Sweet taste enhancer containing ergothioneine as an effective ingredient and food and drink containing the same

Abstract

The technical problem to be solved by this invention is to provide a sweetness enhancer that can enhance the sweetness of food and beverages without increasing the content of sugary sweetness components (sucrose, glucose, or fructose, etc.) or non-sugar sweetness components. Furthermore, this invention provides a food or beverage, such as a food, drink, seasoning, or alcoholic beverage, containing this sweetness enhancer, thereby enhancing the sweetness through its sweetness-enhancing effect. The technical solution to this problem is that the sweetness enhancer of this invention uses ergothioneine as an active ingredient, and it is a sweetness enhancer for food and beverages containing sweetness components; and by containing ergothioneine as a sweetness enhancer, a food or beverage with enhanced sweetness can be obtained.

Description

Technical Field

[0001] This invention relates to sweeteners for food and beverages, and sweetened food and beverages containing the sweetener, such as food, beverages, or seasonings. Background Technology

[0002] Sweetness is a particularly beloved taste among the five primary tastes (bitter, salty, sour, sweet, and umami), but it's not as strongly perceived as bitterness and saltiness. Therefore, excessive intake of high-calorie sugars such as sucrose (refined sugar) and glucose in food and beverages has become a global health problem. Against this backdrop, coupled with increased health awareness in recent years, strategies and demands for limiting sugar intake have become increasingly urgent. However, simply reducing sugar in food and beverages can negatively impact their flavor, leading to a decrease in deliciousness.

[0003] Furthermore, low-calorie or calorie-free sweeteners are widely used as sugar substitutes. However, compared to foods and beverages using these high-sweetness sweeteners, the taste quality is often inferior, with unpleasant bitterness and astringency, or a poor aftertaste due to prolonged sweetness. Therefore, research and development are underway on sweetness-enhancing substances that can maintain the sweetness and original flavor of foods and beverages even with minimal use of sugar substitutes or reduced sugar content.

[0004] As a sweetener, methyl anthranilate is described in Patent Document 1, and it is reported that when added to a sucrose-containing food or beverage with a sucrose concentration of 2% by mass, it exerts a sweetening effect equivalent to increasing the sucrose concentration in the food or beverage by 0.4 to 0.6% by mass. In addition, specific quinoline compounds are described in Patent Document 2, 2-(3-benzyloxypropyl)pyridine in Patent Document 3, and 1-(2-hydroxyphenyl)-3-(pyridin-4-yl)propane-1-one in Patent Document 4. However, these sweeteners are all chemically synthesized substances, and their safety as food has not been confirmed.

[0005] Furthermore, sake, shochu, mirin, and sweet sake, which are alcoholic beverages obtained through saccharification and / or brewing using koji (rice malt), use grains, grain koji, and water as raw materials. Therefore, they contain only the sweet components resulting from the decomposition and saccharification of grain starch. Consequently, various methods have been employed to increase the sugar content and sweetness of these beverages. For example, there are reports of manufacturing methods that involve roasting the brewing grains in hot air at 200–400°C for a few seconds to 5 minutes (Patent Document 5); and methods that repeatedly perform the first and second steps more than twice to effectively increase the sugar content of sweet sake. In the first step, rice koji is added to steamed rice after mixing with water for saccharification and fermentation; in the second step, rice koji is added again to the steamed rice after saccharification and fermentation for further saccharification and fermentation (Patent Document 6). However, as the sugar content increases, a stickier texture also increases.

[0006] Existing technical documents

[0007] Patent documents

[0008] Patent Document 1: Japanese Patent No. 6931289

[0009] Patent Document 2: Japanese Patent Publication No. 2013-525278

[0010] Patent Document 3: Japanese Patent Application Publication Nos. 2016-202017

[0011] Patent Document 4: Japanese Patent Publication No. 2011-512790

[0012] Patent Document 5: Japanese Patent Publication No. 5-28591

[0013] Patent Document 6: Japanese Patent Application Publication No. 2011-55749

[0014] Patent Document 7: Japanese Patent No. 6263672

[0015] Patent Document 8: International Publication No. 2019 / 240243

[0016] Patent Document 9: International Publication No. 2023 / 234307

[0017] Non-patent literature

[0018] Non-patent literature 1: Bioscience, Biotechnology and Biochemistry (2019), Vol. 83, No. 1, pp. 181-184.

[0019] Non-patent literature 2: Journal of Bioscience and Bioengineering (2017), Vol. 124, No. 2, pp. 178-183. Summary of the Invention

[0020] The technical problem that the invention aims to solve

[0021] The technical problem to be solved by this invention is to provide a sweetness enhancer that can enhance the sweetness of food and beverages without increasing the content of sugary and non-sugar sweetness components. Furthermore, this invention provides food, beverages, or seasonings containing this sweetness enhancer, which enhances sweetness through its sweetness-enhancing effect; this invention also provides an alcoholic beverage with enhanced sweetness, obtained by saccharifying and / or brewing raw grains using koji (fermentation starter).

[0022] Technical solutions for solving technical problems

[0023] The inventors of this invention have conducted in-depth research on substances that enhance the sweetness of high-calorie sugars such as sugar. Their findings reveal that when food and beverages containing sugary sweeteners such as sucrose, glucose, or fructose contain trace amounts of ergothioneine, which can only be produced by certain microorganisms such as fungi (like mushrooms and Aspergillus), actinomycetes, and cyanobacteria, the sweetness of the food and beverages can be enhanced. Furthermore, it can enhance the sweetness of alcoholic beverages obtained through saccharification and / or brewing of raw grains using koji (fermentation starter). Further research has shown that ergothioneine can also enhance the sweetness of non-sugar sweeteners, thus completing this invention.

[0024] The present invention relates to the sweetness enhancer described in (1) to (3) below, the food and beverage described in (4) to (9), (11), (13) below, the sweet wine described in (10) below, or the mirin described in (12) below.

[0025] (1) A sweetness enhancer that uses ergothioneine as the active ingredient.

[0026] (2) A sweetener that uses ergothioneine as an active ingredient and is a sweetener for food and beverages containing sugar sweeteners.

[0027] (3) The sweetness enhancer as described in (2) above, wherein the sugar sweetness component is selected from one or more sugar alcohols such as sucrose (granulated sugar), glucose, fructose, fructose glucose solution, oligosaccharide, maltose, trehalose, maltose, sorbitol, mannitol, and xylitol.

[0028] (4) A sweetened food or beverage, characterized in that the food or beverage contains a sugar sweetening component and contains ergothioneine as a sweetening agent at a weight of 0.0001% or more relative to the sucrose content, at a weight of 0.0001% or more relative to the glucose content, or at a weight of 0.0003% or more relative to the fructose content.

[0029] (5) The food and beverage as described in (4) above, wherein the food and beverage is food, beverage or condiment.

