Brazilin-containing composition

Abstract

The invention relates to a composition containing burazil. Compositions comprising Brazilin (or analogs thereof) are disclosed, including sweetener compositions, flavor modification compositions, sweetened compositions, flavor modified compositions. Also disclosed are methods of improving the time profile, flavor profile, and / or taste profile of a sweetenable composition, such as a beverage.

Description

[0001] This application is a divisional application of an application with an application date of April 1, 2020, application number 202080039606.X, and invention name “Composition containing brazyne”.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Application No. 62 / 827,487, filed on April 1, 2019, which is incorporated herein by reference in its entirety. Field of the Invention

[0004] Disclosed herein are sweetener compositions, flavor-modifying compositions, and methods of making and using the same. Also disclosed are consumables (eg, beverages) containing such sweetener compositions and / or flavor-modifying compositions. Background of the Invention

[0006] Natural caloric sugar, such as sucrose, fructose and glucose, is used in a large number in beverage, food, medicine and oral hygiene / cosmetics industry because of its pleasant taste. Especially, sucrose gives the taste that consumers prefer. Although sucrose provides excellent sweetness characteristics, it is caloric. Although calories are necessary for normal body function, some consumers prefer alternative calorie-free or low-calorie sweeteners with sugar-like taste. Calorie-free or low-calorie sweeteners have been introduced to meet consumer demand. However, such sweeteners are related to undesirable taste characteristics. Especially, calorie-free or low-calorie sweeteners show time characteristic curves, maximum response, flavor profile curves, mouthfeel and / or adaptation behavior that are different from sugar. When used for consumer goods, the sweetness start of delay, lasting sweet aftertaste, bitter taste, metallic taste, astringent taste, refreshing taste (cooling taste) and / or liquorice-like taste will usually be detected.

[0007] Thus, there remains a need to develop reduced-calorie or non-calorie sweeteners and sweetening compositions suitable for use in consumables that also provide temporal and flavor profiles similar to those of sucrose. SUMMARY OF THE INVENTION

[0009] Disclosed herein are compositions comprising brazzein (or its analogs), and methods of making and using these compositions.

[0010] In a first embodiment, a sweetener composition or a flavor-modifying composition is disclosed that includes (i) brazin (or an analog thereof) and (ii) at least one steviol glycoside or mogroside.

[0011] In a specific embodiment, the steviol glycoside is selected from rebaudioside M ("Reb M"), rebaudioside A ("Reb A"), and A95. In a specific embodiment, the mogroside is siamenoside I.

[0012] In a second embodiment, a sweetener composition or a flavor-modifying composition is disclosed that includes (i) brazil (or an analog thereof) and (ii) high fructose corn syrup (HFCS).

[0013] In a third embodiment, a consumable (eg, a beverage) is provided that includes (i) brazil (or an analog thereof) and (ii) at least one steviol glycoside or HFCS.

[0014] In particular embodiments, brazeein (or an analog thereof) is present in an amount between about 1 ppm and about 50 ppm.

[0015] In a third embodiment, a method is disclosed for imparting a more sugar-like temporal profile, flavor profile, and / or taste profile to a consumable (e.g., a beverage or other ingestible consumable) by adding a sweetener composition or flavor-modifying composition disclosed herein, thereby providing a consumable having a more sugar-like temporal profile, flavor profile, and / or taste profile.

[0016] In one embodiment, the more sugar-like temporal profile has a reduced sweetness lingering compared to a consumable to which no sweetener composition or flavor-modifying composition disclosed herein has been added.

[0017] In another embodiment, the more sugar-like flavor profile has an improved mouthfeel compared to a consumable to which no sweetener composition or flavor-modifying composition disclosed herein has been added. In a particular embodiment, the improved mouthfeel is an increased body or richness.

[0018] In another embodiment, the more sugar-like taste profile has reduced bitterness compared to a consumable to which no sweetener composition or flavor-modifying composition disclosed herein has been added.

[0019] The method may further include adding other sweeteners, additives, functional ingredients, and combinations thereof.

[0020] The brazynes used in the compositions disclosed herein (e.g., sweetener compositions, flavor-modifying compositions, or consumables comprising the sweetener compositions or flavor-modifying compositions) can be produced by any suitable means (e.g., in vivo or in vitro extraction, chemical synthesis).

[0021] The one or more steviol glycosides can be used in any form. In one embodiment, the steviol glycosides are present in a stevia extract, wherein the steviol glycosides account for about 5% to about 100% of the stevia extract by dry weight. In another embodiment, the steviol glycosides are present in a mixture of steviol glycosides, wherein the steviol glycosides account for about 5% to about 100% of the steviol glycoside mixture by dry weight.

[0022] The compositions disclosed herein may also contain one or more additional sweeteners, including, for example, natural sweeteners, high-potency sweeteners, carbohydrate sweeteners, synthetic sweeteners, and combinations thereof.

[0023] The composition may also contain one or more additives, including, for example, carbohydrates, polyols, amino acids and their corresponding salts, polyamino acids and their corresponding salts, sugar acids and their corresponding salts, nucleotides, organic acids, inorganic acids, organic salts (including organic acid salts and organic base salts), inorganic salts, bitter compounds, flavoring agents and flavoring ingredients, astringent compounds, proteins or protein hydrolysates, surfactants, emulsifiers, flavonoids, alcohols, polymers, and combinations thereof.

[0024] The composition may also contain one or more functional ingredients, such as, for example, saponins, antioxidants, dietary fiber sources, fatty acids, vitamins, glucosamine, minerals, preservatives, hydrating agents, probiotics, prebiotics, weight management agents, osteoporosis management agents, phytoestrogens, long chain primary aliphatic saturated alcohols, plant sterols, and combinations thereof.

[0025] Disclosed consumer products include, for example, pharmaceutical compositions, edible gel mixes and compositions, dental compositions, foods, beverages, and beverage products.

[0026] In certain embodiments, beverages are disclosed that contain the sweetener or flavor modifying composition disclosed herein. The beverage contains a liquid matrix forming its basis, such as, for example, deionized water, distilled water, reverse osmosis water, carbon-treated water, purified water, demineralized water, phosphoric acid, phosphate buffer, citric acid, citrate buffer, and carbon-treated water.

[0027] Also provided are low-calorie and zero-calorie beverages containing the sweetener compositions or flavor-modifying compositions disclosed herein.

[0028] Also provided are tabletop sweetener compositions comprising the sweetener compositions or flavor and / or taste modifying compositions disclosed herein. The tabletop compositions may further comprise at least one bulking agent, additive, anti-caking agent, functional ingredient, and combinations thereof.

[0029] This article also provides the following items:

[0030] 1) A sweetener composition or a flavor-modifying composition comprising (i) brazine and (ii) at least one additional sweetener selected from steviol glycoside sweeteners and mogroside sweeteners.

[0031] 2) The sweetener composition according to item 1), wherein the steviol glycoside sweetener is selected from rebaudioside M ("Reb M"), Reb M80, rebaudioside D ("Reb D"), A95 and rebaudioside A ("Reb A").

[0032] 3) The sweetener composition as described in item 1), wherein the mogroside sweetener is simonoside I.

[0033] 4) A sweetener composition or a flavor-modifying composition comprising brazil and high fructose corn syrup (HFCS).

[0034] 5) A beverage or a beverage product, comprising the sweetener composition or the flavor-modifying composition according to any one of items 1) to 4).

[0035] 6) The beverage or beverage product of item 5), wherein the brazeein is present in an amount between about 1 ppm and about 50 ppm.

[0036] 7) The beverage or beverage product of item 5), wherein the brazeein is present in an amount between about 1 ppm and about 40 ppm.

[0037] 8) A beverage or beverage product as described in item 5), wherein the brazeein is present in an amount selected from the following: between about 1 ppm and about 30 ppm, between about 1 ppm and about 25 ppm, between about 1 ppm and about 20 ppm, or between about 1 ppm and about 15 ppm.

[0038] 9) A beverage or beverage product as described in items 6)-8), wherein the beverage or beverage product has at least one improved sensory characteristic compared to a beverage or beverage product not containing the sweetener composition or flavor modifying composition, wherein the sensory characteristic is selected from the group consisting of: time profile, flavor profile, taste, bitter aftertaste, sweetness retention, mouthfeel or a combination thereof.

[0039] 10) The beverage or beverage product according to item 9), wherein the at least one improved organoleptic property is a reduced bitter aftertaste or an improved mouthfeel.

[0040] 11) A beverage or beverage product as described in item 9), wherein the at least one improved organoleptic property is mouthfeel improvement and the amount of brazeine is between about 1 ppm and about 40 ppm.

[0041] 12) A beverage or beverage product as described in any one of items 6) to 9), wherein the beverage or beverage product is selected from low-calorie or calorie-free beverages or beverage products.

[0042] 13) A beverage or beverage product as described in any one of items 6) to 9), wherein the beverage is selected from the group consisting of: cola, ginger ale, soft drinks, root beer, fruit juice, fruit-flavored juice, vegetable juice, vegetable-flavored juice, sports drink, energy drink, plant protein beverage, near-water beverage (e.g., water containing natural or synthetic flavorings), tea (e.g., black tea, green tea, black tea, oolong tea), coffee, cocoa beverage, beverage containing milk components (e.g., milk beverage, coffee containing milk components, au gratin, milk tea, fruit milk beverage).

[0043] 14) A method for improving at least one sensory property of a beverage or a beverage product, the method comprising adding a sweetener composition or a flavor composition as described in item 1) to a liquid matrix, thereby providing a beverage or a beverage product having at least one improved sensory property.

[0044] 15) The method as described in item 14), wherein the steviol glycoside sweetener is selected from rebaudioside M ("Reb M"), Reb M80, rebaudioside D ("Reb D"), A95 and rebaudioside A ("Reb A").

[0045] 16) The method of item 14) or 15), wherein the improved sensory properties are selected from the group consisting of temporal profile, flavor profile, taste, bitter aftertaste, sweetness lingering, mouthfeel or a combination thereof.

[0046] 17) The method according to item 16), wherein the improved organoleptic property is a reduction in bitter aftertaste or an improvement in mouthfeel.

[0047] 18) The method of item 16), wherein the improved organoleptic property is improved mouthfeel and the amount of brazeine present in the beverage or beverage product is between about 1 ppm and about 40 ppm.

[0048] 19) The method of item 14) or 15), wherein brazein is present in the beverage or beverage product in an amount between about 1 ppm and about 50 ppm.

[0049] 20) A method as described in item 14) or 15), wherein the brazyne is present in the beverage or beverage product in an amount selected from the following: between about 1 ppm and about 40 ppm, between about 1 ppm and about 30 ppm, between about 1 ppm and about 25 ppm, between about 1 ppm and about 20 ppm, or between about 1 ppm and about 15 ppm. DETAILED DESCRIPTION

[0050] definition

[0051] As used herein, the term "analog" refers to a molecule that is not identical but has similar functional or structural characteristics. For example, a polypeptide analog retains the biological activity of the corresponding naturally occurring polypeptide while having certain biochemical modifications that enhance the function of the analog relative to the naturally occurring polypeptide. For example, compared to a wild-type polypeptide, the modification may be a replacement, deletion or insertion of one or more amino acids. In some embodiments, compared to a wild-type peptide or a portion thereof, the analog has no more than 10, 9, 8 or 7 replaced, deleted or inserted amino acids. In some embodiments, compared to a wild-type peptide or a portion thereof, the analog has no more than 6 replaced, deleted or inserted amino acids. In some embodiments, compared to a wild-type peptide or a portion thereof, the analog has no more than 5 or 4 replaced, deleted or inserted amino acids. In some embodiments, compared to a wild-type peptide or a portion thereof, the analog has no more than 3, 2 or 1 replaced, deleted or inserted amino acids. Analogs may include non-natural amino acids.

[0052] As used herein, the term "astringency" refers to a perceived tightening and drying of the palate and is known to gradually increase in intensity and become increasingly difficult to clear from the mouth after repeated exposure. Astringency is a sensation of dryness that occurs in the mouth and is generally explained as a loss of lubricity due to protein precipitation in the salivary membrane that coats and lubricates the oral cavity. Astringency is not confined to a specific area of ​​the mouth, but is a diffuse surface phenomenon characterized by a loss of lubricity.

[0053] As used herein, the term "bitter" or "bitter taste" refers to the perception or taste obtained after detecting a bitter tastant. The following attributes can produce bitter taste: astringency, bitter astringency, metallic taste, bitter metallic taste, as well as off-flavors, aftertastes, and undesirable tastes, including but not limited to freeze burn and cardboard taste and / or any combination of these tastes. It should be noted that in the art, the term "off-flavor" is often synonymous with "bitter taste". The bitter taste of a substance can be compared to the bitterness threshold of 1 for quinine. (Guyton, Arthur C. (1991) Textbook of Medical Physiology. [Medical Physiology Textbook] (8th Edition). Philadelphia: WB Saunders; McLaughlin S., Margolskee RF (1994). "The Sense of Taste [taste]". American Scientist. [American Scientist] 82 (6): 538-545). As described herein, bitter taste can be tested using a panel of subjects, or in vitro, for example, using a taste receptor cell line.

[0054] As used herein, the term "consumer's product" refers to a substance that contacts the mouth of a person or animal, including a substance that is taken into the mouth and subsequently discharged from the mouth and a substance that is drunk, eaten, swallowed or otherwise ingested, and is safe for human or animal consumption when used within a generally acceptable range. Exemplary consumer's products include, but are not limited to, pharmaceutical compositions, edible gel mixtures and compositions, dental compositions, foods (sweets, condiments, chewing gum, cereal compositions, baked goods, dairy products, and tabletop sweetener compositions), beverages, and beverage products. Consumer's products can be sweetened or unsweetened.

[0055] As used herein, the term "degrees Brix" refers to the sugar content of an aqueous solution. One degree Brix is ​​1 gram of sucrose in 100 grams of a solution and represents the strength of the solution in terms of weight percent (% w / w).

[0056] As used herein, the term "expression" refers to the transcription and / or translation process that occurs in a cell. The transcription level of the target nucleic acid sequence in a cell can be determined based on the amount of the corresponding mRNA present in the cell. For example, the mRNA transcribed from the target sequence can be quantified by RT-PCR or Northern blot hybridization (see Sambrook et al., 1999, supra). The polypeptide encoded by the target nucleic acid can be quantified by various methods, such as by ELISA, by measuring the biological activity of the polypeptide, or by adopting an assay independent of this activity (such as Western blot or radioimmunoassay), using an immunoglobulin (see Sambrook et al., 1999, supra) that recognizes and binds to the polypeptide.

[0057] As used herein, the term "expression vector" refers to a nucleic acid that provides all the necessary elements for expressing one or more structural genes contained in a host cell. Typically, an expression plasmid comprises a prokaryotic plasmid propagation unit, such as for Escherichia coli, which comprises an origin of replication and a selective marker, a eukaryotic selection marker, and one or more expression cassettes for expressing one or more target structural genes, each of which comprises a promoter, a structural gene, and a transcription terminator including a polyadenylation signal. Gene expression is usually placed under the control of a promoter, and such a structural gene is referred to as being "operably connected" to a promoter. Similarly, if a regulatory element regulates the activity of a core promoter, then the regulatory element and the core promoter are operably connected.

[0058] As used herein, the term "flavor" or "flavor profile" refers to the sensory perception of the components of taste, smell (aroma) and / or texture. The flavor profile of a composition is a quantitative profile of the relative intensities of all taste attributes exhibited. Such profiles are typically plotted as a histogram or radar plot. In certain embodiments, a flavor profile comprises one or more flavors that contribute to the sensory experience of a subject. In certain embodiments, modifying, altering, or changing the combination of stimuli in a flavor profile can alter the sensory experience of a subject.

[0059] As used herein, the term "flavor-modifying composition" refers to a composition that modulates (including enhancing, multiplying, potentiating, reducing, inhibiting or inducing) the taste, smell, flavor and / or texture of a natural or synthetic tastant, flavoring, taste profile, flavor profile and / or texture profile in a subject to which it is administered.

[0060] As used herein, the term "high intensity sweetener" refers to any synthetic or semi-synthetic sweetener or sweetener found in nature that is many times sweeter than sucrose (e.g., 20 times and more, 30 times and more, 50 times and more, or 100 times sweeter than sucrose).

[0061] As used herein, the term "sweetness" refers to a composition with equivalent sweetness. In general, the sweetness of a given composition is typically measured with reference to a sucrose solution. See "A Systematic Study of Concentration-Response Relationships of Sweeteners", G.E. DuBois, D.E. Walters, S.S. Schiffman, Z.S. Warwick, B.J. Booth, S.D. Pecore, K. Gibes, B.T. Carr and L.M. Brands, Sweeteners: Discovery, Molecular Design and Chemoreception, D.E. Walters, F.T. Orthoefer and G.E. DuBois, eds., American Chemical Society, Washington, D.C. (1991), pp. 261-276.

[0062] As used herein, the term "mouthfeel" refers to the sensory and tactile properties of a consumer product that are felt when the composition contacts the oral cavity and surfaces. Sensory and tactile properties include texture, thickness, consistency, and body.

[0063] As used herein, the term "organoleptic" or "organoleptic characteristics" refers to the sensations perceived by the five senses when consuming a consumable (e.g., a food or beverage). Thus, organoleptic quality relates to the taste and aroma as well as the color and texture of the consumable. Organoleptic properties are subjective and their impact varies from person to person. These organoleptic properties can be evaluated, for example, by a panel of trained or untrained individuals with the necessary sensory skills. Analytical methods can include, for example, differentiation / difference and descriptive analysis. In certain embodiments, the majority of individuals tested experience an improvement in the organoleptic property or therein.

[0064] As used herein, the term "ppm" means parts per million and is a weight relative parameter. Parts per million is micrograms per gram, such that a component present at 10 ppm is present at 10 micrograms of the particular component per 1 gram of aggregate mixture.

[0065] As used herein, the term "purity" refers to a substance that is substantially or essentially free of the components that normally accompany a compound found in its native state. Purity and homogeneity are typically determined using analytical chemistry techniques. In particular, in one embodiment, the compound is at least 85% pure, more preferably at least 90% pure, more preferably at least 95% pure, and most preferably at least 99% pure. In another embodiment, the compound is at least 90% pure, at least 91% pure, at least 92% pure, at least 93% pure, at least 95% pure, at least 95% pure, at least 96% pure, at least 97% pure, at least 98% pure, or at least 99% pure.

