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
  • A23L27/20—Synthetic spices, flavouring agents or condiments
  • A23L27/26—Meat flavours
• • 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
  • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
  • A23L11/50—Fermented pulses or legumes; Fermentation of pulses or legumes based on the addition of microorganisms
• • 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
  • A23L23/00—Soups; Sauces; Preparation or treatment thereof
• • 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
  • A23L27/10—Natural spices, flavouring agents or condiments; Extracts thereof
• • 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
  • A23L27/20—Synthetic spices, flavouring agents or condiments
  • A23L27/202—Aliphatic compounds
  • A23L27/2024—Aliphatic compounds having oxygen as the only hetero atom
• • 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
  • A23L27/20—Synthetic spices, flavouring agents or condiments
  • A23L27/205—Heterocyclic compounds
  • A23L27/2052—Heterocyclic compounds having oxygen or sulfur as the only hetero atoms
• • 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
  • A23L27/20—Synthetic spices, flavouring agents or condiments
  • A23L27/21—Synthetic spices, flavouring agents or condiments containing amino acids
• • 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
  • A23L27/20—Synthetic spices, flavouring agents or condiments
  • A23L27/24—Synthetic spices, flavouring agents or condiments prepared by fermentation
• • 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
  • A23L27/88—Taste or flavour enhancing agents

Definitions

• the present invention relates to flavor and sensorial materials and consumable compositions comprising the same, in particular, meatless, plant-based, animal protein-free flavor materials and compositions imparting a meaty and/or animal protein-like flavor to food stuff to which they are added.
• the present invention in embodiments thereof provides consumable flavor-rich flavor and sensorial materials, as well as compositions and food products containing the same.
• the flavor materials are animal protein-free, yet, advantageously provide to food stuff to which they are added, the complex taste and aroma, typically associated with animal protein (e.g. meat, milk, butter, cream, cheese, fish, etc.), products derived from animal protein, or foods prepared with animal protein or such products.
• the herein disclosed consumable flavor materials and compositions serve to create and enhance the experience of "deliciousness" in general and in animal protein analogues in particular, and to elevate the taste experience of consuming such products and make it as close as possible to the total experience of consuming animal protein or materials prepared with animal protein, as opposed to currently available flavor solutions which focus on imitating particular attributes of meat / animal protein.
• the herein disclosed consumable flavor and sensorial materials and compositions containing same are characterized by helping plant-based food taste delicious in the way that foods prepared with animal protein are delicious; putting the pleasure of consuming food at the center.
• flavor materials and compositions that in it is possible to obtain a plant-based food stuff that provides a pleasure-inducing taste-experience characteristic to animal protein or foods prepared with animal protein, by adding to such food stuff flavor materials and/or compositions which abide to certain criteria.
• criteria include for example the presence of at least one volatile substance at an odor activity value (OAV) of at least 10 when the flavor material(s) are concentrated so that their Brix value is approximately 60 ⁇ Bx to 80 ⁇ Bx and/or their water content (moisture level) is approximately 40% to 60%.
• OAV odor activity value
• Such criteria also include for example a non-volatile taste distribution characterized by about 50-85% (for example, about 60-80%) non-volatile taste substances having a sweet, and/or about 3-25% (for example, about 3-7%) non-volatile taste substances having a sour taste, and/or about 4-25% (for example, about 4-7%) non-volatile taste substances having a bitter taste and/or about 2- 10% (for example, about 8-10%) of non-volatile taste substances having an umami taste, (all out of the total amount of non-volatile taste substances).
• a non-volatile taste distribution characterized by about 50-85% (for example, about 60-80%) non-volatile taste substances having a sweet, and/or about 3-25% (for example, about 3-7%) non-volatile taste substances having a sour taste, and/or about 4-25% (for example, about 4-7%) non-volatile taste substances having a bitter taste and/or about 2- 10% (for example, about 8-10%)
• a consumable flavor rich flavor material comprising: at least one volatile substance at an odor activity value (OAV) of at least 10, the volatile substances may be selected from: 2,3-Octanedione, 3-methyl-Butanal, 2- methyl-Butanal, 2,6-dimethyl-Pyrazine, a-ethylidene-Benzeneacetaldehyde, dimethyl Trisulfide, lH-indole, 2-Propenal, Hexanal and Hexadecanoic acid, ethyl ester; and a nonvolatile taste distribution of: about 50-85% non-volatile taste substances having a sweet or sour taste, about 3-25% non-volatile taste substances having a sour taste; about 4-25% nonvolatile taste substances having a bitter taste; and about 2-10% of non-volatile taste substances having an umami taste out of a total amount of non-volatile taste substances; wherein the flavor material
• the at least one volatile substance may further include one or more of: lH-indole, Ethyl linoleate, Acetaldehyde phenyl-dimethyl acetal, and 2,3-Butanediol. Each possibility is a separate embodiment.
• the taste substances having a sweet taste comprises alanine, proline, fructose, rhamnose, asparagine, serine, mannitol, sucrose, glutamine, threonine, myo-inositol, glycine, glucose and raffinose.
• the consumable flavor rich flavor material comprises isomaltose at an IRR of at least 40.
• the taste substances having a sweet taste comprises glucose.
• the taste substances having a sour taste comprises lactic acid, citric acid, maleic acid, succinic acid and tartaric acid. Each possibility is a separate embodiment.
• the taste substances having a sour taste comprises lactic acid.
• the consumable flavor rich flavor material comprises lactic acid at an IRR of at least 40.
• the taste substances having a bitter taste comprises an amino acid including, for example, arginine, cystine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, tryptophan, tyrosine and/or valine.
• an amino acid including, for example, arginine, cystine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, tryptophan, tyrosine and/or valine.
• the taste substances having a bitter taste comprises arginine.
• the taste substances having an umami taste include an amino acid, such as, for example, glutamate, aspartate, and/or betaine.
• the umami taste substance may be selected from uridine 5'-monophosphate (5'-Ump), aspartic acid, glutamic acid, adenosine 5'- monophosphate (Amp), guanosine 5'-monophosphate (Gmp), or combinations thereof.
• the umami taste substance comprises glutamate.
• the consumable flavor rich flavor material has a meaty flavor.
• the consumable flavor rich flavor material is obtained by solid state or submerged (liquid state) fermentation of one or more legumes, grains, oilseeds, vegetables, nuts, agro-industrial waste streams or any combinations thereof.
• the legume may be selected from, but not limited to: chickpeas, soybeans, black beans, white beans, green beans, kidney beans, red beans, broad beans, lima beans, blacked-eyed peas, navy beans, adzuki beans, mung beans, lupin, green lentils, black lentils, red / yellow lentils, fava beans, broad beans, Cranberry ( Borlotti) Beans, green peas, yellow peas, peanuts, bambara nut, and the like.
• a consumable composition comprising the herein disclosed flavor rich flavor material.
• the consumable composition further comprises a flavor base.
• the flavor base is plant based (vegeta ria n/vegan).
• a method for providing meaty flavorto a meatless food product comprising the addition of the herein disclosed consumable flavor rich flavor material or the herein disclosed composition to the meatless food product.