[0030] (6) The food and beverage as described in (4) above, wherein the food and beverage is not soy sauce for steamed or boiled food, steamed or boiled food containing soy sauce, beverages with enhanced sourness, or food and beverage containing fermented biomass or supernatant of ergot-producing fungi that is colored with pigments.

[0031] (7) The food or beverage as described in any one of (4) to (6) above, wherein the food or beverage is liquid sugar, prepared soy milk, milk or alcohol.

[0032] (8) The food and beverage as described in (7) above, wherein the alcoholic beverage is sake, shochu, mirin or sweet wine obtained by saccharifying and / or brewing raw grains using koji.

[0033] (9) The food or beverage as described in any one of (4) to (8) above is manufactured using a high-yield Aspergillus strain of ergothionein.

[0034] (10) A sweet wine containing more than 30 ppm of ergothioneine.

[0035] (11) The food and beverage as described in (4) above, wherein the food and beverage is not a sweet wine.

[0036] (12) A mirin containing more than 2.5 ppm of ergothioneine.

[0037] (13) The food and beverage as described in (4) above, wherein the food and beverage is not mirin.

[0038] In addition, the present invention relates to the sweetness enhancer described in (14) and (15) below, the food and beverage described in (16) and (17) below, or the method for enhancing sweetness described in (18) below.

[0039] (14) A sweetener that uses ergothioneine as an active ingredient and is a sweetener for food and beverages containing non-sugar sweeteners.

[0040] (15) The sweetness enhancer as described in (14) above, wherein the non-sugar sweetness component is selected from one or more of the following as high-sweetness sweeteners: aspartame, acesulfame potassium, sucralose, stevia, monk fruit, stevia, licorice, and sweet amino acids.

[0041] (16) A sweetened food and beverage, characterized in that the food and beverage contains a non-sugar sweetening component and contains ergothioneine as a sweetening enhancer at a weight of 0.0005% or more relative to the content of a high-sweetness sweetener, or contains ergothioneine as a sweetening enhancer at a weight of 0.0004% or more relative to the content of sweet amino acids.

[0042] (17) The food and beverage as described in (16) above, wherein the food and beverage is not soy sauce for steamed or boiled food, steamed or boiled food containing soy sauce, beverages with enhanced sourness, or food and beverage containing fermented biomass or supernatant of ergot-producing fungi that is colored with pigments.

[0043] (18) A method for enhancing the sweetness of a food or beverage by making the food or beverage contain ergothioneine.

[0044] In addition, the present invention relates to the use of ergothioneine as described in (19) to (23) below.

[0045] (19) Use of ergothioneine for enhancing the sweetness of food and beverages.

[0046] (20) Use of ergothioneine for enhancing the sweetness of a food or beverage containing sugar sweeteners.

[0047] (21) The use of ergothioneine for enhancing sweetness as described in (20) above, wherein the sugar sweetness component is selected from one or more sugar alcohols such as sucrose (granulated sugar), glucose, fructose, fructose glucose solution, oligosaccharides, maltose, trehalose, maltose, sorbitol, mannitol, and xylitol.

[0048] (22) Use of ergothioneine for enhancing the sweetness of food and beverages containing non-sugar sweeteners.

[0049] (23) The use of ergothioneine for enhancing sweetness as described in (22) above, wherein the non-sugar sweetener is selected from one or more of aspartame, acesulfame potassium, sucralose, stevia, monk fruit, stevia, licorice, and sweet amino acids as high-sweetness sweeteners.

[0050] Invention Effects

[0051] By using the sweetener of the present invention, the content of sweet components in food and beverages can be reduced and sweetness can be enhanced by adding only a very small amount, thus naturally enhancing sweetness without compromising flavor. Furthermore, consuming seasonings, foods, beverages, and other foods containing this substance can reduce sugar intake and prevent excessive sugar consumption.

[0052] Furthermore, since it can bring out the original sweetness of the alcohol without increasing the sugar content, it not only naturally enhances the sweetness without compromising the flavor, but also, when the alcohol is mirin, the addition of ergothioneine enhances both sweetness and umami. Additionally, if the sweet wine is made using a high-yield Aspergillus strain of ergothioneine, it not only enhances the sweetness but also reduces koji (3-octanol) odor, resulting in a pleasant aroma and fewer large particles, thus creating a smoother, more palatable drink. Detailed Implementation

[0053] This invention relates to a sweetener that uses ergothioneine as an active ingredient, and is a sweetener for food and beverages containing sugary sweeteners and / or non-sugar sweeteners; the invention also relates to a sweetened food and beverage containing ergothioneine as a sweetener.

[0054] Additionally, in this invention, when the unit is not explicitly stated, "%" refers to "weight %", and 1 ppm is 0.0001 weight.

[0055] Ergothioneine (hereinafter also referred to as "ERG"), a sulfur-containing amino acid isolated from *Claviceps purpurea*, has been confirmed to exist in both plant and animal organisms. However, plants and animals cannot synthesize ergothioneine, suggesting that the ergothioneine in organisms originates from ergothioneine synthesized by microorganisms such as basidiomycetes. It is known to be present in some edible mushrooms, including oyster mushrooms, shiitake mushrooms, maitake mushrooms, and king oyster mushrooms, particularly abundant in *Pleurotus ostreatus*. Additionally, *Aspergillus* is known to produce ERG.

[0056] Regarding ERG, a histidine-based biosynthetic pathway is known in microorganisms, with sulfur atoms supplied by cysteine. ERG has been reported to possess high antioxidant properties and also exhibits elastase and tyrosinase inhibitory effects, attracting particular attention in the cosmetic and food industries, particularly in areas such as skin whitening and anti-wrinkle treatments. Furthermore, ERG's involvement in the body's oxidative defense system is well-established, and its applications in the medical field are being explored. ERG exhibits high thermal and pH stability, maintaining its antioxidant activity even at high temperatures. This suggests its potential for formulation into food for physiological effects and its use as an antioxidant in food products.

[0057] Current methods for manufacturing ERG involve extraction from basidiomycetes such as *Pleurotus ostreatus*, chemical synthesis, and fermentation using microorganisms. However, extraction from basidiomycetes is time-consuming and unsuitable for large-scale production. While chemical synthesis is suitable for large-scale production, it requires expensive synthetic reagents. Therefore, research is currently being conducted on manufacturing methods utilizing fermentation techniques such as: fermentation using C1 compound-assimilating bacteria or yeast; fermentation using microorganisms that overexpress the ERG biosynthetic gene (Patent Document 7); and solid-state culture using *Aspergillus* that has been introduced to overexpress the ERG biosynthetic gene (Non-Patent Document 1).

[0058] In the sweetness enhancer of the present invention, commercially available ERG synthesized chemically can be used, or extracts or refined products derived from ERG-containing mushrooms, ERG-producing Aspergillus and their cultures, or sake lees produced using Aspergillus can be used. Furthermore, for food and beverage products using Aspergillus as a raw material, instead of adding ERG, by using Aspergillus with a high ERG yield, the amount of ERG in the food and beverage can be increased, thereby enhancing the sweetness.