[0066] As used herein, "sensory experience" refers to a subject's sensory perception of a taste, taste profile, flavor, flavor profile, or texture profile.

[0067] As used herein, the term "acid" or "sour taste" refers to a taste that detects acidity. It is caused by hydrogen atoms, or ions. The more atoms there are in a food, the more sour it tastes. The sour taste of a substance is rated relative to dilute hydrochloric acid, which has a sourness index of 1. In contrast, the sourness index of tartaric acid is 0.7, the index of citric acid is 0.46, and the index of carbonic acid is 0.06. The reduction in sour taste can be expressed as a percentage of sour taste inhibition. In one embodiment, the taste modification composition of the present invention reduces the sour taste of a consumer product (e.g., a beverage) by at least about 5%, at least about 10%, at least about 15%, at least about 20%, or at least about 25% or more relative to a consumer product that does not contain the taste modification composition.

[0068] As used herein, the term "one or more steviol glycosides" refers to glycosides of steviol, including but not limited to naturally occurring steviol glycosides, such as rebaudioside A (Reb A), rebaudioside B (Reb B), rebaudioside C (Reb C), rebaudioside D (Reb D), rebaudioside E (Reb E), rebaudioside F (Reb F), rebaudioside G (Reb G), rebaudioside H (Reb H), rebaudioside I (Reb I), rebaudioside J (Reb J), rebaudioside K (Reb K), rebaudioside L (Reb L), rebaudioside M (Reb M), rebaudioside N (Reb N), rebaudioside O (Reb O), rebaudioside Q (Reb Q), rebaudioside R (Reb R), rebaudioside S (Reb S), rebaudioside T (Reb T), rebaudioside U (Reb U), rebaudioside V (Reb V), rebaudioside W (Reb W), rebaudioside Y (Reb Y), stevioside, steviol bioside, dulcoside A and rubusoside, etc., or synthetic or biosynthesized steviol glycosides, such as enzymatically glycosylated steviol glycosides, steviol glycoside products from bioconversion of steviol glycosides by biocatalysts, steviol glycosides fermented from recombinant microbial hosts capable of synthesizing steviol glycosides from scratch, and combinations thereof. In a specific embodiment, the steviol glycoside is a rebaudioside analog. In terms of sweetness, steviol glycosides are 40 to 300 times sweeter than sucrose, and are also characterized by being thermally stable, pH stable, and non-fermentable.

[0069] As used herein, the term "sucrose equivalence" refers to the sweetness of a non-sucrose composition relative to a sucrose reference. Typically, taste panelists are trained to detect the sweetness of a reference sucrose solution containing between 1%-15% sucrose (w / v). Other non-sucrose sweeteners are then tasted at a series of dilutions to determine the concentration of the non-sucrose sweetener that is as sweet (i.e., isosweet) as a given percentage of a sucrose reference.

[0070] The term "sugar-like characteristics" refers to any characteristics similar to those of sucrose, and includes, but is not limited to, maximum response, flavor profile, taste profile, time profile, adaptation behavior, mouthfeel, concentration / response function, tastants / and flavor / sweetness interactions, spatial pattern selectivity, and temperature effects. These characteristics are aspects in which the taste of sucrose is different from the taste of other compounds. As used herein, the term "sweetener composition" means a composition containing at least one sweet component and at least one other substance, such as a combination of another sweetener or additive.

[0071] As used herein, the term "sweetenable composition" means a substance that contacts the mouth of a human or animal, including a substance that is taken into the mouth and subsequently discharged from the mouth and a substance that is drunk, eaten, swallowed or otherwise ingested, and is safe for human or animal consumption when used within a generally acceptable range. For example, a beverage without a sweetener component is a class of sweetenable compositions. A sweetener composition comprising Reb X and erythritol can be added to an unsweetened beverage to provide a sweetened beverage. In another example, a beverage containing Reb M or Reb A is a class of sweetenable compositions. A sweetener composition comprising brazin can be added to the beverage containing Reb M or Reb A to provide a sweetened beverage.

[0072] As used herein, the term "sweetened composition" means a material containing a sweetenable composition and a sweetener or a sweetener composition.

[0073] As the term is generally used herein, the "sweetness recognition threshold concentration" is the lowest known concentration of a sweet compound that can be perceived by the human palate, typically about 1.0% sucrose equivalent (1.0% SE). The sweetness recognition threshold concentration can be readily determined by taste testing of increasing concentrations of a given enhancer in a given beverage matrix up to greater than 1.0% sucrose equivalent. A concentration that provides about 1.0% sucrose equivalent is considered the sweetness recognition threshold.

[0074] As used herein, the term "synergistic effect" refers to a situation where a combination of substances or compounds produces an activity (chemical and / or biological) that is greater than the sum of their individual activities. In one embodiment, the activity is sweetness.

[0075] As used herein, the term "taste" refers to a sensation or perception caused by activation or inhibition of receptor cells in a subject's taste buds. Taste buds are able to distinguish different tastes by detecting the interaction of different molecules or ions. Taste is considered to include sweet, sour, salty, bitter (the so-called "basic tastes"), as well as kokumi and umami.

[0076] As used herein, "texture profile" or "mouthfeel" refers to the physical and chemical interactions of a composition in the mouth. The texture profile of a composition can include one or more textures such as, for example, but not limited to, savory, lubricious, smooth, astringent, hardness, cohesiveness, stickiness, elasticity, viscosity, brittleness, chewiness, gummyness, moisture content, grittiness, smoothness, oiliness, and greasiness. In certain embodiments, a texture profile can include one or more texture characteristics of the same or different intensity.

[0077] Sweetener compositions / flavor modifying compositions

[0078] Disclosed herein are sweetener compositions and flavor-modifying compositions containing brazil (or its analogs) in each case. In certain embodiments, the sweetener compositions and flavor-modifying compositions alter (e.g., improve) one or more sensory experiences of a subject consuming the sweetener compositions and flavor-modifying compositions.

[0079] The flavor-modifying composition can modify (e.g., enhance, inhibit, or change) the taste, aroma, and / or texture of a given composition (e.g., a consumable). In certain embodiments, the flavor-modifying composition modifies (e.g., enhances, inhibits, or changes) one or more specific flavors. In another embodiment, the flavor-modifying composition modifies (e.g., enhances, inhibits, or changes) a given texture. In certain embodiments, the flavor-modifying composition modifies (e.g., enhances, inhibits, or changes) one or more given flavors, as well as texture.

[0080] The flavor-modifying composition can be sweetened or unsweetened. Thus, in some embodiments, the addition of a flavor-modifying composition can both serve as the addition of a flavor modifier and further provide sweetness to the composition selected for taste adjustment. The addition of a sweetened flavor-modifying composition can be used to supplement or replace the addition of another sweetening composition.

[0081] The sweetener compositions and flavor modification compositions disclosed herein contain bulazines, variants or their analogs. In certain embodiments, bulazines (or their analogs) are the only sweetening component in the sweetener composition or flavor modification composition. In certain embodiments, the sweetener composition or flavor modification composition further comprises one or more additional sweetening components. In specific embodiments, the one or more sweetening components include steviol glycosides (e.g., Reb M, Reb A) and HFCS.

[0082] Brazzein is a small sweet protein originally isolated from the fruit of the West African plant Baillon Pentadiplandra brazzeana. (Ming D et al., FEBS Lett. [Communication of the Federation of European Biochemical Societies] (1994) 355:106-108). It is a monomeric protein with a molecular weight of 6.5 kd. As a member of the Csβα fold family, it contains four disulfide bonds, which make its structure highly thermally and pH stable. In particular, in the pH range of 2.5-8, the sweet taste of brazzein still exists after incubation at 98°C for 2h and at 80°C for 4.5h. It is also water-soluble (>50mg / mL). It is a highly soluble protein (greater than 50g / L) with an isoelectric point of 5.4

[0083] Although only two forms are present in ripe fruit, at least three forms of the protein are known. The major form (pGlu-brazein) (about 80%) contains a pyroglutamic acid (pGlu) residue at its N-terminus, and the minor form (des-pGlu1-brazein, commonly referred to as "brazein") (about 20%) lacks this residue.

[0084] The 53-amino acid sequence of wild-type brazelain (minor form) is shown in SEQ ID NO: 1. The compositions disclosed herein may contain wild-type brazelain or a naturally occurring variant or a recombinant analog thereof.

[0085] The minor form is almost twice as sweet as the major form. In particular, bulazein is about 500 to 2000 times more potent than sucrose. Its taste is more similar to sucrose than the taste of thaumatin, another sweet protein. In particular embodiments, a sweetener composition or flavor-modifying composition disclosed herein comprises a minor form of wild-type bulazein.

[0086] Brazin is thought to be involved in a multi-point binding interaction with the human sweet taste receptor, a heterodimeric G protein-coupled receptor composed of type 1 taste receptor 2 (T1R2) and type 1 taste receptor 3 (T1R3) subunits. (Assadi-Porter FM, et al., J Mol. Bio. (2010) 14; 398(4): 584-99). The brazin binding site is distinct from the sucrose binding site.

[0087] In a specific embodiment, the sweetener composition and flavor modification composition disclosed herein include a bulaziyin analog. In one embodiment, a bulaziyin analog is different from a wild-type bulaziyin at at least one amino acid position, more particularly, at one, two, three or more amino acid positions. In a specific embodiment, the wild-type amino acid sequence of a bulaziyin analog is conserved at site 1 (cyclic residue R43), site 2 (N-terminal and C-terminal regions / residues E36 and cyclic 33) or site 3 (cyclic residues 9-19). In a specific embodiment, a bulaziyin analog is selected from the group comprising: Asp40Ala, Asp40Ly, Glu41Ala, Lys42Ala. Asp50Lys, Tyr54Trp, Asp29Ala / Glu41Lys, Asp29Asn / Glu41Lys, Asp29Lys / Glu41Lys.

[0088] In one embodiment, the sweetener compositions and flavor-modifying compositions disclosed herein comprise a brazelaine analog having a sweetness equal to or greater than wild-type brazelaine.

[0089] In one embodiment, the sweetener compositions and flavor-modifying compositions disclosed herein comprise a brazelaine analog having a stability equal to or greater than that of wild-type brazelaine.

[0090] Brazia (or its analog) suitable for use in the compositions disclosed herein (e.g., sweetener compositions, flavor and / or taste modification compositions, consumer products) can be produced in any suitable manner. Representative production methods include extraction, chemical synthesis (i.e., solid state synthesis) or recombinant production (i.e., in vivo production or in vitro production).

[0091] In one embodiment, the brazein used in the compositions disclosed herein is isolated from the edible fruit of the Baillon tree, as described, for example, in WO 97 / 94 / 19467. The brazein content in the pulp between the pericarp and the seeds is between about 0.2% and 0.05% by weight.

[0092] In another embodiment, the brazyne (or its analogue) used in the compositions disclosed herein is produced in vivo. In one embodiment, the nucleic acid coding sequence (optionally optimized) of the minor form of brazyne (or its analogue) isolated from the fruit of the Burrata agrocybe is introduced into a suitable vector and then cloned into a host cell in an appropriate growth system / environment to express the protein in a recombinant manner. In one embodiment, the host cell is a prokaryotic cell or a eukaryotic cell.

[0093] In certain embodiments, bulazines (or their analogs) are produced in a cell-free system, i.e., synthesized in vitro. A cell-free system is a system capable of translating polynucleotides into peptides, polypeptides, and / or proteins that do not occur in intact cells. Cell-free systems that can be used to produce bulazines for use in the compositions described herein include, but are not limited to, protein expression components from eukaryotic, prokaryotic, and / or viral sources. For example, as used herein, a cell-free system can include mammalian and / or bacterial protein expression systems derived from mammalian and / or bacterial lysates.

[0094] In certain embodiments, the recombinant braaziein is produced by the transgenic mammal, ie, in milk.

[0095] In certain embodiments, brazepine (or its analogs) can be expressed as a fusion protein. In certain embodiments, the fusion protein comprises brazepine and a fusion tag, such as an affinity tag (e.g., a His tag) or a solubility enhancing tag (e.g., a GST tag).

[0096] The expression of brazil (or its analogs) can be stable or transient. In a stable expression system, the foreign DNA is integrated into the chromosome or as an episome (a separate nuclear DNA fragment) and passed on to the progeny of the host cell.

[0097] In a cell-based system, the first step in the protein purification process is to extract the protein from the cells by lysing or opening the cells. Any suitable cell lysis method can be used, for example, mechanical disruption, chemical decomposition, freeze-thaw cycles, or enzymatic digestion. The protein can then be purified by any suitable protein purification method, for example, affinity chromatography, ion exchange chromatography, filtration, electrophoresis, hydrophobic interaction chromatography, gel filtration chromatography, reverse phase chromatography, concanavalin A chromatography, chromatofocusing, and differential precipitation or dissolution.

[0098] The yield of in vivo production of bulazein (or its analog) may vary. In a specific embodiment, bulazein comprises at least about 1% of total cellular protein. In a specific embodiment, bulazein comprises between about 1% and about 5% of total cellular protein. In another embodiment, bulazein comprises between about 5% and about 10% of total cellular protein. In another embodiment, bulazein comprises between 10% and about 20% of total cellular protein. In certain embodiments, bulazein comprises more than 20% of total cellular protein.

[0099] In another specific embodiment, bulaziein is purified to provide a yield between about 1 mg / mL and about 200 mg / mL, more particularly, a yield between about 20 mg / mL and about 180 mg / mL, between about 40 mg / mL and about 160 mg / mL, between about 60 mg / mL and about 140 mg / mL, or between about 80 mg / mL and about 120 mg / mL. In one embodiment, bulaziein is purified to provide a yield of about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 125 mg / mL, about 150 mg / mL, about 175 mg / mL, or about 200 mg / L or more. Optionally, bulaziein is produced as a fusion protein that further comprises a tag, and the above yields reflect both purification and removal of the tag.

[0100] In certain embodiments, brazein (or its analog) is substantially pure. In one embodiment, brazein (or its analog) is at least about 80% pure, at least about 85% pure, at least about 90% pure, at least about 95% pure, or at least about 99% pure. In another embodiment, brazein (or its analog) is about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% pure.

[0101] In specific embodiments, the bulazines (or their analogs) used in the compositions disclosed herein are produced in a yeast expression system (i.e., yeast-derived bulazines or their analogs), for example, in Kluyveromyces (e.g., K. lactis), Lactococcus (e.g., L. lactis), Lactobacillus, Saccharomyces (e.g., S. cerevisiae), Pischia (e.g., P. pastoris), Hansenula (e.g., H. polymorpha), or Yarrowia (e.g., Y. lipolytica).

[0102] In another specific embodiment, the bulazein (or its analog) used in the compositions disclosed herein is produced in a bacterial expression system (i.e., bulazein or its analog derived from bacteria), for example, in Escherichia coli or Bacillus subtilis. In one embodiment, bulazein (or its analog) is not produced in Escherichia coli.

[0103] In another specific embodiment, the brazepine (or its analog) used in the compositions disclosed herein is produced in an insect expression system (i.e., insect-derived brazepine or its analog), for example, in baculovirus-infected or unlysed insect cells (e.g., sf9, Sf21).

[0104] In another embodiment, the brazyne (or its analog) used in the compositions disclosed herein is produced in a fungal expression system (i.e., brazyne or its analog derived from a fungus), for example, in Chrysosporium, Thielavia, Talaromyces, Thermomyces, or Thermoascus.

[0105] In another embodiment, brazepine (or its analog) is produced in an algal expression system (ie, brazepine or its analog is derived from algae).

[0106] In another embodiment, brazepine (or its analog) is produced in a plant expression system (i.e., brazepine or its analog is derived from a plant), for example, in maize, tobacco, potato, strawberry or sugar cane. In one embodiment, the plant expression system is a plant cell culture expression system.

[0107] In a specific embodiment, brazein (or its analog) is produced in maize, and more specifically, maize seeds. According to this embodiment, brazein (or its analog) can be used as brazein-containing germ meal.

[0108] In yet another embodiment, brazepine (or its analog) is produced in a mammalian expression system (i.e., brazepine or its analog derived from a mammal), for example, in Chinese hamster ovary (CHO) cells, human embryonic kidney (HEK), COS and baby hamster kidney (BHK) cells.

[0109] Alternatively, brazyne (or its analogs) used in the compositions disclosed herein can be produced in vitro using a cell-free expression system (eg, E. coli S30 extract).

[0110] The amount of brazein (or its analog) in the sweetener compositions and flavor-modifying compositions disclosed herein can vary. In one embodiment, brazein (or its analog) is present above its sweetness threshold concentration.

[0111] In one embodiment, brazil (or its analog) is present in the sweetener composition or flavor-modifying composition in any amount to impart the desired sweetness when the sweetener composition or flavor-modifying composition is added to a consumable (e.g., a beverage) either alone or in combination with one or more additional sweet components (e.g., steviol glycosides, HFCS) present in the sweetener composition or flavor-modifying composition (i.e., before such composition is added to the consumable).

[0112] In particular embodiments, the desired sweetness of the consumable is isosweet to a sucrose sweetened consumable having a sweetness of at least about 8 degrees Brix, such as, for example, about 9 degrees Brix, about 10 degrees Brix, about 11 degrees Brix, about 12 degrees Brix, about 13 degrees Brix, about 14 degrees Brix, or about 15 degrees Brix.

[0113] In another embodiment, the desired sweetness of the consumable is isosweet to a sucrose sweetened consumable having a sweetness from about 10 to about 15 Brix, such as, for example, from about 10 to about 14 Brix, from about 10 to about 13 Brix, from about 10 to about 12 Brix, from about 10 to about 11 Brix, from about 11 to about 15 Brix, From about 11 degrees Brix to about 14 degrees Brix, from about 11 degrees Brix to about 13 degrees Brix, from about 11 degrees Brix to about 12 degrees Brix, from about 12 degrees Brix to about 15 degrees Brix, from about 12 degrees Brix to about 14 degrees Brix, from about 12 degrees Brix to about 13 degrees Brix, from about 13 degrees Brix to about 15 degrees Brix, from about 13 degrees Brix to about 14 degrees Brix and from about 14 degrees Brix to about 15 degrees Brix.

