WO2025215649A1 - Sweet proteins and uses thereof - Google Patents

Sweet proteins and uses thereof

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Publication number
WO2025215649A1
WO2025215649A1 PCT/IL2025/050330 IL2025050330W WO2025215649A1 WO 2025215649 A1 WO2025215649 A1 WO 2025215649A1 IL 2025050330 W IL2025050330 W IL 2025050330W WO 2025215649 A1 WO2025215649 A1 WO 2025215649A1
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WO
WIPO (PCT)
Prior art keywords
seq
ppm
flavor
protein
examples
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/IL2025/050330
Other languages
English (en)
French (fr)
Inventor
Neta BEN-BASSAT
Inbar ZUKER
Shmuel Marko
Ilan Samish
Naama KOPELMAN
Marina KASCHDAN
Keren GORZALCZANY
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Amai Proteins Ltd
Original Assignee
Amai Proteins Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Amai Proteins Ltd filed Critical Amai Proteins Ltd
Publication of WO2025215649A1 publication Critical patent/WO2025215649A1/en
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/30—Artificial sweetening agents
    • A23L27/31—Artificial sweetening agents containing amino acids, nucleotides, peptides or derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Preparation or treatment thereof
    • A23L2/52—Adding ingredients
    • A23L2/60—Sweeteners
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C07K14/43—Sweetening agents, e.g. thaumatin, monellin

Definitions

  • the present disclosure relates to sweet proteins and uses thereof.
  • Taste modifiers are substances that can alter or modulate the perception of taste by interacting with taste receptors on the tongue. These modifiers can enhance, diminish, or change the taste of food and beverages and are used in the food industry for various purposes, such as enhancing desirable flavors and masking unpleasant tastes.
  • Some common examples of taste modifiers include sweet enhancers, butter blockers, salt enhancers and umami enhancers.
  • salt enhancers can amplify or intensify the perception of saltiness in food, allowing for the reduction of actual salt content while maintaining a desired level of salty taste.
  • a method for modifying at least one flavor in a comestible product comprises adding a flavor modifying concentration of at least one sweet protein to the comestible product, wherein the flavor modifying concentration, when added to the comestible product, results in a sweetness level lower than that achieved by 1.5% sucrose, wherein the at least one sweet protein selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO 466, SEQ ID NO:350, SEQ ID NO:480, SEQ ID NO:481, a variant thereof or a combination thereof.
  • a method for modifying at least one flavor in a comestible product comprises adding a flavor modifying concentration of at least one protein to the comestible product, wherein the flavor modifying concentration of the at least one protein is between about 1 ppm and about 300 ppm, wherein the at least one protein selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NON, SEQ ID N0:8, SEQ ID N0:9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO 466, SEQ ID NO:350, SEQ ID NO:480, SEQ ID NO:481, a variant thereof or a combination thereof.
  • a comestible product comprising at least one protein, the at least one protein is present in a concentration that produces a sweetness level lower than that achieved by 1.5% sucrose, wherein the at least one protein selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NON, SEQ ID NON, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NON, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO 466, SEQ ID NO:350, SEQ ID NO:480, SEQ ID NO: 481, a variant thereof or a combination thereof.
  • a comestible product comprising at least one protein, the at least one protein is present at a concentration of between about 1 ppm and about 300 ppm, wherein the at least one protein is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO 466, SEQ IDNO:350, SEQ IDNON80, SEQ ID NO:481, avariant thereof or a combination thereof.
  • the present disclosure relates to sweet proteins and their use as flavor-modifying proteins (FMPs). More specifically, the disclosure is directed to the use of one or more sweet proteins added to comestible products at concentrations sufficiently low such that the sweetness induced by the protein(s) at this concentration is perceived as less than that of 1.5% (w/w) sucrose solution. This concentration is referred to herein as the “flavormodifying concentration” or “FMP concentration ”
  • the present disclosure provides a method for modifying at least one flavor attribute in a comestible product.
  • the method comprises adding at least one sweet protein to the comestible product at a flavor modifying concentration.
  • the flavor-modifying concentration of a given sweet protein was determined by comparing the sweetness intensity of samples or products containing varying concentrations of the protein(s), in the absence of added sweeteners, to that of matched control samples or products containing approximately 1.5% sucrose and no sweet protein. The highest protein concentration that produced a sweetness perception lower than that of control samples or products was designated as the flavor-modifying concentration.
  • an FMP concentration or FMP concentration refers to a concentration of one or more sweet proteins that induces a sweetness perception lower than that of a 1.5% (w/w) sucrose solution.
  • Sweetness perception may be assessed using any suitable method known in the art. For example, sensory evaluation is performed by trained panelists using methods such as the two-alternative forced choice (2 -AFC) test, as further detailed in the Examples.
  • the 1.5% sucrose solution may serve as a reference standard for minimal perceptible sweetness.
  • the term “at least one protein” refers to one or more sweet proteins, which may be the same or different.
  • the term “flavor-modifying concentration” or “FMP concentration” refers to the concentration of a single sweet protein or the total concentration of a combination of sweet proteins that exhibit flavor-modifying properties.
  • the present disclosure further provides food and beverage products containing one or more sweet proteins at a flavor-modifying concentration.
  • sweet proteins at such low levels are effective in modulating flavor perception — enhancing or reducing specific flavor notes — without contributing significant sweetness.
  • sweet proteins are readily digestible and, due to their extremely low usage levels, contribute to negligible caloric value, functioning as effectively zero-calorie ingredients.
  • certain sweet proteins exhibit heat stability, making them suitable for thermally processed products. Their stability may also support extended shelf-life applications, enabling their use in packaged products with long storage durations.
  • the present disclosure provides a method for modifying at least one flavor attribute in a comestible product.
  • the method comprises adding at least one sweet protein to the comestible product at a flavor modifying concentration, wherein the flavor modifying concentration is selected such that the sweetness induced by the concentration is lower than the sweetness produced by a sucrose solution at a concentration between about 0.3% and about 3% (w/w), and in some embodiments, lower than that produced by about 1.5% sucrose solution.
  • the method comprises adding at least one sweet protein to the comestible product at a flavor modifying concentration, wherein the flavor modifying concentration is selected such that the sweetness induced by the concentration is lower than the sweetness produced by a sucrose solution at a concentration of about 1.5% sucrose.
  • food and beverage products containing the at least one sweet protein at a flavor-modifying concentration exhibited changes (modifications) — such as enhancement or suppression — in various flavor attributes when compared to corresponding products without the sweet protein.
  • the present disclosure provides a comestible product comprising at least one sweet protein at a flavor-modifying concentration.
  • the present disclosure provides a comestible product comprising at least one sweet protein at a concentration that induces a level of sweetness lower than that produced by up to about 5% (w/w) sucrose in the same comestible product.
  • the comestible product comprises at least one sweet protein at a concentration that induces a level of sweetness lower than that produced by up to about 4% (w/w), at times up to about 3% (w/w), at times up to about 2% (w/w) sucrose in the same comestible product.
  • the comestible product comprises at least one sweet protein at a concentration that induces a level of sweetness lower than that produced by up to about 1.5% (w/w) sucrose in the same comestible product, at times up to 1% sucrose in the same comestible product.
  • the comestible product comprises at least one sweet protein at a concentration that induces a level of sweetness lower than that produced by about 4% (w/w), at times about 3% (w/w), at times about 2% (w/w) sucrose in the same comestible product.
  • the comestible product comprises at least one sweet protein at a concentration that induces a level of sweetness lower than that produced by about 1.5% (w/w) sucrose in the same comestible product, at times about 1% sucrose in the same comestible product.
  • the flavor-modifying concentration of the at least one sweet protein may depend, for example, on its sweetness potency.
  • the flavor-modifying concentration of this sweet protein may be between about 8 ppm and about 20 ppm, and in some cases between about 8 ppm and about 15 ppm.
  • the protein concentration may be at most about 8 ppm, at most about 10 ppm, at most about 15 ppm, or at most about 20 ppm.
  • the flavor modifying concentration is at most about 300 ppm, at most about 250 ppm at most about 150 ppm, at times at most about lOOppm, at times between about 1 ppm and about 100.
  • the flavor modifying concentration may be at most about 300ppm, at times at most about 250 ppm, at times at most about 200 ppm, at times at most about 150 ppm, at most about 100 ppm, at times at most about 95 ppm, at times at most about 90 ppm, at times at most about 85 ppm, at times at most about 80 ppm, at times at most about 75 ppm, at times at most about 70 ppm, at times at most about 65 ppm, at times at most about 60 ppm, at times at most about 55 ppm, at times at most about 50 pp, at times at times at most about 45 ppm, at most about 40 ppm, at times at most about 35 ppm, at times at most about 30 ppm, at times at most about 25 ppm, at times at most about 20 ppm, at times at most about 15 ppm, at times at most about 10 ppm, at times at most about 5 ppm.
  • the flavor modifying concentration may be between about 1 ppm and about 300 ppm, at times between about 10 ppm and about 300 ppm, at times between about 30 ppm and about 300 ppm, at times between about 50 ppm and about 300 ppm, at times between about 70 ppm and about 300 ppm, at times between about 100 ppm and about 300 ppm.
  • the flavor modifying concentration may be between about 1 ppm and about 100 ppm, at times between about 5 ppm and about 100 ppm, at times between about 10 ppm and about 100 ppm.
  • the flavor modifying concentration may be between about 10 ppm and about 90 ppm, at times between about 10 ppm and about 80 ppm, at times between about 10 ppm and about 70 ppm, at times between about 10 ppm and about 60 ppm, at times between about 10 ppm and about 50 ppm, at times between about 10 ppm and about 40 ppm, at times between about 10 ppm and about 30 ppm.
  • the flavor modifying concentration may be between about 20 ppm and about 90 ppm, at times between about 20 ppm and about 80 ppm, at times between about 20 ppm and about 70 ppm, at times between about 20 ppm and about 60 ppm, at times between about 20 ppm and about 50 ppm, at times between about 20 ppm and about 40 ppm, at times between about 20 ppm and about 30 ppm.
  • the flavor modifying concentration may be between about 1 ppm and about 90 ppm, at times between about 1 ppm and about 85 ppm, at times between about 1 ppm and about 80 ppm, at times between about 1 ppm and about 75 ppm, at times between about 1 ppm and about 70 ppm, at times between about 1 ppm and about 65 ppm, at times between about 1 ppm and about 60 ppm, at times between about 1 ppm and about 55 ppm, at times between about 1 ppm and about 50 ppm, at times between about 1 ppm and about 45 ppm, at times between about 1 ppm and about 40 ppm, at times between about 1 ppm and about 35 ppm, at times between 1 ppm and about 30 ppm, at times between about 1 ppm and about 30 ppm, at times between about 1 ppm and about 30 ppm, at times between about 1 ppm and about 25 ppm, at times between about 1 ppm and about
  • the flavor modifying concentration may be between about 1 ppm and about 40 ppm, at times between about 3 ppm and about 40 ppm, at times between 3 ppm and about 35 ppm, at times between about 5 ppm and about 35 ppm, at times between about 5 ppm and about 30 ppm, at times between about 5 ppm and about 25 ppm, at times between about 5 ppm and about 20 ppm.
  • the flavor modifying concentration may be about 1 ppm, at times about 2 ppm, at times about 3 ppm, at times about 4 ppm, at times about 5 ppm, at times about 6 ppm, at times about 7 ppm, at times about 8 ppm, at times about 9 ppm, at times about 10 ppm, at times about 11 ppm, at times about 12 ppm, at times about 13 ppm, at times about 14 ppm, at times about 15 ppm, at times about 16 ppm, at times about 17 ppm, at times about 18 ppm, at times about 19 ppm, at times about 20 ppm, at times about 21 ppm, at times about 22 ppm, at times about 23 ppm, at times about 24 ppm, at times about 25 ppm, at times about 27 ppm, at times about 30 ppm, at times about 32 ppm, at times about 35 ppm, at times about 40ppm, at times about 45ppm, at times about
  • the protein having FMP is a sweet protein.
  • the present disclosure is not limited to a specific sweet protein.
  • the sweet protein is identical to a naturally occurring protein or is a naturally occurring protein.
  • the sweet protein is found in plants such as tropical plants.
  • Non-limiting examples of a plant include at least one of capparis masaikai, oobli, serendipity berry, katemfe, miracle fruit berry, or lemba.
  • the sweet protein is found in the honey truffle.
  • the sweet protein is Monellin, Thaumatin, Miraculin, Curculin, Brazzein, Mabinlin, a sweet protein from honey truffle.
  • the sweet protein is Monellin.
  • Monellin comprises chain A Monellin (GenBank Entry No. P02881; denoted herein by SEQ ID NO: 1) and chain B Monellin (GenBank Entry No. P02882; denoted herein by SEQ ID NO:2).
  • Monellin comprises chain A Monellin (GenBank Entry No. 1KRL A; denoted herein by SEQ ID NO:480) and chain B Monellin (GenBank Entry No. 3M0N_B; denoted herein by SEQ ID NO:481).
  • the sweet protein is Thaumatin.
  • the sweet protein is Thaumatin-1 (GenBank Entry No.
