Beverage, method for producing same, and method for masking unpleasant odor of beverage

Abstract

This beverage contains beneficial bacteria. The beverage is characterized in that the dead bacteria concentration of the beneficial bacteria is at least 5 hundred million / L, and the concentration of pyrazines is at least 1 ppb.

Description

Beverage, its manufacturing method, and method for masking off-flavors in beverages

[0001] The present invention relates to a beverage, a method for producing the same, and a method for masking off-flavors in beverages.

[0002] Essential ingredients contained in foods and beverages, or additive ingredients added with the intention of achieving some effect, often have a characteristic odor, and various studies have been conducted to mask such characteristic odors (see, for example, Patent Documents 1 to 3).

[0003] Furthermore, growing health consciousness has led to an increasing need for functional foods. In particular, beneficial bacteria, especially lactic acid bacteria, are expected to have health benefits such as improving the intestinal flora and enhancing immune function, and various foods containing added lactic acid bacteria have been developed. However, lactic acid bacteria have an odor and flavor due to volatile compounds produced during the fermentation process, and some consumers find these odors and flavors to be unpleasant and off-flavorful, which could impair the palatability of the products.

[0004] To solve this problem of off-taste, for example, a masking agent for the off-taste resulting from lactic acid fermentation in lactic acid bacteria-containing foods and beverages has been investigated in Patent Document 4. Specifically, a masking agent for the off-taste of lactic acid bacteria-containing foods and beverages has been proposed, which contains a triglyceride containing a medium-chain fatty acid as an active ingredient.

[0005] Furthermore, Patent Document 5 studies an off-taste masking agent for masking the off-taste caused by lactic acid bacteria in a beverage containing lactic acid bacteria. Specifically, the off-taste masking agent is proposed, which contains calcium and / or magnesium as an active ingredient.

[0006] JP 2022-73194 A JP 2021-90371 A International Publication No. 2012 / 011402 JP 2021-023294 A JP 2017-209090 A

[0007] However, conventional methods for masking off-flavors lack a universal effect, and it has been necessary to develop a specific masking method for each type of off-flavor. Furthermore, the effectiveness is limited, and further improvements are needed. Therefore, the present invention aims to provide a beverage in which the off-flavor specific to beneficial bacteria is more effectively masked.

[0008] The present inventors have conducted extensive research to solve the above problems, and have newly discovered that when killed beneficial bacteria are added at a specific concentration, the effect of the off-odor of the beneficial bacteria in a beverage is enhanced, and that the coexistence of specific aroma components at specific concentrations can effectively mask the off-odor of the beneficial bacteria in a beverage, thereby completing the present invention.

[0009] The present invention aims to advantageously solve the above-mentioned problems, and provides a beverage comprising [1] killed beneficial bacteria, characterized in that the concentration of the killed beneficial bacteria is 500 million cells / L or more and the beverage contains pyrazines at a concentration of 1 ppb or more. In a beverage with such a composition, the unpleasant odor characteristic of the beneficial bacteria is masked.

[0010] [2] Here, it is preferable that the drink of [1] above has a dead cell concentration of the beneficial bacteria of 2,000 billion / L or less.

[0011] [3] In the beverage of [1] or [2] above, the beneficial bacteria are preferably one or more species selected from the group consisting of bacteria of the genus Lactobacillus and bacteria of the genus Lactococcus.

[0012] [4] In any of the beverages [1] to [3] above, the beneficial bacteria are preferably one or more species selected from the group consisting of Lactobacillus rhamnosus CRL1505, Lactococcus lactis subsp. lactis JCM5805, and Lactobacillus paracasei KW3110.

[0013] [5] In any of the beverages [1] to [4] above, the concentration of pyrazines is preferably 40,000 ppb or less.