[0059] As for mushrooms used for ERG extraction, those with high ERG content are preferably listed as Pleurotus cornucopiae var. citrinopileatus, Pleurotus ostreatus, Lentinula edodes, and Grifola frondosa, which belong to the Pleurotaceae family and the Pleurotus genus. One of these can be used, or multiple can be used in combination.

[0060] Furthermore, any Aspergillus species belonging to the genus Aspergillus can be used as the Aspergillus for ERG production. Examples include: *Aspergillus oryzae*, *Aspergillus sojae*, *Aspergillus niger*, *Aspergillus luchuensis*, and *Aspergillus stamarii*. For the Aspergillus strain, strains obtainable from preservation institutions, such as *Aspergillus oryzae* strain RIB326, can be used; strains contained in commercially available koji starters; or strains isolated from food and beverage manufacturing environments such as sake and soy sauce breweries.

[0061] In addition to the wild-type strains mentioned above, Aspergillus used for ERG production can also be used to isolate mutant strains with high ERG yields using common mutation introduction methods. Examples of mutation introduction methods include: physical DNA damage through irradiation with ultraviolet (UV) or X-rays, and chemical DNA damage through treatment with alkylating agents such as N-methyl-N'-nitro-N-nitrosoguanidine (NTG) or ethyl methanesulfonate (EMS).

[0062] Furthermore, high-yield Aspergillus strains of ERG can be used by introducing genes related to ERG biosynthesis through gene recombination. Examples include: Aspergillus mutant strains with the egtA gene introduced (see Patent Document 8 and Non-Patent Document 1); Aspergillus mutant strains with increased ERG expression by introducing specific gene mutations such as the G428S mutation or C459F mutation, or the insertion of specific gene mutations such as the deletion of 1 to 112 amino acids in the W404 to Q515 regions (the encoded amino acid sequence: sequence number 1) and its orthologs, such as the AO090005000664 gene specific to the genus Aspergillus (see Patent Document 9), etc., but are not limited to these.

[0063] In this specification, a high-yield Aspergillus strain for ERG production refers to a strain that, compared to strains that have not undergone mutation or transformation, produces ERG that is 5 to 10 times higher, preferably 20 times higher, when cultured in liquid medium (5% Pafmin SM (Kikkoman Co., Ltd.) without pH adjustment) at 20–30°C for 3–14 days. Examples include Aspergillus oryzae strains that forcibly express the egtA gene, or Aspergillus oryzae mutant strains containing amino acid mutations or deletions within the W404–Q515 region of the mutated AO090005000664 gene and its orthologous genes.

[0064] Furthermore, in the case of food and beverage products made from Aspergillus fermentation products, such as soy sauce, sake, sweet sake, mirin, and miso, even without adding ERG during manufacturing, the ERG content in these products can be increased by using Aspergillus strains with high ERG yield for koji production. Especially when using Aspergillus strains with high ERG yield in the manufacture of fermented foods, fermented beverages, and fermented seasonings, they can have effects beyond just enhancing sweetness compared to adding ERG.

[0065] Alcoholic beverages made from Aspergillus fermentation products include sake, shochu, mirin, sweet wine, alcoholic sweet flavorings, fermented flavorings, or mirin-flavored flavorings. Additionally, the sweet wine of this invention also includes rice koji sweet wine without alcohol content.

[0066] Sake is made from rice. In addition to the existing raw material processing, which includes polishing brown rice, washing rice, soaking, and steaming, it is also allowed to use dried rice after roasting. However, only rice, rice koji, and water are raw materials.

[0067] The production of sake includes raw material processing (including koji making), feeding, fermentation, loading onto fermentation tanks, and refining. The production of mirin includes raw material processing, feeding, saccharification, aging, loading onto fermentation tanks, and refining. In addition, the production of sweet sake includes raw material processing, feeding, and saccharification.

[0068] Sweet sake is a sweet beverage made from rice that has existed in Japan since ancient times. Known varieties include rice koji sweet sake, obtained by saccharifying rice using rice koji; sweet sake made by dissolving sake lees and adjusting the flavor with sugar; and mixtures of both. Rice koji sweet sake is typically produced through saccharification and fermentation (adding rice koji to steamed rice and maintaining a temperature of around 50°C, during which saccharifying enzymes break down the starch in white rice to produce glucose), and does not contain alcohol. Sweet sake made from sake lees contains less than 1% alcohol. The sweet sake of this invention also includes alcohol-free sweet sake. It has been reported that commercially available sweet sake contains less than 10 ppm of ergothioneine, even at high levels (Non-Patent Literature 2).

[0069] In addition, mirin is a seasoning widely used in food cooking. It is made by mixing raw rice such as steamed glutinous rice with rice koji and alcohol such as shochu, using Aspergillus to saccharify the raw rice, causing the components in the raw rice to decompose and dissolve, and then processing the matured mash by pressing, removing residue, filtering, and heating.

[0070] The sweetening components of this invention include sucrose (granulated sugar), glucose, fructose, fructose-glucose liquid, oligosaccharides, maltose, trehalose, maltose, sorbitol, mannitol, xylitol and other sugar alcohols, maple syrup, or honey, etc.

[0071] In addition, the non-sugar sweeteners of the present invention include acesulfame potassium, aspartame, sucralose, stevia, monk fruit, licorice and other high-sweetness sweeteners, as well as sweet amino acids such as glycine, alanine, threonine, serine, lysine, and proline.

[0072] The food and beverage of the present invention contains one or more sugary sweeteners and / or one or more non-sugar sweeteners.

[0073] The term "sweetness enhancement" refers to the ability to perceive the sweetness more strongly compared to the absence of the sweetness enhancer of this invention. Specifically, it means that by adding the sweetness enhancer, even with a reduction of sugar (e.g., 1-50%), the same level of sweetness can be perceived. Furthermore, it also means that by adding the sweetness enhancer, the same level of sweetness can be perceived as if the amount of sugar (e.g., 1.1-2 times) were present. Alternatively, it means that in situations where a small amount of sugar (e.g.) was imperceptible, the addition of the sweetness enhancer allows the initial perception of sweetness.

[0074] The amount of ERG (electroretic rhein) that enhances sweetness relative to the sucrose content of food and beverages is as follows: 0.01% by weight or more in a sucrose solution of 0.5% or more but less than 1%; 0.0025% by weight or more in a sucrose solution of 1% or more but less than 10%; and 0.0001% by weight or more in a sucrose solution of 10% or more. Since the sweetness is almost imperceptible in food and beverages with a sucrose content of less than 1%, it is generally necessary to contain 0.0001% by weight or more, or 0.0025% by weight or more, of ERG relative to the sucrose content to achieve the desired sweetness enhancement effect.

[0075] As described above, the sweetness enhancer of the present invention contains 0.0001% by weight or more, preferably 0.0025% by weight or more of ERG relative to the sucrose content in the food and beverage, thereby exerting a sweetness-enhancing effect on the food and beverage.