  • the sweet protein is Thaumatin-2 (GenBank Entry No. P02884; denoted herein by SEQ ID NO:4).
  • the sweet protein is Miraculin (GenBank Entry No. P13087; denoted herein by SEQ ID NO:5).
  • the sweet protein is Curculin-1 GenBank Entry No. P19667; denoted herein by SEQ ID NO:6) or Curculin-2 (GenBank Entry No. Q6F495; denoted herein by SEQ ID NO: 7).
  • the sweet protein is Brazzein (also known as: Defensin-like protein) (GenBank Entry No.
  • the sweet protein is Mabinlin I/Sweet protein mabinlin-1 (GenBank Entry No. P80351; denoted herein by SEQ ID NO:9), Mabinlin II (also known as Sweet protein mabinlin-2) (GenBank Entry No. P30233; denoted herein by SEQ ID NO: 10), Mabinlin III (also known as Sweet protein mabinlin-3) (GenBank Entry No. P80352; denoted herein by SEQ ID NO: 11), Mabinlin IV (also known as Sweet protein mabinlin-4) (GenBank Entry No.
  • the sweet protein is derived from the honey truffle, or Mattirolomyces terfezioides. In some embodiments, the sweet protein is a sweet truffle protein or mycodulcein (denoted herein by SEQ ID NO: 466).
  • the sweet protein is or comprises one or more proteins denoted by SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 466, SEQ ID NO:480, SEQ ID NO:481, a variant thereof or any combinations thereof.
  • SEQ ID NO: amino acid listed by the referred SEQ ID NO:.
  • variant refers to a sequence that contains at least one amino acid modification (e.g. substitution, deletion, or insertion) as compared to a reference protein.
  • the present disclosure also encompasses a modified nucleic acid sequence, including corresponding substitution, deletion or insertion of codons such that the modified nucleic acid sequence encodes the protein variant.
  • the present disclosure provides a method for modifying at least one flavor attribute in a comestible product.
  • the method comprises adding at least one sweet protein to the comestible product at a flavor modifying concentration, wherein the at least one sweet protein selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NON, SEQ ID NO:5, SEQ ID NON, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NON, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO 466, SEQ ID NO:480, SEQ ID NO:481 a variant thereof or any combination thereof.
  • a method for modifying at least one flavor in a comestible product comprises adding at least one protein to the comestible product, wherein the concentration of the at least one protein is between about 1 ppm and about 300 ppm, and wherein the at least one protein selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NON, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO 466, SEQ ID NO:480, SEQ ID NO:481, a variant thereof or any combination thereof.
  • a comestible product comprising at least one sweet protein at a flavor-modifying concentration, wherein the at least one protein selected from the group consisting of SEQ ID NO: 1, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO 466, SEQ ID NO:480 SEQ ID NO:481, a variant thereof or any combination thereof.
  • a comestible product comprising at least one sweet protein at a concentration of between about Ippm and about 300ppm, wherein the at least one protein selected from the group consisting of SEQ ID NO: 1, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NON, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO 466, SEQ ID NO:480 SEQ ID NO:481, a variant thereof or any combination thereof.
  • the sweet protein is a non-natural protein.
  • the sweet protein is a synthetic protein or an engineered protein.
  • the synthetic sweet protein may comprise the entire or part of the amino acid sequence of the naturally occurring protein (all or part of the protein's polypeptide chains) or part thereof.
  • the sweet protein may comprise a bond modification of a naturally occurring protein resulting in single polypeptide chain which corresponds to a naturally occurring protein, such that the at least two polypeptide chains of the wild type protein are covalently attached by other amino acids.
  • the sweet protein is a modified Monellin protein known as MNEI. In some embodiments, the sweet protein is MNEI or a variant thereof.
  • the sweet protein is a single chain Monellin (MNEI) protein.
  • the sweet protein is MNEI as denoted by/comprising the amino acid listed in SEQ ID NO: 14 or a variant thereof.
  • the sweet protein is MNEI as denoted by SEQ ID NO: 350 or a variant thereof.
  • the protein is or comprises one or more proteins comprising an amino acid listed as SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NON, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO 350, SEQ ID NO:466, SEQ ID NO:480, SEQ ID NO:481 or a variant thereof or any combination thereof.
  • the reference protein is denoted by SEQ ID NO: 14, by SEQ ID NO: 350, any variants thereof or any combinations thereof.
  • the reference protein is MNEI as denoted by SEQ ID NO: 14 or by SEQ ID NO: 350.
  • the present disclosure provides a method for modifying at least one flavor attribute in a comestible product.
  • the method comprises adding at least one sweet protein to the comestible product at a flavor modifying concentration, wherein the at least one sweet protein selected from the group consisting of SEQ ID NO: 14, by SEQ ID NO: 350, any variants thereof or any combinations thereof.
  • the sweet protein that is a variant of a MNEI protein is characterized by a sweetness threshold of at most about 0.05 mg/ml, at times at most about 0.1 mg/ml, at times at most about 0.2 mg/ml, at times at most about 0.3 mg/ml, at times at most about 0.5 mg/ml, at times at most about 1 mg/ml, at times at most about 2 mg/ml, at times at most about 3 mg/ml, at times at most about 5 mg/ml.
  • the sweet protein that is a variant of a MNEI protein is characterized by a sweetness threshold of about 0.01 mg/ml, at times about 0.03 mg/ml, at times about 0.05 mg/ml, at times about 0. 1 mg/ml, at times about 0.2 mg/ml, at times about 0.3 mg/ml, at times about 0.5 mg/ml, at times about 1 mg/ml, at times about 2 mg/ml, at times about 3 mg/ml, at times about 5 mg/ml.
  • the sweet protein that is a variant of a MNEI protein is characterized by a sweetness that is at least about 500 times as compared to sucrose, at least about 800 times as compared to sucrose, at least about 1000 times as compared to sucrose, at least about 1500 times as compared to sucrose, at least about 2000 times as compared to sucrose, at least about 3000 times as compared to sucrose, at least about 4000 times as compared to sucrose, at least about 5000 times as compared to sucrose, at least about 6000 times as compared to sucrose, at least about 7000 times as compared to sucrose, at least about 8000 times as compared to sucrose, at least about 9000 times as compared to sucrose, at least about 10000 times as compared to sucrose, at least about 11000 times as compared to sucrose, at least about 12000 times as compared to sucrose, at least about 13000 times as compared to sucrose, at least about 14000 times as compared to sucrose, at least about 15000 times as compared to
  • the sweet protein that is a variant of a MNEI protein is characterized by a thermostability of at least about 70°C, at times at least about 73°C, at times at least about 75°C, at times at least about 80°C, at least about 85°C, at times at least about 90°C, at times at least about 94°C, at times at least about 95°C, at times at least about 97°C, at times at least about 98°C, at times at least about 99°C, at times at least about 100°C, at times at least about 101°C, at times at least about 102°C, at times at least about 103°C, at times at least about 104°C, at times at least about 105°C, at times at least about 110°C, at times at least about 115 °C.
  • the sweet protein that is a variant of a MNEI protein is characterized by a thermostability of about 70°C, at times about 73°C, at times about 75°C, at times about 80°C, at times about 85°C, at times about 90°C, at times about 94°C, at times about 95 °C, at times about 97°C, at times about 98°C, at times about 99°C, at times about 100°C, at times about 101°C, at times about 102°C, at times about 103°C, at times about 104°C, at times about 105°C, at times about 110°C, at times about 115°C.
  • thermostability refers to a protein's ability to maintain one or more of structure and function at the recited temperature. Thermostability can be measured through various techniques known in the art. For example, thermostability can be measured by Differential Scanning Calorimetry (DSC) or Differential Scanning Fluorimetry (DSF).
  • DSC Differential Scanning Calorimetry
  • DSF Differential Scanning Fluorimetry
  • the sweet protein is a variant of a MNEI protein (SEQ ID NO: 14 and/or SEQ ID NO:350).
  • the sweet protein has an amino acid sequence denoted by SEQ ID NO: 16 (denoted herein as DM08). In some embodiments, the sweet protein has an amino acid sequence denoted by SEQ ID NO: 17 (denoted herein as DM09). In some embodiments, the sweet protein has an amino acid sequence denoted by SEQ ID NO:39 (denoted herein as DM31). In some embodiments, the sweet protein has an amino acid sequence denoted by SEQ ID NO:53 (denoted herein as DM69). In some embodiments, the sweet protein has an amino acid sequence denoted by SEQ ID NO: 84 (denoted herein as DM330).
  • the sweet protein has an amino acid sequence denoted by SEQ ID NO: 104 (denoted herein as DM137). In some embodiments, the sweet protein has an amino acid sequence denoted by SEQ ID NO: 186 (denoted herein as DM165). In some embodiments, the sweet protein has an amino acid sequence denoted by SEQ ID NO:217 (denoted herein as DM685). In some embodiments, the sweet protein has an amino acid sequence denoted by SEQ ID NO:257 (denoted herein as DM532). In some embodiments, the sweet protein has an amino acid sequence denoted by SEQ ID NO:263 (denoted herein as DM528). In some embodiments, the sweet protein has an amino acid sequence denoted by SEQ ID NO:351 (denoted herein as DM170).
  • the reference protein is one or more proteins denoted by SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO:39, SEQ ID NO:53, SEQ ID NO:84, SEQ ID NO: 104, SEQ ID NO: 186, SEQ ID NO:217, SEQ ID NO:257, SEQ ID NO:263, SEQ ID NO:351 or any combinations thereof.
  • the present disclosure provides a method for modifying at least one flavor attribute in a comestible product.
  • the method comprises adding at least one sweet protein to the comestible product at a flavor modifying concentration, wherein the at least one sweet protein selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO:39, SEQ ID NO:53, SEQ ID NO:84, SEQ ID NO: 104, SEQ ID NO: 186, SEQ ID NO:217, SEQ ID NO:257, SEQ ID NO:263, SEQ ID NO:351, a variant thereof or any combinations thereof.
  • the present disclosure provides a comestible product comprising at least one sweet protein at a flavor-modifying concentration, wherein the at least one protein selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 39, SEQ ID NO:53, SEQ ID NO:84, SEQ ID NO: 104, SEQ ID NO: 186, SEQ ID NO:217, SEQ ID NO:257, SEQ ID NO:263, SEQ ID NO:351, a variant thereof or any combinations thereof.
  • the sweet protein is a variant of a protein denoted by SEQ ID NO: 16 (denoted herein as DM08). In some embodiments, the sweet protein is a variant of a protein denoted by SEQ ID NO: 17 (denoted herein as DM09). In some embodiments, the sweet protein is a variant of a protein denoted by SEQ ID NO:39 (denoted herein as DM31). In some embodiments, the sweet protein is a variant of a sweet protein denoted by SEQ ID NO:53 (denoted herein as DM69). In some embodiments, the sweet protein has an amino acid sequence denoted by SEQ ID NO:84 (denoted herein as DM330).
  • the sweet protein is a variant of a protein denoted by SEQ ID NO: 104 (denoted herein as DM137). In some embodiments, the sweet protein is a variant of a protein denoted by SEQ ID NO: 186 (denoted herein as DM165). In some embodiments, the sweet protein is a variant of a protein denoted by SEQ ID NO:217 (denoted herein as DM685). In some embodiments, the sweet protein is a variant of a protein denoted by SEQ ID NO:257 (denoted herein as DM532). In some embodiments, the sweet protein is a variant of a protein denoted by SEQ ID NO:263 (denoted herein as DM528). In some embodiments, the sweet protein is a variant of a sweet protein denoted by SEQ ID NO: 351 (denoted herein as DM170).
  • the sweet protein is a variant of a protein, for example MNEI protein
  • the protein includes modifications as described herein as compared to the reference protein, for example MNEI, being the reference protein.
  • the present disclosure is not limited to specific number of amino acid modifications made in the reference protein that eventually results in the formation of the protein described herein.
  • a modification as used herein refers to a modification in one or more amino acids of the reference protein and encompasses amino acid substitution (replacement), amino acid deletion, amino acid insertion or any combination thereof.
  • a modification may be an amino acid insertion.
  • the protein comprises an amino acid sequence having at least one, at least two, at least three, at least four, at least five amino acid insertions as compared to the reference protein.
  • a modification may be an amino acid deletion.
  • the sweet protein comprises an amino acid sequence having at least one, at least two, at least three, at least four, at least five amino acid deletions as compared to the reference protein.
  • a modification may be an amino acid substitution.
  • the sweet protein comprises an amino acid sequence having at least one, at least two, at least three, at least four, at least five amino acid substitutions as compared to the reference protein.
  • the sweet protein is a variant of a reference protein including one or more of (i) one or more amino acids replacements, (ii) one or more amino acids deletions, (iii) one or more amino acids insertions or (iv) any combination thereof.
  • the sweet protein is a modified reference protein including one or more amino acids replacements. In some embodiments, the sweet protein is a modified reference protein including one or more amino acids deletions. In some embodiments, the sweet protein is a modified reference protein including one or more amino acids insertions.
  • the sweet protein comprises one or more amino acid modifications as compared to MNEI protein having an amino acid sequence denoted by SEQ ID NO: 14. In some embodiments, the sweet protein comprises one or more amino acid modifications as compared to MNEI protein having an amino acid sequence denoted by SEQ ID NO:350.
