Off-flavor reducing agent containing ergothioneine as an active ingredient

Abstract

Task: To provide an off-flavor reducing agent that reduces the off-flavors of bitter substances or seasonings, or to provide food and beverages in which off-flavors are reduced by the addition of an off-flavor reducing agent. Solution: As an off-flavor reducing agent with ergothioneine as an active ingredient, food and beverages containing ergothioneine as a bitterness reducing agent have reduced bitterness, seasonings with irritating flavor have reduced irritating flavor, and soybean odors such as soy milk can also be reduced.

Description

Technology Field

[0001] The present invention relates to an off-flavor reducer for bitter substances or seasonings, and to food and beverages with reduced off-flavor. Background Technology

[0002] Ergothioneine is found as a hydrophilic sulfur-containing amino acid isolated from the rye fungus *Claviceps purpurea*, and it has been confirmed to exist in the bodies of plants and animals. However, plants and animals cannot synthesize ergothioneine, and the ergothioneine in the body is believed to originate from ergothioneine synthesized by microorganisms such as basidiomycetes.

[0003] Ergothioneine has high antioxidant activity and has also been reported to have elastase and tyrosinase inhibitory effects, and is receiving particular attention in the beauty and food fields, such as for brightening and wrinkle prevention. In addition, it has been found that ERG is involved in the body's oxidative defense system, and applications in the medical field are also being attempted.

[0004] Health functional foods containing ergothioneine are useful for maintaining or promoting health, but ergothioneine has a unique aroma, particularly a bitter taste. It has been reported that in order to suppress this bitter taste, an oral composition can be obtained in which the bitter taste derived from ergothioneine is suppressed by combining ergothioneine with vitamin E (Patent Document 1), arachidonic acid (Patent Document 2), or theanine (Patent Document 3).

[0005] Meanwhile, potassium chloride is used as a substitute for sodium chloride, but its bitter taste is a problem. In koji-fermented beverages such as Japanese sake, the bitterness of arginine is a challenge, and attempts are being made to reduce it. Furthermore, in soybean hydrolysates, the bitterness caused by soybean peptides is a challenge. Prior art literature

[0006] International Publication No. 2023 / 120370 International Publication No. 2023 / 120378 International Publication No. 2023 / 120367 International Publication No. 2019 / 240243 International Publication No. 2023 / 234307 The problem to be solved

[0007] The problem to be solved by the present invention is to provide an off-flavor reducing agent that reduces the off-flavor of bitter substances or seasonings, or to provide a food or beverage in which the off-flavor is reduced by the addition of an off-flavor reducing agent. means of solving the problem

[0008] As a result of conducting diligent research, the inventors discovered that by including a small amount of ergothioneine, which has a bitter taste, in food and beverages containing bitter substances, the bitterness of the bitter substances is reduced, thereby reducing the bitterness of the food and beverages. They also discovered that ergothioneine can reduce the stimulating taste of seasonings and the soybean smell of soy milk, and thus completed the present invention.

[0009] The present invention relates to an off-flavor reducing agent as described in (1) to (4) below.

[0010] (1) An off-flavor reducing agent for bitter substances, having ergothioneine as an active ingredient.

[0011] (2) The off-flavor reduction agent described in (1), wherein the off-flavor reduction is the reduction of the bitterness of soybean peptide or mycoprotein, which is a bitter substance.

[0012] (3) An off-flavor reducing agent for beverages containing legumes as raw materials, having ergothioneine as an active ingredient.

[0013] (4) The off-flavor reduction agent described in (3) above, which is the reduction of the off-flavor of soy milk.

[0014] In addition, the present invention relates to the food and beverage described in (5) and (6) below, the soy milk or soy milk fermentation product described in (7) and (8), the mycoprotein described in (9), or the method for reducing off-flavors of the food and beverage described in (10), and other (11) to (16).

[0015] (5) A food and beverage containing a bitter substance, characterized by containing 0.005% by weight or more of ergothioneine relative to the soybean peptide content, with reduced bitterness of the soybean peptide.

[0016] (6) Food and beverages described in (5) that are not food and beverages, including soy sauce for retort food, retort food containing soy sauce, sourness-enhanced beverage, and ergothioneine-producing fungal fermentation biomass or its supernatant colored with a food coloring agent.

