Umami and / or saltiness enhancers and food products containing the same

Abstract

Objective: To provide an umami and / or saltiness enhancer capable of enhancing the umami and / or saltiness of a food product. Additionally, to provide a food product, such as a food, beverage, or seasoning, which includes the said umami and / or saltiness enhancer and has enhanced umami and / or saltiness due to the umami enhancement effect. Solution: An umami and / or saltiness enhancer for food products containing ergothioneine as an active ingredient, and by including ergothioneine at a content of 0.005 weight % or more relative to the content of one umami component in the food product, a food product with enhanced umami and / or saltiness can be obtained.

Description

Technology Field

[0001] The present invention relates to an umami and / or saltiness enhancer for enhancing the umami and / or saltiness of a food product, and to a food product, such as a food, beverage, or seasoning, that has enhanced umami and / or saltiness containing said umami and / or saltiness enhancer. Background Technology

[0002] Recently, there has been a general demand throughout society to reduce the intake of table salt (sodium chloride) and prevent diseases such as hypertension, kidney disease, and heart disease, leading to a trend toward low-salt consumption in food products in general. However, simply reducing the salt content from conventional products results in a bland taste, and using substitute salts such as potassium chloride degrades the taste due to savory flavors like bitterness. Although a low-salt method is known to satisfy taste by compensating for the reduction in saltiness by increasing the umami flavor of broths, this method is limited to food products where umami is an important flavor, and in such cases, it is necessary to significantly increase the umami components.

[0003] For this reason, research and development is underway on saltiness-enhancing substances that can enhance the saltiness of food products by increasing the saltiness even when the amount of salt is reduced, and umami-enhancing substances that can enhance the umami of food products even with low salt content.

[0004] As such salty taste enhancing substances, Patent Document 1 describes a salty taste enhancing agent having methional, 4-hydroxy-3(2H)-planones, or 3-hydroxy-2(5H)-planones as active ingredients, and Patent Document 2 describes a salty taste enhancing agent containing niacin, salicin, or helicin, but most of these salty taste enhancing substances are chemically synthesized substances and include those whose safety as food has not been confirmed.

[0005] In addition, regarding umami-enhancing substances, Patent Document 3 describes an umami-enhancing agent having N-(1-deoxy-D-fructose-1-yl)-pyroglutamic acid, N-(1-deoxy-D-fructose-1-yl)-valine, or N-(1-deoxy-D-fructose-1-yl)-methionine as active ingredients, and Patent Document 4 describes an umami-enhancing agent having pyroglutamyl dipeptide as an active ingredient. The components that contribute to umami are mainly amino acids and nucleic acids, and it is known that some nucleic acids have an umami-enhancing effect when combined with glutamic acid.

[0006] In addition, Patent Document 5 describes a salty and / or umami flavor enhancer having trehalose as an active ingredient.

[0007] Sodium reduction is no exception even for soy sauce, which is indispensable in Japanese cuisine, and technology development is underway to produce soy sauce that has a good salty taste and flavor even with a low salt content.

[0008] For example, Patent Document 6 describes a soy sauce in which the umami flavor of glutamic acid is enhanced by containing methional, and Patent Document 7 reports a soy sauce with excellent saltiness and soy sauce characteristics containing 0.5 to 5% (w / v) salt and 0.2 to 4% (w / v) ammonium ions and a pH of 3.2 to 5.0.

[0009] In addition, Patent Document 8 describes that by using a salty taste and / or umami taste enhancer having trehalose as an active ingredient, the salty taste and umami taste of sodium chloride can be enhanced, and as a result, a food product with a rich and excellent taste can be produced. Prior art literature

[0010] Patent Document 1: Japanese Published Patent No. 2012-223147 Patent Document 2: Japanese Published Patent No. 2023-135861 Patent Document 3: Japanese Published Patent No. 2012-29615 Patent Document 4: Japanese Published Patent No. 2012-29616 Patent Document 5: Japanese Published Patent / Hei 10-66540 Patent Document 6: Japanese Published Patent No. 2014-18146 Patent Document 7: Japanese Patent No. 5812864 Patent Document 8: Japanese Published Patent / Hei 10-66540 Patent Document 9: International Publication No. 2019 / 240243 Patent Document 10: International Publication No. 2023 / 234307

[0011] Non-patent literature 1: BIOSCIENCE BIOTHECHNOLOGY AND BIOSCHEMISTRY(2019) Vol. 83, No. 1, p. 181-184 The problem to be solved

[0012] The problem to be solved by the present invention is to provide an umami and / or saltiness enhancer capable of enhancing the umami and / or saltiness of a food product. In addition, the invention provides a food product, such as a food, beverage, or seasoning, which includes the said umami and / or saltiness enhancer and has enhanced umami and / or saltiness due to the umami and / or saltiness enhancing effect. means of solving the problem

[0013] The inventors of the present invention conducted keen research on substances capable of enhancing umami and / or saltiness, and discovered that ergothioneine, which can be produced only by certain microorganisms such as mushrooms, fungi such as koji mold, actinomycetes, and cyanobacteria, can enhance umami and / or saltiness when included in food products containing umami and / or saltiness components, thereby completing the present invention.

[0014] The present invention relates to an umami and / or saltiness enhancer described in [1] below, or to food products described in [2] to [7].

[0015] [1] An umami and / or salty flavor enhancer containing ergothioneine as an active ingredient.

[0016] [2] An umami-enhanced food product containing 0.005% by weight of ergothioneine, an umami enhancer, relative to the content of one umami component in the food.

[0017] [3] A food product with an enhanced salty taste, containing ergothioneine, a salty taste enhancer, in any one of the following concentrations in a food product containing salt.

[0018] (1) Among food products with a salt content of 0.1% by weight or more and less than 5% by weight, 0.5 ppm or more of ergothioneine

[0019] (2) 0.1 ppm or more of ergothioneine in food products with a salt content of 5 weight % or more

[0020] [4] Food products described in [2] or [3] above that are soy sauce, miso, fish sauce, mirin, bonito broth, liquid extract, umami seasoning, fermented umami seasoning, ketchup, sauce, or mycoprotein, which are foods with enhanced umami and / or salty flavors.

[0021] [5] A food product described in any one of [2] to [4] that is manufactured using a high-yielding ergothioneine yeast strain.

[0022] [6] Food products with enhanced umami and / or saltiness containing 10 ppm or more of ergothioneine.

[0023] [7] Soy sauce for retort food containing 10 ppm or more of ergothioneine, retort food containing soy sauce, and food products that are not food products containing ergothioneine-producing fungal fermentation biomass or the supernatant thereof colored with a food coloring, umami and / or saltiness enhanced food products.

[0024] In addition, the present invention relates to soy sauce described in [8] and [9] below, food products described in

[10] and

[11] , ergothioneine-rich koji fermented product described in

[12] , a method for enhancing umami and / or saltiness described in

[13] , or the use of ergothioneine described in

[14] .

[0025] [8] Soy sauce or soy sauce-containing seasoning that is not retort food soy sauce containing 10 ppm or more of ergothioneine.

[0026] [9] Soy sauce as described in [8] above, which is low-sodium soy sauce.

[0027]

[10] The food product described in [5] above is a rice koji that is a koji product containing 0.3g of ergothioneine and 1kg or more of dried koji product, obtained by culturing a high-ergothioneine koji strain in a medium composed of raw materials with a protein content of less than 5~10%.

[0028]

[11] The food product described in [5] above is a food product that is a mycoprotein or soy sauce koji containing 0.4g of ergothioneine / 1kg of dry koji fermented product, which is a koji fermented product containing ergothioneine high content, obtained by culturing a high-ergothioneine koji strain in a medium composed of raw materials with a protein content of 10% or more.

[0029]

[12] A high-ergothioneine fermented product containing 50 to 50,000 ppm of ergothioneine.

[0030]

[13] A method to enhance the umami and / or saltiness of food by incorporating ergothioneine into the food.