• the food product is a ready-to-cook or ready-to-eat food product.
• the meatless food product is a ready-to-cook or ready-to-eat meat-less food product.
• a food product comprising the herein disclosed consumable, flavor rich flavor material or the herein disclosed composition.
• the food product is a plant-based food product.
• the food product is a sauce and/or a condiment.
• the food product is a prepared food such as a casserole, stew, legume or grain-based dish, soup, or the like.
• the food product is a stock, or bullion, fish-sauce or "dashi" analogue.
• the stock, bullion, and/or fish-sauce analogue is concentrated.
• the food product is a taste modulator or taste intensifier/enhancer.
• the flavor material is capable of at least partially masking beany off-flavors.
• the flavor material is capable of at least partially masking bitterness.
• the flavor material is capable of at least partially masking off-flavors typical of alternative sweeteners, such as stevia.
• the flavor material can at least partially increase the perception of fattiness including the mouthfeel typical of increased fattiness.
• the flavor material may at least partially function as a "sweetness modulator", for example by modulating and improving the mouthfeel of a product with a reduced sugar content so that the mouthfeel is more pronounced of that of a product prepared with a customary quantity of sugar.
• the flavor material may improve creamy and/or milk-fatty mouthfeel.
• the flavor material can increase the complexity of taste in reduced salt food products, including, without limitation, in low pH food products.
• the flavor material is capable of at least partially “masking” and/or “blocking” astringency.
• the flavor material is at least partially capable of "masking” and/or “blocking” sour taste.
• the flavor material is capable of any of the above attributes in a food product which is plant-based, or a food product which is not essentially plant based.
• Certain embodiments of the present disclosure may include some, all, or none of the above advantages.
• One or more technical advantages may be readily apparent to those skilled in the art from the figures, descriptions and claims included herein.
• specific advantages have been enumerated above, various embodiments may include all, some or none of the enumerated advantages.
• FIG. 1 presents a schematic block diagram of flavor materials production process, according to some embodiments.
• FIG. 2 shows a hierarchical clustering dendrogram of volatile and non-volatile compounds combined in the herein disclosed flavor material, meat products as well as a variety of nonmeat flavorings (here shown: Industrial "meaty” flavoring, Imitation meat bouillon). Analysis was done in JMP software using Maximum Likelihood method. Distance between clusters was calculated using Ward method. Similar products are clustered into one trunk.
• FIG. 3 presents known volatile materials represented in meat jus, an example of herein disclosed flavor material and non-meat flavorings (here shown: Industrial "meaty” flavoring, Imitation meat bullion). (Visual analysis created using Venny 2.1).
• FIGs. 4A - 4C present the concentration of different volatile component represented in meat jus, flavor material and non-meat flavorings (here shown: Industrial "meaty” flavoring, Imitation meat bouillon).
• FIG. 5 presents a stacked column chart showing the distribution of umami substances in the herein disclosed flavor material as well as in representative meat products and known non-meat flavorings (here shown: Industrial "meaty” flavoring, Imitation meat bouillon).
• FIG. 6 presents a stacked column chart showing the distribution of sour substances in the herein disclosed flavor materials as well as in representative meat products and known meat- free flavorings (here shown: Industrial "meaty” flavoring, Imitation meat bouillon).
• FIG. 7 presents a stacked column chart showing the distribution of sweet substances in the herein disclosed flavor materials as well as in representative meat products and known meat-free flavorings (here shown: Industrial "meaty” flavoring, Imitation meat bouillon).
• FIG. 8 presents a stacked column chart showing the distribution of bitter substances in the herein disclosed flavor materials as well as in representative meat products and known non-meat flavorings (here shown: Industrial "meaty” flavoring, Imitation meat bouillon).
• FIG. 9 presents a stacked column chart showing the distribution of koku (also known as “kokumi”) substances in the herein disclosed flavor materials as in representative meat products and known meat-free flavorings (here shown: Industrial “meaty” flavoring, Imitation meat bouillon).
• koku also known as "kokumi”
• FIG. 10A presents a column chart showing a paired comparison test (2-AFC) for fat perception of the reference sample "plant-based burger" with and without the herein disclosed flavor materials.
• FIG. 10B presents a graphical display (spider graphs) of sensory attributes based on Quantitative Descriptive Analysis (QDA) test results of "plant burger” and of “plant burger” including the herein disclosed flavor materials for the taste sensations: salty, sweet, bitter, sour, and astringent.
• QDA Quantitative Descriptive Analysis
• FIG. 11A presents a column chart showing a paired comparison test (2-AFC) for bitter perception of the reference sample of a commercial tonic water with and without the herein disclosed flavor materials.
• FIG. 11B presents a graphical display (spider graph) of sensory attributes based on Quantitative Descriptive Analysis (QDA) test results of a commercial tonic water with and without the herein disclosed flavor materials for the taste sensations: salty, sweet, bitter, sour, and astringent.
• QDA Quantitative Descriptive Analysis
• FIG. 12A presents a column chart showing the paired comparison test (2-AFC) for acidity perception of a reference sample of lemon juice with and without the herein disclosed flavor materials.
• FIG. 12B presents a graphical display (spider graphs) of sensory attributes based on Quantitative Descriptive Analysis (QDA) test results of lemon juice with and without the herein disclosed flavor materials for the taste sensations: salty, sweet, bitter, sour, and astringent.
• QDA Quantitative Descriptive Analysis
• FIG 13A presents a column chart showing the paired comparison test (2-AFC) for beany off-flavor perception of the reference sample 6% pea flour in water with and without the herein disclosed flavor materials.
• FIG. 13B presents a graphical display (spider graphs) of sensory attributes based on Quantitative Descriptive Analysis (QDA) test results of 6% pea flour in water with and without the herein disclosed flavor materials for the taste sensations: salty, sweet, bitter, sour, and astringent.
• QDA Quantitative Descriptive Analysis
• FIG 14A presents a column chart showing the paired comparison test (2-AFC) for astringency perception of a reference sample of cranberry juice with and without the herein disclosed flavor materials.
• FIG. 14B presents a paired comparison test (2-AFC) for stevia off-flavor perception of a reference sample containing 3% stevia in water with and without the herein disclosed flavor materials.
• FIG. 14C presents a paired comparison test (2-AFC) for mouthfeel perception of a reference sample of milk 1% fat with and without the herein disclosed flavor materials.
• a consumable flavor-rich, plant-based flavor material which includes 2,3 octanedione at an odor activity value (OAV) of at least 10 (as defined herein) and having a non-volatile taste distribution of: about 50%-85% (for example, about 75-85%) taste substances having a sweet or sour taste, about 3-25% (for example, about 3.5-10%) non-volatile taste substances having a sour taste, about 4-25% (for example, about 4.5-8%) non-volatile taste substances having a bitter taste; and about 2-12% (for example, about 8 -10%) of non-volatile taste substances having an umami taste out of a total amount of non-volatile taste substances.
• the flavor material may further include non-volatile koku substances.