  • the sweet protein comprises one or more amino acid modifications as compared to one or more proteins denoted by SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO:39, SEQ ID NO:53, SEQ ID NO: 84, SEQ ID NO: 104, SEQ ID NO: 186, SEQ ID NO:217, SEQ ID NO:257, SEQ ID NO:263, SEQ ID NO:350, SEQ ID NO:351 or any combinations thereof.
  • the sweet protein comprises one or more amino acid modifications in one or more residue G1,E2, E4, 18, Fl 1, T12, N14, G16, K17, F18, A19, V20, D21, E23, K25, 126, G27, Q28, R31, T33, N35, K36, C41, E50, F52, R53, E59, Q61, L62, V64, Y65, S67, D68, L70, A73, D74, 175, S76, E77, R84, F89 or any combination thereof as compared to the reference protein.
  • the sweet protein comprises one or more amino acid replacement as compared to MNEI protein having an amino acid sequence denoted by SEQ ID NO: 14 or SEQ ID NO:350. In some embodiments, the sweet protein comprises one or more amino acid replacements as compared to one or more proteins denoted by SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO:39, SEQ ID NO:53, SEQ ID NO:84, SEQ ID NO: 104, SEQ ID NO: 186, SEQ ID NO:217, SEQ ID NO:257, SEQ ID NO:263, SEQ ID NO:350, SEQ ID NO:351 or any combinations thereof.
  • the sweet protein comprises one or more amino acid replacement in one or more residue G1,E2, E4, 18, Fl 1, T12, N14, G16, K17, F18, A19, V20, D21, E23, K25, 126, G27, Q28, R31, T33, N35, K36, C41, E59, Q61, L62, V64, Y65, S67, D68, L70, A73, D74, 175, S76, E77, R84, F89 or any combination thereof as compared to the reference protein, for example MNEI protein.
  • the sweet protein comprises one or more amino acid deletions as compared to one or more proteins denoted by SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO:39, SEQ ID NO:53, SEQ ID NO: 84, SEQ ID NO: 104, SEQ ID NO: 186, SEQ ID NO:217, SEQ ID NO:257, SEQ ID NO:263, SEQ ID NO:350, SEQ ID NO:351 or any combinations thereof.
  • the sweet protein comprises one or more amino acid deletions in one or more amino acid E50, F52, R53 as compared to the reference protein. In some embodiments, the sweet protein comprises one or more amino acid deletions in one or more amino acid E50, F52, R53 as compared to MNEI protein having an amino acid sequence denoted by SEQ ID NO: 14. In some embodiments, the sweet protein comprises one or more amino acid deletions in one or more amino acid E50, F52, R 3 as compared to MNEI protein having an amino acid sequence denoted by SEQ ID NO:350.
  • the sweet protein comprises one or more amino acid replacements in one or more amino acid S76, E23, Q28, E2, Y65, D68 as compared to MNEI protein having an amino acid sequence denoted by SEQ ID NO: 14. In some embodiments, the sweet protein comprises one or more amino acid replacements in one or more amino acid S76, E23, Q28, E2, Y65, D68 as compared to MNEI protein having an amino acid sequence denoted by SEQ ID NO: 350.
  • the sweet protein has an amino acid sequence denoted by one or more of SEQ ID NO: 14-SEQ ID NO:465, SEQ ID NO:467- SEQ ID NO:479 or any combination thereof.
  • the sweet protein is any one of the proteins defined in Table 40 shown below. It should be noted that at times, each row in Table 40 represents a different embodiment of the present application. It should be noted that the present disclosure encompasses a combination of two or more sweet proteins, at times three or more, at times four or more, at times five or more sweet proteins selected from SEQ ID NO: 14-SEQ ID NO:465, SEQ ID NO:467- SEQ ID NO:479 or any combination thereof.
  • the protein is or comprises a combination of two or more proteins, wherein one of the proteins is MNEI or any variant thereof.
  • the protein comprises a combination of two or more proteins, wherein at least one of the two or more proteins is a protein denoted by one or more of SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO:39, SEQ ID NO:53, SEQ ID NO:84, SEQ ID NO: 104, SEQ ID NO: 186, SEQ ID NO:217, SEQ ID NO:257, SEQ ID NO:263, SEQ ID NO:350, SEQ ID NO:35 lor any combinations thereof.
  • the protein comprises a combination of three or more proteins, wherein at least one of the two or more proteins is a protein denoted by one or more of SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO:39, SEQ ID NO:53, SEQ ID NO:84, SEQ ID NO: 104, SEQ ID NO: 186, SEQ ID NO:217, SEQ ID NO:257, SEQ ID NO:263, SEQ ID NO:350, SEQ ID NO:351 or any combinations thereof.
  • the protein comprises a combination of four or more proteins, wherein at least one of the two or more proteins is a protein denoted by one or more of SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO:39, SEQ ID NO:53, SEQ ID NO:84, SEQ ID NO: 104, SEQ ID NO: 186, SEQ ID NO:217, SEQ ID NO:257, SEQ ID NO:263, SEQ ID NO:350, SEQ ID NO:351 or any combinations thereof.
  • the protein comprises a combination of a protein denoted by SEQ ID NO: 39 (denoted herein as DM31) and one or more of a protein denoted by SEQ ID NO:217 (denoted herein as DM685), a protein denoted by SEQ ID NO:263 (denoted herein as DM528), a protein denoted by SEQ ID NO: 104 (denoted herein as DM137), a protein denoted by SEQ ID NO: 17 (denoted herein as DM09), a protein denoted by SEQ ID NO: 351 (denoted herein as DM170), a protein denoted by SEQ ID NO:53 (denoted herein as DM69), a protein denoted by SEQ ID NO: 186 (denoted herein as DM165), a protein denoted by SEQ ID NO:84 (denoted herein as DM330), a protein denoted by SEQ ID NO:257 (
  • the one or more proteins comprises a combination of a protein denoted by SEQ ID NO: 39 (denoted herein as DM31) and a protein denoted by SEQ ID NO: 8. In some embodiments, the one or more proteins comprises a combination of a protein denoted by SEQ ID NO:39 (denoted herein as DM31) and a protein denoted by SEQ ID NO: 17 (denoted herein as DM09). In some embodiments, the one or more proteins comprises a combination of a protein denoted by SEQ ID NO:39 (denoted herein as DM31) and a protein denoted by SEQ ID NO:53 (denoted herein as DM69).
  • the one or more proteins comprises a combination of a protein denoted by SEQ ID NO:39 (denoted herein as DM31) and a protein denoted by SEQ ID NO:84 (denoted herein as DM330). In some embodiments, the one or more proteins comprises a combination of a protein denoted by SEQ ID NO:39 (denoted herein as DM31) and a protein denoted by SEQ ID NO: 104 (denoted herein as DM137).
  • the one or more proteins comprises a combination of a protein denoted by SEQ ID NO:39 (denoted herein as DM31) and a protein denoted by SEQ ID NO: 186 (denoted herein as DM165). In some examples, the one or more proteins comprises a combination of a protein denoted by SEQ ID NO: 39 (denoted herein as DM31) and a protein denoted by SEQ ID NO:217 (denoted herein as DM685).
  • the one or more proteins comprises a combination of a protein denoted by SEQ ID NO:39 (denoted herein as DM31) and a protein denoted by SEQ ID NO:257 (denoted herein as DM532). In some embodiments, the one or more proteins comprises a combination of a protein denoted by SEQ ID NO:39 (denoted herein as DM31) and a protein denoted by SEQ ID NO:263 (denoted herein as DM528).
  • the one or more proteins comprises a combination of a protein denoted by SEQ ID NO:39 (denoted herein as DM31) and a protein denoted by SEQ ID NO:357 (denoted herein as DM170). In some embodiments, the one or more proteins comprises a combination of a protein denoted by SEQ ID NO:39 (denoted herein as DM31), a protein denoted by SEQ ID NO: 17 (denoted herein as DM09) and a protein denoted by SEQ ID NO: 186 (denoted herein as DM165).
  • the one or more proteins comprises a combination of MNEI or a variant thereof and one or more proteins denoted by SEQ ID NO:3 or SEQ ID NON.
  • the one or more proteins comprises a combination of a protein denoted by SEQ ID NO: 39 (denoted herein as DM31) and a protein denoted by SEQ ID SEQ ID NO: 3 or SEQ ID NON.
  • the sweet protein includes a combination of any one or more of the proteins set forth in SEQ ID NO: 1 to SEQ ID NO:481 and a saccharide.
  • the herein-disclosed saccharide may include monosaccharide, disaccharide, oligosaccharide, or polysaccharide.
  • the sweet protein includes a combination of any one or more of the MNEI variants (i.e., variants of SEQ ID NO: 14 or SEQ ID NO: 350) and the saccharide.
  • the sweet protein includes a combination of any one of the sweet proteins selected from SEQ ID NO: 14-SEQ ID NO:481 or any combination thereof and the saccharide. In some embodiments, the sweet protein includes a combination of any one of the sweet proteins defined in Table 40 shown below and the saccharide.
  • the saccharide can be bound or unbound to the one or more herein-disclosed sweet proteins.
  • the saccharide may be selected from, but not limited to, mannan disaccharide, mannan polysaccharide, mannan oligosaccharide (MOS), betaglucan disaccharide, beta-glucan polysaccharide, beta-glucan-oligosaccharide, alpha- glucan disaccharide, alpha-glucan polysaccharide, alpha-glucan-oligosaccharide, chitin disaccharides, chitin polysaccharide, chito oligosaccharide, galactomannan polysaccharide, galactomanno-oligosaccharide, xyloglucan polysaccharide, xyloglucanoligosaccharide, rhamnomannan polysaccharide, rhamnomanno disaccharide, rhamnomanno-oligosaccharide, phosphomannan disaccharide, phosphomannan polysaccharide, phosphomanno-oligosaccharide
  • the herein-disclosed saccharide may be selected from, but not limited to, mannan disaccharide, mannan polysaccharide, mannan oligosaccharide (MOS), beta-glucan disaccharide, beta-glucan polysaccharide, beta-glucan oligosaccharides, alpha-glucan disaccharide, alpha-glucan polysaccharide, alpha-glucan- oligosaccharide, chitin disaccharide, chitin polysaccharide, chito oligosaccharide, and any combination thereof.
  • the polysaccharide includes mannan polysaccharide and/or mannan oligosaccharide (MOS).
  • the herein-disclosed saccharide may include mannose, glucose, galactose, rhamnose, N-acetylglucosamine, N-acetylgalactosamine, N- acetyhnuramic acidxylose, fucose, arabinose, trehalose, fructose, maltotriose, glucuronic acid, iduronic acid, hyaluronic acid, N-acetylmuramic acid, 3-deoxy-D-manno-oct-2- ulosonic acid, Chondroitin Sulfate, Dermatan Sulfate, Heparan Sulfate, Keratan Sulfate, glycerol, glycerol phosphate, ribitol, ribitol phosphate, L-glycero-D-manno-heptose, mannuronic acid, or any combination thereof.
  • Each possibility is a separate embodiment.
  • the herein-disclosed saccharide may be further modified.
  • the modification of the herein-disclosed saccharide may include, but is not limited to, phosphorylation, methylation, acylation, acetylation, N- acetylation, sulphation, amination, peptide-conjugation, lipid-conjugation, carboxylation, or any combination thereof. Each possibility is a separate embodiment.
  • the sweet protein is one or more sweet proteins.
  • the sweet protein is a combination of 2, 3, 4, 5, or more sweet proteins. Each possibility is a separate embodiment.
  • the combination includes 2, 3, 4, 5, or more sweet protein sequences selected from the sequences set forth in Table 40.
  • a method for modifying at least one flavor in a comestible product comprises adding at least one protein to the comestible product, wherein the concentration of the at least one protein is between about 1 ppm and about 300 ppm, and wherein the at least one protein selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO 350, SEQ ID NO:466, SEQ ID NO:480, SEQ ID NO:481, any variants thereof or any combinations thereof.
  • a method for modifying at least one flavor in a comestible product comprises adding at least one protein to the comestible product, wherein the concentration of the at least one protein is between about 1 ppm and about 300 ppm, and wherein the at least one protein selected from the group consisting of SEQ ID NO: 14, by SEQ ID NO: 350, any variants thereof or any combinations thereof.
  • a method for modifying at least one flavor in a comestible product comprises adding at least one protein to the comestible product, wherein the concentration of the at least one protein is between about 1 ppm and about 300 ppm, and wherein the at least one protein selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO:39, SEQ ID NO:53, SEQ ID NO:84, SEQ ID NO: 104, SEQ ID NO: 186, SEQ ID NO:217, SEQ ID NO:263, SEQ ID NO:257, SEQ ID NO:351, any variants thereof or any combinations thereof.
  • a comestible product comprising at least one protein, wherein the concentration of the at least one protein is between about 1 ppm and about 300 ppm, and wherein the at least one protein selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NON, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO: 8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO 350, SEQ ID NO:466, SEQ ID NO:480, SEQ ID NO:481, any variants thereof or any combinations thereof.