[0017] (7) Soy milk or fermented soy milk containing 10 ppm or more of ergothioneine.

[0018] (8) Soy milk or soy milk fermentation product described in (7) above, which is not an acid-enhanced beverage.

[0019] (9) Mycoprotein containing 300 ppm or more of ergothioneine.

[0020] (10) A method to reduce off-flavors in food and beverages by incorporating ergothioneine into the food and beverages that have off-flavors.

[0021] (11) Ergothioneine for use in reducing off-flavors in beverages containing legumes as ingredients.

[0022] (12) A fermented product containing ergothioneine for use in reducing off-flavors in beverages containing legumes as raw materials.

[0023] (13) Use of ergothioneine to reduce off-flavors in beverages containing legumes as ingredients.

[0024] (14) Use of a fermented product containing ergothioneine to reduce off-flavors in beverages containing legumes.

[0025] (15) Use of ergothioneine in the manufacture of an off-flavor reducing agent for a beverage containing legumes as a raw material.

[0026] (16) Use of a koji fermentation product containing ergothioneine in the manufacture of an off-flavor reducing agent for a beverage containing legumes as a raw material. Effects of the invention

[0027] According to the off-flavor reducing agent of the present invention, by adding ergothioneine, the bitterness of bitter substances such as caffeine, potassium chloride, arginine, or soybean peptides can be reduced, or the stimulating sensation of seasonings such as sauces or the soybean smell such as soy milk can be reduced, so by adding a very small amount of ergothioneine to food and beverages containing these bitter substances, the taste of these food and beverages can be improved. Specific details for implementing the invention

[0028] The present invention relates to an off-flavor reducing agent having ergothioneine as an active ingredient, and specifically, to a bitterness reducing agent for bitter substances such as caffeine, potassium chloride, arginine, or soybean peptides, or to a stimulating taste reducing agent for seasonings such as sauces, or to a soybean odor reducing agent such as soy milk. Furthermore, the present invention relates to a food and beverage in which off-flavors are reduced by containing ergothioneine, which is an off-flavor reducing agent.

[0029] Also, in the present invention, “%” means “weight%” even if the unit is not specified, and 1 ppm is 0.0001 weight%.

[0030] Ergothioneine (hereinafter also referred to as "ERG") is contained in some basidiomycetes, such as oyster mushrooms, shiitake mushrooms, maitake mushrooms, and king oyster mushrooms, and is particularly abundant in yellow oyster mushrooms. It is also known that koji molds produce ERG. Methods for producing ERG include extraction from basidiomycetes such as yellow oyster mushrooms, chemical synthesis, and fermentation using microorganisms.

[0031] Mushrooms used for extracting ERG include oyster mushrooms and mushrooms of the genus Pleurotus, such as Pleurotus cornucopiae var. citrinopileatus, Pleurotus ostreatus, Lentinula edodes, and Grifola Frondosa, which are preferably examples due to their high ERG content, and one or a combination of these types may be used.

[0032] Extraction using basidiomycetes such as the yellow oyster mushroom is not suitable for mass production because it is time-consuming to obtain raw materials. While chemical synthesis is suitable for mass production, it requires the use of expensive synthetic reagents. For this reason, research is being conducted on production methods by fermentation, such as fermentation using C1 compound-associating bacteria or yeasts, fermentation using microorganisms that overexpress ERG biosynthetic genes, or solid culture using koji mold that overexpresses ERG by introducing ERG biosynthetic genes.

[0033] As the koji mold that produces ERG, any strain belonging to the genus Aspergillus may be used. Examples include Aspergillus oryzae, Aspergillus sojae, Aspergillus niger, Aspergillus luchuensis, and Aspergillus tamarii. In addition, as the koji mold that produces ERG, a mutant strain with high ERG production may be isolated and used by using a general mutation introduction method in addition to the above wild strain.

