about 5 ppm.Sweeteners

[0169] In some cases, a sweetener can be added to the beverage product (e.g., a lemonade, soda, sparkling juice, fruit punch, mocktail mixer, and / or cocktail mixer) described herein. For example, a sweetener added to a beverage product described herein can include a plant-based sugar composition (also referred to herein as a plant-sugar). In some cases, the plant-based sugar composition is a low glycemic index sweetener. A low glycemic index sweetener can be added to the compositions described herein, for example, to improve organoleptic properties, such as a faster onset of sweetness compared to a composition without the low glycemic index sweetener. The glycemic index is a value assigned to foods and beverages based on how slowly or how quickly those foods or beverages cause increases in blood glucose level. Low glycemic index foods and beverages can achieve a more beneficial effect on blood glucose control, for example, in people with diabetes mellitus, and may also provide metabolic benefits for the general population. Methods for testing the glycemic index of a beverage have been described. See, for example, Wolever, et al. Nutrition Research 23:621-629, 2003. Low glycemic index beverages, including low glycemic index sweeteners have a glycemic index of 55 or less.

[0170] Plant-based sugar compositions can be a low glycemic index sweetener and / or a natural sweetener. In some cases, the plant-based sugar composition includes only nature identical ingredients. Non-limiting exemplary plant-based sugar compositions include, agave, agave syrup, Jerusalem artichoke inulin, yacon, maple syrup, molasses, lucuma fruit, and monk fruit. In some cases, a plant-based sugar composition includesfructose, glucose, and combinations thereof. In some cases, the plant-based sugar compositions used for beverages described herein exclude stevia, erythritol, xylitol, rebaudiosides or any combinations thereof.

[0171] The plant-based sugar compositions can be present in any of the beverage products, such as, lemonade, fruit punch, soda, sparkling juice, mocktail mixer, and / or cocktail mixer described herein. In some cases, the plant-based sugar composition is present in an amount between about 0.1% and about 10% volume / total volume, or any range included therein. For example, the plant-based sugar composition can be present in an amount between about 0.1% and about 5% vol / total vol, about 0.2% and about 5% vol / total vol, about 0.3% and about 5% vol / total vol, about 0.4% and about 5% vol / total vol, about 0.5% and about 5% vol / total vol, about 0.6% and about 5% vol / total vol, about 0.7% and about 5% vol / total vol, about 0.8% and about 5% vol / total vol, about 0.9% and about 5% vol / total vol, about 1% and about 5% vol / total vol, about 1% and about 4.9% vol / total vol, about 1% and about 4.8% vol / total vol, about 1% and about 4.7% vol / total vol, about 1% and about 4.6% vol / total vol, about 1% and about 4.6% vol / total vol, about 1% and about 4.5% vol / total vol, about 1% and about 4.4% vol / total vol, about 1% and about 4.3% vol / total vol, about 1% and about 4.2% vol / total vol, about 1% and about 4.1% vol / total vol, about 1% and about 4.0% vol / total vol, about 1% and about 3.9% vol / total vol, about 1% and about 3.8% vol / total vol, about 1% and about 3.7% vol / total vol, about 1% and about 3.6% vol / total vol, about 1% and about 3.5% vol / total vol, about 1% and about 3.4% vol / total vol, about 1% and about 3.3% vol / total vol, about 1% and about 3.2% vol / total vol, about 1% and about 3.1% vol / total vol, about 1% and about 3% vol / total vol.

[0172] In some cases, any of the beverage products described herein (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) do not include sugar, such as cane sugar. In some cases, any of the beverage products described herein do not include any natural sweetener and / or any natural plant-based sugar composition. In some cases, the beverage products described herein do not include any high intensity sweeteners. Non-limiting examples of high intensity sweeteners include rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, rebaudioside I; Rebaudioside J, rebaudioside K, rebaudioside M,rebaudioside N, rebaudioside O, rebaudioside Q, rebaudioside R, dulcoside A, dulcoside C; Rubusoside, steviol monoside, steviol bioside, stevioside, mogroside V, and saccharin. In some cases, the beverage products described herein, do not include allulose, aspartame, or sugar alcohols.