[0114] In one embodiment, brazil (or its analog) is present in a sweetener composition or flavor-modifying composition in an amount that enhances the sweetness of the consumable to which it is added by about 1.0% (w / v) sucrose equivalence (SE) or more, either alone or in combination with one or more additional sweet components (e.g., steviol glycosides, HFCS) present in the sweetener composition or flavor-modifying composition (i.e., before such composition is added to the consumable).

[0115] In particular embodiments, brazil (or its analog) is present in a sweetener composition in an amount that enhances the sweetness of the consumable to which it is added from about 1.0% to about 3.0% (w / v) sucrose equivalence (SE), such as, for example, about 1.1%, about 1.2%, about 1.3%, about 1.4%, about 1.5%, about 1.6%, about 1.7%, about 1.8%, about 1.9%, about 2.0%, about 2.1%, about 2.2%, about 2.3%, about 2.4%, about 2.5%, about 2.6%, about 2.7%, about 2.8%, about 2.9% or about 3.0% sucrose equivalence, either alone or in combination with one or more additional sweet components (e.g., steviol glycosides, HFCS) present in a sweetener composition or a flavor-modifying composition (i.e., before such composition is added to a consumable).

[0116] In another specific embodiment, brazil (or its analog) is present in a sweetener composition or a flavor-modifying composition in an amount that enhances the sweetness of the consumable to which it is added by about 3.0% to about 5% (w / v) sucrose equivalence (SE), e.g., about 3.1%, about 3.2%, about 3.3%, about 3.4%, about 3.5%, about 3.6%, about 3.7%, about 3.8%, about 3.9%, about 4.0%, about 4.1%, about 4.2%, about 4.3%, about 4.4%, about 4.5%, about 4.6%, about 4.7%, about 4.8%, about 4.9%, or about 5.0%, either alone or in combination with one or more additional sweet components (e.g., steviol glycosides, HFCS) present in a sweetener composition or a flavor-modifying composition (i.e., before such composition is added to a consumable).

[0117] The sweetness of a given composition is usually measured with reference to a sucrose solution. See generally "A Systematic Study of Concentration-Response Relationships of Sweeteners," GE DuBois, DE Walters, SS Schiffman, ZS Warwick, BJ Booth, SD Pecore, K. Gibes, BTC Carr, and LM Brands, in Sweeteners: Discovery, Molecular Design and Chemoreception, DE Walters, FT Orthoefer, and GE DuBois, eds., American Chemical Society, Washington, DC (1991), pp. 261-276.

[0118] The amount of sucrose in a reference solution can be described in degrees Brix (°Bx). One degree Brix is ​​1 gram of sucrose in 100 grams of solution and expresses the strength of the solution (strictly speaking, by mass) as a percentage by weight (% w / w).

[0119] In one embodiment, a sweetener composition is provided, the sweetener composition containing brazein (or its analogue), when added to a consumable alone or with one or more sweet components (e.g., steviol glycosides, HFCS) present in a sweetener composition or a flavor-modifying composition or a consumable, the amount of brazein (or its analogue) is effective to provide a sweetness equivalent to sugar of from about 1 to about 12 degrees Brix (e.g., from about 2 to about 9 degrees Brix, from about 3 to about 8 degrees Brix, from about 4 to about 7 degrees Brix, or about 5 degrees Brix). In another embodiment, brazein (or its analogue) is present in an amount effective to provide a sweetness equivalent to about 10 degrees Brix when added to a sweetenable composition alone or with one or more sweet components (e.g., steviol glycosides, HFCS) present in a sweetener composition, a flavor-modifying composition, or a consumable to which it is added.

[0120] The sweetness of non-sucrose sweeteners can also be measured relative to a sucrose reference by determining the sucrose equivalence of the non-sucrose sweetener. Typically, taste panelists are trained to detect the sweetness of reference sucrose solutions containing between 1%-15% sucrose (w / v). Other non-sucrose sweeteners are then tasted at a series of dilutions to determine the concentration of the non-sucrose sweetener that is as sweet as a given percentage of the sucrose reference. For example, if a 1% solution of a sweetener is as sweet as a 10% sucrose solution, the sweetener is said to be 10 times more potent than sucrose.

[0121] In one embodiment, the amount of brazein (or its analog) present in the sweetener composition or flavor modification composition disclosed herein is any amount that contributes to one or more improved sensory properties of the consumable (e.g., beverage) to which the sweetener composition or flavor modification composition is added. In a particular embodiment, the improved sensory properties are related to taste. In one embodiment, improving one or more sensory properties can improve the taste profile. The overall taste profile of the composition is the interaction of several different tastes (e.g., sweetness, sourness, etc.).

[0122] Examples of improved organoleptic properties may include, for example, reduced bitterness, reduced astringency and licorice, slowed sweetness onset, reduced lingering sweetness, reduced lingering bitterness, reduced bitter aftertaste, reduced metallic aftertaste, reduced chemical and synthetic aftertaste, and combinations thereof. In particular embodiments, the term "improved organoleptic properties" means that a sweetened or flavor-modified composition (e.g., a beverage) will have one or more improved organoleptic properties for most users. The improvement may be expressed qualitatively or quantitatively, for example, as a percentage improvement.

[0123] Improved sensory properties can be measured by or using technology such as a taste sensing system (TSS), a term that refers to an analytical sensory array unit (e.g., electrochemical, gravimetric, optical, or biosensor) that can detect specific substances. Sliwi′nska, M et al. J. Agric. Food Chem. (2014), 62, 1423-1448.

[0124] In certain embodiments, brazil (or its analog) is present in a sweetener composition or flavor-modifying composition alone or together with one or more sweet components (e.g., steviol glycosides, HFCS) in a sweetener composition or flavor-modifying composition (i.e., before it is added to a consumable) in any amount that reduces, inhibits, or masks the bitter taste of a consumable (e.g., a beverage) to which the sweetener or flavor-modifying composition is added, as compared to a consumable to which the sweetener or flavor-modifying composition has not yet been added.

[0125] In a particular embodiment, when combined with one or more sweet components (e.g., steviol glycosides, HFCS) in a sweetener composition or flavor modification composition (i.e., before adding it to a consumable), brazil (or its analog) is present in a sweetener composition or flavor modification composition in an amount that reduces the bitterness of the consumable (e.g., beverage) to which it is added by at least about 5%, at least about 10%, at least about 15%, at least about 20%, or at least about 25% or more. In one embodiment, most subjects experienced a reduction in bitterness. Compared to a consumable to which a sweetener or flavor modification composition has not yet been added.

[0126] In a specific embodiment, when combined with one or more sweet components (e.g., steviol glycosides, HFCS) in a sweetener composition or a flavor modification composition (i.e., before it is added to a consumer's product), bulazines (or its analogs) are present in a sweetener composition or a flavor modification composition in any amount to reduce the bitter aftertaste of the consumer's product (e.g., beverage) to which a sweetener or a flavor modification composition has been added. In a specific embodiment, bulazines (or its analogs) are present in a sweetener composition or a flavor modification composition in an amount that reduces the bitter aftertaste of the consumer's product (e.g., beverage) to which it is added by at least about 5%, at least about 10%, at least about 15%, at least about 20%, or at least about 25% or more. In one embodiment, most subjects have experienced a reduction in bitter aftertaste. Compared with a consumer's product to which a sweetener or a flavor modification composition has not yet been added.

[0127] In another embodiment, bulazie (or its analog) is present in the sweetener composition or the taste modifier composition with any amount of the sweetness retention of the consumer's goods (for example, beverage) to which the sweetener or taste modifier composition has been added. Sucrose exhibits sweetness, wherein when swallowing food or beverage, the maximum response is quickly perceived, and the perceived sweetness disappears relatively quickly. In contrast, the sweetness of substantially all high-efficiency sweeteners reaches its maximum response more slowly than sucrose, and their subsequent intensity decline is also slower than sucrose. This decline in sweetness is commonly referred to as "sweetness retention", and is the main limitation of the high-efficiency sweeteners including NHPS. The slow start of sweetness is also a problem. However, in general, the retention of sweetness is a more significant problem. Therefore, preferred embodiments of the present invention exhibit a significant reduction in the retention of sweetness.

[0128] In a specific embodiment, alone or in combination with one or more sweet components (e.g., steviol glycosides, HFCS) in a sweetener composition or a flavor modification composition (before adding it to a consumer's product), brazil (or its analog) is present in a sweetener composition or a flavor modification composition in an amount that reduces the sweetness retention of the consumer's product (e.g., beverage) to which it is added by at least about 5%, at least about 10%, at least about 15%, at least about 20%, or at least about 25% or more. In one embodiment, most subjects perceive a reduction in the sweetness retention. In a specific embodiment, compared with a consumer's product to which a sweetener composition or a flavor composition has not yet been added.

[0129] In certain embodiments, sweetener composition or local flavor and / or taste modification composition contain one or more other sweeteners.In one embodiment, other sweetener exists higher than its sweetness threshold concentration.In certain embodiments, the sweetener composition containing Braqiyin and these one or more other sweeteners synergistically enhances the sweetness of the consumer's goods having added sweetener composition.In one embodiment, the sweetness of consumer's goods is enhanced in a manner unexpected by those skilled in the art.

[0130] The additional sweetener can be any type of sweetener, for example, a natural sweetener, a non-natural sweetener, or a synthetic sweetener.

[0131] As used herein, the term "high intensity sweetener" refers to any synthetic or semi-synthetic sweetener or a sweetener found in nature. High intensity sweeteners are compounds or mixtures of compounds that are sweeter than sucrose. High intensity sweeteners are typically many times sweeter than sucrose (e.g., 20 times and more, 30 times and more, 50 times and more, or 100 times or more sweeter than sucrose).

[0132] In at least one embodiment, the at least one additional sweetener is selected from natural sweeteners other than stevia sweeteners. In another embodiment, the at least one additional sweetener is selected from synthetic high-potency sweeteners (SHPS).

[0133] In certain embodiments, the one or more additional sweeteners may be natural high-potency sweeteners (NHPS). Suitable natural high-potency sweeteners include, but are not limited to, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, dulcoside A, dulcoside B, sweet leaf rubusoside, stevia, stevioside, mogroside IV, mogroside V, Luo Han Guo (Luo Han Guo), and styracifolia. Guo) sweetener, simonoside, monatin and its salts (monatin SS, RR, RS, SR), curculin, glycyrrhizic acid and its salts, thaumatin, monellin, mabinlin, brazin, hernandulcin, phyllotoxin, phloroglucin, phloroglucin, trilobatin, baiyunoside, osladin, polypodoside A, pterocaryoside A, pterocaryoside B, mukurozioside, phlomisoside I, periandrin I, abrusoside A, stevioside and cyclocarioside I. Natural high potency sweeteners can be provided as pure compounds or alternatively as part of an extract. For example, rebaudioside A can be provided as a separate compound or as part of a stevia extract.

[0134] In one embodiment, the one or more additional sweeteners are selected from the group consisting of rebaudioside M, rebaudioside A, simonoside I and mogroside V.

[0135] In certain embodiments, the sweetener compositions and / or flavor-modifying compositions of the present invention comprise brazyne (or an analog thereof) and simonoside I.

[0136] In another particular embodiment, the sweetener composition and / or flavor modifying composition of the present invention comprises brazyne (or an analog thereof) and mogroside V.

[0137] In another embodiment, the one or more additional sweeteners are selected from the group consisting of rebaudioside D, rebaudioside N, rebaudioside O, rebaudioside E, stevia monoside, steviol bioside, rubusoside, dulcoside B, dulcoside A, rebaudioside B, rebaudioside G, stevioside, rebaudioside C, rebaudioside F, rebaudioside I, rebaudioside H, rebaudioside L, rebaudioside K, rebaudioside J, rebaudioside M2, rebaudioside D2, rebaudioside S, rebaudioside T, rebaudioside U, rebaudioside V, rebaudioside W, rebaudioside Z1, rebaudioside Z2, rebaudioside IX, enzymatically glucosylated steviol glycosides, and combinations thereof.

[0138] In another embodiment, the one or more additional sweeteners are selected from the group consisting of mogroside IA, mogroside IE, 11-oxomogroside IA, mogroside II, mogroside II A, mogroside II B, mogroside II E, 7-oxomogroside II E, mogroside III, mogroside IIIe, 11-deoxymogroside III, mogroside IV, 11-oxomogroside IV, 11-oxomogroside IV A, 11-deoxymogroside V, 7-oxomogroside V, 11-oxomogroside V, isomogroside V, mogroside VI, mogroside alcohol, 11-oxomogroside alcohol, the 1,6-alpha isomer of simanoside I, monk fruit extract, and combinations thereof.

[0139] In particular embodiments, the one or more additional sweeteners are rebaudioside M (13-[2-O-β-D-glucopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy] ent-kaur-16-ene-19-oleic acid-[2-O-β-D-glucopyranosyl-3-O-β-D-pyrro ...

[0140]

[0141] Reb M can be provided in purified or unpurified form (i.e., as part of a naturally occurring mixture containing Reb M). In one embodiment, Reb M can be obtained from a Stevia extract by any suitable purification method. Suitable purification methods are known in the art and include, but are not limited to, column chromatography, recrystallization, phase separation, extraction, high performance liquid chromatography, and combinations thereof.

[0142] In another embodiment, the additional sweetener is a steviol glycoside composition. An exemplary steviol glycoside composition is A95, which contains primarily reb D and reb M and a small amount of one or more of the following: Reb E, Reb O, Reb N, Reb A, stevioside, Reb C, and Reb B. Methods for obtaining A95 are provided in WO 2017 / 059414, which is incorporated herein by reference. An exemplary A95 blend is provided in Example 7 herein.

[0143] The amount of Reb M in a sweetener composition or taste-modifying composition can vary. In one embodiment, Reb M is present in the sweetener composition in any amount that imparts a desired sweetness when the sweetener composition is added to a consumable (e.g., a beverage). In a particular embodiment, the desired sweetness of the consumable is greater than about 10 degrees Brix.

[0144] In one embodiment, the sweetener composition contains Reb M in an amount effective to provide a sweetness equivalent to from about 1 to about 12 degrees Brix when added to a consumable (e.g., a beverage), such as, for example, from about 2 to about 9 degrees Brix, from about 3 to about 8 degrees Brix, from about 4 to about 7 degrees Brix, or about 5 degrees Brix.

[0145] In particular embodiments, Reb M is present in an effective amount to provide a sucrose equivalency (SE) of about 8 or less, such as, for example, about 7, about 6.5, about 6, about 5.5, or about 5 SE.

[0146] In another specific embodiment, Reb M is present in an effective amount to provide a sucrose equivalence value of about 8 or greater, such as, for example, about 9, about 9.5, about 10, about 10.5, about 11, about 11.5, about 12, about 12.5, about 13, about 13.5, about 14, about 14.5, or about 15.

[0147] In one embodiment, Reb M is present in the flavor-modifying composition in any amount that imparts a desired flavor when the flavor-modifying composition is added to a flavor-modifiable composition (e.g., a beverage). In a particular embodiment, the desired flavor is a more sugar-like temporal or taste profile.

[0148] In one embodiment, Reb M and brazil produce a synergistic effect, such as synergistic sweetness, i.e., the sweetness of the combination is greater than the sum of the individual sweeteners. In a particular embodiment, Reb M and brazil produce an effect that would be unexpected to one skilled in the art.

[0149] Reb M can be provided in purified form or as a component of a mixture containing Reb M and one or more additional components. In one embodiment, Reb X is provided as a component of a mixture. In a particular embodiment, the mixture is a Stevia extract. The Stevia extract can contain Reb M in an amount ranging from about 5% to about 100% by dry weight, such as, for example, from about 10% to about 100%, from about 20% to about 100%, from about 30% to about 100%, from about 40% to about 100%, from about 50% to about 100%, from about 60% to about 100%, from about 70% to about 100%, from about 80% to about 100%, and from about 90% to about 100%. In still further embodiments, the Stevia extract contains Reb M in an amount greater than about 90% by dry weight, e.g., greater than about 91%, greater than about 92%, greater than about 93%, greater than about 94%, greater than about 95%, greater than about 96%, greater than about 97%, greater than about 98%, and greater than about 99%.

[0150] In one embodiment, Reb M is provided as a component of a steviol glycoside mixture (i.e., a mixture of steviol glycosides), wherein the remainder of the mixture other than the Reb M portion consists entirely of steviol glycosides. The properties of steviol glycosides are known in the art and include, but are not limited to, steviol, steviol monoside, rubososide, steviol bioside, stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, and dulcoside A. The steviol glycoside mixture may contain from about 5% to about 100% Reb M by dry weight. For example, the steviol glycoside mixture may contain from about 10% to about 100%, from about 20% to about 100%, from about 30% to about 100%, from about 40% to about 100%, from about 50% to about 100%, from about 60% to about 100%, from about 70% to about 100%, from about 80% to about 100%, and from about 90% to about 100% Reb M by dry weight. In still other embodiments, the steviol glycoside mixture may contain greater than about 90%, e.g., greater than about 91%, greater than about 92%, greater than about 93%, greater than about 94%, greater than about 95%, greater than about 96%, greater than about 97%, greater than about 98%, and greater than about 99% Reb M by dry weight.

[0151] Reb M80 refers to a stevia extract or steviol glycoside composition having about 80% Reb M by weight.

[0152] In particular embodiments, the one or more additional sweeteners is rebaudioside A.

[0153] Reb A can be provided in purified or unpurified form (i.e., as part of a naturally occurring mixture containing Reb A). In one embodiment, Reb A can be obtained from a Stevia extract by any suitable purification method. Suitable purification methods are known in the art and include, but are not limited to, column chromatography, recrystallization, phase separation, extraction, high performance liquid chromatography, and combinations thereof.