[0034] In addition, ERG-high-producing koji strains can be used by introducing genes related to ERG biosynthesis through genetic recombination. Examples include a koji strain with the egtA gene introduced (see Patent Document 4), or a koji strain with increased ERG expression by introducing the AO090005000664 gene (coding amino acid sequence: SEQ ID NO. 1) characteristic of the genus Aspergillus, which introduces specific gene mutations such as introducing the G428S mutation or the C459F mutation, or deleting 1 to 112 amino acids in the W404 to Q515 region, and the ortholog gene thereof (see Patent Document 5), but are not limited to these.

[0035] In this specification, the ERG-producing koji strain indicates that 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 Corporation) is added to water and suspended, pH unadjusted) compared to a strain that is not mutated or transformed. Examples include the Aspergillus oryzae strain in which the above egtA gene is forcibly expressed, or the Aspergillus oryzae mutant strain containing amino acid mutations or amino acid deletions in the W404 to Q515 regions of the above mutant AO090005000664 gene and its ortholog gene.

[0036] The ERG of the present invention may be a commercially available ERG produced by chemical synthesis, or may be an extract or purified product thereof derived from mushrooms containing ERG, koji molds that produce ERG and their cultures, or sake lees produced using koji molds. Furthermore, for food and beverages using koji molds as raw materials, the amount of ERG in the food and beverage can be increased by using koji molds that produce a high amount of ERG without adding ERG.

[0037] The bitter substances targeted for bitterness reduction in the present invention include caffeine, potassium chloride, arginine, or soy peptides, but the bitter substances are not limited to these.

[0038] In addition, the seasoning subject to the reduction of irritating taste according to the present invention is a seasoning having an irritating taste including spices, and includes, for example, sauces such as Worcestershire sauce or medium-thick sauce, but is not limited thereto.

[0039] In addition, the beverage subject to the soybean odor reduction of the present invention is a beverage such as soy milk that has soybeans as at least part of its raw materials.

[0040] The food and beverage of the present invention is a food, beverage, or seasoning and contains one or more bitter substances. As a food, beverage, or seasoning, it can be used in a food, beverage, or seasoning containing one or more bitter substances, but is not limited to.

[0041] The reduction of off-flavor means that the off-flavor is perceived as weaker compared to the case where there is no off-flavor reducing agent in the food or beverage. For example, it means that by adding a bitterness reducing agent, even if the caffeine, etc. in the food or beverage is increased by, for example, 1 to 50%, an equivalent bitter taste is perceived. Furthermore, it means that by adding a bitterness reducing agent, it is possible to perceive a bitter taste equivalent to that of, for example, containing 0.5 to 0.9 times the amount of caffeine, etc.

[0042] In the process of manufacturing food and beverages, when ERG is added to the food and beverage, 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 required amount is included in the final food and beverage product.

[0043] As a bitter substance, for food and beverages containing caffeine, ERG is included in an amount of 0.1% by weight or more, for example, 0.1, 0.2, 0.3, or 0.5% by weight or more relative to the caffeine content, and for food and beverages containing potassium chloride or arginine, ERG is included in an amount of 0.01% by weight or more, for example, 0.01, 0.02, 0.03, or 0.05% by weight or more relative to the potassium chloride or arginine content, thereby exhibiting an effect of reducing the bitterness of the food and beverage.

[0044] In addition, by including ERG at a weight of 0.005% or more relative to the soybean peptide content, for example, 0.005, 0.01, 0.05, or 0.08% or more, the bitterness reduction effect of food and beverages is exhibited.

[0045] In this way, when food and beverages contain one or more of the bitter substances caffeine, potassium chloride, arginine, or soy peptides, the bitterness reduction effect is achieved by including 0.005 to 0.1 weight percent or more of ERG relative to the content of any one of them.

[0046] In addition, since the bitterness reduction effect is not enhanced even if a large amount of ERG is included, considering economic feasibility, ERG can be included in an amount of 15% by weight or less, for example, 12% or 9% by weight or less, or 6% or 3% by weight or less, with respect to the content of bitter substances in food and beverages.