• flavor material As used herein, the terms “flavor material”, “flavor materials”, “FM”, “FMs”, “MFL flavor material”, “MFL” and “flavor product(s)” may be used interchangeably.
• the terms refer to a product (such as a flavor concentrate, etc.) prepared by a process of fermentation of legumes and/or grains and/or materials prepared from such, and/or waste streams from the processing of such.
• the flavor material may be a solution, a viscous liquid, a paste, a powder, or a dry granulated substance among others, containing essential constituents of a complex material, such as plants.
• the flavor material is devoid of any synthetic and/or added natural and/or artificial flavor or aroma substances.
• the flavor material is devoid of any materials commonly considered by regulatory authorities to be additives, flavors, aromas, "E- numbers", or other such ingredients.
• the flavor material is comprised only of commonly recognizable food ingredients that an average consumer might have in their kitchen at any given time ("kitchen cupboard” ingredients, also commonly known as “foodstuff” ingredients).
• a flavor material may include a combination or mixture of distinct flavor or sensorial materials.
• the term “meatless”, “meat free” and “animal protein free” may be used interchangeably and refer to products which contain no meat, meat substances or other animal protein or animal protein substances, and which are produced in an animal protein-free process.
• the flavor material is vegetarian.
• the flavor material is vegan.
• the flavor material is non-diary.
• the term "taste sensation” refers to the sensation that results when taste receptors in the oral cavity (for example, mouth, on the tongue and/or throat), and/or elsewhere throughout the digestive system to convey information about the chemical composition of a soluble stimulus.
• the term “aromatic sensation” refers to a sense of smells (or odor) or olfaction. It occurs when an odor binds to a receptor within the nasal cavity or elsewhere, transmitting a signal through the olfactory system.
• mouthfeel refers to the sensation or sum of sensations other than taste or odor created by food or drink in the mouth.
• taste experience refers to a holistic experience involving taste sensation, mouthfeel and aromatic sensation.
• the taste experience refers to a "cross- modal" taste experience, or multidimensional sensorial attributes, meaning the experience whereby there is an "explosion of flavor” in the mouth, when substances/products are tasted in many places at once, in contrast to a one-dimensional experience.
• Koku refers to a sensation which can be described as a perceived richness, complexity of taste and flavor, viscosity, mouth-coating, and roundness that enhances thickness, continuity, complexity, freshness, and mouthfeel-ness of foods and beverages.
• the herein disclosed flavor materials are characterized by providing a long-lasting pleasure and satisfaction that is typical to the experience of consuming foods that contain and/or are prepared with animal protein but is not present in the consumption of plant-based products, in particular so called "meat analogues", and products prepared with industrial flavor solutions attempting to emulate the characteristics of animal protein.
• this may be due to a combination of (a) emotional experiences, (b) the fact that taste receptors exist throughout the human digestive system such that in essence, one continues to taste and experience the food that has been eaten long after finishing eating, and (c) the complex, mostly heretofore unexplored interactions between taste and aroma materials and the human microbiome.
• the herein disclosed flavor materials may have "masking” and/or “blocking” capability, i.e. the ability to mask bitter and/or "off” flavors such as “beany” flavors and/or to block taste receptors so that bitterness and/or beany off flavors are perceived at a lower level than would be expected otherwise, a quality which is advantageous as unpleasant bitter and/or "off", and or "beany” flavors are often found in plant-based meat replacement and/or other animal protein replacement products as well as many other foodstuffs.
• the herein-disclosed flavor materials can further advantageously mask, decrease or block various off-flavor sensation when used in a variety of food products; and/or has the ability to facilitate taste modulation.
• the flavor materials possess the ability to mask, decrease and/or block the off-flavor typical to alternative sweeteners such, as stevia, in variety of reduced sugar food products (applications).
• Stevia extracts for example, contain steviol glycosides, which are considered to be high-intensity sweeteners (with sweetness of about 250-300 times that of sucrose) and have been used as a sweetener for a range of food products. All steviol glycosides are bitter at different levels. Stevia delivers a sweet taste profile below 200-300 ppm and becomes bitter above 300 ppm and in beverage applications, it might have astringent and metallic properties, thus affecting mouthfeel.
• the disclosed flavor materials have the ability to facilitate sweetness modulation which improves a sugar-like mouthfeel in reduced sugar applications and also has the ability to mask the bitter and off-flavor of steviol glycosides.
• the herein-disclosed flavor materials may have "masking” and/or “blocking” capability of "astringency".
• Astringency is a sensation in the mouth that is usually described as a shrinking, drying-out, stretching, and wrinkling feeling. Astringency can be felt in tasting un-ripen, fruits as well as in soy-based yogurt, other plant-based applications, and other types of food products.
• the herein-disclosed flavor materials may have "masking" and/or "blocking" capability of sour taste. Sourness is an important taste in many foods and drinks. Acidulants are added in many food products (applications) not only as a flavor but also as an antiseptic agent, antibacterial, preservative, and/or buffering activity However, the addition of an acidulant may cause a decrease in pH and a change in the taste balance, in addition to unpleasant sourness. Without being bound to any theory or mechanism, the herein-disclosed flavor material contains a large number of acids and a complex collection of sugars, this complexity having the capability of masking sourness, as further exemplified hereinbelow.
• the herein-disclosed flavor materials have the ability to return the complexity and deeper taste in reduced salt food products.
• the herein disclosed flavor materials are rich in umami materials and other, amino acids, and in koku peptides and can consequently be useful when added to reduced salt food products, including without limitation condiments.
• the herein-disclosed flavor materials may in some embodiments provide a "mouthfeel" which is reminiscent of the mouthfeel provided by some foodstuffs containing animal protein.
• This mouthfeel may be achieved by delivering a perception of fat / fattiness / the presence of collagen resembling that of meat and/or fat and/or other animal protein, and/or— when claimed flavor materials are added to a foodstuff— by creating the impression that the said foodstuff contains a higher degree of fat and/or collagen than it actually does ("fat perception modulation"). All this while in fact the flavor material itself contains less than 5% and in some embodiments less than 2% or less than 1% fat.
• the herein-disclosed flavor materials can improve creaminess and milk fatty mouthfeel. Creaminess plays a unique role in the consumption experience due to its multidimensional textural attribute sensed during the consumption of some foods and beverages generated by mouthfeel. Thus, the herein-disclosed flavor materials can improve creaminess perception in plant-based milk and related food products, including nutritional drinks, without increasing fat percentages.
• the terms “substance” and “compound” may be used interchangeably and refer to a species of matter of definite chemical composition. According to some embodiment, the substance is an organic compound or molecule.
• non-volatile substance and “NVS” may be used interchangeably and refer to substances that do not evaporate or sublimate at temperatures below 40°C. Each possibility is a separate embodiment. Non-volatile substances exhibit a low vapor pressure and a high boiling point. Sugars and salts are examples of non-volatile substance.