[0031]

[14] Use of ergothioneine to enhance the flavor and / or saltiness of food and beverages. Effects of the invention

[0032] According to the umami and / or saltiness enhancer of the present invention, umami and / or saltiness can be produced by adding a minute amount of ergothioneine, so umami and / or saltiness can be naturally enhanced without impairing the flavor. When added to food products with umami to enhance the flavor, the saltiness can be felt more even if the salt content in the food product is reduced; therefore, when consuming food products such as seasonings, foods, and beverages with enhanced flavor by adding this, the taste of the food is maintained, and the amount of salt intake can be reduced without difficulty.

[0033] Of course, by adding a tiny amount of ergothioneine to food products containing umami ingredients such as L-glutamate monosodium, sodium inosinate, or sodium guanylate, the flavor can be enhanced without including salt.

[0034] Soy sauce containing a very small amount of ergothioneine has enhanced saltiness and umami, so even if the salt content of the soy sauce is reduced, it can bring out the natural soy sauce flavor without compromising the flavor, making it particularly effective for low-salt soy sauce, and the sweetness of the soy sauce is also enhanced by ergothioneine. In addition, soy sauce produced using a high-yielding ergothioneine koji strain has the advantage of not only improving the polishing of the taste but also improving the compressibility and workability during the filtration process of soy sauce production.

[0035] Furthermore, the koji fermented product produced using a high-ergothioneine koji strain has a high ergothioneine content and can be used as a food, food material, or food ingredient that allows for sufficient ergothioneine intake when consumed as is. Due to the high ergothioneine content, it becomes a more delicious food with enhanced flavors such as umami, sweetness, and saltiness. Additionally, the koji fermented product of the present invention has a good aroma because it has less of the characteristic odors of koji fermented products, such as koji smell, rice bran smell, odorlessness, and oxidation smell. Specific details for implementing the invention

[0036] The present invention relates to an umami and / or saltiness enhancer for food products having ergothioneine as an active ingredient, and to a food product with enhanced umami and / or saltiness containing ergothioneine, which is an umami and / or saltiness enhancer.

[0037] Also, in the present invention, "%" means "weight %" even if the unit is not specified, and 1 ppm is 0.0001 weight %.

[0038] Ergothioneine (hereinafter also referred to as "ERG") was discovered as a sulfur-containing amino acid isolated from the rye ergot fungus (Claviceps purpurea) and was confirmed to exist in the bodies of plants and animals. However, plants and animals cannot synthesize ergothioneine, and it is believed that ergothioneine in the body originates from ergothioneine synthesized by microorganisms such as basidiomycetes. It is found in some edible mushrooms of basidiomycetes, such as oyster mushrooms, shiitake mushrooms, maitake mushrooms, and king oyster mushrooms, and is known to be particularly abundant in yellow oyster mushrooms. It is also known that Aspergillus niger produces ERG.

[0039] The biosynthetic pathway of ERG from histidine in microorganisms is known, and sulfur atoms are supplied from cysteine. ERG possesses high antioxidant activity, and since elastase and tyrosinase inhibitory effects have also been reported, it is receiving particular attention in the beauty and food industries, such as for skin whitening and wrinkle prevention. Furthermore, it has been established that ERG is involved in the body's oxidative defense system, and applications in the medical field are also being explored. ERG exhibits high thermal and pH stability and can maintain its antioxidant activity even at high temperatures; therefore, in addition to being incorporated into food formulations for its physiological effects, its use as a food antioxidant is also anticipated.

[0040] As methods for producing ERG, extraction from basidiomycetes such as yellow oyster mushrooms, chemical synthesis, and fermentation using microorganisms are being carried out. Extraction from basidiomycetes such as yellow oyster mushrooms takes time to obtain raw materials and is not suitable for mass production. Chemical synthesis is suitable for mass production, but it requires the use of expensive synthetic reagents. For this reason, research is being conducted on production methods by fermentation, such as fermentation using bacteria or yeast capable of assimilating C1 compounds, fermentation using microorganisms that overexpress ERG biosynthetic genes, and solid culture using koji mold that overexpresses ERG by introducing ERG biosynthetic genes (Non-patent Literature 1).

[0041] As an umami and / or saltiness enhancer of the present invention, commercially available ERG produced by chemical synthesis may be used, or extracts or purified products thereof from mushrooms containing ERG, koji molds that produce ERG and their cultures, or brewing products made using koji molds may be used. In addition, for food products using koji molds as raw materials, the amount of ERG in the food product can be increased by using koji molds that produce a high amount of ERG without adding ERG.

[0042] Mushrooms used for extracting ERG include, preferably, mushrooms of the Pleurotus family such as Pleurotus cornucopiae var. citrinopileatus, Pleurotus ostreatus, Lentinula edodes, and Grifolia Frondosa that have a high ERG content, and one or a combination of these types can be used.

[0043] In addition, any strain belonging to the genus Aspergillus may be used as the koji mold that produces ERG. Examples include Aspergillus oryzae, Aspergillus sojae, Aspergillus niger, Aspergillus luchuensis, and Aspergillus tamarii. As for the strain of the koji mold, strains available from deposit institutions, such as Aspergillus oryzae RIB326 strain, strains included in commercially available starter cultures, or strains isolated from food production environments such as sake or soy sauce breweries may be used.

[0044] In addition, as a koji bacterium that produces ERG, a mutant strain with high ERG production can be isolated and used by using a general mutation introduction method in addition to the above wild strain. Mutation introduction methods include, for example, ultraviolet (UV) or X-ray irradiation, which physically damage DNA to introduce mutations, and treatment with alkylating agents such as N-methyl-N'-nitro-N-nitrosoguanidine (NTG) or ethylmethanesulfonic acid (EMS), which chemically damage DNA to introduce mutations.

[0045] In addition, ERG-high-production koji strains can be used by introducing genes related to ERG biosynthesis through genetic recombination. Examples include, but are not limited to, koji strains with introduced egtA genes (see Patent Document 9 and Non-Patent Document 1), or specific gene mutations such as introducing G428S mutations or C459F mutations, or deleting 1 to 112 amino acids in the W404 to Q515 region, or the AO090005000664 gene (coding amino acid sequence: sequence number 1) characteristic of Aspergillus traits, and Aspergillus trait strains with increased ERG expression by introducing the ortholog gene thereof (see Patent Document 10).

[0046] In this specification, a high-ERG-producing koji strain refers to a strain in which the produced ERG is 5 to 10 times, preferably 20 times or more, in liquid culture (a solution in which 5 to 10% of Papmin SM (manufactured by Kikkoman Co., Ltd.) is added to water and suspended) at 20 to 30°C for 3 to 14 days compared to a strain that is not mutated or transformed. Examples include Aspergillus soya strains in which the above egtA gene is forcibly expressed, or Aspergillus or oryzae mutant strains containing amino acid mutations or amino acid deletions in the regions of the above mutant AO090005000664 gene and its ortholog genes W404 to Q515.

[0047] The AO090005000664 gene is cloned from the Aspergillus oryzae RIB40 strain, and there are multiple conserved regions that are highly conserved compared to other strains. In particular, the region corresponding to amino acid positions 411–470 of sequence number 1 can be cited as a conserved region, and by introducing a substitution mutation at the position corresponding to position 428 and / or position 459 within this conserved region, the expression of ERG in the mutant strain increases by more than 5–10 times. In addition, even if a deletion mutation is introduced in which 1 to 112 amino acids are deleted in the region corresponding to positions 404–515, the expression of ERG increases by more than 5–10 times in the same way.

[0048] In addition, in the case of food products made from koji mold fermentation products, such as soy sauce, sake, sweet rice wine, mirin, and soybean paste, the ERG content in the food product can be increased by using a high-ERG-producing koji strain to make the koji, even without adding ERG during production. In particular, when a high-ERG-producing koji strain is used in the production of fermented foods, fermented beverages, and fermented seasonings, effects other than the enhancement of umami and / or saltiness are also exhibited compared to when ERG is added.

[0049] The enhancement of umami and / or saltiness means that the umami and / or saltiness resulting from these ingredients is perceived more strongly compared to the case where the umami and / or saltiness enhancer of the present invention is not present. Specifically, it means that by adding the umami and / or saltiness enhancer, an equivalent umami and / or saltiness can be perceived even if the umami and / or saltiness components are reduced by, for example, 1 to 50%. Furthermore, it means that by adding the umami and / or saltiness enhancer, an equivalent umami and / or saltiness can be perceived as if, for example, 1.1 to 2 times the amount of umami and / or saltiness components are contained. Alternatively, it means that in cases where a small amount of umami components and / or saltiness is contained but cannot be perceived, the umami and / or saltiness can be perceived for the first time by adding the umami and / or saltiness enhancer.