[0047] That is, for food and beverages containing caffeine, ERG is contained in an amount of 0.1% by weight or more, for example, 0.1, 0.2, 0.3, 0.5% by weight or more, and 15% by weight or less, for example, 12, 9, 6, or 3% by weight or less, with respect to the caffeine content; and for food and beverages containing potassium chloride or arginine, ERG is contained in an amount of 0.01% by weight or more, for example, 0.01, 0.02, 0.03, 0.05% by weight or more, and 15% by weight or less, for example, 12, 9, 6, or 3% by weight or less, with respect to the potassium chloride or arginine content, thereby exhibiting a bitterness reduction effect in food and beverages with all combinations of these lower and upper limit values.

[0048] In addition, by including ERG at a content of 0.005% by weight or more, for example, 0.005, 0.01, 0.05, 0.08% by weight or more, and 15% by weight or less, for example, 12, 9, 6, or 3% by weight or less, with respect to the soybean peptide content, the bitterness reduction effect of food and beverages is exhibited with content amounts of all combinations of these lower and upper limits.

[0049] For sauces such as Worcestershire sauce or medium-thick sauce, which are seasonings containing the spices of the present invention that have an irritating taste, the irritating taste is reduced by containing ERG at a concentration of 5 ppm or more, for example, 10 ppm or 15 ppm or more, or 20 ppm, 25 ppm, 30 ppm or more, 35 ppm, 40 ppm, 45 ppm, or 50 ppm or more. In addition, since the effect of reducing the irritating taste is not enhanced even if a large amount of ERG is contained, considering economic feasibility, the ERG may be contained at a concentration 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.

[0050] In addition, in soy milk or fermented soy milk products containing soybean peptides, which are bitter substances, by including ergothioneine, not only the bitter taste but also off-flavors such as the smell of soybeans can be reduced.

[0051] In the present embodiment, "soy milk" refers to a beverage containing an oily soy milk liquid obtained by extracting proteins and other components from soybeans (whole soybeans, defatted processed soybeans, powdered soybeans, etc.) using hot water, etc., and removing fibers. Examples include soy milk products (soy milk, prepared soy milk, soy milk beverages) listed in the quality labeling standards for soy milk products notified by the Consumer Affairs Agency, and soy milk products other than those listed in the quality labeling standards for soy milk products are also included. Furthermore, the soy milk in the present embodiment may include other auxiliary ingredients (e.g., animal and vegetable oils, sweeteners, seasonings such as salt, fruit juice, vegetable broth, flavor ingredients such as coffee and cocoa, emulsifiers, thickeners, flavorings, etc.). In addition, so-called soybean liquid preparations prepared by suspending, dissolving, or homogenizing powder of hulled soybeans or whole soybeans in water, or soybean beverages made from these raw materials, are also included in the soy milk of the present embodiment. The soy milk may also be soy milk that has been sterilized for long-term preservation and sealed in a state where commercial sterility is maintained.

[0052] There are no specific restrictions on the method of manufacturing soy milk, but it can be manufactured using methods known to those skilled in the art, such as soaking soybeans in water overnight and then grinding them with a grinder. Soybeans may also be used after being pretreated by roasting, etc. Soy milk that is generally commercially available may also be used.

[0053] If the final product is soy milk other than unadjusted soy milk, various additives used in general soy milk beverages may be added. Examples of additives include sugars such as sucrose, salt, calcium lactate, emulsifiers, thickeners such as carrageenan, flavorings, colorings, oils and fats, preservatives, antioxidants, emulsifiers, spices, various extracts and pastes, proteins and their hydrolysates, organic acids, organic acids, starch, stabilizers, etc.

[0054] Regarding the soybean solid content, the Japanese Agricultural and Forestry Standard defines soy milk as having a soybean solid content of 8.0% or more, adjusted soy milk as having a soybean solid content of 6% or more, and soy milk beverage as having a soybean solid content of 4% or more, but all of these can be used as soy milk in the present embodiment.

[0055] A fermented soy product refers to a fermented product obtained by fermenting soy products using koji mold. As for the soy products used as raw materials for the production of fermented soy products, soy products classified as soy milk, prepared soy milk, and soy milk beverages under Japanese agricultural standards may be used, and there are no particular limitations as long as they are soy products obtained by conventional methods.