• IRR internal standard respective ratio
• the term "internal standard respective ratio (IRR) refers to the respective ratio of the peak area of a given substance (when measured using mass spectrometry-based analytical chemistry technology) to the peak area of the internal standard 13-sorbitol (in non-volatiles analysis), or Isobutylbenzene (for volatile analysis) measured when the flavor material(s) is concentrated such that their Brix value is approximately 60 ⁇ Bx to 80 ⁇ Bx or a Brix value of 65 ⁇ Bx to 75 ⁇ and/or their water content (moisture level) is approximately 40% to 60% or between approximately 45% to 55%).
• volatile substance volatile organic substance
• VOC volatile organic substance
• odor activity value and “OAV” may be used interchangeably and refer to a measure of the importance of a specific substance to the odor of a sample (e.g. food). It is calculated by dividing the concentration of each individual substance by its odor detection threshold, i.e. the lowest concentration of odor that can be detected in the sample.
• the OAV refers to a measure obtained when the flavor material(s) is concentrated such that their Brix value is approximately 605 Bx to 805 Bx and/or their water content (moisture level) is approximately 40% to 60%.
• a substance having an OAV above 10 (when the flavor material(s) is/are concentrated such that their Brix value is approximately 605 Bx to 805 Bx or a Brix value of 65 ⁇ Bx to 755 and/or their water content (moisture level) is approximately 40% to 60% or between approximately 45% to 55%) is referred to herein as a "key odorant" i.e. a substance that significantly contributes to the aroma of a given product.
• Substances having an OAV when the flavor material(s) is concentrated such that their Brix value is approximately 605 Bx to 805 Bx or a Brix value of 655 Bx to 755 and/or their water content (moisture level) is approximately 40% to 60% or between approximately 45% to 55%) between 1-10 are referred to herein as "notes", i.e. substances that are not key odorants, but which can be sensed.
• all referral to values refer to values obtained when the flavor material(s) is concentrated such that their Brix value is approximately 605 Bx to 805 Bx or a Brix value of 655 Bx to 755 or a Brix value of about 055 Bx to 555, and/or the water content (moisture level) is between about 40% to 60% or between about 45% to 55%.
• the term "at least one" with regards to volatile and non-volatile substances may refer to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more substances. Each possibility is a separate embodiment.
• the flavor material may include both Hexadecanoic acid and 2,3-Octanedione.
• the consumable flavor material may include one or more of the volatile substances 2,3-Octanedione, 3-methyl-Butanal, 2-methyl-Butanal, 2,6-dimethyl- Pyrazine, a-ethylidene-Benzeneacetaldehyde, dimethyl Trisulfide, lH-indole, 2-Propenal, Hexanal and/or Hexadecanoic acid, ethyl ester at an odor activity value (OAV) of at least 10.
• the volatile substances 2,3-Octanedione 3-methyl-Butanal, 2-methyl-Butanal, 2,6-dimethyl- Pyrazine, a-ethylidene-Benzeneacetaldehyde, dimethyl Trisulfide, lH-indole, 2-Propenal, Hexanal and/or Hexadecanoic acid, ethyl ester at an odor activity value (OAV) of at least 10.
• OAV
• the consumable flavor material may include one or more of the volatile substances 2,3-Octanedione, lH-indole, 2-Propenal, Hexanal and/or Hexadecanoic acid, ethyl ester at an odor activity value (OAV) of at least 10 or at least 20.
• OAV odor activity value
• the consumable flavor material may include one or more of the volatile substances 2,3-Octanedione, 3-methyl-Butanal, 2-methyl-Butanal, 2,6-dimethyl- Pyrazine, a-ethylidene-Benzeneacetaldehyde, and/or dimethyl Trisulfide at an odor activity value (OAV) of at least 10, at least 20 or at least 40.
• OAV odor activity value
• the at least one volatile substance may be selected from 2,3-Octanedione, lH-indole, Ethyl linoleate, Acetaldehyde phenyl-dimethyl acetal, 2,3- Butanediol, Hexanal, (2E)-5-methyl-2-phenyl-2-hexenal, Benzaldehyde, Hexadecanoic acid, ethyl ester, Benzoic Acid, 2-Propenal, Hexanal, 2-phenylpropenal, Propanoic acid, methyl ester, 3-methyl-Butanal, 2-methyl-Butanal, 2,6-dimethyl-Pyrazine, a-ethylidene- Benzeneacetaldehyde, dimethyl Trisulfide, or any combination thereof.
• Each possibility is a separate embodiment.
• At least one -volatile substance may be selected from 2,3-Octanedione, lH-indole, 2-Propenal, Hexanal, Hexadecanoic acid, ethyl ester, Ethyl linoleate, Acetaldehyde phenyl-dimethyl acetal, 2,3-Butanediol, 3-methyl-Butanal, 2-methyl- Butanal, 2,6-dimethyl-Pyrazine, a-ethylidene-Benzeneacetaldehyde, dimethyl Trisulfide, or any combination thereof.
• 2,3-Octanedione lH-indole, 2-Propenal, Hexanal, Hexadecanoic acid, ethyl ester, Ethyl linoleate, Acetaldehyde phenyl-dimethyl acetal, 2,3-Butanediol, 3-methyl-Butan
• least one volatile substance having a OAV above 5 may be selected from 2,3-Octanedione, lH-indole, Hexanal, Benzaldehyde, (2E)-5-methyl-2- phenyl-2-hexenal, Hexadecanoic acid, ethyl ester, Benzoic Acid, 2-Propenal, Propanoic acid, methyl ester, 3-methyl-Butanal, 2-methyl-Butanal, 2,6-dimethyl-Pyrazine, a-ethylidene- Benzeneacetaldehyde and dimethyl Trisulfide.
• the at least one volatile substance is 2,3- Octanedione.
• the at least one volatile substance may have an odor activity value (OAV) of at least 5, at least 10, at least 20, at least 50, at least 100, at least 200, at least 500, or at least 1000.
• OAV odor activity value
• each substance may have a different OAV.
• the flavor material may include 2,3- Octanedione at an OAV of at least 10, at least 20, at least 40 or at least 50.
• OAV odor activity value
• the OAV of the 2,3-octanedione is at least 40.
• the flavor material comprises at least about 60%, at least about 65%, at least about 70% or at least about 80% non-volatile taste substances having a sweet taste out of the total amount of non-volatile taste substances (i.e., sweet, bitter, sour and umami).
• non-volatile taste substances i.e., sweet, bitter, sour and umami.
• the flavor material comprises at least about 5% w/w, at least about 10% w/w or at least about 15% w/w non-volatile taste substances having a sweet taste.
• the flavor material comprises at least about 5% w/w, at least about 10% w/w or at least about 15% w/w non-volatile taste substances having a sweet taste.
• the taste distribution of sweet substances is in a range of 150-450 IRR, preferably in a range of 200-350 IRR.
• the taste substances having a sweet taste may be selected from alanine, proline, fructose, rhamnose, asparagine, serine, mannitol, sucrose, glutamine, threonine, myo-inositol, glycine, glucose and raffinose or any combination thereof.
• the taste substances having a sweet taste are selected from isomaltose, xylulose, sorbitol, mannitol, myo Inositol, fructose and sorbose or any combination thereof. Each possibility is a separate embodiment.