[0050] The umami component in the present invention is L-glutamate monosodium, inosinate sodium, or guanylate sodium, and the content of one umami component in the present invention refers to the content of any one of these three components included in the food product.

[0051] In a food product containing an umami component, the umami flavor of the food product is enhanced by including ergothioneine, an umami flavor enhancer, in an amount of 0.005 weight % or more relative to the content of one umami component. Specifically, the umami flavor is enhanced by including 1 ppm or more of L-glutamate monosodium, an umami component, in a 2% solution, or 1 ppm or more in a 5% solution of sodium inosinate or sodium guanylate. In a food product containing two or more umami components, it is sufficient to include 0.005 weight % or more of ERG relative to the content of the minimum umami component.

[0052] In addition, the umami flavor enhancer of the present invention exhibits an umami flavor enhancing effect in a food product by containing 10 ppm or more, for example, 10 ppm or more, 15 ppm or more, 20 ppm or more, or 30 ppm or more, or 40 ppm or more, or 50 ppm or more with respect to a general food product.

[0053] Since the umami enhancement effect does not increase even when the umami enhancer of the present invention is contained in large quantities, considering economic feasibility, the food product may contain ERG in an amount of 0.15 weight % or less, for example, 0.12 or 0.09 weight % or less, or 0.06 or 0.03 weight % or less. Regarding the umami component content of the food product, the food product may contain ERG in an amount of 15 weight % or less, for example, 12 or 9 weight % or less, or 6 or 3 weight % or less. The umami enhancer of the present invention exhibits an umami enhancement effect for various umami components in the food product at content amounts of all combinations of these lower and upper limits.

[0054] The salty component in the present invention is table salt (NaCl), and the salty flavor enhancer of the present invention exhibits a salty flavor enhancing effect in food products by containing 1 ppm or more, for example, 1, 5, 10, 15, 20, or 30 ppm or more, or 40 ppm or 50 ppm or more, with respect to general food products.

[0055] Specifically, in food products containing salt, the saltiness of the food product is enhanced by including ergothioneine, a saltiness enhancer, at a concentration of 0.5 ppm or more in food products with a salt content of 0.1 weight % or more and less than 5 weight %, or at a concentration of 0.1 ppm or more in food products with a salt content of 5 weight % or more.

[0056] In addition, since the saltiness enhancing effect is not increased even when the saltiness enhancing agent of the present invention is contained in large quantities, considering economic feasibility, ERG may be contained in the food product in an amount of 0.15 weight % or less, for example, 0.12 or 0.09 weight % or less, or 0.06 or 0.03 weight % or less. The saltiness enhancing agent of the present invention exhibits a saltiness enhancing effect in the food product with respect to various saltiness components in the food product, with a content amount of all combinations of these lower and upper limits.

[0057] In addition, the concentration of ERG is measured by LCMS under the following conditions.

[0058] (HPLC conditions)

[0059] HPLC analysis is performed under the following conditions.

[0060] Device: HPLC Device: HPLC: Nexera Series Formula (Shimazu Corporation)

[0061] Column: COSMOSIL 2.5 HILIC Column 3.0×150 2.5㎛ 3.0mm I. D.×15.cm (Nakarai Co., Ltd.)

[0062] Flow rate: 0.5 mL / min

[0063] Temperature: 40℃

[0064] Mobile phase: A) 0.1% (v / v) aqueous formic acid, B) 0.1% (v / v) acetonitrile formate

[0065] Isocrative: 0-10 minutes (B: 80%)

[0066] Injection volume: 5μL

[0067] (Mass Spectrometry)

[0068] Device: LCMS-2020 (Shimazwaza)

[0069] (Mass Spectrometry (LC-MS) Analysis Conditions)

[0070] Ionization condition: ESI+

[0071] MS Condition: SIM ERG: m / z230.1([M+H] ＋ )

[0072] Duration: Around 5 minutes

[0073] The food product of the present invention is a food, beverage, or seasoning.

[0074] The food can be used in any food, preferably one containing umami ingredients, and is not limited to. Examples include retort pouch foods such as curry, stew, and meat sauce; processed vegetable products such as salad and pickles; seafood such as salmon, tuna, sardines, shrimp, skipjack tuna, saury, squid, scallops, crabs, and oysters; processed seafood and livestock products such as fish cakes, hamburgers, ham, and sausages; dairy products; cooked rice products such as rice balls, pilaf, fried rice, porridge, rice with tea, chicken and egg rice bowls, Chinese rice bowls, and katsudon; cooked foods such as spring rolls, shumai, dumplings, curry, stews, and fried foods; mycoproteins; pet food; and medical food. Examples of mycoproteins include proteins derived from yeast, proteins derived from filamentous fungi, and proteins derived from fungi such as mushrooms.

[0075] Beverages are not limited to specific types and examples include soft drinks, nutritional drinks, fruit drinks, fermented drinks, sports drinks, soups such as egg soup, seaweed soup, Chinese soup, consommé soup, and potage, liquid juices, miso soup, alcoholic beverages such as beer, soju, and cocktails, and nutritional drinks classified as quasi-drugs. In addition to liquid forms, gel-like and semi-solid forms such as jelly drinks are also included.

[0076] Seasonings are not limited and examples include soy sauce, low-sodium soy sauce, mirin, miso, fish sauce, cooking wine, liquid extracts, bonito broth, kelp broth, umami seasonings, fermented umami seasonings, sushi vinegar, seasoning sauces, sauces, oyster sauce, and various other types of sauces, ketchup, dressings, mayonnaise, etc.

[0077] Umami seasoning refers to a chemical seasoning obtained by refining compounds that possess umami flavor; specifically, examples include amino acid-based seasonings and nucleic acid-based seasonings. Examples of amino acid-based seasonings include glutamic acid and its salts (such as sodium glutamate). Examples of nucleic acid-based seasonings include sodium inosinate and sodium guanylate. Fermented umami seasoning refers to enzymatic hydrolysates of proteins such as wheat gluten and soybeans.

[0078] In addition, in the process of manufacturing the above food product, when the umami flavor enhancer of the present invention is added to the food product, the timing of the addition may be at any stage of the process, and the method of addition is not limited as long as the necessary amount is included in the final food product.

[0079] Soy sauce is typically brewed soy sauce produced by heat-treating plant-based raw materials such as soybeans or wheat, cultivating koji mold thereon, and then fermenting and aging it in brine, with a salt concentration generally of 15 to 18 weight %. In addition to this, there are other types of soy sauce, such as chemical soy sauce produced by decomposing plant-based raw materials with acid or enzymes, fish sauce produced by fermenting seafood, and meat sauce produced by fermenting livestock meat, but the soy sauce of the present invention is preferably brewed soy sauce using koji mold.

[0080] Brewed soy sauce is classified into types based on differences in production methods, such as the ratio of soybeans to wheat in the raw materials, the method of processing the raw materials, and the salt concentration. Examples include dark soy sauce, light soy sauce, tamari, shiro, and ongsimi. Additionally, there is low-salt soy sauce with the salt content of regular soy sauce reduced by nearly half, and the soy sauce of the present invention also includes low-salt soy sauce.

[0081] The ERG of the present invention is contained in soy sauce at a concentration of 5 ppm or more, for example, 10 ppm, 15 ppm, 25 ppm, 30 ppm or more, or 35 ppm, 40 ppm, 45 ppm, 50 ppm or more, thereby enhancing the salty taste and umami taste of the soy sauce. In addition, the salty taste and umami taste of the low-sodium soy sauce are similarly enhanced by containing 5 ppm or more, for example, at a concentration greater than the above concentration, in the low-sodium soy sauce.

[0082] In addition, for soy sauce-containing seasonings containing soy sauce, flavors such as saltiness and umami are enhanced by including 5 ppm or more of ERG.