[0154] Reb A can be provided in purified form or as a component of a mixture containing Reb A and one or more additional components. In one embodiment, Reb A is provided as a component of a mixture. In a particular embodiment, the mixture is a Stevia extract. The Stevia extract can contain Reb A in an amount ranging from about 5% to about 100% by dry weight, such as, for example, from about 10% to about 100%, from about 20% to about 100%, from about 30% to about 100%, from about 40% to about 100%, from about 50% to about 100%, from about 60% to about 100%, from about 70% to about 100%, from about 80% to about 100%, and from about 90% to about 100%. In yet further embodiments, the Stevia extract contains Reb A in an amount greater than about 90% by dry weight, e.g., greater than about 91%, greater than about 92%, greater than about 93%, greater than about 94%, greater than about 95%, greater than about 96%, greater than about 97%, greater than about 98%, and greater than about 99%.

[0155] In one embodiment, Reb A is provided as a component of a steviol glycoside mixture (i.e., a mixture of steviol glycosides), wherein the remainder of the mixture other than the Reb A portion consists entirely of steviol glycosides. The steviol glycoside mixture may contain from about 5% to about 100% Reb A by dry weight. For example, the steviol glycoside mixture may contain from about 10% to about 100%, from about 20% to about 100%, from about 30% to about 100%, from about 40% to about 100%, from about 50% to about 100%, from about 60% to about 100%, from about 70% to about 100%, from about 80% to about 100%, and from about 90% to about 100% Reb A by dry weight. In yet other embodiments, the steviol glycoside mixture may contain greater than about 90%, e.g., greater than about 91%, greater than about 92%, greater than about 93%, greater than about 94%, greater than about 95%, greater than about 96%, greater than about 97%, greater than about 98%, and greater than about 99% Reb A by dry weight.

[0156] The amount of Reb A in the sweetener composition or taste-modifying composition can vary. In one embodiment, Reb A is present in the sweetener composition in any amount that imparts the desired sweetness when the sweetener composition is added to a sweetenable composition. In a particular embodiment, the desired sweetness of the sweetened composition is greater than about 10 degrees Brix.

[0157] In one embodiment, Reb A and brazil produce a synergistic effect, such as synergistic sweetness, i.e., the sweetness of the combination is greater than the sum of the individual sweeteners. In certain embodiments, Reb A and brazil produce effects that would be unexpected to one skilled in the art.

[0158] In particular embodiments, Reb A is present in an effective amount to provide a sucrose equivalence (SE) of about 8 or less, such as, for example, about 7, about 6.5, about 6, about 5.5, or about 5 SE.

[0159] In another specific embodiment, Reb A is present in an effective amount to provide a Sucrose Equivalence (SE) of about 8 or greater, such as, for example, about 9, about 9.5, about 10, about 10.5, about 11, about 11.5, about 12, about 12.5, about 13, about 13.5, about 14, about 14.5, or about 15.

[0160] In another specific embodiment, Reb A is present in an effective amount to provide a sucrose equivalence value of greater than about 8 SE, e.g., about 9 SE, about 9.5 SE, about 10 SE.

[0161] In one embodiment, Reb A is present in the flavor-modifying composition in any amount that imparts a desired taste when the taste-modifying composition is added to a taste-modifiable composition (e.g., a beverage). In a particular embodiment, the desired taste is a sugar-like taste.

[0162] In certain embodiments, braazein and Reb A produce a synergistic effect. In one embodiment, braazein and Reb A produce an effect that would be unexpected to one skilled in the art.

[0163] In another embodiment, the one or more additional sweeteners may be carbohydrate sweeteners. Non-limiting examples of suitable carbohydrate sweeteners include sucrose, fructose, glucose, erythritol, maltitol, lactitol, sorbitol, mannitol, xylitol, D-tagatose, trehalose, galactose, rhamnose, cyclodextrins (e.g., α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin), ribulose, threose, arabinose, xylose, lyxose, allose, altrose, mannose, idose, lactose, maltose, invert sugar, isotrehalose, neotrehalose, palatinose or isomaltulose, erythrose, deoxyribose, gulose, idose, talose, erythrulose, xylulose, psicose, turanose, cellobiose, glucosamine, mannosamine, fucose, fucose, glucuronic acid, gluconic acid, gluconolactone, abequose, galactose, Glycosamine, xylooligosaccharides (xylotriose, xylobiose, etc.), gentiooligosaccharides (gentiobiose, gentiotriose, gentiotetraose, etc.), galacto-oligosaccharides, sorbose, ketotriose (dihydroxyacetone), aldotrose (glyceraldehyde), nigrooligosaccharides, fructooligosaccharides (kestose, nysitose, etc.), maltotetraose, maltotriol, tetrasaccharides, manno-oligosaccharides, malto-oligosaccharides (maltotriose, maltotetraose, maltopentaose, maltohexaose sulfate, maltoheptaose, etc.), dextrin, lactulose, melibiose, raffinose, rhamnose, ribose, isomerized liquid sugars such as high fructose corn / starch syrup (HFCS / HFSS) (e.g., HFCS55, HFCS42 or HFCS90), coupling sugars, soybean oligosaccharides, glucose syrup, and combinations thereof.

[0164] In other embodiments, the at least one additional sweetener is a synthetic sweetener. As used herein, the phrase "synthetic sweetener" refers to any composition that is not naturally found in nature and has a sweetness effect greater than sucrose, fructose or glucose in terms of characteristics, and has lower calories. Non-limiting examples of synthetic high-efficiency sweeteners suitable for embodiments of the present disclosure include sucralose, acesulfame potassium, aspartame, alitame, saccharin, neohesperidin dihydrochalcone, cyclamates, neotame, advantame, glucosylated steviol glycosides (GSG), and combinations thereof.

[0165] Sweetener compositions can be tailored to obtain a desired calorie content. For example, sweetener compositions can be "high calorie" such that they impart a desired sweetness when added to a sweetenable composition such as, for example, a beverage and have about 120 calories per 8 ounce serving.

[0166] Sweetener compositions can be tailored to obtain a desired calorie content. For example, sweetener compositions can be "mid-calorie" such that they impart a desired sweetness when added to a sweetenable composition such as, for example, a beverage and have about 80 calories per 8 ounce serving.

[0167] For example, the sweetener compositions can be "low calorie" such that they impart a desired sweetness when added to a sweetenable composition (such as, for example, a beverage) and have less than 40 calories per 8 ounce serving.

[0168] In other embodiments, the sweetener compositions may be "zero calorie" such that they impart a desired sweetness when added to a sweetenable composition (such as, for example, a beverage) and have less than 5 calories per 8 ounce serving.

[0169] additive

[0170] In addition to bulazines (or its analogs) and optionally, one or more additional sweeteners (e.g., one or more steviol glycosides), the sweetener compositions or flavor modification compositions disclosed herein may optionally include additional additives described in detail below. In some embodiments, the sweetener compositions contain additives, including but not limited to carbohydrates, polyols, amino acids and their corresponding salts, polyamino acids and their corresponding salts, sugar acids and their corresponding salts, nucleotides, organic acids, inorganic acids, organic salts (including organic acid salts and organic base salts), inorganic salts, bitter compounds, flavoring agents and flavoring ingredients, astringent compounds, proteins or protein hydrolysates, surfactants, emulsifiers, flavonoids, alcohols, polymers, and combinations thereof. In some embodiments, additives are used to improve the time profile and flavor profile of the sweetener to provide a sweetener composition having a taste similar to sucrose.

[0171] In one embodiment, the sweetener composition or flavor modification composition contains one or more polyols. As used herein, the term "polyol" refers to a molecule containing more than one hydroxyl group. Polyols can be diols, triols or tetraols containing 2, 3 and 4 hydroxyls respectively. Polyols can also contain more than 4 hydroxyls, such as pentahydric alcohols, hexahydric alcohols, heptahydric alcohols etc. containing 5, 6 or 7 hydroxyls respectively. In addition, polyols can also be sugar alcohols, polyhydroxy alcohols or polyols as a reduced form of carbohydrates, in which the carbonyl (aldehyde or ketone, reducing sugar) has been reduced to a primary hydroxyl group or a secondary hydroxyl group.

[0172] In some embodiments, non-limiting examples of polyols include erythritol, maltitol, mannitol, sorbitol, lactitol, xylitol, isomalt, propylene glycol, glycerol (glycerine), threitol, galactitol, palatinose, reduced isomalt-oligosaccharides, reduced xylo-oligosaccharides, reduced gentio-oligosaccharides, reduced maltose syrup, reduced glucose syrup, and sugar alcohols or any other carbohydrates that can be reduced without adversely affecting the taste.

[0173] Suitable sweet taste improving amino acid additives include but are not limited to aspartic acid, arginine, glycine, glutamic acid, proline, threonine, theanine, cysteine, cystine, alanine, valine, tyrosine, leucine, arabinose, trans-4-hydroxyproline, isoleucine, asparagine, serine, lysine, histidine, ornithine, methionine, carnitine, aminobutyric acid (α-isomer, β-isomer and / or δ-isomer), glutamine, hydroxyproline, taurine, norvaline, sarcosine and its salt forms such as sodium salt or potassium salt or acid salt. Sweet taste improving amino acid additives can also be in D-configuration or L-configuration and in the monobasic, dibasic or ternary form of the same or different amino acids. In addition, if appropriate, amino acid can be α-, β-, γ- and / or δ-isomer. In certain embodiments, the combination of the above amino acid and its corresponding salt (for example, its sodium salt, potassium salt, calcium salt, magnesium salt or other alkali metal salt or alkaline earth metal salt, or acid salt) is also a suitable sweet taste improving additive. Amino acids can be natural or synthetic.Amino acids can also be modified.Modified amino acids refer to any amino acids (e.g., N-alkyl amino acids, N-acyl amino acids or N-methyl amino acids) in which at least one atom has been added, removed, substituted or a combination thereof.Non-limiting examples of modified amino acids include amino acid derivatives, such as trimethylglycine, N-methyl-glycine and N-methyl-alanine.As used herein, modified amino acids encompass both modified amino acids and unmodified amino acids.As used herein, amino acids also encompass peptides and polypeptides (e.g., dipeptides, tripeptides, tetrapeptides and pentapeptides), such as glutathione and L-alanyl-L-glutamine. Suitable sweet taste improving polyamino acid additives include poly-L-aspartic acid, poly-L-lysine (e.g., poly-L-α-lysine or poly-L-ε-lysine), poly-L-ornithine (e.g., poly-L-α-ornithine or poly-L-ε-ornithine), poly-L-arginine, other polymer forms of amino acids, and salt forms thereof (e.g., calcium salts, potassium salts, sodium salts or magnesium salts, such as monosodium L-glutamate). Sweet taste improving polyamino acid additives may also be in D- or L-configuration. In addition, if appropriate, the polyamino acid may be an α-, β-, γ-, δ- and ε-isomer. In some embodiments, a combination of the above polyamino acids and their corresponding salts (e.g., sodium salts, potassium salts, calcium salts, magnesium salts or other alkali metal salts or alkaline earth metal salts, or acid salts) is also a suitable sweet taste improving additive. The polyamino acids described herein may also include copolymers of different amino acids. The polyamino acids may be natural or synthetic. The polyamino acid may also be modified such that at least one atom is added, removed, substituted, or a combination thereof (e.g., N-alkyl polyamino acids or N-acyl polyamino acids). As used herein, polyamino acids encompass both modified polyamino acids and unmodified polyamino acids.For example, modified polyamino acids include, but are not limited to, polyamino acids having different molecular weights (MW), such as poly-L-α-lysine having a MW of 1,500, a MW of 6,000, a MW of 25,200, a MW of 63,000, a MW of 83,000, or a MW of 300,000.

[0174] Suitable sugar acid additives include, but are not limited to, aldonic acid, uronic acid, aldaric acid, alginic acid, gluconic acid, glucuronic acid, glucaric acid, galactaric acid, galacturonic acid, and salts thereof (e.g., sodium salt, potassium salt, calcium salt, magnesium salt or other physiologically acceptable salts), and combinations thereof.

[0175] Suitable nucleotide additives include, but are not limited to, inosine monophosphate ("IMP"), guanosine monophosphate ("GMP"), adenosine monophosphate ("AMP"), cytosine monophosphate (CMP), uracil monophosphate (UMP), inosine diphosphate, guanosine diphosphate, adenosine diphosphate, cytosine diphosphate, uracil diphosphate, inosine triphosphate, guanosine triphosphate, adenosine triphosphate, cytosine triphosphate, uracil triphosphate, alkali metal salts or alkaline earth metal salts thereof, and combinations thereof. Nucleotides described herein may also include nucleotide-related additives, such as nucleosides or nucleic acid bases (e.g., guanine, cytosine, adenine, thymine, uracil). In particular embodiments, nucleotides are present in the sweetener composition in an amount from about 5ppm to about 1,000ppm.

[0176] Suitable organic acid additives include any compound comprising a -COOH moiety, such as, for example, C2-C30 carboxylic acids, substituted hydroxy C2-C30 carboxylic acids, benzoic acid, substituted benzoic acid (e.g., 2,4-dihydroxybenzoic acid), substituted cinnamic acid, hydroxy acids, substituted hydroxybenzoic acids, substituted cyclohexyl carboxylic acids, tannic acid, lactic acid, tartaric acid, citric acid, gluconic acid, glucoheptonic acid, adipic acid, hydroxycitric acid, malic acid, fruitaric acid (a blend of malic acid, fumaric acid, and tartaric acid), fumaric acid, maleic acid, succinic acid, chlorogenic acid, salicylic acid, creatine, caffeic acid, bile acid, acetic acid, ascorbic acid, alginic acid, isoascorbic acid, polyglutamic acid, glucono delta lactone, and alkali metal salts or alkaline earth metal salt derivatives thereof. In addition, the organic acid additive may also be in the D-configuration or the L-configuration.

[0177] Suitable organic acid additive salts include, but are not limited to, sodium, calcium, potassium, and magnesium salts of all organic acids, such as citrates, malates, tartrates, fumarates, lactates (e.g., sodium lactate), alginates (e.g., sodium alginate), ascorbates (e.g., sodium ascorbate), benzoates (e.g., sodium benzoate or potassium benzoate), and adipates. The example of the sweet taste improving organic acid additive can be optionally substituted by at least one group selected from the following: hydrogen, alkyl, alkenyl, alkynyl, halogen, haloalkyl, carboxyl, acyl, acyloxy, amino, amido, carboxyl derivatives, alkylamino, dialkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfo, thiol, imine, sulfonyl, alkylthio, sulfinyl, sulfamoyl, carboxyalkoxy, formamido, phosphonyl, phosphinyl, phosphoryl, phosphino, thioester, thioether, anhydride, oxime, hydrazine, carbamoyl, phosphorus or phosphonate. In a particular embodiment, the organic acid additive is present in the sweetener composition in an amount from about 10ppm to about 5,000ppm.

[0178] Suitable inorganic acid additives include, but are not limited to, phosphoric acid, phosphorous acid, polyphosphoric acid, hydrochloric acid, sulfuric acid, carbonic acid, sodium dihydrogen phosphate, and alkali metal or alkaline earth metal salts thereof (eg, phytate Mg / Ca).

[0179] Suitable bitter compound additives include, but are not limited to, caffeine, quinine, urea, bitter orange oil, naringin, quassia, and salts thereof.

[0180] Suitable flavoring agents and flavoring ingredient additives include, but are not limited to, vanillin, vanilla extract, mango extract, cinnamon, citrus, coconut, ginger, viridiflorol, almonds, menthol (including mint-free menthol), grape skin extract, and grape seed extract. "Flavoring agent" and "flavoring ingredient" are synonymous and may include natural or synthetic substances or combinations thereof. Flavoring agents also include any other substances that impart flavor and may include natural or non-natural (synthetic) substances that are safe for humans or animals when used within the generally accepted range. Non-limiting examples of proprietary flavoring agents include Natural flavoring sweetness enhancer K14323( Darmstadt, Germany), Symrise TM Sweetness Natural Flavor Masking Agents 161453 and 164126 (Symrise TM , Holzminden, Germany), Natural Advantage TM Bitterness Blockers 1, 2, 9 and 10 (Natural Advantage TM, Freehold, New Jersey, USA) and Sucramask TM (Creative Research Management, Stockton, California, USA).

[0181] Suitable polymer additives include, but are not limited to, chitin, pectin, pectic acid, polyuronic acid, polygalacturonic acid, starch, food hydrocolloids or crude extracts thereof (e.g., Senegalese gum arabic, Fibergum TM ), gum arabic, carrageenan), poly-L-lysine (e.g., poly-L-α-lysine or poly-L-ε-lysine), poly-L-ornithine (e.g., poly-L-α-ornithine or poly-L-ε-ornithine), polypropylene glycol, polyethylene glycol, poly(ethylene glycol methyl ether), polyarginine, polyaspartic acid, polyglutamic acid, polyethyleneimine, alginic acid, sodium alginate, propylene glycol alginate, and sodium polyethylene glycol alginate, sodium hexametaphosphate and its salts, and other cationic polymers and anionic polymers.

[0182] Suitable protein or protein hydrolysate additives include, but are not limited to, bovine serum albumin (BSA), whey protein (including fractions or concentrates thereof, such as 90% instant whey protein isolate, 34% whey protein, 50% hydrolyzed whey protein, and 80% whey protein concentrate), soluble rice protein, soy protein, protein isolate, protein hydrolysate, reaction products of protein hydrolysate, glycoprotein and / or proteoglycans containing amino acids (e.g., glycine, alanine, serine, threonine, asparagine, glutamine, arginine, valine, isoleucine, leucine, norvaline, methionine, proline, tyrosine, hydroxyproline, etc.), collagen (e.g., gelatin), partially hydrolyzed collagen (e.g., hydrolyzed fish collagen), and collagen hydrolysate (e.g., porcine collagen hydrolysate).

[0183] Suitable surfactant additives include, but are not limited to, polysorbates (e.g., polyoxyethylene sorbitan monooleate (polysorbate 80), polysorbate 20, polysorbate 60), sodium dodecylbenzenesulfonate, dioctyl sulfosuccinate or sodium dioctyl sulfosuccinate, sodium lauryl sulfate, cetylpyridinium chloride (cetylpyridinium chloride), cetyltrimethylammonium bromide, sodium cholate, carbamoyl, choline chloride, sodium glycocholate, sodium taurodeoxycholate, lauroyl arginine ester, sodium stearoyl lactylate, sodium taurocholate, lecithin, sucrose oleate, sucrose stearate, sucrose palmitate, sucrose laurate, and other emulsifiers.