[0014] [6] In any of the beverages [1] to [5] above, the concentration of pyrazines and the concentration of dead beneficial bacteria preferably satisfy the following relational expression (1): pyrazine concentration (ppb) ≥ 0.0009 × dead beneficial bacteria concentration (100 million / L) + 0.9 (1)

[0015] [7] In the beverage according to any one of [1] to [6] above, the pyrazines preferably include at least one of 2-methylpyrazine, 2,5-dimethylpyrazine, and 2,6-dimethylpyrazine.

[0016] [8] Any of the beverages described in [1] to [7] above is preferably any of a coffee beverage, a green tea beverage, a black tea beverage, and a cereal tea beverage.

[0017] [9] It is preferable that any one of the beverages [1] to [8] above is a packaged beverage.

[0018]

[10] The present invention also provides a method for producing a beverage containing killed beneficial bacteria, the method comprising a blending step of blending the killed beneficial bacteria and pyrazines so that the concentration of the killed beneficial bacteria is 500 million / L or more and the concentration of the pyrazines is 1 ppb or more. This method allows for the production of a beverage in which the unpleasant odor of the beneficial bacteria is effectively masked.

[0019]

[11] Furthermore, in the beverage manufacturing method of the above

[10] , it is preferable to further include a heat treatment step of heat treating the blend obtained in the blending step.

[0020]

[12] The present invention provides a method for masking an off-odor in a beverage containing killed beneficial bacteria, the off-odor masking method including a blending step of blending the killed beneficial bacteria and the pyrazines so that the concentration of the killed beneficial bacteria is 500 million / L or more and the concentration of the pyrazines is 1 ppb or more. This off-odor masking method can effectively mask the off-odor of the beneficial bacteria.

[0021]

[13] Here, it is preferable that the off-odor masking method of

[12] above includes a heat treatment step of heat-treating the compound obtained in the compounding step.

[0022] According to the present invention, a beverage can be provided in which the unpleasant odor specific to beneficial bacteria is masked.

[0023] (Beverage) The beverage of the present invention is a beverage containing killed beneficial bacteria. The beverage of the present invention is characterized by having a killed beneficial bacteria concentration of 500 million cells / L or more and containing pyrazines at a concentration of 1 ppb or more. Furthermore, the beverage of the present invention has an excellent flavor because the unpleasant odor characteristic of the beneficial bacteria is masked.

[0024] In the present invention, "drinks containing killed useful bacteria" include "lactic acid bacteria beverages" as defined in the "Ministerial Ordinance on the Compositional Standards of Milk and Dairy Products" (hereinafter referred to as "Milk, etc. Ordinance"). However, the "drinks containing killed useful bacteria" in the present invention are not limited to lactic acid bacteria beverages that comply with the definition of the Milk, etc. Ordinance, and may contain useful bacteria other than lactic acid bacteria. For example, it may be a milk beverage that complies with the definition of the Milk, etc. Ordinance, or a soft drink such as fruit juice, vegetable juice, soy milk, tea beverage, or coffee beverage, but more preferably a coffee beverage, green tea beverage (preferably a roasted tea beverage), black tea beverage, or grain tea beverage (preferably a barley tea beverage), more preferably a coffee beverage, roasted tea beverage, or barley tea beverage, and even more preferably a coffee beverage.

[0025] In this specification, the term "offensive odor specific to beneficial bacteria" refers to a specific odor resulting from the addition of beneficial bacteria.

[0026] <Killed Useful Bacteria> In the present invention, killed useful bacteria are used. Examples of useful bacteria include, but are not limited to, bacteria of the genus Oenococcus, Bifidobacterium, Weissella, Tetragenococcus, Lactococcus, Leuconostoc, Pediococcus, Streptococcus, Enterococcus, Lactobacillus, acetic acid bacteria, and Bacillus.