• the taste substances having a sweet taste are selected from alanine, glycine, glucose, raffinose, rhamnose, myo-inositol and sucrose or any combination thereof. Each possibility is a separate embodiment.
• the taste substances having a sweet taste comprise glucose and raffinose. According to some embodiments, the taste substances having a sweet taste comprise glucose.
• the sweet taste substance comprises glucose with a concentration of at least about 2% w/w, at least about 3% w/w, at least about 4% w/w or at least about 4.5% w/w.
• concentration of at least about 2% w/w, at least about 3% w/w, at least about 4% w/w or at least about 4.5% w/w.
• the taste distribution of sour substances is in a range of 30-100 IRR, preferably in a range of 40-90 IRR.
• the flavor material comprises at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 10% or at least about 12% non-volatile taste substances having a sour taste out of the total amount of non-volatile taste substances (i.e., sweet, bitter, sour and umami).
• non-volatile taste substances i.e., sweet, bitter, sour and umami
• the flavor material comprises no more than about 30%, no more than about 25%, no more than about 20%, no more than about 10%, no more than about 7% or no more than about 5% non-volatile taste substances having a sour taste out of the total amount of non-volatile taste substances (i.e., sweet, bitter, sour and umami).
• non-volatile taste substances i.e., sweet, bitter, sour and umami.
• the flavor material comprises at least about 0.2% w/w, at least about 0.3% w/w, at least about 0.4% w/w, or at least about 0.45% w/w nonvolatile taste substances having a sour taste.
• the taste substances having a sour taste may comprise one or more of lactic acid 3-phenyl, Quinic acid-lactic acid, citric acid, malic acid, succinic acid and tartaric acid, or any combination thereof. Each possibility is a separate embodiment.
• the taste substances having a sour taste includes lactic acid.
• the flavor material includes lactic acid at an IRR of at least 25, at least 40 or at least 50. Each possibility is a separate embodiment.
• the taste substances having a sour taste comprises lactic acid, and/or lactic acid 3-phenyl.
• the sour taste substance comprises lactic acid at a concentration of at least about 0.2% w/w, at least about 0.3% w/w or at least about 0.4% w/w.
• concentration of at least about 0.2% w/w, at least about 0.3% w/w or at least about 0.4% w/w.
• the taste distribution of bitter substances is in a range of 30-100 IRR, in a range of 40-100 IRR, in a range of 40-60 IRR or in a range of 60-100 IRR.
• Each possibility is a separate embodiment.
• the flavor material comprises at least about 3%, at least about 5%, abt least about 7%, at least about 10%, at least about 12% , at least about 15%, non-volatile taste substances having a bitter taste out of the total amount of non-volatile taste substances (i.e., sweet, bitter, sour and umami).
• the flavor material comprises no more than about 30%, no more than about 25% no more than about 20%, no more than about 10%, no more than about 7%, no more than about 6% or no more than about 5% non-volatile taste substances having a bitter taste out of the total amount of non-volatile taste substances (i.e., sweet, bitter, sour and umami).
• sweet, bitter, sour and umami i.e., sweet, bitter, sour and umami
• the flavor material comprises at least about 0.2%, at least about 0.3% w/w, at least about 0.4% w/w or at least about 0.5% w/w non-volatile taste substances having a bitter taste.
• the bitter taste substance is an amino acid.
• the taste substances having a bitter taste may comprise one or more of: arginine, cystine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, tryptophan, tyrosine, valine, and any combination thereof. Each possibility is a separate embodiment.
• the taste substances having a bitter taste comprise arginine, cystine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, tryptophan, tyrosine and valine and any combination thereof.
• the taste substances having a bitter taste comprise arginine.
• the bitter taste substance comprises arginine with concentration of at least about 0.1% w/w, at least about 0.15% w/w or at least about 0.2% w/w.
• concentration of at least about 0.1% w/w, at least about 0.15% w/w or at least about 0.2% w/w.
• the taste distribution of umami substances is in a range of 7-50 IRR or in a range of 10-40 IRR.
• the flavor material comprises at least about 3%, at least about 5%, at least about 6%, at least about 8%, at least about 10% or at least about 12% non-volatile taste substances having an umami taste out of the total amount of non-volatile taste substances (i.e., sweet, bitter, sour and umami).
• the flavor material comprises no more than 20%, no more than 15% no more than 12%, no more than 10%, no more than 9%, no more than 8% or no more than 7% non-volatile taste substances having a bitter taste out of the total amount of non-volatile taste substances.
• Each possibility is a separate embodiment.
• the flavor material comprises at least about 0.6% w/w, at least about 0.7% w/w or at least about 0.8% w/w non-volatile taste substances having an umami taste.
• the umami taste substance is an amino acid.
• the umami taste substance comprises aspartic acid
• the umami taste substance comprises glutamate.
• the umami taste substance comprises glutamate at a concentration of at least about 0.4% w/w, at least 0.5% w/w or at least 0.6% w/w. Each possibility is a separate embodiment.
• the flavor material comprises aspartic acid at an IRR of at least 10, at least 15 or at least 20.
• IRR an IRR of at least 10, at least 15 or at least 20.
• the koku sensation substances may comprise one or more of Aspartate-Proline, Cysteine-Methionine-Threonine, y-Glutamyl-Aspartate, y-[Glu]3- Methionine, y-[Glu]2-Phenylalanine, y-Glutamyl-Glutamate, y-Glutamyl-Tyrosine and Glycine- Histidine-Glycine-Aspartate, or any combination thereof.
• Each possibility is a separate embodiment.
• a consumable composition that has a meaty flavor, and/or is able to impart such flavors to other food stuff, the composition comprising the flavor rich flavor material disclosed herein.
• the composition further comprises a flavor base in which the flavor material is blended with additional materials in order to create particular flavor profiles, which may be geared towards particular food applications.
• the flavor base is plant-based (vegetarian/vegan).
• flavor base refers to a combination of all the usual art- recognized ingredients required for the particular consumable composition including the composition detailed above, in additional to other consumable materials such as fruit and/or vegetable concentrates, wine concentrate, yeast products, yeast extracts, natural flavor materials, spices, extracts, botanicals, salt, and the like. According to some embodiments, there is provided a method for producing a meatless or animal-protein free food product, the method comprising the addition of the consumable flavor-rich flavor material or the composition disclosed herein, to the meatless food product.
• a meatless food product comprising the consumable flavor rich flavor material and/or the product bases (flavor bases) disclosed herein.
• the meatless/animal protein free food product may be a ready-to-cook food product.
• the ready-to-cook food product may be an analogue of a form of animal product, including without limitation meatless ground “meat” (also referred to as “mince”), meatless burgers, meatless “meat” balls, a meatless “chicken” product, such as nuggets, tenders, breasts, strips, cutlets, etc.
• a meatless "beef”, “pork”, “lamb” "fish” or other animal product a product which is a plant-based analogue of a product which traditionally is derived from animal protein, a traditional meat analogue such as tempeh, tofu, instantan, yuba, and so on, a plant-based product which may be traditionally prepared with some form of animal protein which imparts typical flavors and sensorial experiences to said product, a plant-based stock, broth, bouillon, or similar which aims to be a plant-based analogue of similar products derived from animal sources (e.g.