[0173] In some embodiments, a beverage product of the present disclosure (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) can include between 0% and 3% sugar; such as, between 0% and 0.1% sugar, between 0% and 0.2% sugar, between 0% and 0.3% sugar, between 0% and 0.4% sugar, between 0% and 0.5% sugar, between 0% and 0.6% sugar, between 0% and 0.7% sugar, between 0% and 0.8% sugar, between 0% and 0.9% sugar, between 0% and 1% sugar, between 0% and 1.1% sugar, between 0% and 1.2% sugar, between 0% and 1.3% sugar, between 0% and 1.4% sugar, between 0% and 1.5% sugar, between 0% and 1.6% sugar, between 0% and 1.7% sugar, between 0% and 1.8% sugar, between 0% and 1.9% sugar, between 0% and 2% sugar, between 0% and 2.1% sugar, between 0% and 2.2% sugar, between 0% and 2.3% sugar, between 0% and 2.4% sugar, between 0% and 2.5% sugar, between 0% and 2.6% sugar, between 0% and 2.7% sugar, between 0% and 2.8% sugar, or between 0% and 2.9% sugar. For example, a beverage product of the present disclosure (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) can include about 0% sugar, about 0.1% sugar, about 0.2% sugar, about 0.3% sugar, about 0.4% sugar, about 0.5% sugar, about 0.6% sugar, about 0.7% sugar, about 0.8% sugar, about 0.9% sugar, about 1% sugar, about 1.1% sugar, about 1.2% sugar, about 1.3% sugar, about 1.4% sugar, about 1.5% sugar, about 1.6% sugar, about 1.7% sugar, about 1.8% sugar, about 1.9% sugar, about 2% sugar, about 2.1% sugar, about 2.2% sugar, about 2.3% sugar, about 2.4% sugar, about 2.5% sugar, about 2.6% sugar, about 2.7% sugar, about 2.8% sugar, about 2.9% sugar, or about 3% sugar. In some embodiments, a beverage product of the present disclosure (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) contains no sugar (e.g., contains no added sugar and / or contains 0% added sugar).Acid Components

[0174] In some cases, any of the beverage products described herein (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) can include preservative, an acid, and / or a pH adjuster. In some cases, the beverage product can include maleic acid, citric acid, lemon juice, and combinations thereof. Citric acid can be an additional flavoring agent, a preservative, an acid, a pH adjuster, or any combination thereof. In some instances, any of the beverage products described herein have a pH between about 2.5 and about 4.0; such as, between about 2.5 and about 3.9, between about 2.5 and about 3.8, between about 2.5 and about 3.7, between about 2.5 and about 3.6, between about 2.5 and about 3.5, between about 2.5 and about 3.4, between about 2.5 and about 3.3, between about 2.5 and about 3.2, between about 2.5 and about3.1, between about 2.5 and about 3.0, between about 2.5 and about 2.9, between about 2.5 and about 2.8, between about 2.5 and about 2.7, between about 2.7 and about 4.0, between about 2.7 and about 3.9, between about 2.7 and about 3.8, between about 2.7 and about 3.7, between about 2.7 and about 3.6, between about 2.7 and about 3.5, between about 2.7 and about 3.4, between about 2.7 and about 3.3, between about 2.7 and about3.2, between about 2.7 and about 3.1, between about 2.7 and about 3.0, between about 2.7 and about 2.9, between about 2.7 and about 2.8, between about 2.8 and about 3.4, between about 2.9 and about 3.3, or between about 3.0 and about 3.2. For example, any of the beverage products described herein can have a pH of about 4.0, about 3.9, about 3.8, about 3.7, about 3.6, about 3.5, about 3.4, about 3.3, about 3.2, about 3.1, about 3.0, about 2.9, about 2.8, about 2.7, about 2.6, or about 2.5.BRIX