  • a comestible product comprising at least one protein, wherein the concentration of the at least one protein is between about 1 ppm and about 300 ppm, and wherein the at least one protein selected from the group consisting of SEQ ID NO: 14, by SEQ ID NO: 350, any variants thereof or any combinations thereof.
  • a comestible product comprising at least one protein, wherein the concentration of the at least one protein is between about 1 ppm and about 300 ppm, and wherein the at least one protein selected from the group consisting of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO:39, SEQ ID NO:53, SEQ ID NO:84, SEQ ID NO: 104, SEQ ID NO: 186, SEQ ID NO:217, SEQ ID NO:263, SEQ ID NO:257, SEQ ID NO:351, any variants thereof or any combinations thereof.
  • the protein may modify one or more flavors and hence may be considered as modifying flavor properties.
  • modifying flavor properties refers to the ability of the one or more sweet proteins for example, a sweet protein denoted by SEQ ID NO: 14, SEQ ID NO:350 variants thereof or any combinations thereof to alter the taste perception of food or beverage product without necessarily introducing a distinct flavor of their own.
  • the flavor-modifying properties include the ability to increase, decrease, suppress or otherwise modify the perception of specific flavor profile or an overall flavor profile.
  • the modified flavor is distinct from sweetness.
  • flavor profile encompasses one or more sensory attributes of a comestible product, including taste, aroma, mouthfeel (texture), aftertaste, and flavor intensity.
  • the sweet proteins at FMP concentrations may enhance or suppress individual flavor components.
  • the sweet proteins at FMP concentrations may alter the duration or onset of specific flavor perceptions. In some examples, the sweet proteins at FMP concentrations may reduce sweet lingering.
  • the sweet proteins at FMP concentrations may modify the perception of basic tastes. In some examples, the sweet proteins at FMP concentrations may modify sweetness, sourness, bitterness, saltiness, and umami (savory).
  • the sweet proteins at FMP concentrations may modify complex flavor sensations. In some examples, the sweet proteins at FMP concentrations may modify fruity, nutty, creamy, meaty, or roasted notes.
  • the sweet proteins at FMP concentrations may modify an existing overall flavor profile of a product. In some examples, the sweet proteins at FMP concentrations may increase an existing overall flavor profile of a product. In some examples, the sweet proteins at FMP concentrations may decrease an existing overall flavor profile of a product. In some examples, the sweet proteins at FMP concentrations may change the flavor profile of a food product. In some examples, the sweet proteins at FMP concentrations may affect the duration of the perception of specific characteristics of the flavor profile. Increasing flavor as used herein refers to amplify/increase/intensify/enhance, for example, in an existing (specific) flavor or flavor profile by increasing strength, or perceptibility of a particular flavor or flavor profiles.
  • Decreasing flavor refers to a decrease/reduction/diminish strength, or perceptibility of a flavor, for example, in an existing (specific) flavor or flavor profile, by decreasing strength or perceptibility of a particular flavor or flavor profile.
  • the sweet protein at an FMP concentration modifies one or more tastes of sweetness, sourness, bitterness, saltiness, umami or a combination thereof.
  • the sweet protein at an FMP concentration modifies flavors which are combinations of tastes and odors, such as but not limited to strawberry flavor, vanilla flavor etc.
  • the sweet protein at an FMP concentration modifies or alters sweetness. In some examples, the sweet protein at an FMP concentration increases sweetness. In some examples, the sweet protein at an FMP concentration reduces sweetness.
  • the sweet protein at an FMP concentration reduces sweet linger. In some examples, the sweet protein at an FMP concentration improves mouthfeel.
  • the sweet protein at an FMP concentration modifies or alters sourness. In some examples, the sweet protein at an FMP concentration reduces or eliminates sourness. In some examples, the sweet protein at an FMP concentration reduces or eliminates sour taste or flavor. In some examples, the sweet protein at an FMP concentration alters sourness taste or flavor.
  • the sweet protein at an FMP concentration modifies or alters saltiness. In some examples, the sweet protein at an FMP concentration reduces or eliminates saltiness. In some examples, the sweet protein at an FMP concentration reduces or eliminates saltiness taste. In some examples, the sweet protein at an FMP concentration modifies or alters bitterness. In some examples, the sweet protein at an FMP concentration reduces or eliminates bitterness. In some examples, the sweet protein at an FMP concentration reduces or eliminates bitter taste. In some examples, the sweet protein at an FMP concentration reduces or eliminates bitter linger.
  • the sweet protein at an FMP concentration modifies or alters umami. In some examples, the sweet protein at an FMP concentration increases umami. In some examples, the sweet protein at an FMP concentration decreases umami. In some examples, the sweet protein at an FMP concentration changes savory taste.
  • the sweet protein at an FMP concentration modifies or alters astringency. In some examples, the sweet protein at an FMP concentration reduces or eliminates astringency.
  • the sweet protein at an FMP concentration modifies or alters one or more of sweetness flavor, sourness flavor or saltiness flavor and at least one additional flavor.
  • the sweet protein at an FMP concentration modifies or alters one or more of sweetness, sourness or saltiness and at least one additional flavor.
  • the sweet protein at an FMP concentration modifies or alters one or more of sweetness, sourness, bitterness or saltiness and at least one additional flavor.
  • the sweet protein at an FMP concentration increases one or more flavors.
  • the sweet protein at an FMP concentration alters or modifies one or more of the following sensory attributes: sweetness, sourness, bitterness, saltiness, flavor intensity, seasoning intensity, fruity flavor, fragrance intensity, astringency, body.
  • the sweet protein at an FMP concentration increases one or more of sweetness, fruity flavor, ripe strawberry flavor, strawberry flavor intensity, apple flavor, apple flavor/taste intensity, milk taste, oat milk intensity, oat flavor intensity, vanilla taste, tomato concentrate flavor, peanut butter taste, lemon taste, caramelized flavor, dark chocolate taste, coffee taste, meaty taste, species flavor, herb flavor, nutty flavor, roasted flavor, creamy flavor, smoky flavor or any combination thereof.
  • the sweet protein at an FMP concentration increases sweetness and one or more of fruity flavor, ripe strawberry flavor, strawberry flavor intensity, apple flavor/taste, apple flavor/taste intensity, milk flavor, oat milk intensity, oat flavor intensity, vanilla flavor, tomato concentrate flavor, peanut butter flavor, lemon flavor, caramelized flavor, dark chocolate flavor, coffee flavor, meaty flavor, species flavor, herb flavor, nutty flavor, roasted flavor, creamy flavor, smoky flavor or any combination thereof.
  • the sweet protein at an FMP concentration decreases or eliminates sweetness and increases (enhances) one or more of fruity flavor, ripe strawberry flavor, strawberry flavor intensity, apple flavor/taste, apple flavor intensity, milk flavor, oat milk intensity, oat flavor, vanilla flavor, tomato concentrate flavor, peanut butter flavor, lemon flavor or any combination thereof.
  • the sweet protein at an FMP concentration increases (enhances) sweetness and one or more of fruity flavor, ripe strawberry flavor, strawberry flavor intensity, apple flavor, apple flavor/taste intensity, milk flavor, oat milk flavor intensity, oat flavor, vanilla flavor, tomato concentrate flavor, peanut butter flavor, lemon flavor or any combination thereof.
  • the sweet protein at an FMP concentration increase one or more of fruity flavor, ripe strawberry flavor, strawberry flavor intensity, apple flavor, apple taste intensity, milk flavor, oat milk flavor intensity, oat flavor, vanilla flavor, tomato concentrate flavor, peanut butter flavor, lemon flavor, caramelized flavor, dark chocolate flavor, coffee flavor, meaty taste, species flavor, herb flavor, nutty flavor, roasted flavor, creamy flavor, smoky flavor or any combination thereof, without increasing sweetness.
  • the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 10% of the added sweeteners.
  • the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 10% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 15% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 20% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 25% of the added sweeteners.
  • the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 30% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 35% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 40% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 45% of the added sweeteners.
  • the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 50% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 55% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 60% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 65% of the added sweeteners.
  • the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 70% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 75% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 80% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 85% of the added sweeteners.
  • the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 90% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 95% of the added sweeteners. In some examples, the sweet protein at an FMP concentration increases (enhances) the perceived sweetness of sugar or other sweeteners, allowing for the reduction of at least 97% of the added sweeteners.
  • the sweet protein at an FMP concentration increases fruity flavor.
  • the fruity flavor is one or more of strawberry flavor, raspberry flavor, apple flavor, banana flavor, pine-apple flavor, peach flavor, mango flavor cherry flavor or citrus flavor.
  • the sweet protein at an FMP concentration increases ripe strawberry flavor and/or strawberry flavor intensity. In some examples, the sweet protein at an FMP concentration increases apple taste, apple flavor and/or apple taste intensity. In some examples, the sweet protein at an FMP concentration increases oat milk flavor intensity. In some examples, the sweet protein at an FMP concentration increases vanilla flavor. In some examples, the sweet protein at an FMP concentration increases peanut butter flavor. In some examples, the sweet protein at an FMP concentration increases caramelized flavor. In some examples, the sweet protein at an FMP concentration increases tomato concentrate flavor. In some examples, the sweet protein at an FMP concentration increases lemon flavor. In some examples, the sweet protein at an FMP concentration increases dark chocolate flavor. In some examples, the sweet protein at an FMP concentration increases coffee flavor. In some examples, the sweet protein at an FMP concentration increases meaty taste.
  • the sweet protein at an FMP concentration increases species flavor.
  • the sweet protein at an FMP concentration increases a herb flavor. In some examples, the sweet protein at an FMP concentration increases seasoning flavor. In some examples, the species or herb flavor is one or more of cinnamon flavor, pepper flavor, cumin flavor, garlic flavor, maple flavor or basil flavor.
  • the sweet protein at an FMP concentration increases a nutty flavor. In some examples, the sweet protein at an FMP concentration increases roasted flavor.
  • the sweet protein at an FMP concentration increases a creamy flavor.
  • the sweet protein at an FMP concentration increases a smoky flavor.
  • the sweet protein at an FMP concentration reduces one or more flavors.
  • the sweet protein at an FMP concentration reduces one or more saltiness enhancement, sourness, astringency, bitterness, citrusy flavor, tart flavor, acidic flavor, hoppy flavor or any combination thereof.
  • the sweet protein at an FMP concentration reduces or eliminates one or more saltiness enhancement, sourness, astringency, bitterness or any combination thereof. In some examples, the sweet protein at an FMP concentration reduces or eliminates saltiness enhancement. In some examples, the sweet protein at an FMP concentration increases saltiness enhancement. In some examples, the sweet protein at an FMP concentration reduces or eliminates bitterness or bitter linger. In some examples, the sweet protein at an FMP concentration reduces or eliminates citrusy flavor. In some examples, the sweet protein at an FMP concentration reduces or eliminates tart flavor. In some examples, the sweet protein at an FMP concentration reduces or eliminates acidic flavor. In some examples, the sweet protein at an FMP concentration reduces or eliminates reduces hoppy flavor. In some examples, the sweet protein at an FMP concentration reduces or eliminates metallic notes.
  • the sweet protein at an FMP concentration is characterized by being capable of reducing (decreasing) flavor off-notes.
  • flavor off-notes refers to undesirable or unpleasant flavor or aroma present in a food product or beverage product.
  • the off note can be for example due to the ingredients used in the product, the processing methods, or the storage conditions of the product.
  • reducing flavor off notes comprises reducing one or more of biterness, sourness, acidity, rancidity, metallic taste, chemical taste or a combination thereof.
  • Biterness may result from the presence of certain compounds in a product, such as alkaloids or tannins or from over-roasting or overcooking. Sourness may result from presence of acidic compounds beyond the desired level. Rancidity may result when fats or oils in the product undergo oxidative degradation, resulting in the formation of unpleasant odors or flavors. Metallic or chemical taste may be caused by the presence of metallic ions in the product (e.g. iron or copper), as well as by contamination with chemical substances from processing equipment or packaging materials.
  • metallic ions e.g. iron or copper
  • the sweet protein maintains FMP after been heated and/or added to warm/heated food products.
  • the sweet protein maintains FMP after heating for about at least 5 min at about 60°C and/or when added food products have a temperature of about 60°C. According to some embodiments, the sweet protein maintains FMP after heating for at least 5 min at about 80°C and/or when added food applications have a temperature of about 80°C. According to some embodiments, the sweet protein maintains FMP after heating for at least 5 min at 90°C and/or when added food applications have a temperature of about 90°C. According to some embodiments, the sweet protein maintains FMP after heating for at least 5 min at 100°C and/or when added food applications have a temperature of about 100°C.
  • the present application is not limited to a specific comestible product and is applicable to any comestible product.
  • comestible product refers to an item or substance intended for human consumption.
  • the sweet protein is added to a comestible product at a concentration at which it does not act as a sweetener per se.
  • the comestible product may be essentially free of a sweetener. In some examples, the comestible product is essentially free of any added sweetener. In some examples, the comestible product is essentially free of any artificial sweetener. In some examples, the comestible product is essentially free of any naturally occurring sweetener.
  • essentially free of sweeteners means that the comestible product does not contain any added caloric or non-caloric sweeteners, such as sucrose, glucose, fructose, high-fructose com syrup, stevia, erythritol, sucralose, or the like, except for a sweet protein present at a PMF concentration.