[0184] Suitable flavonoid additives are classified as flavonols, flavones, flavanones, flavan-3-ols, isoflavones, or anthocyanidins. Non-limiting examples of flavonoid additives include, but are not limited to, catechins (e.g., green tea extracts such as Polyphenon TM 60. Polyphenon TM 30 and Polyphenon TM 25 (Mitsui Norin Co., Ltd., Japan), polyphenols, rutin (e.g., enzyme-modified rutin Sanmelin TM AO (San-fi Gen FFI, Inc., Osaka, Japan), neohesperidin, naringin, neohesperidin dihydrochalcone, etc.

[0185] Suitable alcohol additives include, but are not limited to, ethanol.

[0186] Suitable astringent compound additives include, but are not limited to, tannic acid, europium chloride (EuCl3), gadolinium chloride (GdCl3), terbium chloride (TbCl3), alum, tannic acid, and polyphenols (e.g., tea polyphenols). In a particular embodiment, the astringent additive is present in an amount from about 10 ppm to about 5,000 ppm.

[0187] In certain embodiments, the sweetener composition or flavor modification composition comprises brazein (or its analog), optionally in combination with: one or more steviol glycosides (e.g., Reb M, Reb A); a polyol selected from erythritol, maltitol, mannitol, xylitol, sorbitol, and combinations thereof; and optionally at least one additional sweetener and / or functional ingredient. In certain embodiments, the polyol is erythritol. As described above, steviol glycosides (e.g., Reb M, Reb A) can be provided as a pure compound or as part of a stevia extract or steviol glycoside mixture. Steviol glycosides (e.g., Reb M, Reb A) can be present in a steviol glycoside mixture or a stevia extract in an amount from about 5% to about 100% by dry weight.

[0188] In certain embodiments, the sweetener composition or flavor modification composition comprises brazein (or its analog), optionally in combination with: one or more steviol glycosides (e.g., Reb M, Reb A); a carbohydrate sweetener selected from sucrose, fructose, glucose, maltose, and combinations thereof; and optionally at least one additional sweetener and / or functional ingredient. As described above, steviol glycosides (e.g., Reb M, Reb A) can be provided as a pure compound or as part of a stevia extract or steviol glycoside mixture. Steviol glycosides (e.g., Reb M, Reb A) can be present in a steviol glycoside mixture or a stevia extract in an amount from about 5% to about 100% by dry weight.

[0189] In certain embodiments, a sweetener composition or a flavor-modifying composition comprises brazein (or an analog thereof), optionally in combination with: one or more steviol glycosides (e.g., Reb M, Reb A); an amino acid selected from glycine, alanine, proline, and combinations thereof; and optionally at least one additional sweetener and / or functional ingredient. As described above, steviol glycosides can be provided as a pure compound or as part of a stevia extract or steviol glycoside mixture. Steviol glycosides (e.g., Reb M, Reb A) can be present in a steviol glycoside mixture or a stevia extract in an amount from about 5% to about 100% by dry weight.

[0190] In certain embodiments, a sweetener composition or a flavor-modifying composition comprises brazein (or an analog thereof), optionally in combination with: one or more steviol glycosides (e.g., Reb M, Reb A); a salt selected from sodium chloride, magnesium chloride, potassium chloride, calcium chloride, and combinations thereof; and optionally at least one additional sweetener and / or functional ingredient. As described above, steviol glycosides (e.g., Reb M, Reb A) can be provided as a pure compound or as part of a stevia extract or steviol glycoside mixture.

[0191] Functional Ingredients

[0192] The sweetener compositions or flavor-modifying compositions disclosed herein may also contain one or more functional ingredients that provide actual or perceived health benefits to the compositions. Functional ingredients include, but are not limited to, saponins, antioxidants, dietary fiber sources, fatty acids, vitamins, glucosamine, minerals, preservatives, hydrating agents, probiotics, prebiotics, weight management agents, osteoporosis management agents, phytoestrogens, long-chain primary aliphatic saturated alcohols, phytosterols, and combinations thereof.

[0193] Examples of suitable antioxidants for use in embodiments of the present invention include, but are not limited to, vitamins, vitamin cofactors, minerals, hormones, carotenoids, carotenoid terpenes, non-carotenoid terpenes, flavonoids, flavonoid polyphenols (e.g., bioflavonoids), flavonols, flavones, phenols, polyphenols, phenolic esters, polyphenolic esters, non-flavonoid phenols, isothiocyanates, and combinations thereof. In some embodiments, the antioxidant is vitamin A, vitamin C, vitamin E, ubiquinone, the mineral selenium, manganese, melatonin, alpha-carotene, beta-carotene, lycopene, lutein, zeanthin, crypoxanthin, reservatol, eugenol, quercetin, catechins, gossypol, hesperidin, curcumin, ferulic acid, thymol, hydroxytyrosol, turmeric, thyme, olive oil, lipoic acid, glutathione, gutamine, oxalic acid, compounds derived from tocopherol, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediaminetetraacetic acid (EDTA), tert-butylhydroquinone, acetic acid, pectin, tocotrienols, tocopherols, coenzyme Q10, zeaxanthin, astaxanthin, canthaxanthin, saponins, limonin, kaempfedrol, myricetin, isorhamnetin, proanthocyanidins, quercetin, rutin, luteolin, apigenin, tangeritin, hesperidin, naringenin, erodictyol, flavan-3-ols (e.g., anthocyanidins), epigallocatechin gallate, epicatechin and its gallate forms, epigallocatechin and its gallate forms (ECGC), theaflavine In some embodiments, the antioxidant is a phytoestrogens, ...Other sources of suitable antioxidants for use with embodiments of the present invention include, but are not limited to, fruits, vegetables, tea, cocoa, chocolate, spices, herbs, rice, organ meats from livestock, yeast, whole grains or cereal grains.

[0194] Specific antioxidants belong to the class of phytonutrients known as polyphenols (also known as "polyphenols"), which are a group of chemicals found in plants characterized by the presence of more than one phenolic group per molecule. Suitable polyphenols for use in embodiments of the present invention include catechins, proanthocyanidins, proanthocyanidins, anthocyanidins, quercetin, rutin, resveratrol, isoflavones, curcumin, punicalagins, ellagitannins, hesperidin, naringin, citrus flavonoids, chlorogenic acid, other similar materials, and combinations thereof.

[0195] In certain embodiments, the antioxidant is a catechin, such as, for example, epigallocatechin gallate (EGCG). Suitable sources of catechins for embodiments of the present invention include, but are not limited to, green tea, white tea, black tea, oolong tea, chocolate, cocoa, red wine, grape seeds, red grape skins, purple grape skins, red grape juice, purple grape juice, berries, pycnogenol, and red apple peels.

[0196] In some embodiments, the antioxidant is selected from proanthocyanidins, proanthocyanidins, or a combination thereof. Suitable sources of proanthocyanidins and proanthocyanidins for embodiments of the present invention include, but are not limited to, red grapes, purple grapes, cocoa, chocolate, grape seeds, red wine, cocoa beans, cranberries, apple peels, plums, blueberries, black currants, choke berries, green tea, sorghum, cinnamon, barley, red kidney beans, black pinto beans, hops, almonds, hazelnuts, pecans, pistachio fruits, pycnogenol, and colored berries.

[0197] In a particular embodiment, the antioxidant is anthocyanidin. Suitable sources of anthocyanidins for embodiments of the present invention include, but are not limited to, red berries, blueberries, bilberries, cranberries, raspberries, cherries, pomegranates, strawberries, elderberries, rowan berries, red grape skins, purple grape skins, grape seeds, red wine, black currants, red currants, cocoa, plums, apple skins, peaches, red pears, red cabbage, red onions, red oranges, and blackberries.

[0198] In some embodiments, the antioxidant is selected from quercetin, rutin or a combination thereof. Suitable sources of quercetin and rutin for embodiments of the present invention include, but are not limited to, red apples, onions, kale, blueberries, bilberries, rowan berries, cranberries, blackberries, blueberries, strawberries, raspberries, blackcurrants, green tea, black tea, plums, apricots, parsley, leeks, broccoli, red peppers, berry wine and ginkgo.

[0199] In some embodiments, the antioxidant is resveratrol. Suitable sources of resveratrol for use in embodiments of the present invention include, but are not limited to, red grapes, peanuts, cranberries, blueberries, bilberries, mulberries, Japanese Itadori tea, and red wine.

[0200] In particular embodiments, the antioxidant is an isoflavone. Suitable sources of isoflavones for embodiments of the present invention include, but are not limited to, soybeans, soybean products, beans, alfalfa sprouts, chickpeas, peanuts, and red clover.

[0201] In some embodiments, the antioxidant is curcumin. Suitable sources of curcumin for embodiments of the present invention include, but are not limited to, turmeric and mustard.

[0202] In particular embodiments, the antioxidant is selected from punicalagins, ellagitannins, or combinations thereof. Suitable sources of punicalagins and ellagitannins for embodiments of the present invention include, but are not limited to, pomegranates, raspberries, strawberries, walnuts, and aged red wine.

[0203] In some embodiments, the antioxidant is a citrus flavonoid, such as hesperidin or naringin. Suitable sources of citrus flavonoids, such as hesperidin or naringin, for embodiments of the present invention include, but are not limited to, orange, grapefruit, and citrus juice.

[0204] In a particular embodiment, the antioxidant is chlorogenic acid. Suitable sources of chlorogenic acid for embodiments of the present invention include, but are not limited to, green coffee, yerba mate, red wine, grape seeds, red grape skins, purple grape skins, red grape juice, purple grape juice, apple juice, cranberries, pomegranates, blueberries, strawberries, sunflowers, echinacea, pycnogenol, and apple peels.

[0205] Suitable dietary fibers include, but are not limited to, non-starch polysaccharides, lignin, cellulose, methylcellulose, hemicellulose, beta-glucan, pectin, gums, mucilages, waxes, inulin, oligosaccharides, fructooligosaccharides, cyclodextrins, chitin, and combinations thereof.

[0206] Food sources of dietary fiber include, but are not limited to, cereals, beans, fruits and vegetables. Cereals that provide dietary fiber include, but are not limited to oats, rye, barley and wheat. Beans that provide fiber include, but are not limited to peas and kidney beans such as soybeans. Fruits and vegetables that provide fiber sources include, but are not limited to apples, oranges, pears, bananas, berries, tomatoes, green beans, broccoli, cauliflower, carrots, potatoes, celery. Plant foods such as bran, nuts and seeds (such as linseed) are also sources of dietary fiber. Plant parts that provide dietary fiber include, but are not limited to stems, roots, leaves, seeds, pulp and skin.

[0207] Fatty acids are any straight chain monocarboxylic acids and include saturated fatty acids, unsaturated fatty acids, long chain fatty acids, medium chain fatty acids, short chain fatty acids, fatty acid precursors (including ω-9 fatty acid precursors) and esterified fatty acids. As used herein, "long chain polyunsaturated fatty acids" refer to any polyunsaturated carboxylic acid or organic acid with a long aliphatic tail. Suitable ω-3 fatty acids include, but are not limited to, linolenic acid, α-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, octadecatetraenoic acid, eicosatetraenoic acid and combinations thereof.

[0208] Suitable ω-6 fatty acids include, but are not limited to, linoleic acid, γ-linolenic acid, dihomo-γ-linolenic acid, arachidonic acid, eicosadienoic acid, docosadienoic acid, adrenal acid, docosapentaenoic acid, and combinations thereof. Suitable esterified fatty acids for embodiments of the present invention include, but are not limited to, monoacylglycerols containing ω-3 and / or ω-6 fatty acids, diacylglycerols containing ω-3 and / or ω-6 fatty acids, or triacylglycerols containing ω-3 and / or ω-6 fatty acids, and combinations thereof.

[0209] Suitable vitamins include vitamin A, vitamin D, vitamin E, vitamin K, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12, and vitamin C. Various other compounds have been classified as vitamins by some officials. These compounds can be referred to as pseudovitamins, and include but are not limited to compounds such as ubiquinone (coenzyme Q10), pancreatic acid, dimethylglycine, taestrile, amygdalin, flavonoids, para-aminobenzoic acid, adenine, adenylic acid, and s-methylmethionine. As used herein, the term vitamin includes pseudovitamins.

[0210] Minerals are selected from bulk minerals, trace minerals or combinations thereof. Non-limiting examples of bulk minerals include calcium, chlorine, magnesium, phosphorus, potassium, sodium and sulfur. Non-limiting examples of trace minerals include chromium, cobalt, copper, fluorine, iron, manganese, molybdenum, selenium, zinc and iodine. Although iodine is usually classified as a trace mineral, it requires a larger amount than other trace minerals and is often classified as a bulk mineral.

[0211] In other specific embodiments of the invention, the mineral is a trace mineral considered essential for human nutrition, non-limiting examples of which include bismuth, boron, lithium, nickel, rubidium, silicon, strontium, tellurium, tin, titanium, tungsten, and vanadium.

[0212] The preservative is selected from an antimicrobial, an antioxidant, an antienzyme or a combination thereof. Non-limiting examples of antimicrobials include sulfites, propionates, benzoates, sorbates, nitrates, nitrites, bacteriocins, salts, sugars, acetic acid, dimethyl dicarbonate (DMDC), ethanol and ozone. Sulfites include but are not limited to sulfur dioxide, sodium bisulfite and potassium bisulfite. Propionates include but are not limited to propionic acid, calcium propionate and sodium propionate. Benzoates include but are not limited to sodium benzoate and benzoic acid. Sorbic acids include but are not limited to potassium sorbate, sodium sorbate, calcium sorbate and sorbic acid. Nitrates and nitrites include but are not limited to sodium nitrate and sodium nitrite. In another specific embodiment, the at least one preservative is a bacteriocin, such as nisin. In another specific embodiment, the preservative is ethanol. In another specific embodiment, the preservative is ozone. Antienzyme agents suitable for use as preservatives in certain embodiments of the invention include ascorbic acid, citric acid, and metal chelators such as ethylenediaminetetraacetic acid (EDTA).

[0213] The hydration product can be an electrolyte, non-limiting examples of which include sodium, potassium, calcium, magnesium, chloride, phosphate, bicarbonate, and combinations thereof. Suitable electrolytes for use in specific embodiments of the present invention are also described in U.S. Patent No. 5,681,569, the disclosure of which is expressly incorporated herein by reference. Non-limiting examples of salts for use in specific embodiments include chlorides, carbonates, sulfates, acetates, bicarbonates, citrates, phosphates, hydrogen phosphates, tartrates, sorbates, citrates, benzoates, or combinations thereof.

[0214] In a specific embodiment of the present invention, hydration products are carbohydrates for replenishing the energy storage burned by muscle. Suitable carbohydrates for use in a specific embodiment of the present invention are described in U.S. Patent Nos. 4,312,856, 4,853,237, 5,681,569 and 6,989,171, and the disclosure of the patent is expressly incorporated herein by reference. The limiting examples of suitable carbohydrates include monose, disaccharides, oligosaccharides, complex polysaccharides or their combinations. The limiting examples of monose for the suitable type used in a specific embodiment include triose, tetrose, pentose, hexose, heptose, octose and nonose. The limiting examples of suitable monosaccharides of a particular type include glyceraldehyde, dihydroxyacetone, erythrose, threose, erythrulose, arabinose, lyxose, ribose, xylose, ribulose, xylulose, allose, altrose, galactose, glucose, gulose, idose, mannose, talose, fructose, psicose, sorbose, tagatose, mannoheptulose, sedoheltulose, octolose and sialose. The limiting examples of suitable disaccharides include sucrose, lactose and maltose. The limiting examples of suitable oligosaccharides include sucrose, maltotriose and maltodextrin. In other specific embodiments, carbohydrates are provided by corn syrup, beet sugar, cane sugar, juice or tea. In another specific embodiment, hydration is the flavanol that provides cell rehydration. Non-limiting examples of suitable flavanols for use in particular embodiments of the invention include catechin, epicatechin, gallocatechin, epigallocatechin, epicatechin gallate, epigallocatechin 3-gallate, theaflavins, theaflavins 3-gallate, theaflavins 3'-gallate, theaflavins 3,3' gallate, thearubigins, or combinations thereof. In particular embodiments, the hydration product is a glycerol solution that enhances athletic endurance.

[0215] Probiotics include microorganisms that are beneficial to health when consumed in effective amounts. Probiotics may include, but are not limited to, bacteria, yeasts, and fungi. Examples of probiotics include, but are not limited to, bacteria of the genus Lactobacilli, the genus Bifidobacteria, the genus Streptococci, or a combination thereof. In a particular embodiment of the invention, the at least one probiotic is selected from the genus Lactobacilli. Lactobacilli (i.e., bacteria of the genus Lactobacillus, hereinafter referred to as "L."). Non-limiting examples of lactobacillus species found in the human gastrointestinal tract include Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus saliva roes, Lactobacillus brevis, Lactobacillus leichmannii, Lactobacillus plantarum, Lactobacillus cellobiosus, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus GG, Lactobacillus bulgaricus, and Lactobacillus thermophilus. According to other specific embodiments of the present invention, the probiotic is selected from the genus Bifidobacterium.Non-limiting species of Bifidobacteria found in the human gastrointestinal tract include B. angulata, B. animalis, B. asteroides, B. bifidum, B. boum, B. breve, B. catenulatum, B. choerinum, B. coryneforme, B. cuniculi, B. dentium, B. gallicum, B. gallinarum, B. chrysanthemi, B. scutellariae ... In some embodiments, the probiotics are selected from the genus Streptococcus. Streptococcus thermophilus is a gram-positive facultative anaerobic bacterium. Other non-limiting probiotic species of this bacteria include Streptococcus salivarus and Streptococcus cremoris.

[0216] Prebiotics are compositions that promote the growth of beneficial bacteria in the intestine. Prebiotics include but are not limited to mucopolysaccharides, oligosaccharides, polysaccharides, amino acids, vitamins, nutrient precursors, proteins and combinations thereof. According to specific embodiments, prebiotics are selected from dietary fiber, including but not limited to polysaccharides and oligosaccharides. The non-limiting examples of oligosaccharides classified as prebiotics according to specific embodiments of the present invention include fructooligosaccharides, inulin, isomalt oligosaccharides, lactitol (lactilol), oligolactose, lactulose, pyrodextrin, soybean oligosaccharides, oligomeric transgalactose and oligoxylose. According to other specific embodiments, prebiotics are amino acids.