[0027] The Lactobacillus bacteria of the present invention include bacteria that were classified into the genus Lactobacillus before the reclassification of the genus Lactobacillus. For example, with the reclassification of the genus Lactobacillus, the following genera have been newly added: Acetilactobacillus, Agrilactobacillus, Amylolactobacillus, Apilactobacillus, Bombilactobacillus, Companilactobacillus, Dellaglioa, Fructilactobacillus, Furfurilactobacillus, Holzapfelia, Lacticaseibacillus, Lactiplantibacillus, and Lapidilactobacillus. This includes bacteria classified into the genera Lapidilactobacillus, Latilactobacillus, Lentilactobacillus, Levilactobacillus, Ligilactobacillus, Limosilactobacillus, Liquorilactobacillus, Loigolactobacillus, Paralactobacillus, Paucilactobacillus, Schleiferilactobacillus, Secundilactobacillus, and the like.

[0028] Among the above, the useful bacteria are preferably Oenococcus, Bifidobacterium, Lentilactobacillus, Weissella, Tetragenococcus, Lactococcus, Leuconostoc, Pediococcus, Enterococcus, Lactobacillus, and Lactiplantibacillus.Moreover, it is more preferable that the useful bacteria include one or more species selected from the group consisting of Lactobacillus and Lactococcus.

[0029] Examples of the Oenococcus include Oenococcus oeni, etc. Specific examples of the Oenococcus include Oenococcus oeni JCM6125, etc.

[0030] Examples of the Bifidobacterium bacteria include Bifidobacterium animalis subsp. lactis and Bifidobacterium longum subsp. infantis. Specific examples of the Bifidobacterium bacteria include Bifidobacterium animalis subsp. lactis JCM10602 and Bifidobacterium longum subsp. infantis JCM1222.

[0031] Examples of the Weissella genus include Weissella paramesenteroides and Weissella viridescens. Specific examples of the Weissella genus include Weissella paramesenteroides JCM9890 and Weissella viridescens JCM1174.

[0032] Examples of the Tetragenococcus bacteria include Tetragenococcus halophilus, etc. Specific examples of the Tetragenococcus bacteria include Tetragenococcus halophilus NRIC0098, etc.

[0033] Examples of the Lactococcus bacteria include Lactococcus lactis, Lactococcus lactis subsp. lactis, Lactococcus garvieae, Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. hordniae, and Lactococcus plantarum.

[0034] Specific examples of the Lactococcus bacteria include Lactococcus lactis subsp. lactis JCM5805, Lactococcus lactis subsp. lactis NBRC12007, Lactococcus lactis subsp. lactis NRIC1150, Lactococcus lactis subsp. lactis JCM20101, Lactococcus lactis subsp. lactis JCM7638, Lactococcus lactis subsp. lactis ATCC11454, Lactococcus garvieae NBRC100934, Lactococcus lactis subsp. cremoris JCM16167, Lactococcus lactis subsp. cremoris NBRC100676, Lactococcus lactis subsp. holdoniae JCM1180, Lactococcus lactis subsp. holdoniae JCM11040, and Lactococcus plantarum JCM11056.

[0035] Examples of the Leuconostoc bacteria include Leuconostoc carnosum and Leuconostoc lactis. Specific examples of the Leuconostoc bacteria include Leuconostoc carnosum JCM9695 and Leuconostoc lactis NBRC12455.

[0036] Examples of the Pediococcus bacteria include Pediococcus acidilactici, Pediococcus pentosaceus, Pediococcus cellicola, Pediococcus claussenii, Pediococcus damnosus, Pediococcus ethanolidurans, Pediococcus inopinatus, Pediococcus parvulus, and Pediococcus stilesii. Specific examples of the Pediococcus include Pediococcus acidilactici JCM8797, Pediococcus acidilactici K15, and Pediococcus damnosus JCM5886.

[0037] Examples of the genus Streptococcus include Streptococcus thermophilus, etc. Specific examples of the genus Pediococcus include Streptococcus thermophilus SBC8781, etc.

[0038] Examples of the Enterococcus bacteria include Enterococcus alcedinis.

[0039] Examples of the Lactobacillus bacteria include Lactobacillus paracasei, Lactobacillus delbrueckii, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus fructivorans, Lactobacillus hilgardii, Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus bulgaricus, and Lactobacillus parakeefilii. parakefiri), Lactobacillus plantarum, and Lactobacillus pentosus.