[0076] Furthermore, regarding food and beverages containing glucose or fructose, the sweetness-enhancing effect is achieved by containing 0.0001% by weight or more, preferably 0.0005% by weight or more, of ERG relative to the glucose content, and by containing 0.0003% by weight or more of ERG relative to the fructose content. When the food and beverage contain any one or more of sucrose, glucose, or fructose, it is sufficient to contain 0.0001 to 0.0003% by weight or more of ERG relative to the content of any one of them.

[0077] The sweetener of the present invention contains 0.0005% by weight or more of ERG relative to the aspartame content in the food and beverage, thereby exerting a sweetness-enhancing effect on the food and beverage.

[0078] Furthermore, for food and beverage products containing stevia, the sweetness-enhancing effect is achieved by containing 0.05% by weight or more of ERG relative to the stevia content, and 0.0004% by weight or more of ERG relative to the sweet amino acid content. When the food and beverage contain non-sugar sweeteners, it is sufficient to contain 0.0004 to 0.0005% by weight or more of ERG relative to non-sugar sweeteners other than stevia.

[0079] Furthermore, even with a large amount of the sweetener of the present invention, the sweetness-enhancing effect cannot be improved. Therefore, considering economic efficiency, the content of ERG in food and beverages can be 0.15% by weight or less, for example, 0.12 or 0.09% by weight or less, or 0.06 or 0.03% by weight or less. Relative to the content of sweeteners in food and beverages, the content of ERG can be 15% by weight or less, for example, 12 or 9% by weight or less, or 6 or 3% by weight or less.

[0080] In alcoholic beverages obtained by saccharifying and / or brewing the raw grains of this invention using koji, the ERG content is typically 10 ppm or more, for example, 10 ppm or more, 15 ppm or more, 30 ppm or more, or 35 ppm or more, thereby enhancing sweetness. Sweet wines preferably contain 30 ppm. Furthermore, mirin is sweeter than other alcoholic beverages; therefore, an ERG content of 2.5 ppm or more is effective.

[0081] In addition, a high content of ERG does not necessarily enhance the sweetness. Therefore, considering economic factors, the ERG content in alcoholic beverages can be below 0.15% by weight, such as below 0.12% or 0.09% by weight, or below 0.06% or 0.03% by weight.

[0082] Compared to various sweetening components in food and beverages, the sweetness enhancer of the present invention can enhance the sweetness of food and beverages by using the content of all combinations of these lower and upper limits.

[0083] In addition, the concentration of ERG was determined using LCMS under the following conditions.

[0084] (HPLC conditions)

[0085] HPLC analysis was performed under the following conditions.

[0086] Apparatus: HPLC apparatus: one set of Nexera series (manufactured by Shimadzu Corporation);

[0087] Column: COSMOSIL 2.5HILIC column, 3.0 × 150 mm, 2.5 μm, 3.0 mm I.D. × 15.0 cm (manufactured by Nacalai).

[0088] Flow rate: 0.5 mL / min;

[0089] Temperature: 40℃;

[0090] Mobile phase: A) 0.1% (v / v) formic acid aqueous solution, B) 0.1% (v / v) formic acid acetonitrile;

[0091] Isocratic elution: 0-10 minutes (B: 80%);

[0092] Injection volume: 5μL.

[0093] (Mass spectrometry)

[0094] Device: LCMS-2020 (manufactured by Shimadzu Corporation);

[0095] (Mass spectrometry (LC-MS) analysis conditions);

[0096] Ionization conditions: ESI+;

[0097] MS conditions: SIM ERG: m / z230.1 ([M+H] + );

[0098] Hold for approximately 5 minutes.

[0099] The food and beverage containing sugary sweeteners and / or non-sugar sweeteners of the present invention are foods, beverages, or seasonings.

[0100] For food products, there are no restrictions as long as they can be used in all food products. Examples include: frozen desserts such as ice cream and sherbet; sweet desserts such as jelly, pudding, and yogurt; dairy products or dairy substitutes such as yogurt, cheese, and whipped cream; spreads such as nut butter; baked desserts such as cookies, biscuits, and cakes; desserts such as chocolate, chewing gum, and steamed buns; breads such as pastry bread and white bread; jams; marbled soft drinks; flake foods; compressed candies; bacterial protein; pet food; and medical foods.

[0101] There are no limitations on the definition of beverages. Examples include: soft drinks, milk drinks, canned beverage soups, soy milk drinks such as flavored soy milk, sports drinks, fruit juices, sake, shochu, cocktails, sweet wines made from rice saccharides, black tea, coffee, green tea, cocoa, and quasi-drug nutritional drinks. This includes not only liquid beverages but also gel-like and semi-solid beverages such as jelly drinks.

[0102] There are no restrictions on seasonings, and examples include soy sauce, mirin, miso, cooking wine, sauces, dipping sauces, dashi (broth) such as bonito broth, sushi vinegar, condiments, ketchup, liquid sugar, syrup, etc.

[0103] Example

[0104] The following examples illustrate the details of the invention, but the invention is not limited thereto.

[0105] In the embodiments, when referred to as "%", it means "weight %". Additionally, samples at room temperature are used in the sensory evaluation.

[0106] [Experiment 1: The sweetness-enhancing effect of sucrose solution produced by ERG]

[0107] Sucrose was added to pure water to prepare six sucrose samples with concentrations of 0%, 1%, 1.5%, 2%, 2.5%, and 3%. As a seventh sample, a sample was prepared by adding refined ergothioneine (purity greater than 99.5%, Tetrahedron) to 2% sucrose at a final concentration of 60 ppm.

[0108] The sweetness was sensorily evaluated by a panel of five trained professionals who act as subjects of sweetness perception. For the seventh 2% sucrose sample with 60 ppm ERG, the five panel members were asked to select samples of equivalent sweetness from the first through sixth sucrose samples (concentrations not disclosed to the panel members). After tasting each sample, the mouths were thoroughly rinsed with room temperature water before tasting the next sample.

[0109] As a result, all five panel members rated the seventh 2% sucrose sample with added ERG as having the same sweetness as the 2.5% sucrose solution. By adding 60 ppm of ERG, the sweetness of the 2% sucrose solution became comparable to that of the 2.5% sucrose solution, thus confirming the sweetness-enhancing effect of ERG.

[0110] [Experiment 2: Confirmation of the sweetness of ERG itself]

[0111] Similar to Experiment 1, six sucrose samples with concentrations of 0%, 1%, 1.5%, 2%, 2.5%, and 3% were prepared. As a seventh sample, ergothioneine was added to water to achieve a final concentration of 60 ppm. As in Experiment 1, a drinking test was conducted by a panel of five experts. After tasting each sample, the participants rinsed their mouths thoroughly with room temperature water before tasting the next sample, and then their sweetness was evaluated sensorily. For the sweetness of the seventh sample, the five experts were required to select a sample with equivalent sweetness from the first through sixth sucrose samples (concentrations not disclosed to the panel members).