[0083] In addition, since the effect of enhancing umami and saltiness in soy sauce is not enhanced even if ERG is contained in large quantities, considering economic feasibility, ERG may be contained in the soy sauce at 0.15 weight % or less, for example, 0.12, 0.09 weight % or less, or 0.06 weight % or less. The soy sauce of the present invention exhibits an effect of enhancing umami and saltiness with ERG content in all combinations of these lower and upper limits.

[0084] The ERG-rich koji fermented product of the present invention is prepared by fermenting in either a conventional solid medium or a liquid medium for culturing koji. As for the forms of koji used in industry, there are two forms: solid koji obtained by directly growing koji on soybeans, wheat, rice, etc., and liquid koji obtained by growing koji in a liquid medium in which soybeans, wheat, rice, etc. are suspended. The koji fermented product of the present invention comprises solid koji or liquid koji. Conventional conditions used in the art may be used for fermentation.

[0085] Solid culture media used for making solid koji include solid culture media composed of raw materials with a protein content of less than 5 to 10% of the solid content (protein amount in the food nutrition table) and solid culture media composed of raw materials with a protein content of 10% or more of the solid content (protein amount in the food nutrition table). As raw materials for such solid culture media, for example, rice (protein content 6%), which is a raw material with a protein content of less than 5 to 10%, or legumes, fish, meat, algae, and processed products thereof containing 20% ​​or more of protein, which are raw materials containing 10% or more of protein. Processed products containing 20% ​​or more of protein include defatted soybeans, fish meal, meat meal, and spirulina powder. Solid culture media can be prepared by steaming or boiling the aforementioned raw materials in boiling hot water, and in some cases, by crushing them. In addition, raw materials used in the liquid medium for producing liquid koji include wheat flour, rice, defatted soybeans, corn, soybean pulp, yeast extract, dextrin, etc.

[0086] After the fermentation is complete, the ERG-rich koji fermented product is recovered by separating the liquid from the culture medium, and depending on the application, it can be concentrated with an ion exchange resin or the like, dried as is, seasoned as needed, and used as a raw material for other foods as an ERG-containing food ingredient.

[0087] In addition, the ERG-rich koji fermentation product obtained by using a specific material for a solid medium or liquid medium, and by adding seasonings as needed, can be used as a food ingredient, for example, as a food containing mycoprotein, or as a rice koji or soy sauce koji. Thus, the ERG-rich koji fermentation product of the present invention can be used as an intermediate raw material for food or food made from koji fermentation products, for example, fermented food, fermented beverage, or fermented seasoning.

[0088] When an ERG-rich solid fermented product is prepared using the ERG-high-production koji strain of the present invention, for example, an ERG-rich solid fermented product containing 0.3g or 0.4g / 1kg or more of ERG in the solid fermented product, preferably 0.5g / 1kg or more of ERG in the solid fermented product, more preferably 0.6g / 1kg or more of ERG in the solid fermented product, and even more preferably 0.8g / 1kg or more of ERG in the solid fermented product can be prepared.

[0089] Rice koji obtained from a medium composed of rice, a raw material with a protein content of less than 5~10%, becomes a koji fermented product with a high ERG content containing 0.3g of ERG / 1kg or more of dry koji fermented product, and also soy sauce koji obtained from a medium with a protein content of 25% using soybeans and wheat as raw materials, or mycoprotein obtained from a medium composed of raw materials with a protein content of 10% or more, becomes a koji fermented product with a high ERG content containing 0.4g of ERG / 1kg or more of dry koji fermented product.

[0090] When an ERG-high content koji mold liquid fermentation product is prepared using the ERG-high production koji mold strain of the present invention, an ERG-high content koji mold liquid fermentation product containing 50 to 50,000 ppm of ERG can be prepared. For example, a koji mold fermentation product containing 50 ppm or more, 100, 300, 500, or 1,000 ppm or more of ERG, and also containing 5% or less, 4, 3, 2, or 1% or less of ERG is obtained.

[0091] [Example]

[0092] The details of the present invention are described below with reference to examples, but the present invention is not limited thereto.

[0093] Also, in the examples, where "%" is simply indicated, it means "weight %". Additionally, samples at room temperature were used for the sensory evaluation.

[0094] [Test 1: Umami-enhancing effect of sodium glutamate solution by ERG]

[0095] 0.2% NaCl was mixed with pure water. L-sodium glutamate (hereinafter also referred to as "MSG") was added to this, and six MSG samples with concentrations of 0%, 0.75%, 1%, 1.25%, 1.5%, and 1.75% were prepared. As a seventh sample, purified ergothioneine (purity 99.5% or higher, Tetrahedron) was added to 1% MSG to achieve a final concentration of 60 ppm. 0.2% NaCl was added for sensory evaluation because MSG alone would result in a bland taste.

[0096] Umami was sensorially evaluated by a panel of five trained umami-sensitive experts. The five experts were asked to select a sample with equivalent umami taste from the 1st to 6th MSG samples (concentrations were not disclosed to the panelists) for the 7th 1% MSG solution with added 60 ppm ERG. After tasting each sample and before tasting the next sample, the mouth was thoroughly rinsed with room temperature water.

[0097] In the sensory evaluation, for the 7th sample, 4 out of 5 panelists evaluated it as having an umami taste equivalent to a 1.25% MSG solution and 1 panelist evaluated it as having an umami taste equivalent to a 1.5% MSG solution, and as a result, all 5 panelists evaluated that the umami taste was enhanced by the addition of 60 ppm ERG.

[0098] [Test 2: Amount of ERG Added to Enhance the Umami of MSG]

[0099] (1) In the case of a 2% MSG sample (0.2% NaCl added)

[0100] As in Test 1, samples were prepared by adding ergothioneine to a 2% MSG sample mixed with 0.2% NaCl in pure water to achieve final concentrations of 0.5 ppm, 1 ppm, 3 ppm, 5 ppm, 10 ppm, and 20 ppm (ERG 0 to 20 in Table 1 below, and hereinafter "ERGX" refers to a sample with X ppm of ERG added).

[0101] [Table 1]

[0102]

[0103] For 2% MSG samples with each concentration of ERG added (concentrations were not disclosed to the panel), a 2% MSG sample without ergothioneine was used as a control, and a tasting test was conducted by four expert panelists to sensorially evaluate the umami taste. After tasting each sample and before tasting the next sample, the mouth was thoroughly rinsed with room temperature water.

[0104] For the expert panel, pairs of a control 2% MSG sample and 2% MSG samples with ergothioneine added at each concentration were presented, and the panel evaluated which of the presented pairs tasted better using a 2-point discrimination test. The results are shown in Table 2.

[0105] <Evaluation Method of the 2-Point Identification Test>

[0106] Evaluation results of the umami of the test product compared with the control

[0107] ○: The umami flavor is enhanced compared to the contrast.

[0108] ◇: It has the same umami taste as the contrast.

[0109] And, the evaluation method of the 2-point identification test and the display of the evaluation results are the same in other tests.

[0110] [Table 2]

[0111]

[0112] All panelists evaluated the 2% MSG sample with 1 ppm or more of ERG added as having enhanced umami. This is equivalent to adding 0.005 weight % or more of ERG relative to the MSG content, which is an umami component.

[0113] (2) In the case of the 1% MSG sample (0.2% NaCl added)

[0114] Samples (ERG 15–900) were prepared by adding ergothioneine to a 1% MSG sample mixed with 0.2% NaCl in pure water, with final concentrations of 15 ppm, 20 ppm, 40 ppm, 60 ppm, 80 ppm, 100 ppm, 400 ppm, and 900 ppm. For each 1% MSG sample with added ERG at each concentration, a drinking test was conducted by four expert panelists, identical to those in Test 2, and the umami taste was sensorially evaluated. For the four expert panelists, pairs of 1% MSG samples with ERG added at sequentially different concentrations (e.g., a pair of ERG 20 ppm and 40 ppm) were presented, and the panelists evaluated which of the presented pairs they tasted by a two-point discrimination test. The results are shown in Table 3.

[0115] [Table 3]

[0116]

[0117] According to an evaluation by three panelists, the MSG sample with 80 ppm of ERG added had an equivalent level of umami taste to the sample with 60 ppm of ERG added. It was evaluated that even with the addition of 80 ppm of ERG, the degree of umami enhancement was not different from that with the addition of 60 ppm, and it was confirmed that the umami enhancement effect was constant at 60 ppm of ERG.