• prepared foods such as soups, sauces, stews, legume and/or grain based dishes, pastas, etc.
• prepared foods such as soups, sauces, stews, legume and/or grain based dishes, pastas, etc.
• dairy-free products which aim to impart the sensorial experience traditionally provided by dairy products, hot and cold sauces, table sauces, condiments, ready-to cook vegan or vegetarian dishes, chilled or frozen vegetarian/vegan dishes, or the like.
• a food product comprising the consumable flavor rich flavor material and/or the product bases (flavor bases) disclosed herein.
• Such food product may include any type of food product, in any form (for example, liquid, semi liquid, hard, soft, etc.).
• the food product is not a meatless food product.
• the flavor materials disclosed herein are produced by a process of fermentation.
• the flavor materials disclosed herein are produced by a process of fermentation of legumes and/or grains and/or agro-industrial waste streams.
• Fig. 1 is a schematic illustration of a process 100 for the preparation of a flavor material 170, according to some embodiments.
• a first substrate (“substrate A” 102, which can be, for example, a legume and/or grain)
• substrate A which can be, for example, a legume and/or grain
• stage 108 is incubated at stage 108, (optionally after a pre-treatment (106) stage), in the presence of a starter microorganism culture (“starter culture” 104) under specified conditions, to produce biocatalyst 110.
• starter culture starter microorganism culture
• a second substrate (“Substrate B” 122 (which may be similar or different than substrate A)) may undergo a process (which may be similar, identical or different with respect to any of the microorganisms used and/or other process conditions), including incubation with starter culture 124, after optional pretreatment stage 126, to produce an interim product 132.
• the interim product is mixed at stage 136 with fermentate 110 and/or salt 138 (such as, NaCI) and undergoes a period of holding at stage 140 under suitable conditions, to produce a flavor-rich mass 150.
• the resulting mass 150 may then be separated at stage 154 (by any suitable means such as decantation, press, centrifugation, etc.), to a solid residue fraction 156 and to a supernatant fraction 160.
• the supernatant 160 may then be concentrated at stage 162 (by any suitable means, such as freeze concentration, membrane-based concentration, dialysis, concentration columns, evaporation, distillation and the like) and optionally treated by heat or other means (e.g. pasteurized or sterilized) at stage 164, to result in flavor material 170.
• suitable means such as freeze concentration, membrane-based concentration, dialysis, concentration columns, evaporation, distillation and the like
• heat or other means e.g. pasteurized or sterilized
• the substrate may be, for example, legumes and/or grains and/or seeds, nuts, and/or plant, and/or vegetable, and/or fruit, and/or a product produced using one or more of the above, and/or agro-industrial waste produced during the processing of one or more of the above.
• the legumes may include, for example, but not limited to: chickpeas, lentils, peas, black beans, green beans, kidney beans, blacked-eyed, navy beans, lupin, green lentils, black lentils, red/ yellow lentils, green peas, yellow peas, bambara nut, or any combinations thereof. Each possibility is a separate embodiment.
• the grains may be selected from, but not limited to: wheat, einkorn, emmer, farro, Khorasan wheat ("Kamut”), rye, oat, rice, corn (maize), barley, buckwheat, quinoa, teff, amaranth, spelt, freekeh, Sorghum, millet, fonio, or any combinations thereof.
• Kamut Khorasan wheat
• rye oat
• rice corn (maize)
• barley buckwheat, quinoa, teff, amaranth, spelt, freekeh, Sorghum, millet, fonio, or any combinations thereof.
• buckwheat quinoa
• teff amaranth
• spelt freekeh
• Sorghum millet, fonio
• the seeds may be selected from, but not limited to: flaxseeds, chia seeds, hemp seeds, sesame seeds, pumpkin seeds, sunflower seeds, rapeseed, palm kernel, coconut, copra, cottonseed, rapeseed, safflower, olive, and the like, or any combinations thereof.
• the nuts may be selected from, but not limited to: macadamia nuts, brazil nuts, walnuts, hazelnuts, pecan nuts, chestnuts, peanuts, cashew nuts, almonds, and the like, or any combinations thereof. Each possibility is a separate embodiment.
• the plant or vegetable may be selected from, but not limited to: leeks, radish, celeriac, turnip, spinach, chard, kale, pepper, parsnip, onion, tomato, celery, grapes, potatoes, garlic, beet, carrot, and the like, or any combinations thereof. Each possibility is a separate embodiment.
• the fermentation process is performed in the presence of one or more types of microorganisms.
• the microorganism(s) may be selected from, but not limited to various species and strains of fungi and bacteria.
• the fungi may be selected from the fungal division of Ascomycota, fungal division of Basidiomycota, fungal class of Sacchromycetes, or combinations thereof.
• fungi selected from the fungal division of Ascomycota may include any strains of Aspergillus spp. (such as, but not limited to: A. oryzae, A. sojae, A. luchensis, and/or A. niger, any strain of Rhizopus spp, Trichoderma spp, Fusarium spp, Penicillium spp, and/or Neurospora spp. Each possibility is a separate embodiment.
• fungi selected from the fungal division of Basidiomycota may include any strains from the Agaricomycetes class, such as but not limited to Agaricus spp., Amanita spp. Armaillaria spp., Pleurotus spp., Pluteus spp., Grifola spp., Hydnum spp., Hygrophorus spp., Lenulates spp., Lepiota spp., Ramaria spp., Russula spp., Sparassis spp., Tricholomoa spp., Tuber spp., Volvariella spp., Each possibility is a separate embodiment.
• the bacteria may be selected from a bacteria of the Bacilli class (including any non-toxic strains of the Bacillales, Caryophanales, Desulfuribacillales and Lactobacillales orders, and/or any non-toxic strains of Actinomycetaceae, Brevibacteriaceae and Micrococcaceae). In some embodiments, the bacteria may be selected from a bacteria belonging to the Actinomycetia class, or any combinations thereof. Each possibility is a separate embodiment.
• the salt may be selected from sodium chloride, potassium chloride, calcium chloride, sodium bisulfate, copper sulfate, magnesium sulfate, and the like, or any combinations thereof. Each possibility is a separate embodiment.
• the incubation conditions may be selected from: incubation time (length), temperature, humidity, oxygen and/or CO2 concentration, light regime, air flow, presence or absence of agitation, agitation type and speed, characteristics of growth vessels (containers) and agitation devices, and the like.
• the incubation time may be in the range of about 8-126 hours, or any subranges thereof, such as, for example, 10-40 hours, 20-60 hours, 30-100 hours, and the like.
• the incubation temperature may be in the range of about 20- 55°C, or any subranges thereof, such as, for example, 25-35°C, 30-40 °C, 30-50°C and the like.
• the relative humidity may be in the range of about 40-99%, or any subranges thereof, such as, for example, 40-60%, 50-70%, 60-80%, 70-90%, 65-85% and the like.