  • ‘essentially free’ refers to a product in which any non-protein sweeteners are present only as incidental carryover or excipients, in amounts insufficient to contribute perceptible sweetness.
  • the comestible product may comprise one or more sweeteners.
  • the comestible product comprises at least about 1% one or more sweeteners, at times at least about 3% one or more sweeteners, at times at least about 10% one or more sweeteners, at times at least about 15% one or more sweeteners, at times at least about 25% one or more one or more sweeteners, at times at least about 50% one or more sweeteners.
  • the comestible product comprises between about 1% one or more sweeteners and about 50% one or more sweeteners. In some examples, the comestible product comprises between about 1% one or more sweeteners and about 20% one or more sweeteners. In some examples, the comestible product comprises between about 1% one or more sweeteners and about 15% one or more sweeteners.
  • the comestible product comprises stevia or any derivative thereof.
  • the stevia is or comprises one or more of steviol glycoside, Rebaudioside A (Reb A), Rebaudioside M (Reb M), Rebaudioside D (Reb D), Stevioside, Rebaudioside or a combination thereof.
  • the one or more sweeteners is selected from the group consisting of at least one sugar, non-nutritive sweetener, high intensity sweetener, a natural sweetener, a sugar alcohol, a rare sugar, an amino acid or a combination thereof.
  • the at least one sweetener is at least one of (i) at least one high intensity sweetener, (ii) at least one natural sweetener, (iii) at least one sugar alcohol, (iv) at least one rare sugar, (v) at least one sugar, (vi) an amino acid or (vii) a combination thereof.
  • the at least one high intensity sweetener is one or more of potassium (ace-k), aspartame, sucralose, cyclamate, neotame, saccharin, Neohesperidin dihydrochalcone, stevia, monk fruit, fructose, acesulfame, advantame or any combination thereof.
  • the amino acid is glycine.
  • the at least one natural sweetener is at least one of stevia, monk fruit, agava syrup, palatinose, glycyrrhizin, sucrose, fructose, glucose or any combination thereof.
  • the at least one sugar alcohol is at least one of sorbitol, erythritol, isomalt, lactitol, mannitol, tagatose, maltitol, xylitol or any combination thereof.
  • the rare sugar is one or more of allulose, turanose, isomaltulose, isomaltose, tagarose or a combination thereof.
  • the at least one sugar is at least one of palatinose, sucrose, fructose, glucose, lactose sucralose, com syrup, high fructose com symp, invert sugar, dextrose, powder sugar, or a combination thereof, in any form of liquid, powder, or solid.
  • the at least one sugar is sucrose.
  • the natural sweetener is one or more of stevia, sweet truffle protein (mycodulcein), brazzein, monk fmit, honey, agave, maple symp, silan, black treacle, molasses, glycine or a combination thereof.
  • the non-nutritive sweetener is selected from a group consisting of aspartame, sucralose, saccharin, stevia, acesulfame potassium, neotame, allulose, monk fmit extract, erythritol, xylitol, mannitol, sorbitol, cyclamate, and any combination thereof.
  • the comestible product comprises one or more artificial sweetener and/or one or more naturally occurring sweetener.
  • the present disclosure is not limited to a specific comestible product and is applicable to a variety of products.
  • the comestible product is a food product.
  • the comestible product is a beverage. In some examples, the comestible product is one or more of a food product, a beverage or any combination thereof.
  • the comestible product is one or more of a food product, a beverage, a beverage concentrate, a frozen beverage, a frozen dessert, a refrigerated dessert, a dietary supplement, a nutraceutical, a dietary supplement, a botanicals or a combination thereof.
  • the food product is a frozen product. In some examples, the food product is a refrigerated product.
  • the comestible product is one or more of a dairy product, a nondairy product, a frozen dessert, a condiment, a spread or any combination thereof.
  • the comestible product is a food product. In some examples, the comestible product is a solid food product. In some examples, the comestible product is a soft food product.
  • the comestible product has a creamy texture.
  • the food product is a milk product. In some examples, the food product is a dairy (milk) product.
  • the dairy product is one or more of yogurt, a flavored yogurt, a cheese, a flavored cheese, a milk.
  • the dairy product is yogurt.
  • the dairy product is an apple flavored yogurt.
  • the dairy product is a strawberry flavored yogurt.
  • the dairy product is a whey cheese.
  • the dairy product is a flavored whey cheese.
  • the dairy product is milk.
  • the dairy product comprises at least one added salt.
  • the dairy product is essentially free of one or more sweeteners.
  • the dairy product comprises at least one added sweetener. In some examples, the dairy product comprises at least sucrose. In some examples, the dairy product comprises sucrose at a concentration of about 4%. In some examples, the dairy product comprises at least sucrose. In some examples, the dairy product comprises at least one sweetener at a concentration of about 4%. In some examples, the dairy product is protein drink milk. In some examples, the food product is milk or milk-based products.
  • dairy products specifically, yogurt samples with DM31 concentrations below 25 ppm were perceived as less sweet than yogurt with 1.5% sucrose.
  • FMP concentration flavor-modifying concentration
  • the present disclosure provides a method for modifying at least one flavor in a comestible product, the method comprises adding a flavor modifying concentration of at least one protein to the comestible product, wherein the product is a diary product.
  • the present disclosure provides a dairy product comprising at least one sweet protein at FMP concentration.
  • the at least one sweet protein at FMP concentration comprising an amino acid sequence denoted by one or more of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO 466, SEQ ID NO:350, SEQ ID NO:480, SEQ ID NO:481, a variant thereof or any combinations thereof.
  • the at least one sweet protein at FMP concentration comprising an amino acid sequence denoted by SEQ ID NO: 14 or SEQ ID NO:350, a variant thereof or any combinations thereof.
  • the at least one sweet protein at FMP concentration comprising an amino acid sequence denoted by SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO:39, SEQ ID NO:53, SEQ ID NO:84, SEQ ID NO: 104, SEQ ID NO: 186, SEQ ID NO:217, SEQ ID NO:263, SEQ ID NO:257, SEQ ID NO:351 or any combinations thereof.
  • the dairy product comprises at least one sweet protein denoted herein as DM31 (SEQ ID NO:39). In some examples, the dairy product comprises at least one sweet protein denoted herein as DM330 (SEQ ID NO: 84). In some examples, the dairy product comprises a combination of DM31 and DM330.
  • the dairy product comprises the one or more sweet proteins at a concentration of between about 1 ppm and about 100 ppm, at times between about 1 ppm and about 90 ppm, at times between about 1 ppm and about 80 ppm, at times between about 1 ppm and about 70 ppm, at times between about 1 ppm and about 60 ppm, at times between about 1 ppm and about 50 ppm, at times between about 1 ppm and about 40 ppm, at times between about 1 ppm and about 35 ppm, at times between 3 ppm and about 35 ppm, at times between about 5 ppm and about 30 ppm, at times between about 5 ppm and about 25 ppm, at times between about 10 ppm and about 20 ppm.
  • the dairy product comprises the one or more sweet proteins at a concentration of about 1 ppm, at times about 2 ppm, at times about 3 ppm, at times about 4 ppm, at times about 5 ppm, at times about 6 ppm, at times about 7 ppm, at times about 8 ppm, at times about 9 ppm, at times about 10 ppm, at times about 11 ppm, at times about 12 ppm, at times about 13 ppm, at times about 14 ppm, at times about 15 ppm.
  • the dairy product comprises the one or more sweet proteins at a concentration of about 15 ppm. In some examples, the dairy product comprises DM31 at a concentration of about 15 ppm. In some examples, the dairy product comprises DM330 at a concentration of about 15 ppm. In some examples, the dairy product comprises a combination of DM330 and DM31 at a concentration of about 15 ppm.
  • the food product is a non-dairy milk product. In some examples, the food product is oat milk. In some examples, the food product is an imitation dairy. In some examples the imitation dairy is soy yogurt.
  • the present disclosure provides a method for modifying at least one flavor in a comestible product, the method comprises adding a flavor modifying concentration of at least one protein to the comestible product, wherein the product is a non-diary product.
  • the present disclosure provides a non-dairy product comprising at least one sweet protein at FMP concentration.
  • the non-dairy product comprising at least one sweet protein denoted herein as DM31. In some examples, the non-dairy product comprising DM31 at a concentration of about 8 ppm. In some examples, the non-dairy product comprising at least one sweet protein at a concentration of about 8 ppm.
  • the non-dairy product is oat milk. In some examples, the nondairy product is oat milk comprising DM31 at a concentration of about 8 ppm.
  • the non-dairy product comprises at least one added salt. In some examples, the non-dairy product is essentially free of one or more sweeteners. In some examples, the non-dairy product comprises at least one added sweetener. In some examples, the non-dairy product comprises at least sucrose. In some examples, the non- dairy product comprises sucrose at a concentration of 3%.
  • the food product is Malabi.
  • the food product is a refrigerated dessert. In some examples, the food product is a frozen dessert. In some examples, the food product is a dairy delicatesse.
  • frozen products comprising at most I5ppm sweet protein were less sweet as compared to the same frozen products with 1.5% sucrose.
  • addition of the sweet protein at a concentration of 15 ppm to frozen products containing 7% sucrose has the capacity to modify flavors in the products.
  • the present disclosure provides a method for modifying at least one flavor in a comestible product, the method comprises adding a flavor modifying concentration of at least one protein to the comestible product, wherein the product is a frozen product.
  • the present disclosure provides a frozen product comprises at least one sweet protein at FMP concentration.
  • the frozen product comprises at least one sweet protein denoted herein as DM31.
  • the dairy product comprises at least one sweet protein denoted herein as DM330.
  • the dairy product comprises a combination of DM31 and DM330.
  • the frozen product comprises the one or more sweet proteins at a concentration of between about 1 ppm and about 100 ppm, at times between about 1 ppm and about 90 ppm, at times between about 1 ppm and about 80 ppm, at times between about 1 ppm and about 70 ppm, at times between about 1 ppm and about 60 ppm, at times between about 1 ppm and about 50 ppm, at times between about 1 ppm and about 40 ppm, at times between about 1 ppm and about 35 ppm, at times between 3 ppm and about 35 ppm, at times between about 5 ppm and about 30 ppm, at times between about 5 ppm and about 25 ppm, at times between about 10 ppm and about 20 ppm.
  • the frozen product comprises the one or more sweet proteins at a concentration of about 1 ppm, at times about 2 ppm, at times about 3 ppm, at times about 4 ppm, at times about 5 ppm, at times about 6 ppm, at times about 7 ppm, at times about 8 ppm, at times about 9 ppm, at times about 10 ppm, at times about 11 ppm, at times about 12 ppm, at times about 13 ppm, at times about 14 ppm, at times about 15 ppm.
  • the frozen product comprises the one or more sweet proteins at a concentration of about 15 ppm. In some examples, the frozen product comprises DM31 at a concentration of about 15 ppm.
  • the frozen product comprises at least one added salt. In some examples, the frozen product is essentially free of one or more sweeteners. In some examples, the frozen product comprises at least one added sweetener. In some examples, the frozen product comprises at least one added sweetener at a concentration of 7%. In some examples, the frozen product comprises at least sucrose. In some examples, the frozen product comprises sucrose at a concentration of 7%.
  • the frozen product is ice cream. In some examples, the frozen product is vanilla ice cream. In some examples, the frozen product is chocolate ice cream.
  • the food product is a seasoning product.
  • the condiment is thousand island dressing.
  • the food product is a gravy.
  • the food product is one or more of ketchup, mustard, mayonnaise, hot sauce, soy sauce, relish, barbecue sauce, salsa, aioli, pesto, ranch, vinegar, tahini, chutney, honey, jam, pickles,zihurri, wasabi, tzatziki, Worcestershire sauce.
  • the food product is condiment.
  • the food product is a relish.
  • the food product is ketchup.
  • ketchup comprising at most 1 Oppm sweet protein was less sweet as compared to ketchup with 1.5% sucrose.
  • the present disclosure provides a method for modifying at least one flavor in a comestible product, the method comprises adding a flavor modifying concentration of at least one protein to the comestible product, wherein the product is a condiment.
  • the present disclosure provides a condiment comprising at least one sweet protein at FMP concentration.
  • the condiment comprises at least one sweet protein denoted herein as DM31. In some examples, the condiment comprises at least one sweet protein denoted herein as DM330. In some examples, the condiment comprises a combination of DM31 and DM330. In some examples, the condiment comprises a combination of DM31 and DM685. In some examples, the condiment comprises DM685. “In some examples, the condiment comprises a combination of DM31 and Brazzein. In some examples, the condiment comprises a combination of DM31 and Thaumatin. In some examples, the condiment comprises Brazzein. In some examples, the condiment comprises Thaumatin.
  • the condiment comprises the one or more sweet proteins at a concentration of between about 1 ppm and about 100 ppm, at times between about 1 ppm and about 90 ppm, at times between about 1 ppm and about 80 ppm, at times between about 1 ppm and about 70 ppm, at times between about 1 ppm and about 60 ppm, at times between about 1 ppm and about 50 ppm, at times between about 1 ppm and about 40 ppm, at times between about 1 ppm and about 35 ppm, at times between 3 ppm and about 35 ppm, at times between about 5 ppm and about 30 ppm, at times between about 5 ppm and about 25 ppm, at times between about 10 ppm and about 20 ppm.