[0040] Specific examples of Lactobacillus bacteria include Lactobacillus paracasei KW3110, Lactobacillus paracasei MCC1849, Lactobacillus paracasei K71, Lactobacillus rhamnosus GG, Lactobacillus rhamnosus CRL1505, Lactobacillus gasseri SBT2055, Lactobacillus acidophilus L-92, Lactobacillus bulgaricus OLL1073R-1, Lactobacillus parakephili (new classification: lentilactobacillus parakephili) JCM8573, Lactobacillus plantarum (new classification: lactipranchibacillus plantarum) L-137, Lactobacillus pentosus (new classification: lactipranchibacillus pentosus) ONRICb0240, and the like.

[0041] The acetic acid bacteria are not particularly limited, but examples thereof include bacteria of the genus Gluconacetobacter, Acetobacter, and Gluconobacter, preferably bacteria of the genus Gluconacetobacter, more preferably Gluconacetobacter hansenii, and even more preferably Gluconacetobacter hansenii GK-1.

[0042] The Bacillus bacteria are not particularly limited, but include, for example, Bacillus coagulans, etc. Specific examples of Bacillus bacteria include Bacillus coagulans SANK70258 strain, etc.

[0043] Among the above, it is preferable that the useful bacteria include one or more species selected from the group consisting of Lactobacillus rhamnosus CRL1505, Lactococcus lactis subsp. lactis JCM5805, and Lactobacillus paracasei KW3110.

[0044] In the present invention, the killed useful bacteria are not particularly limited and may be dried or non-dried, but are preferably dried from the viewpoint of storage stability of the killed useful bacteria. Of these, the killed useful bacteria are preferably a dried powder of the killed useful bacteria.

[0045] The method for preparing killed useful bacteria is not particularly limited, and examples thereof include a method in which the medium in which the useful bacteria have been cultured is sterilized and then the cells are collected by filtration, centrifugation, etc., or a method in which the medium in which the useful bacteria have been cultured is filtrated, centrifugation, etc., and then the cells are collected and sterilized. Among useful bacteria, for example, lactic acid bacteria can be cultured using a culture medium for lactic acid bacteria known to those skilled in the art, such as MRS (de Man-Rogosa-Sharpe) medium, which contains glucose, protein hydrolysate, yeast extract, etc. Generally, the culture temperature is 30°C to 37°C, the culture period is 2 to 3 days, and the culture can be performed under anaerobic conditions.

[0046] The cells collected after the culture can be further dried and crushed as necessary. The sterilization method is not particularly limited, and conventional methods for killing bacteria, such as heating, ultraviolet light, or gamma-ray irradiation, can be used.

[0047] The concentration of killed useful bacteria contained in the beverage of the present invention must be 500 million / L or more, preferably 1 billion / L or more, and preferably 2,000 billion / L or less, preferably 1,000 billion / L or less, preferably 600 billion / L or less, preferably 300 billion / L or less, preferably 200 billion / L or less, preferably 100 billion / L or less, preferably 50 billion / L or less, and preferably 25 billion / L or less. When the concentration of killed useful bacteria is above the above-mentioned lower limit, the efficacy of the useful bacteria can be fully exerted. Furthermore, when the concentration of killed useful bacteria is below the above-mentioned upper limit, there is no need to add excessive amounts of pyrazines to mask the unique odor derived from the killed useful bacteria, and this can prevent increased costs and deterioration of flavor balance due to excess pyrazines. The concentration of killed useful bacteria contained in the beverage can be controlled by adjusting the amount of killed useful bacteria added to the beverage. Furthermore, the concentration of dead bacteria of beneficial bacteria contained in a beverage can be measured by any known method for measuring the number of beneficial bacteria, without any particular limitation, such as direct microscopy, particle electrical detection zone method, PCR method, or flow cytometry method, with flow cytometry method being preferred.