[0056] As for the soy milk, commercially available soy milk may be used, but a liquid obtained by dissolving whole soybean flour, defatted soybean flour, etc. may also be used. As for the soy milk used as a raw material for the production of the fermented soy milk product, beverages containing soybean-derived protein may be used, such as beverages in which soybeans are liquefied as they are or contain fiber (whole soybean beverages, etc.) without removing the soybean fiber (soybean pulp) as in soy milk, or beverages in which soybean fiber is added to soy milk (soybean pulp beverages). The concentration of the protein derived from soybeans may be freely set as long as the koji mold can grow. In addition, the soy milk used as a raw material for the production of the fermented soy milk product may be, for example, soy milk that has been adjusted so that the concentration of the protein derived from soybeans reaches a predetermined concentration.

[0057] Fermented soy milk products are produced by fermenting soy milk and, if necessary, performing additional treatments other than the fermentation process. As additional treatments, there are conventionally known methods for treating cultures, such as spray drying, retort sterilization, freeze-drying, UHT sterilization, pressurized sterilization, high-pressure steam sterilization, dry heat sterilization, electromagnetic sterilization, high-frequency sterilization, ultraviolet sterilization, and chemical sterilization (alcohol sterilization, electrolytic water treatment).

[0058] By including ERG at a concentration of 10 ppm or more, preferably 20 ppm or more, in soy milk or fermented soy milk, the soybean odor can be reduced and the bitterness of soybean peptides can also be reduced. Since the effect of reducing the soybean odor is not enhanced even if a large amount of ERG is included, considering economic feasibility, ERG may be included in the soy milk, etc. at a concentration of 0.15% by weight or less, 0.12% or 0.09% by weight or less, or 0.06% or 0.03% by weight or less.

[0059] In addition, in the case of mycoproteins containing arginine, a bitter substance, the bitterness can be reduced and the umami taste enhanced by including ergothioneine. Mycoproteins are food products obtained by fermenting koji mold in a solid or liquid medium, and then seasoning them as needed. By including 300 ppm or more of ERG in the mycoproteins, the bitterness of the arginine contained in the mycoproteins can be reduced and the umami taste enhanced at the same time.

[0060] In addition, by producing a koji fermentation product with a high ERG content using an ERG-high-production koji strain without adding ERG, a koji 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.

[0061] In addition, the concentration of ERG is measured by LCMS under the following conditions.

[0062] (HPLC conditions)

[0063] HPLC analysis is performed under the following conditions.

[0064] Device: HPLC Device: HPLC: Nexera Series (Manufactured by Shimazu Corporation)

[0065] Column: COSMOSIL 2.5HILIC Column 3.0×15 2.5㎛ 3.0mm I.D.×15.0cm (Manufactured by Nakarai)

[0066] Flow rate: 0.5 mL / min

[0067] Temperature: 40℃

[0068] Mobile phase: A) 0.1% (v / v) aqueous formic acid, B) 0.1% (v / v) acetonitrile formate

[0069] Isocrates: 0-10 mins B: 80%)

[0070] Injection volume: 5μL

[0071] (Mass Spectrometry)

[0072] Device: LCMS-2020 (Manufactured by Shimadzu Corporation)

[0073] (Mass Spectrometry (LC-MS) Analysis Conditions)

[0074] Ionization condition: ESI+

[0075] MS Condition: SIM ERG: m / z230.1([M+H] ＋ )

[0076] Duration: Around 5 minutes

[0077] [Example]

[0078] The details of the present invention will be specifically explained below by presenting examples, but the present invention is not limited thereto.

[0079] Also, in the examples, where "%" is simply indicated, it means "weight%". In addition, room temperature samples were used for the sensory evaluation.

[0080] [Test 1: Bitterness Reduction Effect of Caffeine, a Bitter Taste Standard Substance, by ERG]

[0081] Caffeine was added to pure water to prepare caffeine samples at four concentrations of 0.01%, 0.03%, 0.05%, and 0.06%, to which ergothioneine was added to achieve final concentrations of 0.05 ppm, 0.1 ppm, 0.25 ppm, and 0.5 ppm (ERG 0.05 to 0.5 in Table 1 below, and hereinafter “ERGX” refers to a sample with X ppm of ERG added).