[0118] From the above, it was evaluated that with a 1% MSG solution, when ERG was added up to 60 ppm, the umami taste was sequentially enhanced as the amount added increased. When exceeding 60 ppm, the umami enhancement effect was no different from that of the 60 ppm addition.

[0119] (3) For the 10% MSG sample (no NaCl)

[0120] Test 2 was performed on 10% MSG (without NaCl) as in (1). Samples (ERG 0~20 in Table 4 below) were prepared by adding ergothioneine to the 10% MSG sample so that the final concentrations were 1 ppm, 3 ppm, 5 ppm, 10 ppm, and 20 ppm.

[0121] For four expert panelists, pairs of a control 10% MSG sample and 10% MSG samples with ergothioneine added at each concentration were presented, and the panelists evaluated which of the presented pairs tasted better using a 2-point discrimination test. The results are shown in Table 5.

[0122] [Table 4]

[0123]

[0124] [Table 5]

[0125]

[0126] [Illegible].

[0127] [Test 3: Umami-enhancing effect of inosinic acid]

[0128] (1) In the case of a 5% inosinic acid sample

[0129] Samples (ERG 0.1~20) were prepared by adding ergothioneine to a 5% inosinic acid sample mixed with 0.2% NaCl in pure water to achieve final concentrations of 0.1 ppm, 0.5 ppm, 1 ppm, 3 ppm, 5 ppm, and 20 ppm.

[0130] For 5% inosinic acid samples with each concentration of ERG added (concentrations were not disclosed to the panel), a 5% inosinic acid sample without ergothioneine was used as a control, and a sensory evaluation of umami was conducted by four expert panelists. After tasting each sample and before tasting the next sample, the mouth was thoroughly rinsed with room temperature water.

[0131] For the expert panel, pairs were presented consisting of a control 5% inosinic acid sample and 5% inosinic acid samples with ergothioneine added at each concentration, and the panel evaluated which of the presented pairs tasted umami using a 2-point identification test. The results are shown in Table 6.

[0132] [Table 6]

[0133]

[0134] Three panelists evaluated the 5% inosinic acid sample with 0.5 ppm of ERG added as having enhanced umami, and all panelists evaluated the 5% inosinic acid sample with 1 ppm or more of ERG added as having enhanced umami.

[0135] [Test 4: Saltiness Enhancement Effect of NaCl Solution by ERG]

[0136] Five NaCl samples with concentrations of 0.3%, 0.35%, 0.4%, 0.45%, and 0.5% were prepared by adding NaCl to pure water, and as a sixth sample, purified ergothioneine (purity 99.5% or higher, Tetrahedron) was added to 0.4% NaCl to make the final concentration 60 ppm.

[0137] The saltiness was sensorially evaluated by a panel of five trained salt-sensitive experts. The five experts were asked to select a sample with an equivalent saltiness from the 1st to 5th NaCl samples (concentrations were not disclosed to the panelists) for the saltiness of the 6th 0.4% NaCl solution with 60 ppm ERG added. This was done by thoroughly rinsing the mouth with room temperature water after tasting each sample and before tasting the next sample.

[0138] As a result of the sensory evaluation, all five panelists evaluated the 6th sample of 0.4% NaCl with 60 ppm ERG added as having a salty taste equivalent to that of the 0.45% NaCl solution, and consequently, all five panelists evaluated that the salty taste was enhanced by the addition of 60 ppm ERG.

[0139] [Test 5: Amount of ERG Added to Enhance the Salty Taste of NaCl]

[0140] (1) For 0.1~20% NaCl samples

[0141] Samples were prepared by adding ergothioneine to 0.1–20% NaCl samples to achieve final concentrations of 0.05 ppm, 0.1 ppm, 0.25 ppm, and 0.5 ppm (ERG 0.05–0.5 in Table 7 below, and hereinafter "ERGX" refers to a sample with X ppm of ERG added).

[0142] [Table 7]

[0143]

[0144] For 0.1–20% NaCl samples with ERG added at each concentration (concentrations were not disclosed to the panel), a drinking test was conducted by four expert panelists using 0.1–20% NaCl samples without ergothioneine as a control. The expert panelists were presented with pairs of the control 0.1–20% NaCl samples and 0.1–20% NaCl samples with ergothioneine added at each concentration, and the panelists evaluated which of the presented pairs tasted salty using a 2-point discrimination test. The results are shown in Table 8.

[0145] <Evaluation Method of the 2-Point Identification Test>

[0146] Evaluation results of the saltiness of the test product compared with the control

[0147] ○: The salty taste is increasing compared to the contrast.

[0148] ◇: It has the same salty taste as the contrast.

[0149] And, the evaluation method of the 2-point identification test and the display of the evaluation results are the same in other tests.

[0150] [Table 8]

[0151]

[0152] The results of the sensory evaluation showed that for NaCl samples with 0.1% or more and less than 0.5%, when 0.5 ppm or more of ERG was added, all were evaluated as having an enhanced salty taste. For NaCl samples with 0.5% or more, when 0.1 ppm or more of ERG was added, all were evaluated as having an enhanced salty taste.

[0153] (2) For 3% NaCl samples

[0154] Samples (ERG 0.5–100) were prepared by adding ergothioneine to 3% NaCl samples at final concentrations of 0.5 ppm, 12 ppm, 24 ppm, 60 ppm, 72 ppm, and 100 ppm. For each 3% NaCl sample with added ERG at each concentration, a drinking test was conducted by four expert panelists to sensorially evaluate the saltiness. The four expert panelists were presented with pairs of 3% NaCl samples with ERG added at sequentially different concentrations (e.g., a pair of ERG 24 ppm and 60 ppm), and the panelists evaluated which of the presented pairs tasted salty using a two-point discrimination test. The results are shown in Table 9.

[0155] [Table 9]

[0156]

[0157] The 3% NaCl sample with 0.5 ppm or more of ERG added was evaluated by the panel as having an enhanced salty taste.

[0158] In addition, regarding the 3% NaCl sample with 72 ppm of ERG added, the degree of saltiness was evaluated by three panelists as being no different from that with 60 ppm of ERG added, and it was confirmed that the saltiness enhancement effect is constant at 60 ppm of ERG.

[0159] From the above, it was evaluated that with a 3% NaCl solution, when ERG was added in amounts ranging from 0.5 ppm to 60 ppm, the salty taste was sequentially enhanced as the amount added increased. When the amount exceeded 60 ppm, the effect of enhancing the salty taste was no different from that of adding 60 ppm.

[0160] [Test 6: Amount of ERG Added to Enhance Soy Sauce Flavor]

[0161] (1) In the case of dark soy sauce

[0162] Samples (ERG 5~200) were prepared by adding ergothioneine to concentrated soy sauce at concentrations of 5 ppm, 10 ppm, 50 ppm, 100 ppm, and 200 ppm.

[0163] For dark soy sauce with each concentration of ERG added (concentrations were not disclosed to the panel), a tasting test was conducted by five expert panelists using soy sauce without ergothioneine as a control. The expert panelists were presented with pairs of the control soy sauce and the soy sauce with each concentration of ergothioneine added, and the panelists evaluated which of the presented pairs they tasted by a 2-point discrimination test. The results are shown in Table 10.

[0164] [Table 10]

[0165]

[0166] Soy sauce with 5 ppm or more of ERG added was evaluated by 4 panelists as having enhanced umami, and soy sauce with 10 ppm or more of ERG added was evaluated by all panelists as having enhanced umami.

[0167] Samples (ERG 5–200) were prepared by adding ergothioneine to regular soy sauce at concentrations of 5 ppm, 10 ppm, 50 ppm, 100 ppm, and 200 ppm, and each was diluted 10-fold with pure water. For the soy sauces with ERG added at each concentration, a tasting test was conducted by the same panel of five experts, and the umami taste was sensorially evaluated. The five expert panelists were presented with pairs of ERG-added soy sauces with sequentially different concentrations (e.g., a pair of ERG 10 ppm and 50 ppm), and the panelists evaluated which of the presented pairs they tasted using a two-point discrimination test. The results are shown in Table 11.