• the oxygen concentration may be in the range of about 0-97%, or any subranges thereof, such as, for example, 0-15%, 10-20%, 25-35%, 50-60%, 70-82%, and the like.
• the holding stage may be in the length of about 1 -365 days, or any subranges thereof, such as, for example 1-100 days, 50-200 days, 100-300 days, and the like.
• the holding stage may be performed at a temperature in the range of about 4 - 70 ⁇ C, or any subranges thereof, such as, for example, 10-50 2 C, 15-45 2 C, 20-40 2 C, and the like.
• the concentration may include the use of concertation columns, centrifugation, dialysis, freeze concentration, membrane separation, filters, evaporation, distillation and the like.
• the sterilization may include pasteurization of the flavor material, for example, by ultra-heat treatment (UHT), UV radiation, high pressure treatments and the like.
• UHT ultra-heat treatment
• UV radiation UV radiation
• high pressure treatments high pressure treatments and the like.
• OAV was calculated by dividing the compound concentration by its detection threshold.
• the detection threshold is the lowest concentration of a certain odor compound that is perceivable by the human sense of smell.
• Compound concentration is presented in parts per billion (ppb), e.g. 1 pg/L.
• the concentration was calculated using the standard curve of the internal standard isobutyl benzene.
• Odor type and OAV were determined using the "Good Scents Company's" information system, which is currently the largest publicly-available body of relevant knowledge concerning the typical aromas of given substances for the flavor, food and fragrance industries. Information on odor was gathered by using the CAS (Chemical Abstracts Service) registry number of each compound. When absent from the Good Scents Company's data, OAV and odor type were determined by using previewed literature. Materials and Methods
• Helium was the carrier gas in a constant pressure mode rate of 1 mL-min-1, and the GC temperature was programmed for 40°C (1 min), and increased to 250°C at 6°C /min.
• Ionization energy was 70 eV with a mass acquisition range of 40-400 m/z, and a scanning rate of 6.34 spectra/s.
• Retention index (Rl) was calculated by running C8-C20 n-alkanes. Data analysis was conducted Wiley 10 with NIST 2014 mass spectral library data using the Mass Hunter software package (version B.08.00, Agilent, USA). Further identification of major compounds was based on a comparison of mass spectra and the retention index. Quantitative evaluation was performed using internal standards; peak areas were normalized to the Internal standard (Isobutylbenzene 0.8 pg per sample.
• Non-volatile analysis was done using Liquid Chromatography-Mass Spectrometry (LC- MS). Samples preparation was conducted as follows: 20 mg (+/- 1-5 mg) of each sample was weighted into a 2ml Eppendorf tube and diluted with 1 ml of extraction mixture (Methanol: Acetonitrile: Water at a ratio of 5:3:2 respectively). The samples were vortexted using Precellys 24 homogenizer (6500 RPM, 3 cycles of 30 seconds with 10 seconds break in between). The samples were centrifuged for 15 minutes at 18,000 RPM, supernatant was collected and diluted further on x20 taking 50 ul of sup and diluting with 950 ul of metabolites extraction mix (same solution).
• LC- MS Liquid Chromatography-Mass Spectrometry
• LC-MS metabolomic analysis was performed as described in Mackay GM, et al. (Analysis of Cell Metabolism Using LC-MS and Isotope Tracers. Methods Enzymol 2015, 561, 171-196, doi:10.1016/bs.mie.2015.05.016); Briefly, Thermo Vanquish Flex ultra-high- performance liquid chromatography (UPLC) system coupled to an Orbitrap Exploris 240 Mass Spectrometer (Thermo Fisher Scien-tific) was used. Resolution was set to 120,000 at 200 mass/charge ratio (m/z) with electrospray ionization and polarity switching mode to enable both positive and negative ions across a mass range of 67-1000 m/z.
• UPLC Thermo Vanquish Flex ultra-high- performance liquid chromatography
• Resolution was set to 120,000 at 200 mass/charge ratio (m/z) with electrospray ionization and polarity switching mode to enable both positive and negative
• UPLC setup consisted of a ZIC-pHILIC column (SeQuant; 150 mm x 2.1 mm, 5 pm; Merck). Five pL of extracts were injected and the compounds were separated using a mobile phase gradient of 15 min, starting at 20% aqueous (20 mmol/L ammonium carbonate adjusted to pH 9.2 with 0.1% of 25% ammonium hydroxide): 80% organic (acetonitrile) and terminated with 20% acetonitrile. The flow rate and column temperature were maintained at 0.2 mL/min and 45°C, respectively, for a total run time of 27 min. All metabolites were detected using a mass accuracy below 1 ppm. Thermo Xcalibur 4.4 was used for data acquisition.
• TraceFinderTM 5.0 was used for data analysis. Peak areas of metabolites were determined by using the exact mass of the singly charged ions. The peak areas of different metabolites were determined using Thermo TraceFinder software, where metabolites were identified by the exact mass of the singly charged ion and by the known retention time, using an i MS library built by running commercial standards of all detected metabolites.
• Sensory assessment is a scientific field covering all techniques for eliciting, measuring, analyzing, and interpreting human reactions to food characteristics perceived by the 5 senses. It's particularly important when assessing the attributes of complex foods such as meat substitutes.
• the herein disclosed flavor materials were sensory evaluated by two techniques:
• 2-AFC alternative forced choice
• 2-AFC alternative forced choice
• panelists were given two samples and directed to choose the one that has more of a given attribute, to assess which sample has the highest intensity on a particular characteristic.
• 10 panelists were given two samples: 1. a reference sample, and 2. the reference sample with the addition of the herein flavor material product(s).
• panellists were asked to choose which sample of the two is more (or less) bitter, sour, astringent, has beany off-flavor, stevia off-flavor, has fat perception or mouthfeel perception. Collected data was analysed using the binomial distribution. The test was performed for three different FM samples. The results presented in Figs. 10-14 show the average values of all three tested FMs.
• QDA Quality of Service
• a descriptive analysis technique in sensory evaluation for describing the intensity of product attributes.
• 5 expert panelists were given 4 samples: a reference sample (as is), and three reference samples, each containing different flavor material product FM1, FM 2 or FM3.
• the panelists quantified, per each sample, the following taste sensations: salty, sweet, bitter, sour, and astringent. Collected data was used to generate Spider Graphs, which presented the average values for the 3 FMs tested.
• FMs volatile fi of Flavor Materials
• a food matrix (substrate A) containing one or more plant materials e.g. grains and/or legumes and/or seeds, nuts, and/or plant, and/or vegetable, and/or fruit, and/or a product produced using one or more of the above, and/or agroindustrial waste produced during the processing of one or more of the above
• plant materials e.g. grains and/or legumes and/or seeds, nuts, and/or plant, and/or vegetable, and/or fruit, and/or a product produced using one or more of the above, and/or agroindustrial waste produced during the processing of one or more of the above
• microorganisms e.g. bacteria and/or fungi
• the microorganism(s) were grown on said substrate for between 12 and 126 hours under controlled humidity, temperature and oxygen availability.