[0175] BRIX or degrees BRIX (°Bx), is a measure of dissolved solids in a liquid and is sometimes used to measure dissolved sugar content of an aqueous solution. One degree BRIX is 1 gram of sucrose in 100 grams of solution. If the solution contains dissolved solids other than pure sucrose, then BRIX only approximates the dissolved solid content. BRIX can be used to quantify the strength of a solution as a percentage by mass. BRIX is typically determined by a refractometer or density meter. The beverage products described herein (e.g., lemonade, soda, sparkling juice, fruit punch, mocktailmixer, or cocktail mixer) can have a BRIX less than soft drinks (e.g., sodas) or juices containing sugar. In some cases, any of the beverage products described herein have a BRIX between about l°Bx and about 6°Bx, between about 2°Bx and 6°Bx, between about 2°Bx and about 4°Bx, between about l°Bx and about 2°Bx, between about l°Bx and about 3°Bx, between about 2°Bx and about 3°Bx, or between about 2.3°Bx and about 3.7°Bx.Specific gravity

[0176] Specific gravity, or relative density, is a measure of the density of a substance to the density of a reference material. For beverages, specific gravity is often the density of the substance in comparison to the density of water, and is sometimes used to describe dissolved sugar content. As specific gravity is a relative measurement, specific gravity does not have units. Specific gravity can be determined by a hydrometer, pycnometer, or an oscillating U tube. In some cases, specific gravity can be calculated from a measurement of BRIX (see e.g., homebrewacademy.com / specific-gravity-to- brix / ). In some cases, any of the beverage product described herein (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) has a specific gravity between 1.000 and 1.020, or between about 1.020 and 1.015.V. Packaging

[0177] In some aspects, beverage products described herein (e.g., lemonade, soda, sparkling juice, fruit punch, mocktail mixer, or cocktail mixer) can be packaged into a container. In some cases, beverage products described herein can be packaged in a container selected from the group consisting of: a box, a tube, ajar, a vial, a bag, a pouch, a drum, a bottle, and a can. The container can contain information describing directions for beverage product consumption.Definitions

[0178] The term “promoter” is a nucleic acid sequence that is operably linked to a nucleic acid sequence encoding a polypeptide (e.g., a recombinant peptide) that can increase the transcription of the nucleic acid sequence encoding the polypeptide (e.g., a peptide described herein). In some cases, a promoter is constitutive. In other cases, apromoter is inducible. Non-limiting examples of promoters are described herein. Additional examples of promoters are known in the art.

[0179] The term “culturing” refers to growing a population of cells, e.g., microbial cells, under suitable conditions for growth, in a liquid or solid medium.

[0180] The term “purifying” means a step performed to isolate a recombinant peptide from one or more other impurities (e.g., bulk impurities) or components present in a fluid containing a recombinant peptide (e.g., liquid culture medium polypeptides or one or more other components (e.g., DNA, RNA, other polypeptides, endotoxins, viruses, etc.) present in or secreted from a mammalian cell).

[0181] The terms “isolated”, “purified”, “separated”, and “recovered” as used herein refer to a material (e.g., a polypeptide, nucleic acid, or cell) that is removed from at least one component with which it is naturally associated, for example, at a concentration of at least 90% by weight, or at least 95% by weight, or at least 98% by weight of the sample in which it is contained. For example, these terms can refer to a material which is substantially or essentially free from components which normally accompany it as found in its native state, such as, for example, an intact biological system, or is substantially or essentially free from other proteins in the system from which it is expressed.

[0182] As used herein, the term “host cell” refers to a cell or cell line into which a recombinant expression vector (e.g, a nucleic acid construct) can be introduced for expression of the peptide in the host cell. A host cell including a recombinant vector can be referred to as a “recombinant host cell.”