  • the condiment comprises the one or more sweet proteins at a concentration of about 1 ppm, at times about 2 ppm, at times about 3 ppm, at times about 4 ppm, at times about 5 ppm, at times about 6 ppm, at times about 7 ppm, at times about 8 ppm, at times about 9 ppm, at times about 10 ppm, at times about 11 ppm, at times about 12 ppm, at times about 13 ppm, at times about 14 ppm, at times about 15 ppm.
  • the condiment comprises the one or more sweet proteins at a concentration of about 10 ppm.
  • the condiment comprises the one or more sweet proteins at a concentration of about 12 ppm.
  • the condiment is ketchup.
  • ketchup is essentially free of one or more sweeteners.
  • ketchup comprises at least one added sweetener.
  • ketchup comprises at least sucrose.
  • ketchup comprises sucrose at a concentration of 10%, of 12%, of 13%.
  • ketchup comprises one or more sweeteners at a concentration of between about 10% and about 13%.
  • the food product is a confection.
  • the confection may be maple flavored syrup.
  • the food product is a sweet sauce.
  • the food product is a spread.
  • the spread is at least one of peanut butter, jam, jelly or chocolate spread.
  • the food product is peanut butter.
  • the food product is marzipan. In some examples, the food product a jam. In some examples, the food product is a jelly. In some examples, the food product is a nut product. In some examples, the nut product is peanut butter, marzipan or a Hazelnuts spread.
  • the peanut butter comprising at most lOppm sweet protein
  • addition of the sweet protein at a concentration of 10 ppm to peanut butter containing 5% sucrose has the capacity to modify flavors in the products.
  • the present disclosure provides a method for modifying at least one flavor in a comestible product, the method comprises adding a flavor modifying concentration of at least one protein to the comestible product, wherein the product is a spread.
  • the present disclosure provides a spread comprising at least one sweet protein at FMP concentration.
  • the spread comprising at least one sweet protein denoted herein as DM31. In some examples, the spread comprising at least one sweet protein denoted herein as DM330. In some examples, the spread comprises a combination of DM31 and DM330.
  • the spread comprises the one or more sweet proteins at a concentration of between about 1 ppm and about 100 ppm, at times between about 1 ppm and about 90 ppm, at times between about 1 ppm and about 80 ppm, at times between about 1 ppm and about 70 ppm, at times between about 1 ppm and about 60 ppm, at times between about 1 ppm and about 50 ppm, at times between about 1 ppm and about 40 ppm, at times between about 1 ppm and about 35 ppm, at times between 3 ppm and about 35 ppm, at times between about 5 ppm and about 30 ppm, at times between about 5 ppm and about 25 ppm, at times between about 10 ppm and about 20 ppm.
  • the spread comprises the one or more sweet proteins at a concentration of about 1 ppm, at times about 2 ppm, at times about 3 ppm, at times about 4 ppm, at times about 5 ppm, at times about 6 ppm, at times about 7 ppm, at times about 8 ppm, at times about 9 ppm, at times about 10 ppm, at times about 11 ppm, at times about 12 ppm, at times about 13 ppm, at times about 14 ppm, at times about 15 ppm.
  • the spread comprises the one or more sweet proteins at a concentration of about 10 ppm. In some examples, the spread comprises the one or more sweet proteins at a concentration of about 12 ppm.
  • the spread is peanut butter.
  • peanut butter comprises at least one added salt.
  • peanut butter is essentially free of one or more sweeteners.
  • peanut butter comprises at least one added sweetener.
  • peanut butter comprises at least sucrose.
  • peanut butter comprises sucrose at a concentration of about 5%.
  • peanut butter comprises at least one added sweetener at a concentration of about 5%.
  • the food product is at least one of a baked good, baking mix, breakfast cereals, cheese, chewing gum, condiment, relishe, confection, frostings, egg product, fat, oil, fish product, frozen dairy, fruit ices, gelatin, pudding, granulated sugar, gravy, agranola/energy bar, abar, hard candy, dairy productjam and jelly, meat product, milk product, nut product, grain product, poultry product, processed fruit, processed vegetable, reconstituted vegetable protein, seasoning, snack food, soft candy, soup, sweet sauce or any combination.
  • the breakfast cereals may be sugar free.
  • the food product is a baked good.
  • the baked goods is a cookie or a cake.
  • the baked good is a granola bar or a cupcake.
  • the food product is a baking mix.
  • the food product is a breakfast cereal.
  • the breakfast cereal may be a puffed rice.
  • the food product is a pudding.
  • the pudding may be a chocolate pudding.
  • the pudding may be a vanilla pudding.
  • the food product is a grain product.
  • the grain product is rice pudding.
  • the food product a granola bar. In some examples, the food product is a chewing gum. In some examples, the food product is a frosting.
  • the food product is an egg product. In some examples, the food product is an egg salad.
  • the food product is one or more of a granola bar, a cupcake, a shandy, a red wine, a white wine, a morning cereal, puffed rice, chewing gum, maple flavored syrup, gummies, egg salad, vinaigrette sauce, mustard, thousand island sauce, ice cream, sherbet, sorbet, ice pop, Malabi, chocolate pudding, BBQ sauce, sweet chili sauce, soy yogurt, iced coffee, protein drink milk, peanut butter, marzipan, hazelnuts spread, rice pudding or fudge.
  • a granola bar a cupcake, a shandy, a red wine, a white wine, a morning cereal, puffed rice, chewing gum, maple flavored syrup, gummies, egg salad, vinaigrette sauce, mustard, thousand island sauce, ice cream, sherbet, sorbet, ice pop, Malabi, chocolate pudding, BBQ sauce, sweet chili sauce, soy yogurt, iced coffee, protein drink milk
  • the food product is at least one high fat product. In some examples, the food product is at least one oil. In some examples, the fat or oil product may be vinaigrette, mustard or thousand island dressing.
  • the food product is at least one fish product. In some examples, the food product is at least one meat product. In some examples, the food product is a poultry product.
  • the food product is a hard candy. In some examples, the food product is gelatin. In some examples, the food product is a processed fruit. In some examples, the food product is a processed vegetable. In some examples, the food product is a reconstituted vegetable protein. In some examples, the food product is a snack food.
  • the food product is at least one soft candy.
  • the food product is a soup.
  • the food product is one or more of a yogurt, cheese, whey cheese, milk, ice cream, peanut butter, marzipan ketchup, an egg salad, a granola bar, Malabi, marzipan, or a thousand island dressing.
  • the food product is one or more of a yogurt, cheese, milk, ice cream, peanut butter, or ketchup.
  • the comestible product is a beverage.
  • the beverage is at least one of non-alcoholic beverage, alcoholic beverage, instant coffee, ice coffee, iced tea, tea or any combination thereof.
  • the beverage is shandy.
  • the alcoholic beverage is wine.
  • the wine is white wine.
  • the wine is red wine.
  • the beverage is a diet beverage. In some examples, the beverage is low calorie. In some examples, the beverage contains at least one non-sucrose sweetener.
  • the beverage contains at least one added salt.
  • the beverage is selected from the group consisting of a sparkling beverage, reduced calorie soda, mid calorie beverage, low calorie beverage, reduced calorie beverage, zero calorie beverage, a fruit juice, a fruit-flavored juice, a juice drink, nectar, concentrated juice, vegetable juice, vegetable-flavoredjuice, sports drink, energy drink, enhanced water drink, enhanced water with vitamin, water drink, coconut water, coconut drink, tea type drink, coffee, cocoa drink, chocolate drink, beverages containing milk component, milk alternative beverage, beverage containing cereal extract and smoothie.
  • the beverage is at least one of fruit juice, flavored water, carbonated flavor water, coffee or tea.
  • the beverage is cranberry juice.
  • the beverage is iced tea or tea beverage.
  • the beverage is iced coffee.
  • the beverage is carbonated flavor water.
  • beverages comprising at most lOppm protein were less sweet as compared to beverages with 1.5% sucrose.
  • addition of the protein at a concentration of at mostlO ppm to beverages containing sucrose has the capacity to modify flavors in the products.
  • the present disclosure provides a method for modifying at least one flavor in a comestible product, the method comprises adding a flavor modifying concentration of at least one protein to the comestible product, wherein the product is a beverage.
  • the present disclosure provides a beverage comprising at least one sweet protein at FMP concentration.
  • the beverage comprising at least one sweet protein denoted herein as DM31. In some examples, the beverage comprising at least one sweet protein denoted herein as DM330. In some examples, the beverage comprises a combination of DM31 and DM330.
  • the beverage comprises the one or more sweet proteins at a concentration of between about 1 ppm and about 100 ppm, at times between about 1 ppm and about 90 ppm, at times between about 1 ppm and about 80 ppm, at times between about 1 ppm and about 70 ppm, at times between about 1 ppm and about 60 ppm, at times between about 1 ppm and about 50 ppm, at times between about 1 ppm and about 40 ppm, at times between about 1 ppm and about 35 ppm, at times between 3 ppm and about 35 ppm, at times between about 5 ppm and about 30 ppm, at times between about 5 ppm and about 25 ppm, at times between about 10 ppm and about 20 ppm.
  • the beverage comprises the one or more sweet proteins at a concentration of about 1 ppm, at times about 2 ppm, at times about 3 ppm, at times about 4 ppm, at times about 5 ppm, at times about 6 ppm, at times about 7 ppm, at times about 8 ppm, at times about 9 ppm, at times about 10 ppm, at times about 11 ppm, at times about 12 ppm, at times about 13 ppm, at times about 14 ppm, at times about 15 ppm.
  • the beverage comprises the one or more sweet proteins is at a concentration of about 10 ppm. In some examples, the beverage comprises the one or more sweet proteins is at a concentration of about 12 ppm.
  • the beverage comprises the one or more sweet proteins is at a concentration of about 5 ppm. In some examples, the beverage comprises the one or more sweet proteins is at a concentration of about 10 ppm. In some examples, the beverage comprising at least one sweet protein denoted herein as DM31.
  • the beverage comprises at least one added salt. In some examples, the beverage is essentially free of one or more sweeteners. In some examples, the beverage comprises at least one added sweetener. In some examples, the beverage comprises at least sucrose. In some examples, the beverage comprises sucrose at a concentration of 6%. In some examples, the beverage comprises sucrose at a concentration of 8%. In some examples, the beverage comprises one or more sweeteners at a concentration of between about 5% and about 10%.
  • the beverage is fruit juice, tea, iced tea, flavor water, or carbonated flavor water.
  • white wine comprising at most lOppm protein
  • addition of the sweet protein at a concentration of 10 ppm or two sweet proteins at a concentration of lOppm to white wine has the capacity to modify flavors in the products. It should be noted that the white wine was essentially free of sucrose.
  • the present disclosure provides a method for modifying at least one flavor in a comestible product, the method comprises adding a flavor modifying concentration of at least one protein to the comestible product, wherein the product is an alcoholic beverage.
  • the present disclosure provides an alcoholic beverage comprising at least one sweet protein at FMP concentration.
  • the alcoholic beverage comprising at least one sweet protein denoted herein as DM31. In some examples, the alcoholic beverage comprising at least one sweet protein denoted herein as DM330. In some examples, the alcoholic beverage comprises a combination of DM31 and DM330.
  • the alcoholic beverage comprises the one or more sweet proteins at a concentration of between about 1 ppm and about 100 ppm, at times between about 1 ppm and about 90 ppm, at times between about 1 ppm and about 80 ppm, at times between about 1 ppm and about 70 ppm, at times between about 1 ppm and about 60 ppm, at times between about 1 ppm and about 50 ppm, at times between about 1 ppm and about 40 ppm, at times between about 1 ppm and about 35 ppm, at times between 3 ppm and about 35 ppm, at times between about 5 ppm and about 30 ppm, at times between about 5 ppm and about 25 ppm, at times between about 10 ppm and about 20 ppm.
  • the alcoholic beverage comprises the one or more sweet proteins at a concentration of about 1 ppm, at times about 2 ppm, at times about 3 ppm, at times about 4 ppm, at times about 5 ppm, at times about 6 ppm, at times about 7 ppm, at times about 8 ppm, at times about 9 ppm, at times about 10 ppm, at times about 11 ppm, at times about 12 ppm, at times about 13 ppm, at times about 14 ppm, at times about 15 ppm.
  • the alcoholic beverage comprises the one or more sweet proteins at a concentration of about 10 ppm. In some examples, the alcoholic beverage comprises the one or more sweet proteins at a concentration of about 12 ppm.
  • the alcoholic beverage comprises the one or more sweet proteins at a concentration of about 5 ppm. In some examples, the beverage comprises the one or more sweet proteins at a concentration of about 10 ppm. In some examples, the beverage comprising at least one sweet protein denoted herein as DM31.
  • the present disclosure provides use of at least one protein to modify at least one flavor in a comestible product.
  • the present disclosure provides use of at least one sweet protein at FMP concentration to enhance sweetness of at least one sweetener in a comestible product.