[0168] [Table 11]

[0169]

[0170] Soy sauce with 200 ppm of ERG added was evaluated by four panelists as having an equivalent level of umami to soy sauce with 100 ppm of ERG added. It was evaluated that even with the addition of 200 ppm of ERG, the degree of umami enhancement was not different from that with the addition of 100 ppm.

[0171] From the above, in dark soy sauce, when ERG was added at a concentration of 5 ppm to 100 ppm, it was evaluated that the umami flavor was sequentially enhanced as the amount added increased. When the concentration exceeded 100 ppm, the umami enhancement effect was no different from that of the 100 ppm addition.

[0172] (2) In the case of special whole soybean soy sauce (Tokusen Marudaizu Shoyu), soup soy sauce, or low-sodium soy sauce

[0173] Samples (ERG 5~200) were prepared by adding ergothioneine to commercially available special whole soybean soy sauce (Kikkoman Co., Ltd.) at concentrations of 5 ppm, 10 ppm, 50 ppm, 100 ppm, and 200 ppm, and a tasting test was conducted by five expert panelists as in (1) above. The expert panelists were presented with pairs of the control soy sauce and the soy sauce with ergothioneine added at each concentration, and the panelists evaluated which of the presented pairs they tasted by a two-point identification test.

[0174] In addition, the same 2-point identification test was also conducted on commercially available soup soy sauce and low-sodium soy sauce.

[0175] As a result, all panelists evaluated that the soy sauce with 5 ppm or more of ERG added to the special whole soybean soy sauce, soup soy sauce, and low-sodium soy sauce had enhanced umami flavor.

[0176] [Test 7: Amount of ERG Added to Enhance Saltiness of Soy Sauce]

[0177] (1) In the case of special whole soybean soy sauce

[0178] Samples (ERG 5-50) were prepared by adding ergothioneine to commercially available specialty whole soybean soy sauce (Kikkoman Co., Ltd.) at concentrations of 5 ppm, 20 ppm, and 50 ppm.

[0179] For special whole soybean soy sauce with each concentration of ERG added (concentrations were not disclosed to the panel), a tasting test was conducted by four expert panelists using soy sauce without ergothioneine as a control. The expert panelists were presented with pairs of the control soy sauce and the soy sauce with each concentration of ergothioneine added, and the panelists evaluated which of the presented pairs tasted saltier using a two-point discrimination test. The results are shown in Table 12.

[0180] [Table 12]

[0181]

[0182] All panelists evaluated the soy sauce with 5 ppm or more of ERG added as having an enhanced salty taste.

[0183] (2) In the case of soup soy sauce

[0184] The same 2-point identification test was also conducted on commercially available soup soy sauce. The results are shown in Table 13.

[0185] [Table 13]

[0186]

[0187] All panelists evaluated the soy sauce with 5 ppm or more of ERG added as having an enhanced salty taste.

[0188] (3) In the case of low-sodium soy sauce

[0189] The same 2-point identification test was also conducted on commercially available low-sodium soy sauce. The results are shown in Table 14.

[0190] [Table 14]

[0191]

[0192] All panelists evaluated the soy sauce with 5 ppm or more of ERG added as having an enhanced salty taste.

[0193] [Test 8: Amount of ERG Added to Enhance the Umami Flavor of Seasoning]

[0194] (1) Yeast solution

[0195] Samples (ERG 0.5–10) were prepared by adding ergothioneine to a 1% solution of yeast extract (Springer 0402 / 20-MG-L) at concentrations of 0.5 ppm, 1 ppm, 3 ppm, 5 ppm, and 10 ppm. The umami and saltiness of the test samples were evaluated by a 2-point discrimination test by three expert panelists in comparison to a control without added ERG. The results of the sensory evaluation are shown in Tables 15 and 16 below.

[0196] [Table 15]

[0197]

[0198] [Table 16]

[0199]

[0200] The panel power was evaluated as having enhanced umami and saltiness of the yeast extract by adding more than 0.5 ppm of ERG.

[0201] [Test 9: Amount of ERG Added to Enhance Salty Taste of Yeast Solution]

[0202] Samples (ERG 0.5–10) were prepared by adding ergothioneine to a 1% solution of yeast extract (Springer 0402 / 20-MG-L) at concentrations of 0.5 ppm, 1 ppm, 3 ppm, 5 ppm, and 10 ppm. The saltiness and umami of the test samples were evaluated by a 2-point discrimination test by three expert panelists in comparison to a control without added ERG. The results of the sensory evaluation are shown in Tables 17 and 18 below.

[0203] [Table 17]

[0204]

[0205] [Table 18]

[0206]

[0207] The salty and umami flavors of the yeast extract were evaluated as being enhanced by adding more than 0.5 ppm of ERG to both panels.

[0208] [Test 10: Flavor-enhancing effect of ERG on each type of food (First)]

[0209] For the test food with ERG added and the control food without ERG added, the taste (umami, sweetness, bitterness) of the test food was evaluated by a 2-point discrimination test by an expert panel in comparison to the control. The results of the sensory evaluation are shown in Tables 19 to 24 below.

[0210] Also, "ERGX" in the test sample refers to the test sample with X ppm of ERG added.

[0211] Sensory evaluation

[0212] ○: The flavor is enhanced compared to the contrast.

[0213] ◇: It tastes like contrast.

[0214] (1) Hon-tsuyu (rich tsuyu)

[0215] After adding 5 ppm of ERG to the original solution of Hon-tsuyu (Kikkoman Co., Ltd.), a sensory test was performed by diluting it four times.

[0216] [Table 19]

[0217]

[0218] The umami and sweetness of Hon-tsuyu were enhanced by the addition of 5 ppm of ERG.

[0219] (2) Zaru Soba Tsuyu

[0220] After adding 5 ppm of ERG to Zaru Soba Tsuyu (Straight Tsuyu: Kikkoman Co., Ltd.), a sensory test was conducted.

[0221] [Table 20]

[0222]

[0223] The umami and sweetness of Zaru Soba were enhanced by the addition of 5 ppm of ERG.

[0224] (3) Soybean paste

[0225] A sensory test was conducted after adding 0.5 ppm of ERG to 18 g of soybean paste (Plus Koji (koji)-free Koji Bijin, Marcome Co., Ltd.) and 200 ml of hot water.

[0226] [Table 21]

[0227]

[0228] The umami and sweetness of the soybean paste were enhanced by the addition of 0.5 ppm of ERG.

[0229] (4) Source

[0230] After adding 5 ppm of ERG to the sauce (Delicious Sauce Tonkatsu, Kikkoman Co., Ltd.), a sensory test was conducted.

[0231] [Table 22]

[0232]

[0233] The umami and sweetness of the sauce were enhanced by the addition of 5 ppm of ERG.

[0234] (5) Salt yeast

[0235] After adding 5 ppm of ERG to salt koji (Plus Koji raw salt koji, Marcome Co., Ltd.), a sensory test was performed.

[0236] [Table 23]

[0237]

[0238] With the addition of 5 ppm of ERG, the umami, saltiness, and sweetness of the salt koji were enhanced.

[0239] (6) Tomato ketchup

[0240] After adding 5 ppm of ERG to tomato ketchup (Del Monte), a sensory test was conducted.

[0241] [Table 24]

[0242]

[0243] The umami and sweetness of tomato ketchup were enhanced by the addition of 5 ppm of ERG.

[0244] [Test 11: Taste-enhancing effect of ERG on each type of food (second)]

[0245] For the test food with ERG added and the control food without ERG added, a sensory evaluation by a panel of experts was conducted to compare the saltiness of the test food with the control. The results of the sensory evaluation are shown in Tables 25 to 29 below.

[0246] Also, "ERGX" in the test sample refers to the test sample with X ppm of ERG added.

[0247] Sensory evaluation

[0248] ○: The salty taste is increasing compared to the contrast.

[0249] ◇: It has the same salty taste as the contrast.

[0250] (1) Zaru Soba Tsuyu

[0251] After adding 5 ppm of ERG to Zaru Soba Tsuyu (Straight Tsuyu: Kikkoman Co., Ltd.), a sensory test was conducted.

[0252] [Table 25]

[0253]

[0254] The saltiness of the zaru soba tsuyu was enhanced by the addition of 5 ppm of ERG.