[0112] As a result, all five members of the panel rated the seventh sample (prepared by adding ERG to water at a final concentration of 60 ppm) as having the same sweetness as a 0% sucrose solution, thus confirming that ERG itself has no sweetness.

[0113] [Experiment 3: ERG addition to enhance the sweetness of sucrose]

[0114] (1) Cases of 1-50% sucrose samples

[0115] Samples were prepared by adding ergothioneine to 1-50% sucrose solutions at final concentrations of 0 ppm, 0.05 ppm, 0.1 ppm, 0.25 ppm, and 0.5 ppm (ERG0-0.5 in Table 1 below, and "ERGX" in the following text refers to samples with added ERG of X ppm).

[0116] [Table 1]

[0117]

[0118] For sucrose samples with 1-50% added ERG at various concentrations (concentrations not disclosed to panel members), 1-50% sucrose samples without added ergothioneine served as a control group. A drinking test was conducted by four professional panel members, different from those in experiments 1 and 2, to evaluate the sweetness. After tasting each sample, the participants rinsed their mouths thoroughly with room temperature water before tasting the next sample. Panel members were provided with sample pairs consisting of the control group's 1-50% sucrose samples and 1-50% sucrose samples with added ergothioneine at various concentrations. Panel members evaluated which sample in the provided sample pair perceived sweetness through a two-point discrimination test.

[0119] The evaluation results are shown in Table 2.

[0120] <Evaluation Methods for Two-Point Discrimination Tests>

[0121] The evaluation results of the sweetness of the test sample compared with the control group.

[0122] 〇: The sweetness was enhanced compared to the control group.

[0123] ◇: Same sweetness as the control group.

[0124] The evaluation method and the representation of the evaluation results for the two-point discrimination test are the same as those for other tests.

[0125] [Table 2]

[0126]

[0127] Table 2 shows the evaluation results of the two-point discrimination test given by four professional panel members A to D, indicating the lower limit of ERG concentration for enhancing the sweetness of 1-50% sucrose solutions.

[0128] When ERG at concentrations of 0.5 ppm or higher was added to a sucrose solution of 0.5% or higher but less than 1%, all members reported an enhanced sweetness. When ERG at concentrations of 1% or higher but less than 10% was added, all members reported an enhanced sweetness. When ERG at concentrations of 10% or higher was added, all members reported an enhanced sweetness.

[0129] The amount of ERG that enhances sweetness relative to the sucrose content of food and beverages is equivalent to the following: in a sucrose solution of 0.5% or more but less than 1%, ERG is 0.01% by weight or more; in a sucrose solution of 1% or more but less than 10%, ERG is 0.0025% by weight or more; and in a sucrose solution of 10% or more, ERG is 0.0001% by weight or more.

[0130] (2) Case of 4% sucrose sample

[0131] Samples containing ergothioneine (ERG5–900) were prepared by adding ergothioneine to 4% sucrose solutions at final concentrations of 5 ppm, 20 ppm, 60 ppm, 100 ppm, 400 ppm, and 900 ppm. For the 4% sucrose samples with each concentration of ERG (the concentration was not disclosed to the panel members), drinking tests were conducted by four professional panel members different from those in tests 1 and 2, and the sweetness was evaluated by sensory evaluation.

[0132] The expert panel members were provided with sample pairs consisting of the control group's 4% sucrose sample from Table 3 below and the 4% sucrose sample with various concentrations of ergothioneine. The panel members evaluated which sample in the provided sample pair tasted sweet by using a two-point discrimination test.

[0133] [Table 3]

[0134]

[0135] Table 3 shows the ERG concentrations at which the sweetness-enhancing effect of 4% sucrose tends to be constant.

[0136] The 4% sucrose sample with 400 ppm ERG showed the same sweetness as the sample with 100 ppm ERG. Therefore, it was concluded that even with an addition of more than 100 ppm ERG, the degree of sweetness enhancement was no different from that with 100 ppm. This confirms that the sweetness enhancement effect tends to be constant at 100 ppm ERG.

[0137] Based on the above, the evaluation is as follows: when adding 5 ppm to 100 ppm of ERG to a 4% sucrose solution, the sweetness increases sequentially with increasing dosage. When the dosage exceeds 100 ppm, the sweetness enhancement effect is no different from that of adding 100 ppm.

[0138] [Experiment 4: Maximum Amount of ERG Addition]

[0139] The ergothioneine was dissolved in water at 40°C to a concentration of 10% (100,000 ppm), and then allowed to stand at 4°C. After one day, confirmation of the solution showed that ERG had precipitated. This indicates that the concentration of ERG in the water was less than 10%.

[0140] [Experiment 5: The sweetness-enhancing effect of ERG on glucose solution]

[0141] Glucose was added to pure water to prepare glucose samples with concentrations of 0%, 13%, 14%, 15%, 16%, and 17%. As a seventh sample, ergothioneine was added to 15% glucose to a final concentration of 60 ppm.

[0142] Similar to Experiment 1, the sweetness was sensorily evaluated by a panel of five trained professionals who were also subject to sweetness assessment. For the sweetness of the seventh 15% glucose sample with 60 ppm ERG, the five professionals were asked to select samples with equivalent sweetness from the first through sixth glucose samples (concentrations not disclosed to the panel members).

[0143] As a result, for the seventh 15% glucose sample with added ERG, three of the five panel members rated its sweetness as equivalent to that of a 16% glucose solution, and two rated it as equivalent to that of a 17% glucose solution. By adding 60 ppm of ERG, the sweetness of the 15% glucose solution became comparable to that of the 16% glucose solution, thus confirming the sweetness-enhancing effect of ERG on glucose.

[0144] [Experiment 6: ERG addition to enhance the sweetness of glucose]

[0145] (1) Case of 50% glucose sample

[0146] Samples ERG0–900 were prepared by adding ergothioneine to 50% glucose samples at final concentrations of 0.5 ppm, 1 ppm, 3 ppm, 5 ppm, 10 ppm, 20 ppm, 60 ppm, 100 ppm, 400 ppm, and 900 ppm (Table 4).

[0147] [Table 4]

[0148]

[0149] Four members of a professional panel were provided with sample pairs consisting of a 50% glucose sample from the control group (without ergothioneine) and 50% glucose samples with various concentrations of ergothioneine. The panel members evaluated which sample in the provided sample pair tasted sweet using a two-point discrimination test. The results are shown in Table 5.

[0150] [Table 5]

[0151]

[0152] Table 5 shows the lower limit of ERG concentration for enhancing the sweetness of 50% glucose solution.

[0153] When ERG at a concentration of 0.5 ppm or higher was added to a 50% glucose solution, all panel members rated it as sweetness enhancement. This is equivalent to the following: ERG with a sweetness-enhancing effect of 0.0001% by weight or higher relative to the glucose content of the beverage.