[0048] <Pyrazines> The beverage of the present invention contains pyrazines. Specifically, the beverage preferably contains at least one of 2-methylpyrazine, 2,5-dimethylpyrazine, and 2,6-dimethylpyrazine as the pyrazine, and more preferably contains 2-methylpyrazine. The pyrazines may be added in the form of a preparation, or may be added as an ingredient from a food containing pyrazines, such as coffee or barley tea. When a food containing pyrazines, such as coffee, is added as a pyrazine source, a mixture of multiple types of pyrazines may be added.

[0049] The concentration of pyrazines must be 1 ppb or more, preferably 5 ppb or more, more preferably 10 ppb or more, more preferably 25 ppb or more, more preferably 50 ppb or more, more preferably 100 ppb or more, even more preferably 200 ppb or more, particularly preferably 500 ppb or more, preferably 40,000 ppb or less, more preferably 30,000 ppb or less, even more preferably 20,000 ppb or less, even more preferably 15,000 ppb or less, even more preferably 10,000 ppb or less, particularly preferably 5,000 ppb or less. If the concentration of pyrazines is above the lower limit, the unpleasant odor specific to useful bacteria can be effectively reduced. If the concentration of pyrazines is below the upper limit, the increase in the metallic odor derived from pyrazines can be suppressed. It is preferable that when the beverage contains only one type of pyrazine among the compounds listed above, the content of the pyrazine satisfies the above range, and when the beverage contains multiple types, the total content of the multiple types of pyrazine satisfies the above range. The concentration of pyrazines can be measured according to the method described in the Examples below.

[0050] In the beverage of the present invention, the concentration of pyrazines and the concentration of dead beneficial bacteria preferably satisfy the following relational expression (1): If the concentration of pyrazines and the concentration of dead beneficial bacteria satisfy the following relational expression (1), the unpleasant odor specific to the beneficial bacteria can be effectively masked: Pyrazine concentration (ppb) ≥ 0.0009 × dead beneficial bacteria concentration (100 million / L) + 0.9 (1)

[0051] <Other Ingredients> The beverage of the present invention may contain one or more additives selected from the group consisting of acidulants, flavorings, colorings, sweeteners, preservatives, thickeners, stabilizers, emulsifiers, dietary fiber, bittering agents, antioxidants, pH adjusters, vitamins, nutritional fortifiers, umami components, dietary fiber, extracts, solvents, minerals, water-soluble functional ingredients, and fat-soluble functional ingredients, to the extent that the effects of the present invention are not impaired. The additives are not particularly limited and commonly used ones can be used, but specific examples include acesulfame K, stevia, and sucralose as sweeteners, citric acid as acidulants, soybean polysaccharides and pectin as stabilizers, and sodium, potassium, magnesium, and calcium as minerals.

[0052] The beverage of the present invention may also be an immunostimulating composition. An immunostimulating composition is a composition having immunostimulating ability (immune activation ability). Immunostimulating ability (immune activation ability) refers to the stimulating effect (activation effect) of the innate immune system on cells or living organisms. The immunostimulating composition according to one embodiment may be a composition for activating dendritic cells, or a composition for activating plasmacytoid dendritic cells (pDCs).

[0053] The beverage of the present invention as an immunostimulating composition may be a pharmaceutical composition or a quasi-drug. The beverage of the present invention as an immunostimulating composition contains an effective amount of killed bacteria of the above-mentioned beneficial bacteria. Here, "effective amount" refers to the content of the beverage of the present invention such that when ingested in a normally consumed amount, the killed bacteria of the beneficial bacteria are ingested to an extent that the effects of immunostimulation, etc. are exerted. When the beverage of the present invention is an immunostimulating composition, the beverage of the present invention may be a health food, functional food, nutritional supplement, health functional food (e.g., specified health food, nutritional functional food, functional food), special purpose food (e.g., food for infants, food for pregnant women, food for sick people), or supplement.