[0217] As used herein, "weight management agents" include appetite suppressants and / or thermogenic agents. As used herein, the phrases "appetite suppressants," "appetite satiety compositions," "satiants," and "satiating ingredients" are synonymous. The phrase "appetite suppressants" describes macronutrients, herbal extracts, exogenous hormones, anorectics, anorexigenics, drugs, and combinations thereof that, when delivered in effective amounts, suppress, inhibit, reduce, or otherwise diminish a person's appetite. The phrase "thermogenic agents" describes macronutrients, herbal extracts, exogenous hormones, anorectics, anorexigenics, drugs, and combinations thereof that, when delivered in effective amounts, stimulate or otherwise enhance a person's thermogenesis or metabolism.

[0218] Suitable weight management agents include macronutrients selected from the group consisting of: protein, carbohydrates, dietary fats, and combinations thereof. Carbohydrates generally include sugars, starches, celluloses, and gums that are converted by the body into glucose for energy. Non-limiting examples of carbohydrates include polydextrose; inulin; polyols such as erythritol, mannitol, xylitol, and sorbitol derived from monosaccharides; alcohols such as isomalt, lactitol, and maltitol derived from disaccharides; and hydrogenated starch hydrolysates. Carbohydrates are described in more detail below. Dietary fats are lipids comprising a combination of saturated fatty acids and unsaturated fatty acids. It has been shown that polyunsaturated fatty acids have a greater satiety capacity than monounsaturated fatty acids. Therefore, the dietary fats presented herein ideally include polyunsaturated fatty acids, and non-limiting examples thereof include triacylglycerols.

[0219] In a particular embodiment, the weight management agent is an herbal extract. Non-limiting examples of plants whose extracts have appetite suppressant properties include plants of the genus Hoodia, Trichocaulon, Caralluma, Stapelia, Orbea, Asclepias, and Camelia. Other embodiments include extracts from Gymnema Sylvestre, Kola Nut, Citrus Aurantium, Yerba Mate, Griffonia Simplicifolia, Guarana, myrrh, guggul Lipid, and black currentseed oil. In certain embodiments, the herbal extract is derived from the genus Hoodia, species of which include H. alstonii, H. currorii, H. dregei, H. flava, H. gordonii, H. jutatae, H. mossamedensis, H. officinalis, H. parviflorai, H. pedicellata, H. pilifera, H. ruschii, and H. triebneri. Hoodia is a fleshy plant native to South Africa.

[0220] In another specific embodiment, the herbal extract is derived from a plant of the genus Caralluma, species of which include C. indica, C. fimbriata, C. attenuate, C. tuberculata, C. edulis, C. adscendens, C. stalagmifera, C. umbellate, C. penicillata, C. russeliana, C. retrospicens, C. Arabica, and C. lasiantha. Plants of the genus Caralluma belong to the same subfamily as the genus Caralluma, namely, Asclepiadaceae.

[0221] In another specific embodiment, the at least one herbal extract is derived from a plant of the genus Alocasia. Plants of the genus Alocasia are succulent plants that are generally native to South Africa, similar to the genus Hoodia, and include T. piliferum and T. officinale. In another specific embodiment, the herbal extract is derived from a plant of the genus Staphylococcus or Obea, species of which include S. gigantean and O. variegate, respectively. Both Staphylococcus and Obea belong to the same subfamily Asclepiadaceae as Hoodia.

[0222] In another specific embodiment, the herbal extract is derived from Asclepiadaceae. Asclepiadaceae also belongs to Asclepiadaceae. Non-limiting examples of Asclepiadaceae include A. incarnate, A. curassayica, A. syriaca, and A. tuberose. Without wishing to be bound by any theory, it is believed that these extracts contain steroidal compounds with appetite suppressant effects, such as pregnane glycosides and pregnane aglycones. In a specific embodiment, the weight management agent is an exogenous hormone with a weight management effect. Non-limiting examples of such hormones include CCK, peptide YY, ghrelin, bombesin and gastrin releasing peptide (GRP), enterostatin, apolipoprotein A-IV, GLP-1, amylin, somastatin and leptin.

[0223] In certain embodiments, the osteoporosis management agent is at least one calcium source, i.e., any compound containing calcium, including salt complexes, dissolved substances and other forms of calcium. Non-limiting examples of calcium sources include amino acid chelated calcium, calcium carbonate, calcium oxide, calcium hydroxide, calcium sulfate, calcium chloride, calcium phosphate, calcium hydrogen phosphate, calcium dihydrogen phosphate, calcium citrate, calcium malate, calcium citrate malate, calcium gluconate, calcium tartrate, calcium lactate, dissolved species thereof, and combinations thereof. According to a specific embodiment, the osteoporosis management agent is a magnesium source, i.e., any compound containing magnesium, including salt complexes, dissolved substances and other forms of magnesium. Non-limiting examples of magnesium sources include magnesium chloride, magnesium citrate, magnesium glucoheptonic acid, magnesium gluconate, magnesium lactate, magnesium hydroxide, magnesium picolate, magnesium sulfate, dissolved species thereof, and mixtures thereof. In another specific embodiment, the magnesium source includes amino acid chelated magnesium or creatine chelated magnesium. In other embodiments, the osteoporosis agent is selected from vitamin D, C, K, its precursors and / or beta-carotene and combinations thereof. Various plants and plant extracts have also been identified as being effective for preventing and treating osteoporosis. Without wishing to be bound by any theory, it is believed that these plants and plant extracts stimulate osteogenic proteins and / or inhibit bone resorption, thereby promoting bone regeneration and strength. Non-limiting examples of suitable plants and plant extracts as osteoporosis management agents include Taraxacum and Amelanchier species as disclosed in U.S. Patent Publication No. 2005 / 0106215, and Lindera, Artemisia, Acorus, Carthamus, Carum, Cnidium, Curcuma, Cyperus, Juniperus, Prunus and species of the genera Prunus, Iris, Cichorium, Dodonaea, Epimedium, Erigonoum, Soya, Mentha, Ocimum, Thymus, Tanacetum, Plantago, Spearmint, Bixa, Vitis, Rosemarinus, Rhus, and Anethum.

[0224] Examples of suitable phytoestrogens for embodiments of the present invention include, but are not limited to, isoflavones, stilbenes, lignans, resorcyclic acid lactone, coumarins, coumestrol, equol and combinations thereof. Isoflavones belong to the group of phytonutrients known as polyphenols. Typically, polyphenols (also referred to as "polyphenols") are a group of chemicals found in plants, characterized in that each molecule has more than one phenol group. Suitable phytoestrogen isoflavones according to embodiments of the present invention include genistein, daidzein, glycitein, biochanin A, formononetin, their respective naturally occurring glycosides and glycoside conjugates, matai resin alcohol, secoisolariciresinol, enterodiol, plant tissue protein and combinations thereof.

[0225] Long-chain aliphatic saturated primary alcohol is an organic compound of different groups.The term long chain refers to the following fact: the number of carbon atoms in these compounds is at least 8 carbons.The limiting examples for the specific long-chain aliphatic saturated primary alcohol used in a particular embodiment of the present invention include 8 carbon atom 1-octanol, 9 carbon 1-nonanol, 10 carbon atom 1-decanol, 12 carbon atom 1-dodecanol, 14 carbon atom 1-tetradecanol, 16 carbon atom 1-hexadecanol, 18 carbon atom 1-octadecanol, 20 carbon atom 1-eicosanol, 22 carbon 1-docosanol, 24 carbon 1-tetracosanol, 26 carbon 1-hexacosanol, 27 carbon 1-heptacosanol, 28 carbon 1-octaconic alcohol (octanosol), 29 carbon 1-nonacoustic alcohol, 30 carbon 1-triacontanol, 32 carbon 1-dotriacontanol and 34 carbon 1-tetratriacontanol.In a particularly desirable embodiment of the present invention, the long-chain aliphatic saturated primary alcohol is policosanol. Policosanol is a term for a mixture of long chain aliphatic saturated primary alcohols consisting primarily of 28 carbon 1-octacosanol and 30 carbon 1-triacontanol and lower concentrations of other alcohols such as 22 carbon 1-docosanol, 24 carbon 1-tetracosanol, 26 carbon 1-hexacosanol, 27 carbon 1-heptacosanol, 29 carbon 1-nonacosanol, 32 carbon 1-dotriacontanol and 34 carbon 1-tetracontanol.

[0226] At least 44 naturally occurring phytosterols have been found, and they are generally derived from plants such as corn, soybeans, wheat and tung oil; however, they can also be produced synthetically to form compositions identical to those of nature or compositions having properties similar to those of naturally occurring phytosterols. According to a specific embodiment of the invention, non-limiting examples of phytosterols well known to those of ordinary skill in the art include 4-demethyl sterols (e.g., β-sitosterol, campesterol, stigmasterol, rapeseed sterol, 22-dehydrobrassicasterol, and Δ5-avenasterol), 4-monomethyl sterols and 4,4-dimethyl sterols (triterpenoid alcohols) (e.g., cycloartenol, 24-methylenecycloartanol and cyclostanol (cyclobranol)).

[0227] According to certain embodiments of the present invention, non-limiting examples of phytostanols include β-sitostanol, campestanol, cycloartanol, and saturated forms of other triterpene alcohols.

[0228] As used herein, phytosterols and phytostanols include multiple isomers such as α and β isomers (e.g., α-sitosterol and β-sitosterol, which respectively include one of the most effective phytosterols and phytostanols for reducing serum cholesterol in mammals). The phytosterols and phytostanols of the present invention may also be in the form of their esters. Non-limiting examples of esters of suitable phytosterols and phytostanols include acetic acid sitosterol ester, oleic acid sitosterol ester, oleic acid stigmasterol ester and its corresponding phytostanol ester. The phytosterols and phytostanols of the present invention may also include derivatives thereof.

[0229] In general, the amount of the functional ingredient in the composition varies widely depending on the specific composition and the desired functional ingredient. One of ordinary skill in the art will readily determine the appropriate amount of the functional ingredient for each composition.

[0230] consumer goods

[0231] Disclosed herein are consumables (eg, beverages) comprising a sweetener composition or a flavor composition disclosed herein.

[0232] Brazeain (or its analogs) or sweetener compositions or flavor-modifying compositions containing brazeain disclosed herein can be incorporated into any known edible material (referred to herein as a "consumer product," "sweetenable composition," or "flavor and / or taste-modified composition"), for example, edible gel mixes and compositions, dental compositions, foods (sweets, condiments, chewing gum, cereal compositions, baked goods, dairy products, and tabletop sweetener compositions), beverages, and beverage products.

[0233] In one embodiment, the consumable is a beverage or a beverage product. In other words, disclosed herein is a beverage or a beverage product comprising brazepine (or its analog) or a sweetener composition or a flavor-modifying composition containing brazepine disclosed herein.

[0234] As used herein, "beverage product" is a ready-to-drink beverage, beverage concentrate, beverage syrup or beverage powder brewed beverage. Suitable ready-to-drink beverages include carbonated beverages and non-carbonated beverages. Carbonated beverages include, but are not limited to, cola, ginger ale, soft drinks and root beer. Non-carbonated beverages include, but are not limited to, fruit juices, fruit-flavored juices, vegetable juices, vegetable-flavored juices, sports drinks, energy drinks, plant protein beverages, near-water beverages (e.g., water containing natural or synthetic flavoring agents), teas (e.g., black tea, green tea, black tea, oolong tea), coffee, cocoa drinks, beverages containing milk components (e.g., milk beverages, coffee containing milk components, cafe au lait, milk tea, fruit milk beverages).

[0235] In particular embodiments, the beverage is a flavored black tea beverage, a zero calorie enhanced water beverage, or an orange flavored sparkling beverage.

[0236] Beverage concentrates and beverage syrups are prepared with an initial volume of liquid matrix (e.g., water) and the desired beverage ingredients. Full strength beverages are then prepared by adding additional volumes of water. Beverage powder infusions are prepared by dry mixing all beverage ingredients in the absence of a liquid matrix. Full strength beverages are then prepared by adding the entire volume of water.

[0237] The beverage contains water as a liquid matrix, i.e., a base component in which the ingredients (including the sweetener or sweetener composition) are dissolved. Beverage quality water can be used, such as, for example, deionized water, distilled water, reverse osmosis water, carbon-treated water, purified water, softened water, and combinations thereof. Other suitable liquid matrices include, but are not limited to, phosphoric acid, phosphate buffer, citric acid, citrate buffer, and carbon-treated water.

[0238] In one embodiment, a beverage or beverage product containing bulazein or an analog thereof is disclosed. In a specific embodiment, the beverage contains a bulazein analog having the same or increased sweetness as a beverage containing the same amount of wild-type bulazein.

[0239] In one embodiment, the concentration of brazein (or its analog) in the beverage is equal to or higher than its sweetness recognition threshold concentration or flavor recognition threshold concentration. In a specific embodiment, the concentration of brazein is at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50% or more higher than its sweetness or flavor recognition threshold concentration.

[0240] In one embodiment, the beverage or beverage product contains brazee (or its analog) in an amount from about 1 ppm to about 50 ppm, such as, for example, from about 5 ppm to about 50 ppm, from about 5 ppm to about 40 ppm, from about 5 ppm to about 30 ppm, from about 5 ppm to about 20 ppm, from about 5 ppm to about 10 ppm, from about 10 ppm to about 50 ppm, from about 10 ppm to about 40 ppm, from about 10 ppm to about 30 ppm, from about 10 ppm to about 20 ppm, from about 20 ppm to about 50 ppm, from about 20 ppm to about 40 ppm, from about 20 ppm to about 30 ppm, from about 30 ppm to about 50 ppm, from about 30 ppm to about 40 ppm, or from about 40 ppm to about 50 ppm.

[0241] In more specific embodiments, the beverage or beverage product contains brazeine (or its analog) in an amount from about 10 ppm to about 25 ppm.

[0242] In another embodiment, the beverage contains brazene (or its analog) in an amount from about 1 ppm to less than about 15 ppm. In one embodiment, the beverage contains brazene (or its analog) in an amount from about 1 ppm to about 5 ppm, about 5 ppm to about 10 ppm, or about 10 ppm to about 15 ppm.

[0243] In another embodiment, the beverage contains brazene (or its analog) in an amount greater than about 15 ppm. In one embodiment, the beverage contains brazene (or its analog) in an amount between about 15 ppm and about 20 ppm, between about 20 ppm and about 25 ppm, between about 25 ppm and about 30 ppm, between about 30 ppm and about 35 ppm, between about 35 ppm and about 40 ppm, between about 40 ppm and about 50 ppm, or about 50 ppm or more.

[0244] In one embodiment, the beverage contains brazeein (or its analog) in an amount of about 1 ppm, about 3 ppm, about 5 ppm, about 10 ppm, about 12 ppm, about 15 ppm, about 18 ppm, about 20 ppm, about 22 ppm, about 25 ppm, about 28 ppm, about 30 ppm, about 35 ppm, about 40 ppm, about 45 ppm, about 50 ppm, about 55 ppm, about 60 ppm, about 65 ppm, about 70 ppm, or about 75 ppm or more.

[0245] The beverage may further comprise at least one additional sweetener.Any sweetener detailed herein may be used, including natural sweeteners, non-natural sweeteners or synthetic sweeteners.

[0246] In a particular embodiment, the at least one additional sweetener is one or more steviol glycosides. In a particular embodiment, the steviol glycoside is Reb M. In another particular embodiment, the steviol glycoside is Reb A. In one embodiment, the beverage contains a steviol glycoside blend, e.g., Reb M and Reb A, in addition to brazil (or its analog).

[0247] The amount of steviol glycosides (e.g., Reb M, Reb A) can vary. In one embodiment, the amount of steviol glycosides provides less than about 8 sucrose equivalents (SE), for example, about 7.5 SE, about 7.0 SE, about 6.5 SE, or about 6.0 SE or less. In another embodiment, the amount of steviol glycosides provides greater than about 8 sucrose equivalents (SE), for example, about 8.5 SE, about 9.0 SE, about 9.5 SE, about 10 SE, about 10.5 SE, about 11 SE, or about 12 SE or more.

[0248] In another embodiment, the amount of steviol glycosides (e.g., Reb M, Reb A) is less than about 450 ppm. In specific embodiments, the amount of steviol glycosides is less than about 400 ppm, less than about 350 ppm, less than about 300 ppm, less than about 300 ppm, less than about 250 ppm, less than about 200 ppm, less than about 150 ppm, or about 100 ppm or less.

[0249] In another embodiment, the amount of steviol glycosides (e.g., Reb M, Reb A) is between about 100 ppm and about 450 ppm, more specifically, between about 100 ppm and about 400 ppm, between about 100 ppm and about 350 ppm, between about 100 ppm and about 300 ppm, between about 100 ppm and about 250 ppm, between about 100 ppm and about 200 ppm, or between about 100 ppm and 250 ppm.

[0250] In another embodiment, the amount of steviol glycosides (e.g., Reb M, Reb A) is between about 100 ppm and about 200 ppm, more specifically, about 110 ppm, about 120 ppm, about 130 ppm, about 140 ppm, about 150 ppm, about 160 ppm, about 170 ppm, about 180 ppm, about 190 ppm, or about 200 ppm.

[0251] In another embodiment, the amount of steviol glycosides (e.g., Reb M, Reb A) is between about 200 ppm and about 300 ppm, and more particularly, about 210 ppm, about 220 ppm, about 230 ppm, about 240 ppm, about 250 ppm, about 260 ppm, about 270 ppm, about 280 ppm, about 290 ppm, or about 300 ppm.

[0252] In another embodiment, the amount of steviol glycosides (e.g., Reb M, Reb A) is between about 300 ppm and about 400 ppm, and more particularly, about 310 ppm, about 320 ppm, about 330 ppm, about 340 ppm, about 350 ppm, about 360 ppm, about 370 ppm, about 380 ppm, about 390 ppm, or about 400 ppm.

[0253] In another embodiment, the amount of steviol glycosides (e.g., Reb M, Reb A) is between about 400 ppm and about 500 ppm, and more particularly, about 410 ppm, about 420 ppm, about 430 ppm, about 440 ppm, about 450 ppm, about 460 ppm, about 470 ppm, about 480 ppm.