[0183] The term "ppm", as used herein, means parts-per-million and is a weight relative parameter. A part-per-million is a microgram per gram, such that a component that is present at 10 ppm is present at 10 micrograms of the specific component per 1 gram of the aggregate mixture.

[0184] Reference to the term “about” has its usual meaning in the context of compositions to allow for reasonable variations in amounts that can achieve the same effect and also refers herein to a value of plus or minus 10% of the provided value. For example, “about 20” means or includes amounts from 18 to and including 22.

[0185] Unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. As used herein, the singular form “a”, “an”, and “the” include plural references unless indicated otherwise. For example, “an” excipient includes one or more excipients.EXAMPLES

[0186] Various aspects of the disclosure are further illustrated by the following non-limiting examples.Example 1. Preparation of Sweet Protein Powders

[0187] Formulations using recombinant brazzein-53 (BRZ 53), recombinant brazzein-54 (BRZ 54) and recombinant monellin (MON) alone and in combination were designed to achieve a sweetness benchmark of lemonade, soda, sparkling juice, fruit punch, and mocktail or cocktail mixer beverages. For these examples, the recombinant sweet proteins were each expressed in Pichia, and purified to about 40-55% purity and stored as a powder that was then used as an ingredient in the formulations exemplified in Examples 2-7. The amounts of recombinant sweet protein used in the Examples and set forth in the tables therein are expressed as ppm of pure protein.Example 2. Lemonade with Sweet Proteins

[0188] A base formulation for lemonade was constructed as shown in Table 2. TABLE 2. Lemonade base

[0189] Sweet protein powders were added to the base formulation to create lemonades with single sweet proteins, as well as combinations. Lemonade formulations were bench-marked for taste against Santa Cruz Organic Lemonade (which contains approximately 9% sugar content). A panel of 34 tasters was used to compare thesweetness profiles and preferences of the sweet protein lemonade formulations to the benchmark. Results are shown in Table 3. The bottom 2 rows show the percentage of tasters that found each formulation to be closest to the benchmark, and the percentage of testers that rated each formulation as their favorite, respectively.

[0190] The tasters found that brazzein-53 generally provided a good flavor balance and acidity for the lemonade with little sweetness linger, brazzein-54 provided a faster onset of sweetness with a very low amount of linger and monellin helped in reducing the acidity and tartness of the lemonade. The combination of the 3 sweet proteins provided the most rounded taste with no delay in the onset of sweetness as compared to the other formulations.TABLE 3. Formulations for tasting panel

[0191] From the results obtained with the tasting panel, ranges of sweet proteins (formulated alone and in combinations) with the lemonade base were generated based on ranges relative to the amounts used with the tasting panel (just about right (JAR)) as shown in Table 3. For the single proteins, acceptable ranges were set with a lower limit - 50% from JAR and an upper limit +100% from JAR. For the combinations, the lower limit was set at 25% of the individual protein JAR, and the upper limit was set at 75% of the individual protein JAR. Ranges are shown in Table 4.

[0192] Based on the formulations, it was found that for a lemonade, the amount of monellin had to be increased regardless of its high sweetness potency, suggesting that the sweetness action of monellin decreases in a product with high acidity and high fruit content.TABLE 4. Sweet protein ranges (in ppm) for lemonadesExample 3. Low Tartness and Lower Sweetness Lemonade with Sweet Proteins

[0193] Formulations for lemonade and for strawberry -lemonade for low tartness and lower sweetness were constructed as shown in Tables 5 and 6, including between 30- 70 ppm of brazzein-53.TABLE 5. LemonadeTABLE 6. Strawberry-lemonade

[0194] The lemonade formulations were bench-marked for taste against low- sugar BROOKLYN BEST Lemonades (which contains approximately 18g sugar per 12 oz can).