[0154] (2) Case of 10% glucose solution

[0155] The same tests as those for the 50% glucose solution in (1) above were performed on the 10% glucose solution using the samples (ERG0-20) shown in Table 6. The results of the sensory evaluation obtained by the two-point discrimination test are shown in Tables 7 and 8.

[0156] [Table 6]

[0157]

[0158] [Table 7]

[0159]

[0160] Table 7 shows the lower limit of ERG concentration for enhancing the sweetness of 10% glucose solution.

[0161] When ERG at a concentration of 0.5 ppm or higher was added to a 10% glucose solution, all panel members rated it as sweetness enhancement. This is equivalent to the following: ERG with a sweetness-enhancing effect of 0.0005% by weight or higher relative to the glucose content of the beverage.

[0162] [Table 8]

[0163]

[0164] Table 8 shows the ERG concentrations at which the sweetness-enhancing effect of 10% glucose solution tends to be constant.

[0165] The 10% glucose sample with 100 ppm ERG showed the same sweetness as the sample with 60 ppm ERG. Therefore, it was concluded that even with an addition of more than 60 ppm ERG, the degree of sweetness enhancement was no different from that with 60 ppm. This confirms that the sweetness enhancement effect tends to be constant at 60 ppm ERG.

[0166] Based on the above, the evaluation is as follows: when adding 0.5 ppm to 60 ppm of ERG to a 10% glucose solution, the sweetness increases sequentially with increasing dosage. When the dosage exceeds 60 ppm, the sweetness enhancement effect is no different from that when adding 60 ppm.

[0167] [Experiment 7: The sweetness-enhancing effect of the fructose solution produced by ERG]

[0168] Fructose was added to pure water to prepare six fructose samples with concentrations of 0%, 4.5%, 5%, 5.25%, 5.5%, and 6%. As a seventh sample, ergothioneine was added to 5% fructose to achieve a final concentration of 60 ppm.

[0169] Similar to Experiment 1, the sweetness was sensorily evaluated by a panel of five trained professionals who were also subject to sweetness assessment. For the sweetness of the seventh sample containing 5% fructose with 60 ppm ERG, the five professionals were asked to select samples with equivalent sweetness from the first through sixth fructose samples (concentrations not disclosed to the panel members).

[0170] As a result, for the seventh sample, three of the five panel members rated its sweetness as equivalent to that of a 5.5% fructose solution, and two rated it as sweet as that of a 5.25% fructose solution. By adding 60 ppm of ERG, the sweetness of the 5% fructose solution became equivalent to that of the 5.25–5.5% fructose solutions, thus confirming the sweetness-enhancing effect of ERG on fructose.

[0171] [Experiment 8: ERG Addition Amount to Enhance Fructose Sweetness]

[0172] Samples (ERG0-10) were prepared by adding ergothioneine to 20% fructose samples at final concentrations of 0.5 ppm, 1 ppm, 3 ppm, 5 ppm, and 10 ppm.

[0173] For the 20% fructose samples with various concentrations of ERG (concentrations not disclosed to the panel members), the 20% fructose samples without added ergothioneine served as a control group. A drinking test was conducted by four professional panel members, and the sweetness was evaluated sensorily. The evaluation results are shown in Table 9.

[0174] In addition, samples (ERG0.5–900) were prepared by adding ergothioneine to 20% fructose samples at final concentrations of 0.5 ppm, 5 ppm, 10 ppm, 20 ppm, 40 ppm, 60 ppm, 80 ppm, 100 ppm, 200 ppm, 400 ppm, and 900 ppm.

[0175] The expert panel members were provided with sample pairs consisting of 20% fructose samples with successively different concentrations of ERG (e.g., a sample pair with ERG at 20 ppm and 40 ppm). The panel members evaluated which sample in the provided sample pair tasted sweet using a two-point discrimination test. The evaluation results are shown in Table 10.

[0176] [Table 9]

[0177]

[0178] [Table 10]

[0179]

[0180] For the 20% fructose sample with added ERG of 0.5 ppm or more, all members of the research group evaluated it as having enhanced sweetness. This is equivalent to the following: relative to the fructose content of the beverage, the ERG with a sweetness-enhancing effect is 0.0003% by weight or more.

[0181] Furthermore, the 20% fructose sample with 100 ppm ERG was evaluated as having the same sweetness level as the sample with 80 ppm ERG. Therefore, even with an addition of more than 80 ppm ERG, the degree of sweetness enhancement was no different from the sample with 80 ppm ERG. This confirms that the sweetness enhancement effect tends to be constant at 80 ppm ERG.

[0182] Based on the above, the evaluation is as follows: When adding 0.5 ppm to 80 ppm of ERG to a 20% fructose solution, the sweetness increases sequentially with increasing dosage. When the dosage exceeds 80 ppm, the sweetness enhancement effect is no different from that when adding 80 ppm.

[0183] [Experiment 9: ERG addition to enhance the sweetness of various sweeteners]

[0184] In Experiment 9 below, the same two-point discrimination test was used as in Experiments 3, 5 and 7, and sensory evaluation was conducted by three members of the professional panel.

[0185] (1) Trehalose

[0186] Samples were prepared by adding ergothioneine to 50% trehalose samples at final concentrations of 0.5 ppm, 1 ppm, 3 ppm, 5 ppm, 10 ppm, and 20 ppm (Table 11, ERG0-20).

[0187] [Table 11]

[0188]

[0189] Regarding the 50% trehalose samples with various concentrations of ERG (concentrations not disclosed to the panel members), the panel members were provided with sample pairs consisting of the control group's 50% trehalose sample and the 50% trehalose samples with various concentrations of ergothioneine. The panel members evaluated which sample in the provided sample pair perceived sweetness through a two-point discrimination test. The test results are shown in Table 12.

[0190] [Table 12]

[0191]

[0192] For the 50% trehalose sample with added ERG of 0.5 ppm or more, all members of the panel rated it as having enhanced sweetness. This is equivalent to the following: relative to the trehalose content of the food, the ERG with a sweetness-enhancing effect is 0.0001% by weight or more.

[0193] (2) Fructose-glucose liquid sugar

[0194] Samples were prepared by adding ERG to 50% liquid sugar samples containing more than 55% fructose (New Fruct 55 (NF55), Showa Sangyo Co., Ltd.) at the concentrations shown in Table 13 (ERG0-20). Sample pairs were provided, consisting of a 50% liquid sugar sample from the control group and 50% liquid sugar samples with ERG added at each concentration. The panel members evaluated which sample in the provided sample pairs was perceived as sweet by a two-point discrimination test. The test results are shown in Table 13.

[0195] [Table 13]

[0196]

[0197] For the 50% liquid sugar sample with added ERG of 0.5 ppm or more, all members of the team commented that the sweetness was enhanced.