[0054] When the beverage of the present invention is provided as an immunostimulating composition, the beverage of the present invention may be, in particular, a health food, functional food, nutritional composition, dietary supplement, health food, food for specified health uses, food with nutrient functions, or food with functional claims, etc. When the beverage of the present invention is an immunostimulating composition, the beverage of the present invention can be labeled, for example, for supporting the maintenance of immune function in healthy people (immune care), for those concerned about a decline in immune function, for suppressing a decline in immune function, for those concerned about sunburn, for those concerned about damage to the skin in daily life, for those concerned about dry skin, for those concerned about hot flashes, for those concerned about erythema of the skin, for those concerned about redness of the skin, for those concerned about rosy face, for those concerned about rough hands, etc.

[0055] Furthermore, when the beverage of the present invention is an immunostimulating composition, the beverage of the present invention can be consumed by a subject in need of immunostimulation, which is not particularly limited and may include, for example, a subject infected with a virus, a subject suffering from a cold, or a subject aged 65 or over.

[0056] (Method for Producing Beverage) The method for producing a beverage of the present invention includes a blending step of blending killed beneficial bacteria and pyrazines at predetermined concentrations. This blending step is a step of blending the killed beneficial bacteria concentration to 500 million / L or more and the pyrazines concentration to 1 ppb or more. The method for producing a beverage of the present invention is not particularly limited as long as it includes the above-mentioned specific blending step. In other words, the beverage can be produced according to a conventionally known method for producing a beverage as long as it includes the above-mentioned blending step.

[0057] A blending process for blending useful bacteria to a killed cell concentration of 500 million cells / L or more and pyrazines to a concentration of 1 ppb or more can include, for example, adding pyrazines to a mixing tank in an amount that results in a concentration of 1 ppb or more, optionally a solvent such as water, and other optional components, and then adding killed useful bacteria to the mixture at a rate of 500 million cells / L or more. Alternatively, the blending process can include simultaneously adding killed useful bacteria, pyrazines, and an optional solvent to the mixing tank. Of course, the addition method and blending order are not limited to those described above.

[0058] The beverage of the present invention does not necessarily have to be a packaged beverage, but is preferably a packaged beverage. Examples of such containers include plastic containers (resin bottles) such as PET bottles, polypropylene bottles, and polyvinyl chloride bottles, as well as glass bottles and cans. The volume of the container is not particularly limited, but may be, for example, 100 mL or more, preferably 185 mL or more, and may be, for example, 1000 mL or less, preferably 680 mL or less. The concentration of pyrazines can be measured by the method described in the Examples of this specification.

[0059] A packaged beverage can be produced by filling the beverage obtained according to the above-described production method of the present invention into a container such as those listed above and sealing it in a known manner.

[0060] The method for producing a beverage of the present invention may also include a heat treatment step in which the blend obtained in the blending step is heat-treated. From the viewpoint of improving shelf life, it is preferable to include a heat treatment step. The method and conditions for the heat treatment step may be, for example, methods and conditions typically used for beverages such as packaged beverages. Preferred are retort sterilization, UHT (Ultra High Temperature) sterilization, HTST (High Temperature Short Time) sterilization, or pasteurizer sterilization.

[0061] (Method for masking off-odors in beverages) The method for masking off-odors in beverages of the present invention is a method for masking off-odors in beverages containing killed beneficial bacteria. Such an off-odor masking method is not particularly limited as long as it includes a blending step of blending killed beneficial bacteria and pyrazines at predetermined concentrations. The blending step is characterized by blending the killed beneficial bacteria at a concentration of 500 million / L or more and the pyrazines at a concentration of 1 ppb or more.

[0062] The blending process for blending so that the concentration of dead beneficial bacteria is 500 million / L or more and the concentration of pyrazines is 1 ppb or more can be the same as the process described in relation to the method for producing the beverage of the present invention.

[0063] The method for masking an off-flavor in a beverage of the present invention may include a heat treatment step of heat-treating the blend obtained in the blending step. The heat treatment step may be the same as the step described in relation to the method for producing a beverage of the present invention.