[0195] Additionally, 2-protein lemonade formulations were constructed containing 24 ppm MON and 11 ppm BRZ 53 or 51 ppm MON and 24 ppm BRZ 53. These formulations were bench-marked for taste against sugar added Country Time lemonade and no sugar Crystal Light lemonade, respectively.Example 4. Soda with Sweet Proteins

[0196] A base formulation for lemon-lime soda was constructed as shown inTable 7TABLE 7. Soda base

[0197] Sweet protein powders were added to the base formulation to create sodas with single sweet proteins, as well as combinations. The soda formulations were bench- marked for taste against SPRITE® (Coca-Cola Company), which contains about 10.7% sugar content.

[0198] A panel of 32 tasters was used to compare the sweetness profiles and preferences of the sweet protein lemon-lime soda formulations to the benchmark. Results are shown in Table 8. The bottom 2 rows show the percentage of tasters that found each formulation to be closest to the benchmark, and the percentage of tasters that rated each formulation as their favorite, respectively. Tasters found that brazzein-53 (BRZ 53) had the fastest onset of sweetness, brazzein-54 (BRZ 54) on its own exhibited some linger in the sweetness and monellin (MON) provided an overall roundness in flavor and sweetness. The combination of the 3 sweet proteins was found to have the most rounded character and no delay in the sweetness onset as compared to the single protein formulations.TABLE 8. Formulations for tasting panel

[0199] From the results obtained with the tasting panel, ranges of sweet proteins (formulated alone and in combinations) with the soda base were generated based on ranges relative to the amounts used with the tasting panel (JAR) as shown in Table 8. For the single proteins, acceptable ranges were set with a lower limit -50% from JAR and an upper limit +100% from JAR. For the combinations, the lower limit was set at 25% of the individual protein JAR, and the upper limit was set at 75% of the individual protein JAR. Ranges are shown in Table 9.TABLE 9. Sweet protein ranges (in ppm) for soda

[0200] Additionally, 2-protein formulations for lemon lime soda were constructed with MON and BRZ 53 sweet proteins. One of the lemon lime soda formulations contained 115 ppm MON and 138 ppm BRZ 53. Another lemon lime soda formulation contained 63 ppm MON and 263 ppm BRZ53.Example 5. Sparkling Juice with Sweet Proteins

[0201] A base formulation for sparkling juice was constructed as shown in Table10TABLE 10. Sparkling juice base

[0202] Sweet protein powders were added to the base formulation to create sparkling juices with single sweet proteins, as well as combinations. The sparkling juice formulations were bench-marked for taste against Izze Sparkling Clementine Juice (which contains approximately 7.66% sugar content, primarily from the apple and grape juice concentrates in the product).

[0203] A panel of 33 tasters was used to compare the sweetness profiles and preferences of the sweet protein sparkling juice formulations to the benchmark. Results are shown in Table 11. The bottom 2 rows show the percentage of tasters that found each formulation to be closest to the benchmark, and the percentage of tasters that rated each formulation as their favorite, respectively. Tasters found that brazzein-53 (BRZ 53) had a fast onset of sweetness that was similar to the benchmark, and tasters observed that the formulation with monellin (MON) also had a fast onset of sweetness with no linger. The combination of the 3 sweet proteins resulted in a sparkling juice with a rounded character, no delay in sweetness onset, and little to no linger.TABLE 11. Formulations for tasting panel

[0204] From the results obtained with the tasting panel, ranges of sweet proteins (formulated alone and in combinations) with the sparkling juice base were generated based on ranges relative to the amounts used with the tasting panel (JAR) as shown in Table 11. For the single proteins, acceptable ranges were set with a lower limit -50% from JAR and an upper limit +100% from JAR. For the combinations, the lower limitwas set at 25% of the individual protein JAR, and the upper limit was set at 75% of the individual protein JAR. Ranges are shown in Table 12.