[0082] [Table 1]

[0083]

[0084] Bitterness was sensorially evaluated by four expert panelists who were trained subjects with bitter taste sensitivity. A drinking test was conducted by four expert panelists on 0.01–0.06% caffeine samples with each concentration of ERG added (concentrations were not disclosed to the panelists), with 0.01–0.06% caffeine samples without ergothioneine added serving as a control group. The expert panelists were presented with pairs consisting of the control group's 0.01–0.06% caffeine samples and 0.01–0.06% caffeine samples with each concentration of ergothioneine added. The panelists evaluated which of the presented pairs they perceived as bitter using a 2-point discrimination test. The results are shown in Table 2.

[0085] <Evaluation Method of the 2-Point Identification Test>

[0086] Evaluation results of the taste (bitter taste) of the test product compared to the control group

[0087] ▽: Compared to the control group, the taste (bitter taste) was reduced.

[0088] ◇: It has the same taste (bitter taste) as the control group.

[0089] ○: The taste (bitter taste) was enhanced compared to the control group.

[0090] Also, the evaluation method of the 2-point identification test and the display of the evaluation results are the same in other tests.

[0091] [Table 2]

[0092]

[0093] The results of the sensory evaluation showed that for caffeine samples with 0.01% or more and 0.05% or less, when ERG was added to a final concentration of 0.1 ppm or more, all panelists evaluated that the bitterness was reduced. For caffeine samples with 0.06%, when ERG was added to a final concentration of 0.5 ppm or more, all panelists evaluated that the bitterness was reduced.

[0094] In a 0.01% caffeine sample, the bitterness is reduced when ERG is added to a final concentration of 0.1 ppm or more, so the amount of ERG that has a bitterness-reducing effect with respect to the caffeine content in food and beverages is equivalent to ERG being 0.1 weight% or more.

[0095] [Test 2: Effect of ERG on Reducing the Bitterness of Potassium Chloride]

[0096] A sample (ERG5) was prepared by adding ergothioneine to a potassium chloride sample with a concentration of 5.0%, to which potassium chloride was added to pure water, so that the final concentration was 5 ppm (Table 3).

[0097] For a panel of five trained bitter taste sensitizers, pairs were presented consisting of a control 5% potassium chloride sample without added ERG (ERG0) and a 5% potassium chloride sample with ergothioneine added to a final concentration of 5 ppm. The panelists evaluated which of the presented pairs tasted bitter or salty using a 2-point discrimination test. The results are shown in Table 4.

[0098] [Table 3]

[0099]

[0100] [Table 4]

[0101]

[0102] The results showed that 4 out of 5 panelists evaluated the 5% potassium chloride solution with 5 ppm ERG added as having reduced bitterness, and all 5 panelists evaluated the 5% potassium chloride solution with 5 ppm ERG added as having increased saltiness.

[0103] In a 5% aqueous potassium chloride solution, the bitterness is reduced when 5 ppm or more of ERG is added, so the amount of ERG that has a bitterness-reducing effect with respect to the potassium chloride content in food and beverages is equivalent to 0.01 weight% or more of ERG.

[0104] [Test 3: Effect of ERG on Bitterness Reduction in Potassium Chloride Products]

[0105] Sel Salt I (potassium chloride bitter taste mask processed product: Mitejima Chemical) prepared a sample (ERG5) by adding ergothioneine to a potassium chloride processed product sample with a concentration of 5.0% so that the final concentration was 5 ppm (Table 5).

[0106] It was evaluated by the same 5-person expert panel as in Test 2 using the same 2-point discrimination test as in Test 2. The results are shown in Table 6.

[0107] [Table 5]

[0108]

[0109] [Table 6]

[0110]

[0111] The results showed that all five panelists evaluated the 5% potassium chloride aqueous solution with ERG added to a final concentration of 5 ppm as having reduced bitterness, and evaluated the 5% potassium chloride aqueous solution with ERG added to a final concentration of 5 ppm as having increased saltiness.

[0112] [Test 4: Bitter taste reduction effect of arginine by ERG]

[0113] A sample (ERG5) was prepared by adding ergothioneine to an arginine sample containing arginine, a bitter amino acid, at a concentration of 5.0%, so that the final concentration was 5 ppm (Table 7).