  • the FMP effect of the sweet protein is retained for a long time period. In some embodiments the FMP effect is retained for at least 1 week. In some embodiments the FMP effect is retained for at least 2 weeks. In some embodiments the FMP effect is retained for at least 1 month. In some embodiments the FMP effect is retained for at least 2 months. In some embodiments the FMP effect is retained for at least 3 months. In some embodiments the FMP effect is retained for at least 4 months. In some embodiments the FMP effect is retained for at least 5 months. In some embodiments the FMP effect is retained for at least 6 months. In some embodiments the FMP effect is retained for at least 9 months. In some embodiments the FMP effect is retained for at least 12 months.
  • the FMP effect is retained for at least 18 months.
  • a flavor modifying composition comprising at least one protein and at least one sweetener.
  • the at least one protein is present in the flavor modifying composition in a FMP concentration.
  • the present disclosure encompasses addition of stevia both as a sweetener or as a flavor modifying agent.
  • the term "about” as used herein indicates values that may deviate up to 1%, more specifically 5%, more specifically 10%, more specifically 15%, and in some cases up to 20% higher or lower than the value referred to, the deviation range including integer values, and, if applicable, non-integer values as well, constituting a continuous range. In some embodiments, the term “about” refers to ⁇ 10%.
  • the panel assessed the sweetness of various sweet proteins at different concentrations in several food products, as well as the sweetness of 1.5% sucrose in the same products. This was done using a 2-AFC (two-alternative forced choice) test — also known as a directional paired comparison — to determine whether two samples differ in a specific sensory attribute (e.g., sweetness).
  • 2-AFC two-alternative forced choice
  • the panelists compared a sample containing the sweet protein(s) at each tested concentration with a sample containing 1.5% sucrose, determining which was sweeter.
  • the two samples were presented simultaneously, and each panelist was asked to identify which sample had the higher sweetness.
  • every panelist decided whether the sweet protein sample at the tested concentration or the 1.5% sucrose sample was sweeter.
  • the maximum concentration of the sweet protein (or the combination) that most tasters perceived as less sweet than 1.5% sucrose in the same food product was then evaluated for its flavor-modifying properties.
  • the purpose of this first stage was to identify the highest concentration at which the sweet protein is not considered a sweetener.
  • test products sucrose and the sweet protein at that concentration
  • control products corresponding products containing sucrose only
  • the sensory expert panel was established by a screening process of potential tasters. Screening tests, conducted according to ISO standard (IS 8586-1), examined the sensory sensitivity, consistency, and sensory memory of the taster. The panel is well trained and calibrated. The selected panel is trained regularly to maintain high performance output. A minimum of 12 tasters tested each application.
  • the sweet protein exhibits a sweetness level lower than that of 1.5% sucrose; therefore, its primary influence in the second stage was related to its flavor-modifying properties (FMP) rather than sweetness.
  • FMP flavor-modifying properties
  • Example 1 DM31 effect in water
  • DM31 was added to water at various concentrations (without sucrose), and a trained sensory panel evaluated the sweetness of each sample relative to water containing 1.5% sucrose.
  • Table 1 summarizes, for each tested DM31 concentration, how many panelists found the DM31-containing sample sweeter than 1.5% sucrose and how many found the 1.5% sucrose sample sweeter. This approach allows identification of the maximum DM31 concentration at which it is no longer perceived as a sweetener. Table 1 Assessment of sweetness in water samples with different DM31 concentrations compared to 1.5% sucrose
  • Example 2A determining DM31 concentration
  • DM31 was added to plain yogurt at different concentrations and a trained sensory expert panel evaluated the sweetness of each yogurt relative to plain yogurt containing 1.5% sucrose.
  • Table 2A summarizes how many panelists found each DM31 -containing yogurt sweeter than the 1.5% sucrose yogurt versus how many found the 1.5% sucrose yogurt sweeter. Table 2A. Assessment of sweetness in plain yogurt with different DM31 concentrations compared to plain yogurt with 1.5% sucrose
  • Example 2B DM31 as an FMP in Strawberry Yogurt
  • DM31 was then tested for flavor-modifying properties (FMP) in strawberry yogurt containing 4% sucrose. Two versions were prepared:
  • Example 3 DM330 as an FMP in strawberry yogurt
  • DM330 as an FMP was tested in a strawberry yogurt with 4% sucrose without DM330 (“contror) and with 15 ppm DM330 ("/ ⁇ .s7").
  • Example 4 DM330 and DM31 as an FMP in strawberry yogurt
  • DM31 in combination with DM33 can be used as FMP in strawberry yogurt and more.
  • Example 5 DM31 as an FMP in apple yogurt
  • DM31 as a flavor-modifying property (FMP) was evaluated in apple yogurt containing 4% sucrose. Two versions were prepared:
  • a trained sensory panel evaluated the flavor-modifying features of both samples, as shown in Table 5.
  • Example 6 DM31 as an FMP in Whey Cheese
  • DM31 was evaluated as a flavor-modifying property (FMP) in whey cheese containing 4% sucrose. Two versions were prepared: • Control: Whey cheese with 4% sucrose (no DM31)
  • Example 7 DM31 as an FMP in apple flavored whey cheese
  • DM31 was evaluated as a flavor-modifying property (FMP) in apple flavored cheese containing 4% sucrose. Two versions were prepared:
  • a trained sensory panel evaluated the flavor-modifying features of both samples, as shown in Table 7.
  • Example 8 DM31 effect in oat milk
  • Example 8A determining DM31 concentration
  • DM31 was added to oat milk at a concentration of 8 ppm.
  • the perceived sweetness was evaluated by a sensory panel in comparison to oat milk containing 1.5% sucrose. Results are shown in Table 8A.
  • DM31 was then tested for flavor-modifying properties (FMP) in oat milk containing 3% sucrose. Two versions were prepared:
  • a Control sample oat milk with 3% sucrose and no DM31 and a Test sample: oat milk with 3% sucrose plus 8 ppm DM31.
  • Multiple sensory attributes were evaluated, as described below. Results of the sensory evaluation are presented in Table 8B.
  • Example 9 DM31 effect in vanilla ice cream
  • Example 9A determining DM31 concentration
  • DM31 was added to ice cream at different concentrations and a trained sensory expert panel evaluated the sweetness of each ice cream relative to an ice cream with 1.5% sucrose.
  • Table 9 A summarizes how many panelists found each DM31 -containing ice cream sweeter than the 1.5% sucrose ice cream versus how many found the 1.5% sucrose ice cream sweeter.
  • Example 9B DM31 as an FMP in ice cream
  • DM31 was then tested for flavor-modifying properties (FMP) in ice cream with 7% sucrose and 3% dextrose.
  • a trained sensory panel evaluated the flavor-modifying features of both samples, as shown in Table 9B.
  • the addition of 15 ppm DM31 to ice cream with 7% sucrose and 3% dextrose affected sensory attributes.
  • the addition of 15 ppm DM31 to ice cream with 7% sucrose and 3% dextrose results in an intensified desired vanilla taste/flavor.
  • the addition of 15 ppm DM31 to ice cream with 7% sucrose and 3% dextrose enhanced the sweetness.
  • Example 10A Determining DM31 concentration
  • DM31 was added to ketchup at different concentrations and a trained sensory expert panel evaluated the sweetness of each ketchup relative to ketchup containing 1.5% sucrose.
  • Table 10A summarizes how many panelists found each DM31 -containing ketchup sweeter than the 1.5% sucrose ketchup versus how many found the 1.5% sucrose ketchup sweeter.
  • Example 10B DM31 as an FMP in ketchup
  • DM31 was then tested for flavor-modifying properties (FMP) in ketchup containing 10% sucrose. Two versions were prepared:
  • a trained sensory panel evaluated the flavor-modifying features of both samples, as shown in Table 10B.
  • DM31 modifies sourness in ketchup was evaluated in a 2-AFC test for sourness.
  • a total of 42 trained panelists completed a 2-AFC test for sourness.
  • Twenty-eight (28) responses indicated the Test Sample is sourer.
  • Twenty-eight (28) responses indicated the Test Sample is sourer.
  • Twenty-eight (28) responses indicated the Control Sample is sourer.
  • (14) responses indicated the Control Sample is sourer.
  • DM31 modified sweetness in ketchup was evaluated in a 2-AFC test for sweetness.
  • DM31 modifies tomato concentrate flavor in ketchup was evaluated in a 2-AFC test for tomato concentrate flavor.
  • a total of 28 trained panelists completed a 2-AFC test for tomato concentrate flavor.
  • Twenty (20) responses indicated the Test Sample has stronger tomato concentrate flavor.
  • Example 11 DM31 as an FMP in Ketchup with extended shelf life
  • DM31 as an FMP is tested in ketchup with 10% sucrose without DM31 (“contrary and with 10 ppm DM31 (’7c.s7"). using an extended shelf life protocol.
  • the FMP is tested as detailed below, determining FMP effect after a long time period (1 week, 2 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months).
  • DM31 modification of sourness in ketchup is evaluated in a 2-AFC test for sourness; DM31 modification of sweetness in ketchup is evaluated in a 2-AFC test for sweetness; DM31 modification of tomato concentrate flavor is evaluated in a 2- AFC test for tomato concentrate flavor. Additional flavors are also examined.
  • Example 12 DM330 as an FMP in ketchup
  • DM330 was then tested for flavor-modifying properties (FMP) in a ketchup with 13% sucrose.
  • a trained sensory panel evaluated the flavor-modifying features of both samples.
  • DM330 may be used as FMP in Ketchup and more.
  • Example 13 DM330 and DM31 as an FMP in ketchup
  • a Test sample a ketchup with 13% sucrose plus 12 ppm of a combination of DM31 and DM330
  • a trained sensory panel evaluated the flavor-modifying features of both samples.
  • Example 14 DM685 as an FMP in Ketchup
  • Sucrose, salt, xanthan gum, ketchup spice are mixed well.
  • Tomato paste, water and vinegar are added to kettle and mixed well. Two mixtures are combined together until a uniform ketchup is obtained. Sweet protein is added and mixed well.
  • Example 15 DM31 and DM685 as an FMP in Ketchup
  • Sucrose, salt, xanthan gum, ketchup spice are mixed well.
  • Tomato paste, water and vinegar are added to kettle and mixed well. Two mixtures are combined together until a uniform ketchup is obtained. Sweet proteins combination is added and mixed well.
  • Sucrose, salt, xanthan gum, ketchup spice are mixed well.
  • Tomato paste, water and vinegar are added to kettle and mixed well. Two mixtures are combined together until a uniform ketchup is obtained. Sweet protein is added and mixed well.
  • Thaumatin may be used as FMP in Ketchup and more.
  • Example 17 DM31 & Thaumatin as an FMP in Ketchup
  • Sucrose, salt, xanthan gum, ketchup spice are mixed well.
  • Tomato paste, water and vinegar are added to kettle and mixed well. Two mixtures are combined together until a uniform ketchup is obtained. Sweet proteins combination is added and mixed well.
  • Thaumatin and DM31 may be used as FMP in Ketchup and more.
  • Brazzein may be used as FMP in Ketchup and more.
  • Example 19 A combination of DM31 and Brazzein as an FMP in Ketchup
  • Sucrose, salt, xanthan gum, ketchup spice are mixed well.
  • Tomato paste, water and vinegar are added to kettle and mixed well. Two mixtures are combined together until a uniform ketchup is obtained. Sweet proteins combination is added and mixed well.
  • Example 20 DM31 effect in peanut butter
  • Example 20A determining DM31 concentration
  • DM31 was added to peanut butter at a concentration of 10 ppm and a trained sensory expert panel evaluated the sweetness of the peanut butter relative to a peanut butter with 1.5% sucrose.
  • Table 20A summarizes how many panelists found 1.5% sucrose peanut butter sweeter each DM31 -containing peanut butter.
  • peanut butter containing 10 ppm DM31 or lower was perceived by the majority of panelists as less sweet than peanut butter with 1.5% sucrose.
  • Example 20B DM31 as an FMP in peanut butter
  • DM31 was then tested for flavor-modifying properties (FMP) in peanut butter with 5% sucrose.
  • a trained sensory panel evaluated the flavor-modifying features of both samples, as shown in Table 20B.
  • the addition of 10 ppm DM31 to peanut butter with 5% sucrose affected sensory attributes.
  • the addition of 10 ppm DM31 to peanut butter with 5% sucrose resulted in enhanced peanut butter flavor while reducing the saltiness of the peanut butter.
  • Example 21 Determining DM31 effect in cranberry juice
  • DM31 was added to cranberry juice at a concentration of 10 ppm and trained sensory expert panel evaluated the sweetness of the cranberry relative to cranberry juice with 1.5% sucrose.
  • Table 21A summarizes how many panelists found 1.5% sucrose cranberry juice sweeter each DM31-containing cranberry juice.
  • Table 21A assessment of sweetness in cranberry juice sample with 10 ppm DM31 as compared to cranberry juice with 1.5% sucrose. As shown in Table 21A, cranberry juice containing 10 ppm was perceived by the majority of panelists as less sweet than a cranberry juice with 1.5% sucrose.
  • Example 21B DM31 as an FMP in cranberry juice
  • DM31 was then tested for flavor-modifying properties (FMP) in cranberry juice with 8% sucrose.