[0198] (3) Highly saccharified reducing syrup (sugar alcohol)

[0199] Samples were prepared by adding ERG to a 50% syrup sample containing a large amount of monosaccharide or disaccharide (ESWEE600, Busan Food Science Co., Ltd.) at the concentrations shown in Table 14 (ERG0–10). Sample pairs were provided, consisting of a 50% syrup sample from the control group and 50% syrup samples with ERG added at each concentration. Panel members evaluated which sample in the provided sample pairs perceived sweetness through a two-point discrimination test. The test results are shown in Table 14.

[0200] [Table 14]

[0201]

[0202] For the 50% syrup sample with added ERG of 0.5 ppm or more, all members of the panel rated it as having enhanced sweetness.

[0203] (4) Oligosaccharides

[0204] Samples were prepared by adding 50% oligosaccharide samples (Fujioligo #450, Nippon Food Chemical Co., Ltd.) to ERG concentrations (ERG0-10) as shown in Table 15. Sample pairs were provided, consisting of a 50% oligosaccharide sample from the control group and 50% oligosaccharide samples with each ERG concentration added. Panel members evaluated which sample in the provided sample pairs perceived sweetness through a two-point discrimination test. The test results are shown in Table 15.

[0205] [Table 15]

[0206]

[0207] For the 50% oligosaccharide sample with added ERG of more than 0.5 ppm, all members of the team commented that the sweetness was enhanced.

[0208] (5) Aspartame

[0209] Samples were prepared by adding 10% aspartame (Pal Sweet Slim Up Sugar, Ajinomoto Co., Ltd.) to a concentration of ERG (ERG0-20) as shown in Table 16. Sample pairs were provided, consisting of a control group of 10% aspartame samples and 10% aspartame samples with each concentration of ERG added. Panel members evaluated which sample in the provided sample pairs perceived sweetness through a two-point discrimination test. The test results are shown in Table 16.

[0210] [Table 16]

[0211]

[0212] For a 10% aspartame sample with added ERG of 0.5 ppm or more, the three panel members evaluated it as having enhanced sweetness. This is equivalent to the following: the amount of ERG with a sweetening effect is 0.0005% by weight or more relative to the aspartame content in the beverage.

[0213] (6) Stevia

[0214] Samples were prepared by adding 0.1% stevia to the ERG concentrations (ERG0–20) shown in Table 17. Sample pairs were provided, consisting of a control group of 0.1% stevia and 0.1% stevia samples with each ERG concentration added. Panel members evaluated which sample in the provided sample pairs perceived a sweet taste through a two-point discrimination test. The test results are shown in Table 17.

[0215] [Table 17]

[0216]

[0217] For the 0.1% stevia sample with added ERG of 0.5 ppm or more, all panel members rated it as having enhanced sweetness. This is equivalent to the following: relative to the stevia content in food and beverages, the ERG with a sweetness-enhancing effect is 0.05% by weight or more.

[0218] (7) Amino acid (glycine)

[0219] Samples were prepared by adding 12.5% ​​glycine to the ERG concentrations (ERG0–10) shown in Table 18. Sample pairs were provided, consisting of a control group of 12.5% ​​glycine samples and 12.5% ​​glycine samples with each ERG concentration added. Panel members evaluated which sample in the provided sample pairs perceived sweetness through a two-point discrimination test. The test results are shown in Table 18.

[0220] [Table 18]

[0221]

[0222] For the 12.5% ​​glycine sample with added ERG at a concentration of 0.5 ppm or higher, all members of the research group evaluated it as having enhanced sweetness. This is equivalent to the following: relative to the glycine content of the beverage, the ERG with a sweetness-enhancing effect was 0.0004% by weight or higher.

[0223] [Experiment 10: The sweetness-enhancing effect of sweet wine]

[0224] Sweet wine making

[0225] Soak 1250g of white rice in water overnight, then drain for 2 hours using a strainer. Afterward, wrap the rice in Tetoron cloth and steam at 100°C for 40 minutes. Cool the steamed rice to 45°C and inoculate with koji. Wrap the rice in Pylen cloth and begin koji making at 35°C and 95% humidity. 19 hours into koji making, break up the koji and wrap it in Pylen cloth again. Unwrap the Pylen cloth and spread the koji evenly when the temperature reaches 40°C. Then, after 40 hours, thoroughly break up the koji and let it stand again. After 42 hours, extract the koji. Add 2.5 times the amount of 60°C hot water to 800g of the obtained koji and initiate a saccharification reaction at 55°C and 200 rpm. After 16 hours, adjust the whiteness to 18% and transfer to a heat-resistant container for sterilization at 85°C for 30 minutes. When adjusting the Brix of sweet wine, water or ergothioneine is added to adjust the ergothioneine concentration in the sweet wine to 12ppm (made by adding water), 30ppm, 45ppm, 80ppm, 145ppm, or 345ppm.

[0226] The sweetness of the wine was evaluated by a panel of five trained professionals who are also subject to sweetness sensitivity.

[0227] Five members of the expert panel were provided with sample pairs of sweet wines containing different concentrations of ERG (ERG12-345) as shown in Table 19. The panel members evaluated which sample in the provided sample pair tasted sweet by using a two-point discrimination test.

[0228] The results are shown in Table 19. For sweet wines with ERG adjusted to 30 ppm or higher, all panel members rated the sweetness as enhanced. For sweet wines with ERG adjusted to 345 ppm, more than half of the panel members rated the sweetness as equal to that of sweet wines with ERG adjusted to 145 ppm, indicating no difference in sweetness enhancement compared to the 145 ppm setting.

[0229] The evaluation is as follows: When sweet wine contains 30ppm to 145ppm of ERG, the sweetness increases sequentially with increasing content. Even when the content exceeds 145ppm, the sweetness enhancement effect is no different from that of adding 145ppm.

[0230] In addition, a strain of *Aspergillus oryzae* with the G428S mutation was introduced into the ortholog of *Aspergillus* strain AO090005000664, a high-yield ergothionein-producing strain, to produce sweet wine under the same conditions as in Experiment 6. The resulting sweet wine had an ERG content of 115 ppm. It was not only highly sweet but also had less koji odor, resulting in a good aroma. Furthermore, it contained fewer large particles, leading to a smooth texture.

[0231] [Table 19]

[0232]

[0233] [Experiment 11: The sweetness-enhancing effect of mirin]

[0234] The alcohol in commercially available mirin (1.6 ppm ergothioneine content) was removed by heating, and ergothioneine was added to it in a manner that resulted in final concentrations of 1.6 ppm, 2.1 ppm, 4.6 ppm, 6.6 ppm, 11.6 ppm, and 21.6 ppm (ERG 1.6 to 21.6) to prepare mirin.

[0235] Regarding mirin with various concentrations of ERG (the concentrations were not disclosed to the panel members), mirin without added ergothioneine (ERG 1.6) served as a control group. Three professional panel members conducted a drinking experiment and performed a sensory evaluation of the sweetness.

[0236] Sample pairs were provided to the expert panel members, consisting of mirin from the control group and mirin with various concentrations of ergothioneine added. The panel members evaluated which sample in the provided sample pair perceived sweetness through a two-point discrimination test. The results are shown in Table 20.