[0255] (2) Soybean paste

[0256] A sensory test was conducted after adding 5 ppm of ERG to soybean paste (Plus Koji-free Koji Beauty, Marcome Co., Ltd.).

[0257] [Table 26]

[0258]

[0259] The saltiness of the soybean paste was enhanced by the addition of 5 ppm of ERG.

[0260] (3) Source

[0261] After adding 5 ppm of ERG to the sauce (Delicious Sauce Tonkatsu, Kikkoman Co., Ltd.), a sensory test was conducted.

[0262] [Table 27]

[0263]

[0264] The saltiness of the sauce was enhanced by the addition of 5 ppm of ERG.

[0265] (4) Salt yeast

[0266] After adding 5 ppm of ERG to salt koji (Plus Koji raw salt koji, Marcome Co., Ltd.), a sensory test was performed.

[0267] [Table 28]

[0268]

[0269] The saltiness of the salt koji was enhanced by the addition of 5 ppm of ERG.

[0270] (5) Tomato ketchup

[0271] After adding 5 ppm of ERG to tomato ketchup (Del Monte), a sensory test was conducted.

[0272] [Table 29]

[0273]

[0274] The saltiness of tomato ketchup was enhanced by the addition of 5 ppm of ERG.

[0275] [Test 12: Sweetness-Enhancing Effect of Soy Sauce]

[0276] (1) Special whole soybean soy sauce

[0277] Soy sauce was prepared by adding ergothioneine to commercially available special whole soybean soy sauce (Kikkoman Co., Ltd.) at concentrations of 10 ppm, 20 ppm, and 50 ppm (EGR 10, 20, 50).

[0278] For soy sauce with each concentration of ERG added (concentrations were not disclosed to the panel), a tasting test was conducted by three expert panelists with soy sauce without ergothioneine (ERG 0) as a control, and sweetness was sensorially evaluated by a 2-point identification test. The results are shown in Table 30.

[0279] <Evaluation Method of the 2-Point Identification Test>

[0280] Sweetness evaluation results of the test product compared with the control

[0281] ○: The sweetness is increasing compared to the contrast.

[0282] ◇: It has a sweet taste similar to contrast.

[0283] And, the evaluation method of the 2-point identification test and the display of the evaluation results are the same in other tests.

[0284] [Table 30]

[0285]

[0286] Soy sauce with more than 10 ppm of ERG added was evaluated by all panel members as having enhanced sweetness.

[0287] (2) Sweet soy sauce

[0288] ERG was added to commercially available sweet soy sauces (Golden Purple (Goruden Murasaki), Fundokin soy sauce (Fundokin Shoyu)), and the same test as in (1) above was performed. The results are shown in Table 31.

[0289] [Table 31]

[0290]

[0291] Sweetened soy sauce with more than 10 ppm of ERG added was evaluated by all panel members as having enhanced sweetness.

[0292] [Exam 13: Preparation of Soy Sauce]

[0293] Equal amounts of steamed and modified defatted soybeans and ground roasted wheat were mixed, a starter culture was inoculated into the mixture, and soy sauce koji was obtained by fermenting the mixture for 42 hours using a general method. Four copies of the egtA gene were introduced into the starter culture, and an Aspergillus sojae mutant strain (Example 1) in which the inserted gene was overexpressed and an Aspergillus oryzae strain (Example 2) in which the G428S mutation was introduced into the AO090005000664 gene were used, respectively. 100 parts by mass of the obtained soy sauce koji were immersed in 130 parts by mass of saline solution (salt concentration 26% (w / v)), and fermented and aged for 150 days at 25–30°C while stirring appropriately, using a general method for flavor management. The obtained aging extract was pressed and filtered to obtain raw soy sauce. The obtained raw soy sauce was heated to 80°C for 1 hour, and then clarified and filtered to obtain soy sauce. The concentration of ERG in the soy sauce obtained by the clarification and filtering was 566 ppm for Example 1 and 250 ppm for Example 2.

[0294] Compared to commercially available products, the soy sauce of the example had enhanced umami, saltiness, and sweetness. In addition, the soy sauce of Example 2 had improved workability due to improved compressibility during the filtration process in its manufacture.

[0295] [Test 14: Preparation of Mirin]

[0296] After washing polished rice with water using a general method and performing raw material treatments such as soaking in water, removing moisture, steaming, and cooling, 0.1% of the weight of the rice was inoculated and mixed with the starter culture, and the starter culture was managed at an appropriate temperature of 32–38°C using a constant temperature and humidity chamber to obtain koji. For the starter culture, an Aspergillus oryzae strain with the G428S mutation introduced into the AO090005000664 ortholog gene (Example) and its wild strain (Comparative Example) were used, respectively. According to the general method for producing mirin, 48g of the aforementioned rice koji (approximately 40g as raw polished non-glutinous rice) was placed in a fermentation container, and 290g of glutinous rice (kakemai) that had been washed, soaked in water, removed moisture, steamed, and cooled using a general method was mixed in. Next, 155 ml of neutral alcohol (alcohol content 35% (v / v)) was saccharified at 30°C for one month to obtain aged mirin seasoning. Subsequently, the aged mirin seasoning was pressed by a conventional method to obtain coarse mirin filtrate. This was heat-sterilized to a temperature reaching 90°C, and then clarified by filtration to obtain mirin.

[0297] The concentration of ERG in the mirin obtained by the above clarification filtration was 11 ppm for the example and 2 ppm for the comparative example.

[0298] Compared to the mirin of the comparative example, the mirin of the example had enhanced sweetness as well as umami.

[0299] [Test 15: Preparation of Mycoprotein]

[0300] Fermentation was carried out in a 30L bottle (Marubishi Bioengine Co., Ltd.). Water was added to 1.5 kg of soybean pulp powder to fill the volume to 15 L, and 2.25 g of the defoaming agent Shin-Etsu Silicone (registered trademark) KM72F (Shin-Etsu Kagaku Kogyo Kabushiki Kaisha Co., Ltd. (manufactured by Shin-Etsu Kagaku Kogyo Co., Ltd.)) was added, followed by sterilization at 123°C for 60 minutes. An enzyme solution filtered through a pore size filter of 0.22 μm was added to achieve a final concentration of arabinase of 0.6 U / ml and a final concentration of cellulase of 0.1 U / ml, and the mixture was stirred at 60°C for 2 hours. After the reaction, the mixture was cooled to 30°C.

[0301] Pre-culture was carried out as follows. For each strain, 5g of soybean pulp powder and 100ml of water were added to one 500ml baffled Erlenmeyer flask, and the mixture was sterilized in an autoclave (121°C, 30 min). For the inoculum, an Aspergillus oryzae strain with the G428S variant introduced into the AO090005000664 ortholog gene (Example 1) and a wild-type Aspergillus oryzae strain with the AO090005000664 ortholog gene (Comparative Example 1) were used, respectively. 0.5–1ml of glycerol stock of the inoculum was added, and the initial spore count of the pre-culture solution was 5×10 5 Inoculation was performed to achieve a volume of / ml. This pre-culture medium was shaken and cultured for 24 hours (30℃, 160rpm). Afterward, this pre-culture medium was inoculated into a 30L bottle and cultured at 30℃ for 3 days. The aeration and stirring speeds started at 0.5 vvm and 250rpm, respectively, and were increased to 1 vvm and 330rpm after 18 hours. After culture, the fermented product was heat-sterilized (80℃, 30 min).

[0302] The above-mentioned heat-sterilized fermented product was dried for 1 minute using a drum dryer (Katsuragi Kogyo Kabushiki Kaisha (Katsuragi Industrial Co., Ltd.), product name "Drum Dryer D-00") set to a pore size of 0.3 mm and a surface temperature of 150°C. This is referred to as fermented soybean pulp. The ergothioneine content was 1.70 g / kg dried koji fermented product (Example 1) and 0.1 g / kg dried koji fermented product (Comparative Example 1).