[0254] In particular embodiments, the beverage comprises brazein (or an analog thereof) in an amount between about 1 ppm and about 100 ppm and at least one steviol glycoside (e.g., Reb M, Reb A), wherein the amount of the at least one steviol glycoside is sufficient to provide less than about 8 SE, e.g., about 7.5 SE, about 7.0 SE, about 6.5 SE, about 6 SE, about 5.5 SE, about 5.0 SE, about 4.5 SE, about 4.0 SE, about 3.5 SE, about 3.0 SE, about 2.5 SE, about 2.0 SE, about 1.5 SE, or about 1.0 SE.

[0255] In another particular embodiment, the beverage comprises brazein (or an analog thereof) and at least one steviol glycoside (e.g., Reb M, Reb A) in an amount between about 1 ppm and about 100 ppm, wherein the at least one steviol glycoside is in an amount of about 450 ppm or less, e.g., 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, or about 100 ppm or less.

[0256] In particular embodiments, the beverage comprises brazeein (or its analog) in an amount between about 1 ppm and about 30 ppm, more particularly, between about 1 ppm and about 15 ppm or between about 15 ppm and about 30 ppm, and Reb M in an amount sufficient to provide less than about 8 SE, such as, for example, about 7.5 SE, about 7.0 SE, about 6.5 SE, about 6 SE, about 5.5 SE, about 5.0 SE, about 4.5 SE, about 4.0 SE, about 3.5 SE, about 3.0 SE, about 2.5 SE, about 2.0 SE, about 1.5 SE, or about 1.0 SE.

[0257] In another specific embodiment, the beverage comprises brazeein (or its analog) in an amount between about 1 ppm and about 30 ppm, more particularly, between about 1 ppm and about 15 ppm or between about 15 ppm and about 30 ppm, and Reb M in an amount of about 450 ppm or less, e.g., 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 or about 100 ppm or less.

[0258] In another specific embodiment, the beverage comprises brazeein (or its analog) in an amount between about 1 ppm and about 30 ppm, more particularly, between about 1 ppm and about 15 ppm or between about 15 ppm and about 30 ppm, and Reb M in an amount of about 450 ppm or less, e.g., 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 or about 100 ppm or less.

[0259] For example, the beverage comprises brazein (or its analog) in an amount from about 1 ppm to about 30 ppm, and Reb M80 in an amount from about 200 ppm to about 400 ppm. In a more specific embodiment, the beverage comprises brazein (or its analog) in an amount from about 20 ppm to about 30 ppm, and Reb M80 in an amount from about 300 to about 400 ppm. In an even more specific embodiment, the beverage comprises brazein (or its analog) in an amount of about 25 ppm, and Reb M80 in an amount of about 315 ppm.

[0260] The beverages of the present invention comprise brazepine (or its analog) in an amount from about 1 ppm to about 50 ppm, and Reb M80 in an amount from about 200 ppm to about 400 ppm. In preferred embodiments, the beverages of the present invention taste similar to beverages sweetened with Reb M or Reb M80 alone, wherein the beverages of the present invention have similar sucrose equivalence to beverages sweetened with Reb M or Reb M80. In more specific embodiments, beverages with a citric acid base comprise brazepine (or its analog) in an amount from about 20 ppm to about 30 ppm, and Reb M80 in an amount from about 300 to about 400 ppm. In still more specific embodiments, beverages with a citric acid base comprise brazepine (or its analog) in an amount from about 25 ppm, and Reb M80 in an amount from about 315 ppm. In another more specific embodiment, a lemon-lime sugar-free carbonated beverage comprises brazepine in an amount from about 30 ppm to about 50 ppm, and Reb M80 in an amount from about 150 ppm to about 350 ppm. In yet more particular embodiments, a lemon-lime diet carbonated beverage comprises brazil in an amount of about 40 ppm, and Reb M80 in an amount of about 210 ppm.

[0261] In particular embodiments, the beverage comprises brazeein (or an analog thereof) in an amount between about 10 ppm and about 25 ppm and Reb M in an amount sufficient to provide less than about 8 SE, e.g., about 7.5 SE, about 7.0 SE, about 6.5 SE, about 6 SE, about 5.5 SE, about 5.0 SE, about 4.5 SE, about 4.0 SE, about 3.5 SE, about 3.0 SE, about 2.5 SE, about 2.0 SE, about 1.5 SE, or about 1.0 SE.

[0262] In another specific embodiment, the beverage comprises brazeein (or its analog) in an amount between about 10 ppm and about 25 ppm and Reb M in an amount of about 450 ppm or less, e.g., 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, or about 100 ppm or less.

[0263] In another particular embodiment, the beverage comprises brazein (or an analog thereof) and at least one additional sweetener (e.g., Reb M, Reb A, simianoside I, mogroside IV) in an amount between about 1 ppm and about 50 ppm, wherein the at least one additional sweetener is in an amount less than about 450 ppm, e.g., 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, or about 100 ppm or less.

[0264] In particular embodiments, the beverage comprises brazeein (or an analog thereof) in an amount between about 1 ppm and about 100 ppm and Reb A in an amount sufficient to provide less than about 8 SE, e.g., about 7.5 SE, about 7.0 SE, about 6.5 SE, about 6 SE, about 5.5 SE, about 5.0 SE, about 4.5 SE, about 4.0 SE, about 3.5 SE, about 3.0 SE, about 2.5 SE, about 2.0 SE, about 1.5 SE, or about 1.0 SE.

[0265] In particular embodiments, the beverage comprises brazeein (or an analog thereof) and Reb A in an amount of between about 1 ppm and about 30 ppm, more particularly, between about 1 ppm and about 15 ppm or between about 15 ppm and about 30 ppm, and an amount of less than about 450 ppm, e.g., 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, or about 100 ppm or less.

[0266] In particular embodiments, the beverage comprises brazeein (or an analog thereof) in an amount between about 10 ppm and about 25 ppm and Reb A in an amount sufficient to provide less than about 8 SE, e.g., about 7.5 SE, about 7.0 SE, about 6.5 SE, about 6 SE, about 5.5 SE, about 5.0 SE, about 4.5 SE, about 4.0 SE, about 3.5 SE, about 3.0 SE, about 2.5 SE, about 2.0 SE, about 1.5 SE, or about 1.0 SE.

[0267] In particular embodiments, the beverage comprises brazeein (or an analog thereof) in an amount between about 10 ppm and about 25 ppm and Reb A in an amount less than about 450 ppm, e.g., 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, or about 100 ppm or less.

[0268] The ppm ratio of bulazines (or its analogs) to the one or more steviol glycosides in the beverage can vary. In one embodiment, the ppm ratio of bulazines (or its analogs) to the one or more steviol glycosides is between about 1:2 and about 1:40, more particularly, between about 1:5 and about 1:35, and even more particularly, between about 1:10 and about 1:25.

[0269] In particular embodiments, the ppm ratio of brazein (or its analogue) to the one or more steviol glycosides is about 1:30 or less, more particularly, about 1:25.

[0270] In one embodiment, the steviol glycoside is Reb M, and the ppm ratio of brazin (or its analogue) to Reb M is between about 1:10 and about 1:20, more specifically, about 1:10, about 1:12, about 1:14, about 1:16, about 1:18, or about 1:20.

[0271] In another embodiment, the steviol glycoside is Reb A, and the ppm ratio of brazin (or its analogue) to Reb A is about 1:40 or less, more specifically, between about 1:20 and about 1:30, even more specifically, about 1:20, about 1:22, about 1:24, about 1:26, about 1:28, or about 1:30.

[0272] In another embodiment, the steviol glycoside is Reb A, and the ppm ratio of brazein (or its analog) to Reb A is about 1:70 or less, more specifically, less than about 1:60, less than about 1:50, or less than about 1:40. In a specific embodiment, the ppm ratio of brazein (or its analog) to Reb A is about 1:35.

[0273] In another embodiment, the steviol glycoside is Reb A and the ppm ratio of brazil (or its analogue) to Reb A is about 1:15 or greater, more particularly, greater than about 1:17, about 1:18, about 1:19 or about 1:20.

[0274] In another embodiment, the steviol glycoside is Reb A and the ppm ratio of brazin (or its analogue) to Reb A is about 1:10 or greater, more particularly, greater than about 1:12, about 1:14, or about 1:16.

[0275] In one embodiment, a beverage comprises brazepine (or an analog thereof) and Reb M, wherein brazepine is present in an amount from about 15 ppm to about 30 ppm, more particularly, from about 20 ppm to about 25 ppm, and Reb M (e.g., Reb M80) is present in an amount from about 300 ppm to about 350 ppm, more particularly, from about 315 ppm to about 325 ppm (i.e., Reb M provides about 8 SE). According to this embodiment, the ratio of brazepine to Reb M is between about 1:10 and about 1:20, more particularly about 1:0, 1:12, about 1:14, about 1:16, about 1:18, or about 1:20. In one embodiment, the amount of Reb M provides about 8 sucrose equivalents (SE).

[0276] In one embodiment, the beverage comprises brazeine (or an analog thereof) and Reb M, wherein brazeine is present in an amount from about 1 ppm to about 20 ppm, more particularly, about 1 ppm to about 15 ppm, and Reb M is present in an amount from about 110 ppm to about 140 ppm, more particularly, about 115 ppm, and even more particularly, about 120 ppm. In a particular embodiment, the ppm ratio of brazeine to Reb M is about 1:5 to about 1:10, more particularly about 1:7.

[0277] In one embodiment, the beverage comprises braazein (or an analog thereof) and Reb M, wherein braazein is present in an amount from about 1 ppm to about 15 ppm, more specifically, about 1 ppm to about 5 ppm, even more specifically, about 3 ppm, and Reb M is present in an amount between about 450 ppm and about 500 ppm, more specifically, about 470 ppm to about 475 ppm, even more specifically, about 472 ppm. In a particular embodiment, Reb M is present in an amount to provide a sucrose equivalence (SE) of greater than about 9.

[0278] In one embodiment, the beverage comprises brazein (or an analog thereof) and Reb A, wherein the brazein is present in an amount from about 10 ppm to about 20 ppm, more particularly, about 15 ppm to about 20 ppm, and Reb A is present in an amount from about 375 ppm to about 425 ppm, more particularly, about 400 ppm. In one embodiment, the amount of Reb A provides about 7 SE. According to this embodiment, a ppm ratio of brazein to Reb A between about 1:20 ppm and about 1:30 ppm (more particularly about 1:20 ppm, about 1:22 ppm, about 1:24 ppm, about 1:26 ppm, about 1:28 ppm, or about 1:30 ppm) has at least one improved organoleptic property (e.g., taste) compared to a certain ppm ratio of brazein to Reb A.

[0279] In one embodiment, the beverage comprises brazein (or an analog thereof) and Reb A, wherein the brazein is present in an amount between about 5 ppm and about 15 ppm, more particularly, about 10 ppm, and Reb A is present in an amount between about 325 ppm and about 375 ppm, more particularly, about 350 ppm. In one embodiment, the amount of Reb A provides a sucrose equivalence (SE) between about 6 and about 7. According to this embodiment, a ppm ratio of brazein to Reb A of about 1:30 has at least one improved organoleptic property compared to a ppm ratio of brazein to Reb A of about 1:70 or about 1:17.

[0280] In one embodiment, a beverage comprising brazeine and Reb A is provided, wherein the amount of brazeine is between about 25 ppm and about 35 ppm, more particularly, about 30 ppm, and the amount of Reb A is between about 220 ppm and about 240 pm, more particularly, about 230 ppm. According to this embodiment, a ppm ratio of brazeine to Reb A of greater than about 1:10 has at least one improved organoleptic property relative to a ppm ratio of brazeine to Reb A of less than about 1:10 ppm. In specific embodiments, a ppm ratio of brazeine to Reb A of about 1:12, about 1:14, about 1:16, or about 1:18 has at least one improved organoleptic property relative to a ppm ratio of brazeine of about 1:8, about 1:6, about 1:4, about 1:2, or about 1:1 ppm.

[0281] In one embodiment, a beverage comprising brazeine and Reb A is provided, wherein the amount of brazeine is between about 50 ppm and about 70 ppm, more particularly about 60 ppm, and the amount of Reb A is between about 100 ppm and about 130 pm, more particularly about 120 ppm. According to this embodiment, the ppm ratio of brazeine to Reb A is about 1:2.

[0282] In one embodiment, the beverage comprises from about 300 ppm to about 350 ppm of Reb A and from about 10 ppm to about 50 ppm of brazeine. In a preferred embodiment, the beverage comprising Reb A and brazeine tastes better and / or has a positive synergistic effect compared to a beverage sweetened with Reb A, wherein the beverage comprising Reb A and brazeine has the same sucrose equivalence value as a beverage sweetened with Reb A.

[0283] In one embodiment, the beverage comprises from about 30 ppm to about 50 ppm of brazein, from about 300 ppm to about 450 ppm of simonoside I, and optionally from about 50 ppm to about 100 ppm of Reb M80. In preferred embodiments, the beverage tastes similar to or better than a simonoside I-sweetened control or a Reb M80-sweetened control, wherein the beverage of the invention and the simonoside I-sweetened and / or Reb M80-sweetened control beverage have the same sucrose equivalence value. In a specific embodiment, the beverage comprises from about 40 ppm to about 50 ppm of brazein, and from about 350 ppm to about 450 ppm of simonoside I, without Reb M80. In a more specific embodiment, the beverage comprises from about 40 ppm to about 45 ppm of brazein, and from about 350 ppm to about 450 ppm of simonoside I, without Reb M80. In yet more specific embodiments, the beverage comprises about 45 ppm of brazein and about 410 ppm of simanoside I, without Reb M80. In another embodiment, the beverage comprises from about 25 to about 30 ppm of brazein, from about 300 ppm to about 400 ppm of simanoside I, and from about 50 ppm to about 100 ppm of Reb M80. In a more specific embodiment, the beverage comprises about 30 ppm of brazein, about 350 ppm of simanoside I, and about 70 ppm of Reb M80.

[0284] In one embodiment, the beverage comprises from about 30 to about 50 ppm of brazepine, and from about 7 to about 10 brix of high fructose corn syrup (HFCS). In a preferred embodiment, the beverage of the present invention tastes similar to or better than a HFCS sweetened control (10 brix). In a more specific embodiment, the beverage comprises from about 35 ppm to about 45 ppm of brazepine and from about 7 to about 9 brix of HFCS. In still more specific embodiments, the beverage comprises about 40 ppm of brazepine and about 8 brix of HFCS.

[0285] In one embodiment, a reduced-calorie beverage comprises from about 5 ppm to about 20 ppm of bulazein and from about 100 ppm to about 200 ppm of Reb M. In a preferred embodiment, the beverage of the present invention has a taste profile similar to a sucrose sweetened control (10 degrees Brix). In a specific embodiment, a reduced-calorie non-carbonated (non-carbonated) beverage comprises from about 10 ppm to about 15 ppm of bulazein and from about 100 ppm to about 150 ppm of Reb M. In a still more specific embodiment, a reduced-calorie non-carbonated beverage comprises from about 10 ppm to about 15 ppm of bulazein and about 115 ppm of Reb M. In another specific embodiment, a reduced-calorie carbonated beverage comprises about 20 ppm of bulazein, and from about 100 to about 150 ppm of Reb M.

[0286] In one embodiment, the beverage comprises from about 5 ppm to about 20 ppm of brazene, and from about 300 ppm to about 500 ppm of A95. In a specific embodiment, the beverage comprises from about 15 ppm to about 20 ppm of brazene, and from about 300 ppm to about 500 ppm of A95. In another embodiment, the beverage comprises about 15 ppm or about 20 ppm of brazene and about 400 ppm of A95.

[0287] In one embodiment, the beverage has an improved temporal profile, flavor profile, and / or taste profile relative to a beverage that does not contain bulazeine or contains bulazeine in an amount of less than about 20 ppm, even more particularly, less than about 15 ppm. In a particular embodiment, the temporal profile, flavor profile, and / or taste profile of the beverage is more sugar-like. In one embodiment, the beverage has reduced sweetness lingering, reduced bitterness, reduced bitter aftertaste, reduced astringency, improved mouthfeel (e.g., greater richness or body), etc.

[0288] Beverages may further include additives, including, but not limited to, carbohydrates, polyols, amino acids and their corresponding salts, polyamino acids and their corresponding salts, sugar acids and their corresponding salts, nucleotides, organic acids, inorganic acids, organic salts (including organic acid salts and organic base salts), inorganic salts, bitter compounds, flavoring agents and flavoring ingredients, astringent compounds, proteins or protein hydrolysates, surfactants, emulsifiers, flavonoids, alcohols, polymers and combinations thereof. Any suitable additive as described herein may be used.

[0289] The beverage may further contain one or more functional ingredients described in detail above. Functional ingredients include, but are not limited to, vitamins, minerals, antioxidants, preservatives, glucosamine, polyphenols and combinations thereof. Any suitable functional ingredient described herein may be used.

[0290] It is contemplated that the pH of a sweetened composition (e.g., a beverage) will not substantially or adversely affect the taste of the sweetener. A non-limiting example of a pH range for a sweetenable composition may be from about 1.8 to about 8. Another example includes a pH range from about 2 to about 5. In a particular embodiment, the pH of the beverage is from about 3 to about 3.25.

[0291] The temperature of the beverage comprising brazeein (or its analogue) may for example range from about 4°C to about 100°C, such as for example from about 4°C to about 25°C.

[0292] The beverage may be a high calorie beverage having about 120 calories per 8 ounce serving.

[0293] The beverage may be a mid-calorie beverage having about 80 calories per 8 ounce serving.

[0294] The beverage may be a low calorie beverage having less than 40 calories per 8 ounce serving.

[0295] The beverage may be a zero calorie beverage having less than 5 calories per 8 ounce serving.

[0296] In one embodiment, the beverage or beverage product has a low glycemic index. The glycemic index is a value assigned to foods based on how quickly those foods cause blood sugar levels to rise. Low glycemic index diets (including but not limited to beverages) are intended to have a more beneficial effect on blood sugar control in diabetic populations and may also provide metabolic benefits to the general population.