[0114] A pair of samples was presented to a panel of four experts, consisting of a control group of 5% arginine samples without added ERG (ERG0) and a 5% arginine sample with added ergothioneine to a final concentration of 5 ppm. The panel evaluated which of the presented pairs of samples tasted bitter and savory using a two-point discrimination test.

[0115] The results are shown in Table 8.

[0116] [Table 7]

[0117]

[0118] [Table 8]

[0119]

[0120] The results were that all four panelists evaluated that the 5% arginine aqueous solution with ERG added to a final concentration of 5 ppm reduced bitterness, and evaluated that the 5% arginine aqueous solution with ERG added to a final concentration of 5 ppm increased umami.

[0121] In a 5% arginine aqueous solution, the bitterness is reduced when ERG is added to a final concentration of 5 ppm or more, so the amount of ERG that has a bitterness-reducing effect relative to the arginine content in food and beverages is equivalent to 0.01 weight% or more of ERG.

[0122] [Test 5: Bitterness Reduction Effect of Soybean Peptides by ERG]

[0123] Samples (ERG0.5–10) were prepared by adding ergothioneine to a soybean peptide sample at a concentration of 1.0% of the soybean peptide Hi-Nut S (Fuji Oil Co., Ltd.) at final concentrations of 0.5 ppm, 1 ppm, 3 ppm, 5 ppm, and 10 ppm (Table 9).

[0124] For four expert panelists trained in bitter taste sensitivity, pairs were presented consisting of a control group of 1% soy peptide samples without added ERG (ERG0) and 1% soy peptide samples with added ergothioneine at final concentrations of 0.5 to 10 ppm. The panelists evaluated which of the presented pairs tasted bitter using a 2-point discrimination test. The results are shown in Table 10.

[0125] [Table 9]

[0126]

[0127] [Table 10]

[0128]

[0129] As a result, all four panelists evaluated that the 1% soybean peptide aqueous solution with ERG added to a final concentration of 0.5 ppm reduced bitterness. Since the bitterness is reduced when ERG is added to a final concentration of 0.5 ppm or higher in the 1% soybean peptide aqueous solution, the amount of ERG that has a bitterness-reducing effect relative to the soybean peptide content in food and beverages corresponds to an ERG of 0.005 wt% or more.

[0130] [Test 6: Effect of ERG on Reducing the Irritating Taste of the Sauce]

[0131] A sample (ERG5) was prepared by adding ergothioneine to a medium-thickness sauce (Kikkoman Delicious Sauce Medium-thickness: Co., Ltd.) to a final concentration of 5 ppm.

[0132] Two expert panelists were presented with pairs of a control medium-thick sauce (ERG0) without added ERG and a medium-thick sauce sample with added 5 ppm ergothioneine, and the panelists evaluated which of the presented pairs had a pungent taste (spicy and sour) using a 2-point discrimination test. The results are shown in Table 11.

[0133] [Table 11]

[0134]

[0135] The medium-thick sauce with ERG added to a final concentration of 5 ppm was evaluated to have reduced irritating taste.

[0136] [Test 7: Effect of ERG on Reducing Soybean Odor in Soy Milk]

[0137] Samples (ERG5–100) were prepared by adding ergothioneine to soy milk at final concentrations of 5 ppm, 10 ppm, 20 ppm, 40 ppm, 60 ppm, and 100 ppm (Table 12).

[0138] For five trained expert panelists, pairs of soy milk were presented, consisting of a control group without added ERG (ERG0) and soy milk with added ergothioneine at a final concentration of 5–100 ppm. The panelists evaluated which of the presented pairs they perceived as having a bean smell using a 2-point discrimination test. The results are shown in Table 13.

[0139] [Table 12]

[0140]

[0141] [Table 13]

[0142]

[0143] All panelists evaluated that the soy milk with ergothioneine added at a concentration of 20 ppm or more had a reduced soy odor. Additionally, two panelists evaluated that the soy milk with ergothioneine added at a concentration of 10 ppm or more had a reduced soy odor.

[0144] [Test Example 8: Bitter taste reduction effect of mycoprotein by ERG]

[0145] Mycoprotein was manufactured using the following manufacturing method.