[0303] Using the fermented soybean pulp obtained in the above sequence, 16.7g of the dried material was used, and the dried material and water were mixed in a weight ratio of 1:2. Additionally, 3% soy sauce and 1% yeast solution were mixed, and the mixture was kneaded for 10 seconds using a dough mixer (manufactured by Taiga Mahobin Kabushikigaiya (Tiger Magic Bottle Co., Ltd.), product name "Microcomputer Food Processor SKF-H101"). Afterward, the kneaded sample was molded to a thickness of 1.8cm and an inner diameter of 7.5cm. 5ml of oil was added to a frying pan, and the molded sample was baked over low heat for 1 minute and 20 seconds on each side, for a total of 2 minutes and 40 seconds. The ergothioneine content was 1g / kg dried koji fermented product (Example 1) and 0.05g / kg dried koji fermented product (Comparative Example 1).

[0304] Regarding the taste of the mycoproteins of Comparative Example 1 and Example 1, a sensory evaluation was conducted by five expert panelists, and all panelists evaluated that the mycoprotein of Example 1 had a strong umami taste and was delicious. In addition, Example 1 is suitable for processing because it has a weaker odor compared to Comparative Example 1.

[0305] [Experiment 16: Preparation of Salt Yeast]

[0306] Rice koji production

[0307] 1250g of white rice was soaked overnight, transferred to a sieve, and drained for 2 hours. Afterward, it was wrapped in a polyester (registered trademark) cloth (Toure Kabushiki Kaisha (Toure Co., Ltd.)) and boiled at 100°C for 40 minutes. The steamed rice was cooled to 45°C and inoculated with a starter culture. For the starter culture, an Aspergillus oryzae strain with the G428S variant introduced into the AO090005000664 ortholog gene (Example 2) and a wild-type Aspergillus oryzae strain with the AO090005000664 ortholog gene (Comparative Example 2) were used, respectively. It was wrapped in polypropylene, and koji production was initiated under conditions of 35°C and 95% humidity. After 19 hours of koji production, the rice koji was removed and wrapped again in polypropylene. When the product temperature reached 40℃, polypropylene was spread to flatten the rice koji. Also, after 40 hours, the rice koji was thoroughly loosened and left to stand again. After 42 hours, the koji was released. The ergothioneine content was 0.35 g / kg dry koji fermented product (Example 2) and 0.03 g / kg dry koji fermented product (Comparative Example 2).

[0308] Salt koji production

[0309] 20g of salt and 100ml of hot water at 50℃ were added to 50g of obtained rice koji, and the reaction was carried out at 50℃ and 130rpm for 14 hours. The ergothioneine content was 0.1g / kg dried koji fermented product (Example 2) and 0.01g / kg dried koji fermented product (Comparative Example 2).

[0310] Regarding the umami taste of Comparative Example 2 and Example 2, a sensory evaluation was conducted by four expert panelists, and all panelists evaluated that the salt koji of Example 2 had a strong umami taste and was delicious.

[0311] [Exam 17: Preparation of Gamju]

[0312] Preparation of Gamju

[0313] 2.5 times the amount of hot water at 60°C was added to 800g of rice koji obtained in Comparative Example 2 and Example 2, and a saccharification reaction was carried out at 55°C and 200 rpm. After 16 hours, the Brix was prepared to 18%, transferred to a heat-resistant container, and sterilized at 85°C for 30 minutes. The ERG content of the obtained sweet rice wine was 115 ppm (Example 3) and 7 ppm (Comparative Example 3). Compared to Comparative Example 3, Example 3 had a stronger sweet taste, a whiter color, a stronger chestnut aroma, and a better aroma because it had less koji or rice bran smell. Additionally, it had a smoother mouthfeel because it had fewer large particles.

[0314] [Experiment 18: Preparation of Soy Sauce-Pickled Rice Koji]

[0315] Preparation of soy sauce-pickled rice koji

[0316] 4g of salt and 75ml of soy sauce were added to 50g of rice koji obtained in Comparative Example 2 and Example 2, and the reaction was carried out at 50℃ and 130rpm for 14 hours. The ergothioneine content was 0.14g / kg dried soy sauce-pickled rice koji (Example 4) and 0.01g / kg dried soy sauce-pickled rice koji (Comparative Example 4).

[0317] As a result of a sensory evaluation by four expert panelists on the taste of the soy sauce-pickled rice koji of Comparative Example 4 and Example 4, all panelists evaluated that the soy sauce-pickled rice koji of Example 4 had a strong and delicious taste.

[0318] [Experiment 19: Preparation of Soy Sauce (Hishiho)]

[0319] Soy sauce manufacturing method

[0320] Equal amounts of steamed modified defatted soybeans and ground roasted wheat were mixed, a starter culture was inoculated into the mixture, and soy sauce koji was obtained by fermenting for 42 hours using a general method. For the starter culture, an Aspergillus oryzae strain with a G428S mutation introduced into the AO090005000664 ortholog gene (Example 5) and an Aspergillus oryzae strain with a wild-type AO090005000664 ortholog gene (Comparative Example 5) were used, respectively. 100 parts by mass of the obtained soy sauce koji were immersed in 130 parts by mass of saline solution (salt concentration 26% (w / v)), and fermented and aged for 150 days at 25–30°C while stirring appropriately, using a general method. The ergothioneine content was 0.23 g / kg soy sauce (Example 5) and 0.04 g / kg soy sauce (Comparative Example 5).

[0321] A sensory evaluation was conducted by four expert panelists on the taste of the soy sauces of Comparative Example 5 and Example 5, and all panelists evaluated that the soy sauce of Example 5 had a strong umami flavor and was delicious. In addition, Example 5 had reduced odor and oxidized odor compared to Comparative Example 5.

[0322] [Test 20: Preparation of Koji Fermented Seasoning Liquid]

[0323] Method for preparing koji-fermented seasoning liquid

[0324] An Aspergillus oryzae strain with the G428S mutation introduced into the AO090005000664 ortholog gene was inoculated into a 2.15% dextrin, 4.5% yeast medium and liquid cultured at 26°C for 7 days. The resulting culture was heated in a tank at 90°C for 30 minutes, and then the culture was filtered. The resulting filtrate was used to adsorb ERG onto ion exchange resin UBK08 (manufactured by Mitsubishi Chemical), and the solution was eluted with anhydrous water to obtain a koji fermented seasoning liquid. The ERG concentration of the obtained koji fermented seasoning liquid was 30,000 ppm.

[0325] The effect of enhancing salty, sweet, and umami flavors using koji mold fermented seasoning liquid has been recognized.

Claims

Claim 1 An umami and / or salty taste enhancer having ergothioneine as an active ingredient. Claim 2 An umami-enhanced food product containing 0.005 weight % or more of ergothioneine, an umami enhancer, relative to the content of umami components in the food product. Claim 3 A salty-flavored food product containing ergothioneine, a salty-flavoring enhancer, at a concentration of either one of the following: (1) ergothioneine at a concentration of 0.5 ppm or more in a food product containing 0.1 weight % or more and less than 5 weight % of salt, or (2) ergothioneine at a concentration of 0.1 ppm or more in a food product containing 5 weight % or more of salt. Claim 4 In paragraph 2 or 3, the food product with enhanced umami and / or saltiness is soy sauce, miso paste, fish sauce, mirin, bonito broth, liquid extract, umami seasoning, fermented umami seasoning, ketchup, sauce, or mycoprotein. Claim 5 A food product manufactured using an ergothioneine-high-producing yeast strain in paragraph 2 or 3. Claim 6 Food products with enhanced umami and / or saltiness containing 10 ppm or more of ergothioneine. Claim 7 Soy sauce or soy sauce-containing seasoning, other than soy sauce for retort foods, containing 10 ppm or more of ergothioneine. Claim 8 In paragraph 7, soy sauce is low-salt soy sauce. Claim 9 According to Article 5, a food product is a rice koji containing 0.3g of ergothioneine / 1kg or more of a dried koji fermented product, obtained by culturing a high-ergothioneine koji strain in a medium composed of raw materials with a protein content of less than 5-10%. Claim 10 A food product described in paragraph 5 that is a mycoprotein or soy sauce koji, which is a koji fermented product containing 0.4g of ergothioneine and 1kg or more of dried koji fermented product, obtained by culturing a koji strain that produces a high amount of ergothioneine in a medium composed of raw materials with a protein content of 10% or more. Claim 11 Ergothioneine-high koji fermented product containing 50 to 50,000 ppm of ergothioneine. Claim 12 A method for enhancing the umami and / or salty taste of food by incorporating ergothioneine into the food.

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