[0237] [Table 20]

[0238]

[0239] Not only for mirin with 2.1 ppm of added ERG (0.5 ppm), but also for mirin containing 4.6 to 21.6 ppm of ERG, the team members all commented that the sweetness was enhanced.

[0240] [Experiment 12: The Production of Mirin]

[0241] After processing polished white rice using common methods, including washing, soaking, draining, steaming, and cooling, 0.1% of the rice weight of starter koji was inoculated and mixed. The mixture was then managed in a constant temperature and humidity chamber at a suitable temperature of 32–38°C to obtain koji. Aspergillus oryzae strains with the G428S mutation introduced into the orthologous gene of AO090005000664 (example) and its wild-type strain (comparative example) were used as starter koji. Following common methods for mirin production, 48g of the above-mentioned rice koji (approximately 40g of raw polished white japonica rice) was added to a container and processed using common methods, including washing, soaking, draining, steaming, and mixing with 290g of cooled glutinous rice. Next, 155ml of neutral alcohol (35% v / v) was added at 30°C for one month to obtain mature mirin mash. Next, the aged mirin mash is pressed using common methods to obtain a coarse mirin filtrate. This filtrate is then heated to 90°C for sterilization and clarified by filtration to obtain mirin.

[0242] The concentrations of ERG in mirin obtained by the above clarification and filtration are as follows: 11 ppm in the example and 2 ppm in the comparative example.

[0243] Compared to the mirin in the comparative example, the mirin obtained in the example not only had enhanced sweetness, but also enhanced umami flavor.

[0244] [Experiment 13: The sweetness-enhancing effect of soy sauce]

[0245] (1) Selected round soybean soy sauce

[0246] Ergothioneine was added to commercially available premium round soybean soy sauce (Kikkoman Co., Ltd.) at concentrations of 10 ppm, 20 ppm, and 50 ppm (EGR10, 20, 50) to prepare soy sauce.

[0247] Regarding soy sauces with added ERG at various concentrations (concentrations were not disclosed to the panel members), soy sauces without added ergothioneine (ERG0) served as a control group. Three members of the professional panel conducted a drinking experiment, and the sweetness was evaluated using a two-point discrimination test. The results are shown in Table 21.

[0248] [Table 21]

[0249]

[0250] For soy sauces containing more than 10 ppm of ERG, all team members commented that the sweetness was enhanced.

[0251] (2) Sweet soy sauce

[0252] ERG was added to commercially available sweet soy sauces (Golden Purple, Fundokin soy sauce), and the same experiment as described in (1) was conducted. The results are shown in Table 22.

[0253] [Table 22]

[0254]

[0255] For soy sauces containing more than 10 ppm of ERG, all team members commented that the sweetness was enhanced.

[0256] [Experiment 14: The effect of enhancing the sweetness of soy milk]

[0257] Ergothioneine was added to commercially available flavored soymilk (Kikkoman Co., Ltd.) at final concentrations of 0.5 ppm, 1 ppm, 3 ppm, 5 ppm, 10 ppm, and 20 ppm to prepare sample flavored soymilk (EGR 0.5–20).

[0258] Regarding the modified soy milk with added ERG at various concentrations (concentrations were not disclosed to the panel members), prepared milk without added ergothioneine (ERG0) served as a control group. Three members of the professional panel conducted a drinking experiment, and the sweetness was evaluated using a two-point discrimination test. The results are shown in Table 23.

[0259] [Table 23]

[0260]

[0261] For the soy milk containing 3 ppm of ERG, two group members commented that it was sweeter; for the soy milk containing 5 ppm of ERG, all group members commented that it was sweeter. Additionally, for the soy milk containing 40 ppm of ERG, the creaminess was increased.

[0262] [Experiment 15: The effect of enhancing the sweetness of milk]

[0263] Ergothioneine was added to commercially available milk at final concentrations of 0.5 ppm, 1 ppm, 3 ppm, 5 ppm, 10 ppm, and 20 ppm to prepare sample milk (EGR 0.5–20).

[0264] Regarding milk with added ERG at various concentrations (concentrations were not disclosed to the panel members), milk without added ergothioneine (ERG0) served as a control group. Three members of the professional panel conducted a drinking experiment, and the sweetness was evaluated using a two-point discrimination test. The results are shown in Table 24.

[0265] [Table 24]

[0266]

[0267] The group members unanimously rated the milk containing 5 ppm of ERG as having an enhanced sweetness.

Claims

1. A sweetness enhancer, characterized in that: Ergothioneine is used as the active ingredient.

2. A sweetness enhancer, characterized in that: Ergothioneine is used as the active ingredient, and this sweetener is a sweetener for food and beverage products containing sugar-based sweeteners.

3. The sweetness enhancer as described in claim 2, characterized in that: The sweetening components of the sugars are selected from one or more sugar alcohols, such as sucrose, glucose, fructose, fructose-glucose syrup, oligosaccharides, maltose, trehalose, maltose, sorbitol, mannitol, and xylitol.

4. A food product with enhanced sweetness, characterized in that: The food or beverage contains a sugary sweetener and contains ergothioneine at a rate of 0.0001% by weight or more relative to sucrose, ergothioneine at a rate of 0.0001% by weight or more relative to glucose, or ergothioneine at a rate of 0.0003% by weight or more relative to fructose.

5. The food and beverage as described in claim 4, characterized in that: Food and beverages refer to food, drinks, or seasonings.

6. The food and beverage as described in claim 4, characterized in that: The beverages include liquid sugar, flavored soy milk, milk, or alcoholic beverages.

7. The food and beverage as described in claim 6, characterized in that: The alcoholic beverages are sake, shochu, mirin, or sweet wine obtained by saccharifying and / or brewing raw grains using koji (a type of starter culture).

8. The food and beverage as described in claim 4, characterized in that: It is made using a high-yield Aspergillus strain of ergothionein.

9. A sweet wine, characterized in that: It contains more than 30 ppm of ergothioneine.

10. A type of mirin, characterized in that: It contains more than 2.5 ppm of ergothioneine.

11. A sweetness enhancer, characterized in that: Ergothioneine is used as the active ingredient, and this sweetener is a sweetener for food and beverage products containing non-sugar sweeteners.

12. The sweetness enhancer as described in claim 11, characterized in that: The non-sugar sweetener is selected from one or more of the following as high-sweetness sweeteners: aspartame, acesulfame potassium, sucralose, stevia, monk fruit, licorice, and sweet amino acids.

13. A food or beverage with enhanced sweetness, characterized in that: The food or beverage contains non-sugar sweeteners and contains ergothioneine at a rate of 0.0005% by weight or more relative to the content of high-sweetness sweeteners, or ergothioneine at a rate of 0.0004% by weight or more relative to the content of sweet amino acids.

14. A method for enhancing the sweetness of food and beverages, characterized in that: Enhancing the sweetness of food and beverages by including ergothioneine in them.

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