• humidity was regulated at between 60 to 99% RH
• the temperature was regulated between 20 and 55 ⁇ C
• oxygen was regulated between 0 and 97%, with or without agitation, which in
• the substrates included legumes such as lentils, chickpeas, peas and beans, but similarly, other substrates may be used.
• the herein disclosed flavor materials 1-4 cluster with meat products such as concentrated beef stock and meat jus as opposed to known non-meat flavors (industrial meaty flavoring and imitation meat bullion).
• meat products such as concentrated beef stock and meat jus
• non-meat flavors industrial meaty flavoring and imitation meat bullion.
• the total amount of known volatile taste substances in the herein disclosed flavor materials resembles that of meat products (113 and 169, respectively) and is significantly higher than that of known non-meat products (here shown industrial meat flavoring (90)).
• 50 out of the 113 (44%) volatile substances found in the herein disclosed flavor materials are common to those of meat products while only 26 (29%) of found in the industrial meat flavoring are shared with that of meat products.
• Table 1 various volatile compounds were identified and their OAV was calculated.
• Table 1 OAV of volatile compounds identified in flavor materials FM-1, FM-2 and FM-4.
• OAV of above 3700 namely a-ethylidene-Benzeneacetaldehyde.
• Other substances were found at an average OAV of above 1200, namely dimethyl Trisulfide, and at an average OAV of above 480, namely 2,6- dimethyl-Pyrazine and at an average OAV of above 110, namely 3-methyl-Butanal.
• Other key odorant substances include 2-methyl-Butanal with an average OAV of above 90 and 2,3- Octanedione, with an average OAV of above 50.
• FM-3 had a distinct volatile profile, set forth in Table 2 below, which shows various volatile compounds identified in FM-3 and their calculated OAVs. Table 2: OAV of volatile compounds identified in flavor material FM3.
• FM1-4 are characterized by a high amount of, glutamate and aspartate.
• the herein disclosed flavor materials are characterized by having high levels of glutamate (about 80% out of the total umami substances), in resemblance to meat products (about 95% out of the total umami substances). Sour taste components distribution
• the "sour fingerprint" (FIG. 6) of the herein disclosed flavor materials shows that the distribution of different sour substances in the herein disclosed flavor materials resembles that of meat products (here shown meat jus).
• the herein disclosed flavor materials contain high levels of lactic acids (about 90% out of the total sour substances), in resemblance to meat products (about 80% out of the total sour substances), and lower levels of malic and succinic acids, as compared to other meat-imitation products.
• sweet molecules found in significant amounts in the herein disclosed flavor materials are set forth in Tables 5A-5B below.
• a distinct "sweetness fingerprint” showing the distribution of different sweet substances in the herein disclosed flavor materials is shown in FIG. 7.
• the high concentration (and their variety, 14-17 different sweet substances) of sweet substances out of the total amount of non-volatile taste substances i.e., sweet, bitter, sour and umami
• the 'bitter fingerprint' (FIG. 8) of the herein disclosed flavor materials shows that the distribution of different bitter substances in the herein disclosed flavor materials resembles that of meat products (here shown meat jus).
• the herein disclosed flavor materials contain high levels of arginine, about 40% out of the total bitter substances, in resemblance to meat products (about 35% out of the total bitter substances).
• the herein disclosed flavor materials contain distinct levels of lysine and methionine (18% and 3.5% out of the total bitter substances, respectively), as compared to other meatimitation products.
• the herein disclosed flavor materials have a koku profile which highly resembles the koku's substance distribution in meat product (meat jus).
• industrial meaty flavoring koku profile consists only one substance, Cysteine-Methionine-Threonine peptide out of the substances detected, and devoid other substances such as Aspartate-Proline which appear in meat jus.
• the distribution of koku substances in imitation meat bouillon different from that of meat product in particular consists high levels of y-Glutamyl-Glutamate (55-75%) which are much lower in meat jus (18-22%).
• sweet taste found in significant percentage (81.2% out of the total sweet, sour, bitter and umami) in the herein disclosed non-volatile taste distribution set forth in Table 8 below.
• the herein disclosed flavor materials have taste distribution consist of an average of 8.8% umami, 5.3% bitter, 4.7% sour and 81.2% sweet out of the total taste substances (i.e., sweet, sour, bitter and umami).
• lOOgr of the herein disclosed flavor materials consists about 8.2 gr (about 8.2% w/w) of the taste substances sweet, sour, bitter and umami, with distribution of: about 0.71% w/w of umami substances (from about 0.62gr to 0.84gr), about 0.42% w/w of bitter substances (from about 0.32gr to 0.53gr), about 0.37% w/w of sour substances (from 0.31gr to 0.45gr), and about 6.6% w/w of sweet substances (from about 5.0gr to 7.0gr).
• a plant-based burger was used as the reference sample.
• 90% of the panelists assessed "plant burger with FM” with higher fat perception than the reference sample.
• bitter and astringency are the dominant taste sensations of "plant burger", quantified at 4 and at 3 respectfully (black polygonal).
• the dominant taste sensations of the average results of the FMs (FM-1, FM-2 and FM-3 )with "plant burger” are salty and sweet, assessed and quantified both at above level 3 (grey polygonal).
• bitterness perception analysis a commercial tonic water was used as the reference sample.
• Fig. 11A 90% of the panelists assessed “Tonic” with higher bitter perception than “Tonic with FM”, indicating that the disclosed FMs have the ability to block and/or mask bitterness perception when added to food products.
• Fig. 11B bitter, astringency and sweet are the dominant taste sensations of "tonic” (black polygonal).
• Bitterness sensation of "Tonic” was quantified above 4.5, whereas in the case of the disclosed FMs "Tonic”, bitterness sensations were assessed in lower intensity levels, in the range of 2- 3 (grey polygonal).
• the disclosed FMs may have the ability to block and/or mask bitterness perception when added to food products.
• pea flour mixed into water at a dosage of 6% (w/w) was used as the reference sample.
• Fig. 13A 90% of the panelists assessed "6% pea flour in water” with higher beany off-flavor perception than "6% pea flour in water with FM", indicating that the disclosed FMs may have the ability to block and/or mask beany off- flavor perception when added to food products.
• Fig. 13B bitter and astringent are the dominant sensations of "6% pea flour in water” and were assessed and quantified at 6 and at 4, respectfully (black polygonal).
• cranberry juice For astringency perception analysis, an unsweetened, commercially available, cranberry juice was used as the reference sample. As shown in Fig. 14A, 80% of the panelists assessed "cranberry juice” with higher astringency perception than "cranberry juice with FM", indicating that the disclosed FMs may have the ability to reduce astringency perception when added to food products.
• the herein presented results demonstrate that the herein disclosed flavor materials have a unique volatile and non-volatile profile, which differentiate them from known flavor materials (such as imitation meat bouillon, and industrial meaty flavors) used to replace meat products and/or impart a meaty flavor to meat-substitutes or other foods, , in particular due to said unique volatile and non-volatile profile as well as their high amount of sweet and sour substances and their distribution.
• known flavor materials such as imitation meat bouillon, and industrial meaty flavors

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• 2022
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