[0205] Based on the formulations, it was found that brazzein-53 worked synergistically with fruit / acidic compounds, and with carbonation. For monellin although addition of this sweet protein provided a roundness in sweetness, a higher amount was required in acidic conditions to provide the optimal sweetness (regardless of its higher sweetness potency).TABLE 12. Sweet protein ranges (in ppm) for sparkling juiceExample 6. Spicy Margarita Mocktail with Sweet Proteins

[0206] A spicy margarita mocktail was formulated using the base formulation shown in Table 13.TABLE 13. Spicy margarita base

[0207] Sweet protein powders were added to the base formulation to create the mocktail with single sweet proteins, as well as combinations. The spicy margarita mocktail formulation did not include any added sugars.

[0208] Ranges of sweet proteins (formulated alone and in combinations) were generated as shown in Table 14 using the commercial product Jose Cuervo® Classic Lime Margarita Mix as a benchmark for sweetness (this commercial product containsapproximately 20% sugar). For the single proteins, acceptable ranges were set with a lower limit -50% from JAR and an upper limit +100% from JAR. For the combinations, the lower limit was set at 25% of the individual protein JAR, and the upper limit was set at 75% of the individual protein JAR.TABLE 14. Sweet protein ranges (in ppm) for spicy margarita mocktailExample 7. Elderflower-cucumber-ginger Mocktail with Sweet Proteins

[0209] An elderflower-cucumber-ginger mocktail was formulated using the base formulation shown in Table 15.TABLE 15. Mocktail base

[0210] Sweet protein powders were added to the base formulation to create the mocktail with single sweet proteins, as well as combinations. The mocktail formulations did not include any added sugars.

[0211] Ranges of sweet proteins (formulated alone and in combinations) were generated as shown in Table 16. For the single proteins, acceptable ranges were set with a lower limit -50% from JAR and an upper limit +100% from JAR. For the combinations, the lower limit was set at 25% of the individual protein JAR, and the upper limit was set at 75% of the individual protein JAR.TABLE 16. Sweet protein ranges (in ppm) for elderflower-cucumber-ginger mocktailExample 8. Fruit Punch with Sweet Proteins

[0212] Formulations for fruit punch were constructed containing an unsweetened Kool-Aid brand tropical punch flavoring packet, monellin (MON) and brazzein-53 (BRZ 53), and citric acid dissolved in water. One fruit punch formulation contained 61 ppm MON and 29 ppm BRZ 53. Another fruit punch formulation contained 136 ppm MON and 64 ppm BRZ 53. The 2-protein fruit punch formulations were bench-marked for taste against no sugar Crystal Light Fruit Punch and Kool-Aid Fruit Punch, respectively.

Claims

WHAT IS CLAIMED IS:

1. A sweetened beverage comprising one or more sweet proteins, wherein at least one of the one or more sweet proteins is present in an amount between 5 ppm and 584 ppm.

2. The sweetened beverage of claim 1, wherein the sweetened beverage comprises brazzein-53.

3. The sweetened beverage of claim 2, wherein the brazzein-53 is present in an amount between 5 ppm and 300 ppm.

4. The sweetened beverage of claim 2 or claim 3, wherein the brazzein-53 is the only sweet protein in the sweetened beverage and the brazzein-53 is present in an amount between 28 ppm and 300 ppm.

5. The sweetened beverage of claim 1, wherein the sweetened beverage comprises brazzein-54.

6. The sweetened beverage of claim 5, wherein the brazzein-54 is present in an amount between 11 ppm and 584 ppm.

7. The sweetened beverage of claim 5 or claim 6, wherein the brazzein-54 is the only sweet protein in the sweetened beverage and the brazzein-54 is present in an amount between 65 ppm and 584 ppm.

8. The sweetened beverage of claim 1, wherein the sweetened beverage comprises monellin.

9. The sweetened beverage of claim 8, wherein the monellin is present in an amount between 8 ppm and 440 ppm.

10. The sweetened beverage of claim 8 or claim 9, wherein the monellin is the only sweet protein in the sweetened beverage and the monellin is present in an amount between 46 ppm and 440 ppm.