  • a trained sensory panel evaluated the flavor-modifying features of both samples, as shown in Table 21B.
  • Example 22 Determining DM31 effect in Iced tea
  • DM31 was added to iced tea at different concentrations and a trained sensory expert panel evaluated the sweetness of each iced tea relative to an iced tea with 1.5% sucrose.
  • Table 22A summarizes how many panelists found each DM31 -containing iced tea sweeter than the 1.5% sucrose iced tea versus how many found the 1.5% sucrose iced tea sweeter.
  • DM31 as compared to iced tea with 1.5% sucrose.
  • Example 22B DM31 as an FMP in iced tea
  • DM31 was then tested for flavor-modifying properties (FMP) in iced tea with 6% sucrose.
  • a trained sensory panel evaluated the flavor-modifying features of both samples, as shown in Table 22B.
  • Table 22B assessment of flavor modifying features in iced tea As shown in Table 22B, the addition of 10 ppm DM31 to iced tea with 6% sucrose, affected multiple sensory attributes. In particular, the addition of 10 ppm DM31 to iced tea with 6% sucrose results in sweetness enhancement and lemon flavor.
  • Example 23 DM31 as an FMP in apple flavored carbonated water
  • DM31 was tested for flavor-modifying properties (FMP) in an apple flavored carbonated water with 6% sucrose.
  • a Test sample an apple flavored carbonated water with 6% sucrose plus 5 ppm of DM31
  • a trained sensory panel evaluated the flavor-modifying features of both samples, as shown in Table 23A.
  • DM31 modify sweetness in apple flavored carbonated water was evaluated in a 2-AFC test for sweetness.
  • Example 24 DM330 as an FMP in apple flavored carbonated water
  • DM330 was tested for flavor-modifying properties (FMP) in an apple flavored carbonated water with 6% sucrose.
  • a Control sample an apple flavored carbonated water with 6% sucrose and no
  • DM330 • A Test sample: an apple flavored carbonated water with 6% sucrose plus 8 ppm ofDM330
  • DM330 may be used as FMP in apple flavored carbonated water and more.
  • Example 25 DM330 and DM31 as an FMP in apple flavored carbonated water
  • a combination of DM330 and DM31 was tested for flavor-modifying properties (FMP) in an apple flavored carbonated water with 6% sucrose.
  • a Test sample an apple flavored carbonated water with 6% sucrose plus 5 ppm of a combination of DM31 and DM330
  • DM31 in combination with DM330 may be used as FMP in apple flavored carbonated water and more.
  • Example 26 DM31 as an FMP in strawberry flavored carbonated water
  • DM31 is tested for flavor-modifying properties (FMP) strawberry flavored carbonated water with 6% sucrose.
  • a trained sensory panel evaluated the flavor-modifying features of both samples, as shown in Table 26.
  • Example 27 DM31 as an FMP in Egg salad
  • DM31 is tested for flavor-modifying properties (FMP) in egg salad with ⁇ 3% sucrose.
  • a Test sample granola bar with egg salad with ⁇ 3% sucrose plus about 20 ppm of a DM31
  • 2-AFC two-alternative forced choice
  • 2-AFC is a difference test used when the objective is to determine whether two samples differ in a specified dimension (such as sweetness, sourness, etc.).
  • each taster in the panel decides which of the two samples has a stronger specific flavor, the food product comprising the DM31 or the same food product without DM31.
  • the two samples are presented to the panelist simultaneously and the panelist is asked to identify the sample that is higher in the specified sensory attribute. It was suggested that DM31 may be used as FMP in egg salad and more.
  • Example 28 DM31 as an FMP in Granola Bars
  • Nuts and oatmeal are roasted at 160°C for 15 minutes. Oatmeal, crisped rice, and nuts are ground. Syrup is prepared by mixing palm fat, glycerol, soy lecithin, salt, powdered sugar and glucose syrup. Syrup is heated on low heat and mixed well until all syrup ingredients are dissolved. DM31 is mixed into the warm syrup. Syrup is mixed with oatmeal, crisped rice, and nuts. Bake for 10 minutes at 90°C.
  • DM31 is tested for flavor-modifying properties (FMP) in granola bar with 7% sucrose.
  • 2-AFC two-alternative forced choice
  • 2-AFC is a difference test used when the objective is to determine whether two samples differ in a specified dimension (such as sweetness, sourness, etc.).
  • each taster in the panel decides which of the two samples has a stronger specific flavor, the food product comprising the DM31 or the same food product without DM31.
  • the two samples are presented to the panelist simultaneously and the panelist is asked to identify the sample that is higher in the specified sensory attribute.
  • DM31 As FMP after the heating which is part of the preparation process. It was suggested that DM31 may be used as FMP in granola bar and more.
  • Nuts are roasted and oatmeal at 160°C for 15 minutes. Oatmeal, crisped rice, and nuts are ground. Syrup is prepared by mixing palm fat, glycerol, soy lecithin, salt, powdered sugar and glucose syrup. The syrup is heated on low heat and mixed well until all syrup ingredients are dissolved. DM31 and DM528 are mixed into the warm syrup. Syrup is mixed with oatmeal, crisped rice, and nuts. Bake for 10 minutes at 90°C.
  • a combination of DM31 and DM528 is tested for flavor-modifying properties (FMP) in granola bar with 7X% sucrose.
  • Test sample granola bar with 7% sucrose plus about 20 ppm a combination of
  • 2-AFC two-alternative forced choice
  • 2-AFC is a difference test used when the objective is to determine whether two samples differ in a specified dimension (such as sweetness, sourness, etc.).
  • a specified dimension such as sweetness, sourness, etc.
  • each taster in the panel decides which of the two samples has a stronger specific flavor, the food product comprising the combination of DM31 and DM528or the same food product without DM31 & DM528.
  • the two samples are presented to the panelist simultaneously and the panelist is asked to identify the sample that is higher in the specified sensory attribute.
  • Example 30A DM31 as an FMP in Malabi
  • Com starch is mixed with water.
  • Mixture is poured into serving glasses and put in refrigerator.
  • Table 30A Formulation DM31 is tested for flavor-modifying properties (FMP) in Malabi with less than 10.6% sucrose.
  • 2-AFC two-alternative forced choice
  • 2-AFC is a difference test used when the objective is to determine whether two samples differ in a specified dimension (such as sweetness, sourness, etc.).
  • each taster in the panel decides which of the two samples has a stronger specific flavor, the food product comprising the DM31 or the same food product without DM31.
  • the two samples are presented to the panelist simultaneously and the panelist is asked to identify the sample that is higher in the specified sensory attribute.
  • Example 31 DM528 as an FMP in Malabi
  • Com starch is mixed with water. Milk, cream, and sucrose are boiled. When solution is boiled, starch mixture is poured while stirring. Temperature is lowered to 60- 65°C and then rose water and DM528 are added and mixed well. Mixture is poured into serving glasses and put in refrigerator.
  • DM528 is tested for flavor-modifying properties (FMP) in Malabi with less than 10.6% sucrose.
  • 2-AFC two-alternative forced choice
  • 2-AFC is a difference test used when the objective is to determine whether two samples differ in a specified dimension (such as sweetness, sourness, etc.).
  • each taster in the panel decides which of the two samples has a stronger specific flavor, the food product comprising the DM528 or the same food product without DM528.
  • the two samples are presented to the panelist simultaneously and the panelist is asked to identify the sample that is higher in the specified sensory attribute.
  • DM528 As FMP after the heating which is part of the preparation process. It was suggested that DM528 may be used as FMP in Malabi and more.
  • Example 32 DM31 as an FMP in Iced coffee
  • DM31 is tested for flavor-modifying properties (FMP) in iced coffee with less than 11% sucrose. Two versions are prepared:
  • a Control sample iced coffee with less than 11% sucrose and no DM31 • A Test sample: iced coffee with less than 11% sucrose plus about 20 ppm DM31
  • 2- AFC two-alternative forced choice
  • 2- AFC is a difference test used when the objective is to determine whether two samples differ in a specified dimension (such as sweetness, sourness, etc.).
  • each taster in the panel decides which of the two samples has a stronger specific flavor, the beverage comprising the DM31 or the beverage product without DM31.
  • the two samples are presented to the panelist simultaneously and the panelist is asked to identify the sample that is higher in the specified sensory attribute. It was suggested that DM31 may be used as FMP in iced coffee and more.
  • Example 33 DM31 as an FMP in Marzipan
  • DM31 is tested for flavor-modifying properties (FMP) in marzipan.
  • 2-AFC two-alternative forced choice
  • 2-AFC is a difference test used when the objective is to determine whether two samples differ in a specified dimension (such as sweetness, sourness, etc.).
  • each taster in the panel decides which of the two samples has a stronger specific flavor, the food product comprising the DM31 or the same food product without DM31.
  • the two samples are presented to the panelist simultaneously and the panelist is asked to identify the sample that is higher in the specified sensory attribute. It was suggested that DM31 may be used as FMP in marzipan and more.
  • Sucrose, salt, xanthan gum and ketchup spice are mixed.
  • Tomato paste, water and vinegar are added to a kettle and mixed well. Two mixtures are combined together until a uniform paste is obtained and then mayonnaise is mixed in. Sweet protein is added and mixed well.
  • DM31 is tested for flavor-modifying properties (FMP) in thousand islands dressing with less than 12% sucrose.
  • 2-AFC two-alternative forced choice
  • 2-AFC two-alternative forced choice
  • a specified dimension such as sweetness, sourness, etc.
  • each taster in the panel decides which of the two samples has a stronger specific flavor, the food product comprising the DM31 or the same food product without DM31.
  • the two samples are presented to the panelist simultaneously and the panelist is asked to identify the sample that is higher in the specified sensory attribute. It was suggested that DM31 may be used as FMP in thousand islands dressing and more.
  • Example 35A Determining DM31 concentration
  • DM31 was added to white wine at a concentration of 10 ppm and a trained sensory expert panel evaluated the sweetness of each white wine relative to a white wine with 1.5% sucrose.
  • Table 35A summarizes how many panelists found the DM31 -containing white wine sweeter than the 1.5% sucrose white wine versus how many found the 1.5% sucrose white wine sweeter.
  • white wine containing 10 ppm DM31 perceived by the majority of panelists to be less sweet than white wine with 1.5% sucrose.
  • Example 36 DM31 as an FMP in white wine
  • DM31 as an FMP was tested in white wine without DM31 (“control”) and with 10 ppm DM31 (“test”). The FMP was tested as detailed below and shown in Table 36. DM31 modifies biterness in white wine was evaluated in a 2-AFC test for biterness.
  • DM31 modified sweetness in white wine was evaluated in a 2-AFC test for sweetness.
  • DM31 modified fruity flavor in white wine was evaluated in a 2-AFC test for fruity flavor.
  • Table 36 assessment of flavor modifying features in white wine As can be seen in Table 18B, addition of 10 ppm DM31 to a white wine, effected multiple sensory attributes. It was suggested that DM31 may be used as FMP in white wine and more, with multiple benefits across different sensory attributes
  • Example 37 DM330 as an FMP in white wine
  • Sweet protein DM330 was added to the wine and mixed well. The list of ingredients is shown in Table 37.
  • DM330 was tested for flavor-modifying properties (FMP) in dry white wine with no sugar.
  • DM330 may be used as FMP in white wine and more.
  • Example 38 A combination of DM330 and DM31 as an FMP in dry white wine
  • a combination of sweet proteins DM330 and DM31 was added to dry white wine and mixed well.
  • a combination of DM330 and DM31 was tested for flavor-modifying properties (FMP) in dry white wine with no sugar.

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PCT/IL2025/050330 2024-04-10 2025-04-10 Sweet proteins and uses thereof Pending WO2025215649A1 (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4562076A (en) * 1982-12-27 1985-12-31 Wm. Wrigley Jr. Company Chewing gum with coating of thaumatin or monellin and method
US6610514B2 (en) * 1996-08-23 2003-08-26 Kirin Beer Kabushiki Kaisha Yeast vector and method of producing proteins using the same
AU2018202660A1 (en) * 2013-03-14 2018-05-10 The Coca-Cola Company Beverages containing rare sugars
US20200107567A1 (en) * 2018-05-08 2020-04-09 Epc Natural Products Co., Ltd. Tasteful natural sweetener and flavor
US20210139546A1 (en) * 2018-05-06 2021-05-13 Amai Proteins Ltd. Taste and flavor-modifier proteins

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4562076A (en) * 1982-12-27 1985-12-31 Wm. Wrigley Jr. Company Chewing gum with coating of thaumatin or monellin and method
US6610514B2 (en) * 1996-08-23 2003-08-26 Kirin Beer Kabushiki Kaisha Yeast vector and method of producing proteins using the same
AU2018202660A1 (en) * 2013-03-14 2018-05-10 The Coca-Cola Company Beverages containing rare sugars
US20210139546A1 (en) * 2018-05-06 2021-05-13 Amai Proteins Ltd. Taste and flavor-modifier proteins
US20200107567A1 (en) * 2018-05-08 2020-04-09 Epc Natural Products Co., Ltd. Tasteful natural sweetener and flavor

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