[0146] Fermentation was carried out in a 30L container (manufactured by Marubishi Bioengine Co., Ltd.). Water was added to 1.5 kg of soybean pulp powder to dilute it to the mark of 15L, and 2.25 g of the defoaming agent Shin-Etsu Silicon (registered trademark) KM72F (manufactured by Shin-Etsu Chemical Industry Co., Ltd.) was added, followed by sterilization at 123°C for 60 minutes. An enzyme solution filtered through a filter with a pore size 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, it was cooled to 30°C.

[0147] 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 autoclaved (121°C, 30 minutes). 0.5ml of glycerol stock of the starter strain Aspergillus oryzae was added, and the initial spore count of the pre-culture solution was 5×10 5 New bacteria were inoculated to a volume of / ml. This pre-culture medium was shaken and cultured for 24 hours (30°C, 160 rpm). Afterward, this pre-culture medium was inoculated into a 30L container and cultured at 30°C for 3 days. The aeration and stirring speeds were started at 0.5 vvm and 250 rpm, and increased to 1 vvm and 330 rpm after 18 hours. After culture, the fermented product was heat-sterilized (80°C, 30 min).

[0148] The above-mentioned heat-sterilized fermented product was dried for 1 minute using a drum-equipped dryer (manufactured by Katsuragi Kogyo 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.

[0149] Fermented soybean pulp obtained by the above-described procedure was used, 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 extract were mixed, and the mixture was kneaded for 10 seconds using a dough mixer (manufactured by Tiger Thermos Co., Ltd., product name "Microcomputer Food Processor SKF-H101"). At this time, a sample with 330 ppm of ergothioneine added (Example) and a sample without ergothioneine added (Comparative Example) were prepared, respectively. The kneaded sample was formed to a size of 1.8 cm in thickness and 7.5 cm in inner diameter. 5 ml of oil was placed in 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.

[0150] A sensory evaluation was conducted to determine the taste of the test product, which is an example, compared to the control group, which is a comparative example.

[0151] ○: The flavor is enhanced compared to the control group.

[0152] ◇: It tastes the same as the control group.

[0153] ▽: Compared to the control group, the taste is decreasing.

[0154] [Table 14]

[0155]

[0156] With the addition of 330 ppm of ERG, the bitterness of mycoprotein was reduced and the umami taste was enhanced.

[0157] [Test Example 9: Effect of ERG on Reducing Soybean Odor in Fermented Soy Milk]

[0158] For the starter culture, an Aspergillus oryzae strain was used in which the G428S mutation was introduced into the AO090005000664 ortholog gene. 60 ml of unadjusted soy milk was placed in a sterilized 300 ml Erlenmeyer flask, the glycerol stock of the starter culture was added, and the initial spore count was 2.5 × 10⁶ 4 The culture medium was inoculated to a volume of 1 / ml. This culture medium was incubated with shaking for 48 hours (26℃, 180rpm) and then sterilized. The ergothioneine content was 37 ppm. Sensory evaluations were conducted on the soy milk before and after fermentation by five expert panelists, and all panelists evaluated that the soy milk after fermentation had a weaker soybean odor.

Claims

Claim 1 An off-flavor reducer for bitter substances, having ergothioneine as an active ingredient. Claim 2 In claim 1, the off-flavor reduction agent for a bitter substance is the reduction of the bitterness of a bitter substance, such as a soybean peptide or a mycoprotein. Claim 3 An off-flavor reducing agent for beverages containing legumes as raw materials, having ergothioneine as an active ingredient. Claim 4 In paragraph 3, the off-flavor reduction agent is the reduction of the soybean odor of soy milk. Claim 5 A food and beverage containing a bitter substance, wherein the bitterness of the soybean peptide is reduced by containing 0.005% by weight or more of ergothioneine relative to the soybean peptide content. Claim 6 Soy milk or fermented soy milk containing 10 ppm or more of ergothioneine. Claim 7 Mycoprotein containing 300 ppm or more of ergothioneine. Claim 8 A method for reducing off-flavors in food and beverages by incorporating ergothioneine into the food and beverages having off-flavors.

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