[0297] In a specific embodiment, the glycemic index of a beverage or a beverage product is at least 10% lower than the glycemic index of a substantially similar product made using a conventional sweetener (e.g., a sweetener based on sucrose). The method for testing the glycemic index of a beverage has been described, e.g., at Wolever, et al. Nutrition Research [nutrition research] 23:621-629, provided in 2003. In a specific embodiment, a beverage or a beverage product has a glycemic index (GI) of about 55 or less. In a specific embodiment, a beverage or a beverage product is a nutritious beverage with a low glycemic index.

[0298] In one embodiment, a beverage comprising between about 1 and about 30 ppm and less than about 500 ppm of one or more steviol glycosides is disclosed, wherein the liquid matrix of the beverage is selected from the group consisting of: water, phosphoric acid, phosphate buffer, citric acid, citrate buffer, carbon-treated water and combinations thereof. The pH of the beverage can be from about 3 to about 3.5. The beverage can further include additives, such as, erythritol. The beverage can further include functional ingredients, such as, vitamins.

[0299] Method for improving temporal profile or flavor profile

[0300] Disclosed is a method for imparting a more sugar-like temporal profile, flavor profile, and / or taste profile to a consumable (e.g., a beverage), the method comprising adding brazepine (or its analogs) or a brazepine-containing sweetener composition or a flavor-modifying composition disclosed herein to the consumable. The consumable may be referred to as a sweetenable composition or a flavor-modifiable composition.

[0301] Without being bound by any particular theory, it is believed that brazyne binds to mucins in the oral cavity. Mucins are the major organic component of mucus, a viscous, viscoelastic material that coats all mucosal surfaces. Structurally, mucins are high molecular weight epithelial glycoproteins with a high content of clustered oligosaccharides O-glycosidically linked to tandem repeating peptides rich in threonine, serine, and proline.

[0302] In the mouth, mucins coat both hard and soft tissues. Several salivary mucins are known, including MUC5B, MUC7 (formerly known as MG2), MUC19, MUC1, and MUC4. Based on macromolecular characteristics, they can be divided into high (>1000 kDa) and low (200-300 kDa) molecular weight forms. Salivary mucins are synthesized by paired mucous acinar cells of the submandibular gland (SMG) and sublingual gland (SLG) and small salivary glands distributed throughout the palate and buccal mucosa.

[0303] The method may further include adding other sweeteners, additives, functional ingredients, and combinations thereof. Any sweetener, additive, or functional ingredient described in detail herein may be used.

[0304] In one embodiment, a method for imparting a more sugar-like temporal profile and flavor profile to a consumable (e.g., a beverage) comprises adding a combination of brazepine (or its analogs) or a sweetener composition or a flavor-modifying composition containing brazepine to the consumable, thereby imparting a more sugar-like temporal profile or flavor profile.

[0305] As used herein, "sugar-like" characteristics include any characteristics similar to those of sucrose and include, but are not limited to, maximum response, flavor profile, temporal profile, taste profile, adaptation behavior, mouthfeel, concentration / response function, tastant / and flavor / sweetness interactions, spatial pattern selectivity, and temperature effect.

[0306] These characteristics are the aspects in which the taste of sucrose differs from the taste of bulazeine (or its analogs). Of these, however, the flavor profile and temporal profile are particularly important. In a single tasting of a sweetened food or beverage, differences may be noted in (1) the attributes that make up the flavor profile of a sweetener and (2) the sweetness onset and dissipation rates (between those observed for sucrose and for bulazeine (or its analogs)) that make up the temporal profile of the sweetener.

[0307] In one embodiment, the method disclosed herein improves one or more organoleptic properties of a consumer's product compared to a consumer's product that does not contain a sweetener composition or a flavor modification composition disclosed herein. In certain embodiments, the improvement is measured by a sensory test. The sensory test can be a taste test, a blind test, or a combination thereof. The sensory test can use one or more different schemes. For example, the sensory test can be a "triangulation method", by ISO requirements, or a combination thereof. The taste test can be the average value of multiple tests.

[0308] Taste testing can be a screening test, a professional taste test, or a market research test. A screening test can be performed by at least 1, 2, 3, 4, 5, 6, 7, 8, or 9 taste testers. A professional taste test can be performed by at least 10, 15, 20, 25, or 30 taste testers. A market research test can be performed by at least 31, 40, 50, 60, 70, 80, 90, 100, 150, 200, 300, 400, or 500 taste testers. In some cases, the taste tester can be a person with average taste perception, a professional taste tester, a person who has passed a tasting test by correctly identifying a food or food component, or a person who can identify the relative amount of a taste or flavor (e.g., correctly determining different amounts of sugar in water).

[0309] In one embodiment, whether the profile is more sugar-like is determined by an expert sensory panel that tastes a composition comprising sugar and a composition comprising brazil (or an analog thereof) optionally in combination with at least one steviol (e.g., Reb M, Reb A) or HFCS, both with and without additives, and provides their impressions of the similarity of the profiles of the sweetener compositions (both with and without additives) to those comprising sugar. Suitable procedures for determining whether a composition has a more sugar-like taste are described in the Examples described herein below.

[0310] In a particular embodiment, a group of assessors is used to measure the reduction of sweetness retention. In short, a group of assessors (typically 8 to 12 individuals) are trained to evaluate sweetness perception and measure sweetness at several time points within 3 minutes from the initial intake of the sample until it is spit out. Statistical analysis is used to compare the results between the sample containing additives and the sample without additives. The reduction in the score of the time point measured after the sample is cleared of the outlet indicates the reduction of sweetness perception.

[0311] The group of assessors may be trained using procedures known to those of ordinary skill in the art. In a particular embodiment, the group of assessors may be trained using Spectrum TM Descriptive analysis methods (Melgaard et al., Sensory Evaluation Techniques , 3rd edition, Chapter 11). Ideally, the focus of the training should be on identifying and measuring basic tastes; in particular, sweetness. To ensure accuracy and reproducibility of the results, each assessor should repeat the measurement of the reduction in sweetness retention about three to about five times per sample, resting for at least five minutes between each repeat and / or each sample and rinsing thoroughly with water to clean the mouth.

[0312] Typically, the method for measuring sweetness includes taking a 10 mL sample and placing it in the mouth, keeping the sample in the mouth for 5 seconds and gently rotating the sample in the mouth, assessing the sweetness intensity perceived within 5 seconds, spitting out the sample (not swallowing after spitting out the sample), rinsing with a mouthful of water (e.g., moving the water vigorously in the mouth, as if rinsing with mouthwash) and spitting out the rinse water, assessing the perceived sweetness intensity immediately after spitting out the rinse water, waiting for 45 seconds and confirming the time of the maximum sweetness intensity perceived while waiting for that 45 seconds and assessing the sweetness intensity at that time (moving the mouth normally and swallowing when necessary), assessing the sweetness intensity after another 10 seconds, assessing the sweetness intensity after another 60 seconds (accumulating 120 seconds after rinsing), and assessing the sweetness intensity after another 60 seconds (accumulating 180 seconds after rinsing). Rest for 5 minutes between samples, rinse thoroughly with water to clean the mouth.

[0313] In some embodiments, the temporal profile, flavor profile, and / or taste profile are evaluated in vitro, for example, in a suitable assay, such as an in vitro system based on a cell model overexpressing sweet and / or bitter receptors. In particular embodiments, the in vitro system comprises TAS1R2 / TAS1R3 receptors.

[0314] In one embodiment, the sweetener compositions or flavor and / or taste modifying compositions disclosed herein are capable of modifying one or more characteristics of a consumable including, but not limited to, mouthfeel, bitterness, bitter aftertaste, or lingering sweetness.

[0315] Mouthfeel refers to the textural aspect of a food or beverage that is responsible for the characteristic tactile sensations felt on the lining of the oral cavity, including the tongue, gums, and teeth. These mouthfeel can include, but are not limited to, astringency, stickiness, smoothness, and mouth-coating. Mouthfeel is a fundamental sensory attribute that, along with taste and smell, determines the overall flavor of a consumer product.

[0316] In one embodiment, brazee (or its analogs) or sweetener compositions and / or flavor modifier compositions containing brazee disclosed herein impart improved mouthfeel to the consumables (e.g., beverages) to which they are added, relative to conventional sweetener compositions or flavor and / or flavor modifier compositions. In one embodiment, the "improved mouthfeel" can be determined by a tasting group drinking the beverage by comparing it with the same beverage without the flavor modifier composition or its active ingredient. In a specific embodiment, the mouthfeel is improved by about 5%, about 10%, about 15%, about 20%, or about 25% or more. In a specific embodiment, the improved mouthfeel is characterized by an increased rich taste, body, or richness. In a specific embodiment, the improved mouthfeel is characterized by a more syrupy feeling.

[0317] In another embodiment, the sweetener composition or flavor modifying composition disclosed herein improves (reduces) the bitterness of the consumable (e.g., beverage) to which it is added relative to a conventional sweetener composition or flavor and / or taste modifying composition. The "improved bitterness" or "reduced bitterness" can be determined by a tasting panel drinking the beverage by comparing it with the same beverage without the taste modifying composition or its active component. In a specific embodiment, the bitterness is reduced by about 5%, about 10%, about 15%, about 20%, or about 25% or more.

[0318] In another embodiment, the sweetener composition or flavor modifying composition disclosed herein reduces the bitter aftertaste of the consumable (e.g., beverage) to which it is added relative to a conventional sweetener composition or flavor and / or taste modifying composition. This "reduced bitter aftertaste" can be determined by a tasting panel drinking the beverage by comparing it with the same beverage without the taste modifying composition or its active component. In a specific embodiment, the bitter aftertaste is reduced by about 5%, about 10%, about 15%, about 20%, or about 25% or more.

[0319] In a specific embodiment, the sweetener composition or flavor modification composition disclosed herein reduces the bitterness of the consumables (e.g., beverages) to which it is added, wherein the reduction is measured by the Metachronic VAS score characteristic curve of bitterness, and more particularly, the reduction is at least about 0.5 points, about 1 point, about 1.5 points, about 2.0 points, about 2.5 points, about 3.0 points, about 3.5 points or about 4.0 points or more. In certain embodiments, the reduction is between about 0.5 and about 4.0 points, about 1.0 and about 3.5 points or about 1.5 and about 3.0 points.

[0320] In another embodiment, the sweetener composition and / or flavor modification composition disclosed herein has improved (reduced) sweetness retention. In one embodiment, this "improved sweetness retention" can be checked by a tasting panel drinking the beverage by comparing with the same beverage without the flavor modification composition or its active component. In a specific embodiment, the sweetness retention is reduced by about 5%, about 10%, about 15%, about 20% or about 25% or more.

[0321] In another embodiment, the flavor-modifying composition disclosed herein provides a reduction in sour taste. In one embodiment, this "reduced sour taste" can be examined by a taste panel drinking the beverage by comparing it to the same beverage without the flavor-modifying composition or its active ingredient. In specific embodiments, the sour taste is reduced by about 5%, about 10%, about 15%, about 20%, or about 25% or more.

[0322] Examples

[0323] Example 1: Brazein + Reb M (sweetener composition)

[0324] Brazein and Reb M80 were added to citric acid solution and lemon / lime diet soda, respectively. The results are shown in Tables I and II below, respectively.

[0325] Table I:

[0326] In citric acid solution

[0327]

[0328] Table II:

[0329] In lemon / lime carbonated drinks

[0330]

[0331] Example 2: Comparison of Reb A and Reb A + Brazein (Sweetener Combinations)

[0332] Reb A was compared to Reb A + Brazein in citric acid solution as described below. The results are shown in Tables III and IV below.

[0333] Table III

[0334] 5-10 Brix Citric Acid Sucrose Comparison

[0335]

[0336] Table IV:

[0337] In citric acid solution:

[0338]

[0339] The results showed that the Reb A + brazyne blend had a positive synergistic effect compared with Reb A alone.

[0340] Example 3: Simagroside I + Brazia (sweetener composition)

[0341] Brazyne, simianoside I and / or Reb M80 were added to a citric acid simulated beverage. The results are shown in Table V below.

[0342] Table V:

[0343]

[0344] The results showed that the blends of simonoside I with brazil and / or Reb M had a positive synergistic effect (sweetness quality) compared to Reb M80 alone.

[0345] Example 4: Brazein + HFCS (sweetener composition)

[0346] Brazein and high fructose corn syrup (HFCS) were added to a citric acid simulated beverage. The results are shown in Table V below:

[0347] Table V

[0348] In citric acid simulated beverages

[0349] Brazien HFCS result Comparison N / A 10 degrees Brix refer to Test 3 30ppm 8 degrees Brix Similar, but still lacks some sweetness Test 4 40ppm 8 degrees Brix Similar to control

[0350] The blend of HFCS and brazil provides a similar taste (sweetness quality) as HFCS alone.

[0351] Example 5: Brazein + Reb M (taste-modifying composition)

[0352] Reb M and brazil were added to various beverages as described below. A full sugar sample was provided as a control (10 Brix).

[0353] Reduce sugar by 50% in still lemon / lime drinks

[0354]

[0355] Reduce sugar by 50% in carbonated lemon / lime drinks

[0356]

[0357] Example 6: Brazein + Reb M (taste-modifying composition)

[0358]

[0359] Brazein (1-15ppm) improves the taste properties of Reb M80 at 472ppm such as sourness, less sweetness, masking bitterness and lingering. Better mouthfeel, richness and body.

[0360] Example 7: Brazein + Reb A95 (taste-modifying composition)

[0361] "A95" refers to a steviol glycoside blend comprising primarily reb D and reb M. A method for obtaining A95 is provided in WO 2017 / 059414, which is incorporated herein by reference. The following is an exemplary A95 blend:

[0362]

[0363]

[0364] Brazein (5ppm to 20ppm) can improve mouthfeel, block bitterness, and persist in A95 at 400ppm.

Claims

1. A sweetener composition comprising (i) brazepine or an analog thereof, and (ii) high fructose corn syrup; wherein the brazepine or an analog thereof is present in an amount such that when the sweetener composition is combined with a liquid matrix, it forms a beverage or beverage product comprising brazepine or an analog thereof in an amount between 1 ppm and 50 ppm.

2. A beverage or beverage product comprising the sweetener composition of claim 1, wherein the beverage comprises a liquid base and the sweetener composition of claim 1, and wherein the beverage product is a beverage concentrate, beverage syrup or beverage powder infused beverage from which a full-strength beverage can be prepared by adding water.

3. The beverage or beverage product of claim 2, wherein the brazeein is present in an amount between 1 ppm and 40 ppm.

4. The beverage or beverage product of claim 2, wherein the brazeein is present in an amount selected from the group consisting of: an amount between 1 ppm and 30 ppm.

5. The beverage or beverage product of any one of claims 2-4, wherein the beverage or beverage product has at least one improved organoleptic characteristic compared to a beverage or beverage product without the sweetener composition, wherein the organoleptic characteristic is selected from the group consisting of: temporal profile, flavor profile, taste, bitter aftertaste, sweetness lingering, mouthfeel, or a combination thereof.

6. The beverage or beverage product of claim 5, wherein the at least one improved organoleptic property is a reduced bitter aftertaste or an improved mouthfeel.

7. The beverage or beverage product of claim 5, wherein the at least one improved organoleptic property is mouthfeel improvement and the amount of brazeine is between 1 ppm and 40 ppm.

8. The beverage or beverage product according to any one of claims 2 to 4, wherein the beverage or beverage product is selected from low-calorie or calorie-free beverages or beverage products.

9. The beverage or beverage product of any one of claims 2-4, wherein the beverage is selected from the group consisting of cola, ginger ale, soft drinks, root beer, fruit juice, fruit flavored juice, vegetable juice, vegetable flavored juice, sports drink, energy drink, plant protein beverage, near water beverage, tea, coffee, cocoa drink and beverage containing a dairy component.

10. A method of improving at least one organoleptic property of a beverage or beverage product, the method comprising adding the sweetener composition of claim 1 to a liquid matrix, thereby providing a beverage or beverage product having at least one improved organoleptic property.

11. The method of claim 10, wherein the improved sensory properties are selected from the group consisting of temporal profile, flavor profile, taste, bitter aftertaste, sweetness lingering, mouthfeel, or combinations thereof.

12. The method of claim 10, wherein the improved organoleptic property is a reduction in bitter aftertaste or an improvement in mouthfeel.

13. The method of claim 10, wherein the improved organoleptic property is mouthfeel improvement and the amount of brazeine present in the beverage or beverage product is an amount between 1 ppm and 40 ppm.

14. The method of claim 10 or 11, wherein brazein is present in the beverage or beverage product in an amount between 1 ppm and 50 ppm.

15. The method of claim 10 or 11, wherein the brazeine is present in the beverage or beverage product in an amount selected from the group consisting of: an amount between 1 ppm and 40 ppm.

16. The sweetener composition of claim 1, wherein the bulazein analog differs from wild-type bulazein by at least one amino acid.

17. The sweetener composition of claim 16, wherein the brazyne analog has at least one of the following: Asp40Lys, Glu41Ala, Lys42Ala, Asp50Lys, Tyr54Trp, Asp29Ala, Asp29Asn / Glu41Lys, and Asp29Lys / Glu41Lys.

18. The sweetener composition of claim 1, wherein the brazyne has an amino acid sequence of SEQ ID NO:

1.

19. The sweetener composition of claim 1, wherein the brazeine is pGlu-brazeine or des-pGlu1-brazeine.

20. The beverage or beverage product of claim 2, further comprising at least one organic acid additive salt which is a sodium, calcium, potassium or magnesium salt of an organic acid.

21. The beverage or beverage product of claim 20, wherein the organic acid is selected from the group consisting of citric acid, malic acid, tartaric acid, fumaric acid, lactic acid, alginic acid, ascorbic acid, benzoic acid, and adipic acid.

22. The method of claim 10, wherein the bulazines analog differs from wild-type bulazines by at least one amino acid.

23. The method of claim 22, wherein the brazyne analog has at least one of the following: Asp40Lys, Glu41Ala, Lys42Ala, Asp50Lys, Tyr54Trp, Asp29Ala, Asp29Asn / Glu41Lys, and Asp29Lys / Glu41Lys.

24. The method of claim 10, wherein the brazyne has an amino acid sequence of SEQ ID NO:

1.

25. The method of claim 10, wherein the brazeine is pGlu-brazeine or des-pGlu1-brazeine.

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