11. The sweetened beverage of claim 1, wherein the sweetened beverage comprises only two sweet proteins.

12. The sweetened beverage of claim 11, wherein the two sweet proteins are brazzein-53 and brazzein-54.

13. The sweetened beverage of claim 12, wherein the brazzein-53 is present in an amount between 7 ppm and 131 ppm and the brazzein-54 is present in an amount between 16 ppm and 256 ppm.

14. The sweetened beverage of claim 11, wherein the two sweet proteins are brazzein-53 and monellin.

15. The sweetened beverage of claim 14, wherein the brazzein-53 is present in an amount between 7 ppm and 131 ppm and the monellin is present in an amount between 11 ppm and 193 ppm.

16. The sweetened beverage of claim 14, wherein the brazzein-53 is present in an amount between 10 ppm and 265 ppm and the monellin is present in an amount between 20 ppm and 140 ppm.

17. The sweetened beverage of claim 11, wherein the two sweet proteins are brazzein-54 and monellin.

18. The sweetened beverage of claim 17, wherein the brazzein-54 is present in an amount between 16 ppm and 256 ppm and the monellin is present in an amount between 11 ppm and 193 ppm.

19. The sweetened beverage of claim 1, wherein the sweetened beverage comprises brazzein-53, brazzein-54 and monellin.

20. The sweetened beverage of claim 19, wherein the brazzein-53 is present in an amount between 5 ppm and 87 ppm, the brazzein-54 is present in an amount between 11 ppm and 169 ppm, and the monellin is present in an amount between 8 ppm and 127 ppm.

21. The sweetened beverage of any one of claims 1-20, wherein the sweetened beverage is carbonated.

22. The sweetened beverage of any one of claims 1-21, wherein pH of the sweetened beverage is between about 2.7 and about 3.5.

23. The sweetened beverage of any one of claims 1-22, wherein the sweetened beverage comprises between 0% and 3% sugar.

24. The sweetened beverage of any one of claims 1-23, wherein the sweetened beverage comprises a fruit juice, a fruit concentrate or a combination thereof.

25. The sweetened beverage of any one of claims 1-24, wherein the sweetened beverage is a lemonade.

26. The sweetened beverage of any one of claims 1-24, wherein the sweetened beverage is a sparkling juice.

27. The sweetened beverage of any one of claims 1-24, wherein the sweetened beverage is a soda.

28. The sweetened beverage of any one of claims 1-24, wherein the sweetened beverage is a fruit punch.

29. The sweetened beverage of any one of claims 1-24, wherein the sweetened beverage is a mocktail mixer or a cocktail mixer.

30. The sweetened beverage of claim 29, wherein the mocktail mixer or the cocktail mixer is savory or spicy.

31. The sweetened beverage of claim 29 or 30, wherein the sweetened beverage does not contain added sugar.

32. The sweetened beverage of any one of claims 1-31, wherein the sweetened beverage does not contain any high intensity sweeteners selected from the group consisting of rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, rebaudioside I; Rebaudioside J, rebaudioside K, rebaudioside M, rebaudioside N, rebaudioside O, rebaudioside Q, rebaudioside R, dulcoside A, dulcoside C; Rubusoside, steviol monoside, steviol bioside, stevioside, mogroside V, and saccharin.

33. The sweetened beverage of any one of claims 1-33, wherein at least one of the one or more sweet proteins is a recombinant protein.

34. The sweetened beverage of claim 33, wherein the recombinant protein is produced by fermentation of a microbial organism.

35. The sweetened beverage of claim 34, wherein the microbial organism is a yeast, optionally Komagataella phaffii.

36. The sweetened beverage of any one of claims 33-35, wherein the recombinant protein is selected from the group consisting of brazzein-53, brazzein-54, monellin and any combination thereof.

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