[0064] The present invention will be specifically described below based on exemplary tests, but the present invention is not limited to these. Various measurements and evaluations were carried out on samples from each test group described below using the following methods.

[0065] <Concentration of Pyrazines> <<Quantification Method>> The pyrazine content in the beverage was measured using gas chromatography with a mass spectrometer (GC / MS). Specifically, this GC / MS analysis was performed as follows. First, the aroma components in the sample were separated using a C18 solid-phase extraction column, and the ethyl acetate eluted fraction was used as the analytical sample. Quantification was performed using the internal standard method, using borneol as the internal standard, which was added to the analytical sample to a concentration of 25 ppb. The conditions for the GC / MS analysis are shown in Table 1 below.

[0066]

[0067] (Test 1) Effect of sterilization conditions of beneficial bacteria on unpleasant odors in beverages The following test was conducted to investigate the relationship between sterilization conditions of beneficial bacteria and unpleasant odors specific to beneficial bacteria in beverages.

[0068] <Sample preparation> 350 billion / g live rhamnosus bacteria powder (including one or more species of Lactobacillus rhamnosus) was diluted 35 times with water, and then heated at 80 ° C for 1 hour to prepare a 10 billion / g killed rhamnosus bacteria aqueous solution. This was diluted with water to a rhamnosus killed bacteria concentration of 12 billion / L to obtain a diluted rhamnosus killed bacteria aqueous solution, which was then filled into a can to prepare a sample (beverage) for test area 1. In addition, the same diluted rhamnosus killed bacteria aqueous solution as above was heated under the conditions listed in Table 2 to prepare samples for test areas 2 to 3.

[0069] <Sensory Evaluation> Six trained panelists with sensory discrimination abilities evaluated the off-flavors of the beverages obtained in the above test plots, which had been prepared at approximately 20°C, in increments of 0.5 points according to the criteria established in Test 2 described below, and the average scores of the six panelists were calculated. The standard error of the average scores of all panelists was 0.2 or less. The results are shown in Table 2.

[0070] (Test 2) Effect of the concentration of dead beneficial bacteria on the unpleasant odor of beverages The following test was carried out to investigate the relationship between the concentration of dead beneficial bacteria and the unpleasant odor specific to beneficial bacteria in beverages.

[0071] <Sample Preparation> A 350 billion / g live rhamnosus cell bulk powder (containing one or more species of Lactobacillus rhamnosus) was diluted 35 times with water and then heated for 1 hour at 80°C to prepare an aqueous solution of 10 billion / g killed rhamnosus cells. This was diluted with water to the killed rhamnosus cell concentration shown in Table 3, filled into cans, and heated at 121°C for 5 minutes to prepare samples (beverages) corresponding to test plots 4 to 12.

[0072] <Sensory Evaluation> Six trained panelists with sensory discrimination abilities rated the beverages obtained in the above test plots, which were prepared at approximately 20°C, using a fixed score of 0 for samples without added beneficial bacteria (Rhamnosus) and 5 for samples with 2,000 billion added beneficial bacteria. The 0-5 point interval was divided equally to set a standard for each score of 1 point, and based on this standard, the beverages were rated on a scale of 0 to 5 in 0.5 increments (the lower the score, the better). The average scores of the six panelists were calculated. The results are shown in Table 3. The standard error of the average scores for all panelists was less than 0.2. Off-flavors, including buttery odors, were evaluated, including off-flavors derived from lactic acid bacteria such as Rhamnosus.

[0073] (Test 3) Effect of pyrazine concentration on reducing the unpleasant odor of beverages with a dead beneficial bacteria concentration of 1 billion / L The following test was conducted to investigate the relationship between the pyrazine concentration and the unpleasant odor specific to beneficial bacteria in beverages with a dead beneficial bacteria concentration of 1 billion / L.

[0074] <Sample Preparation> 350 billion / g live rhamnosus bacteria powder (containing one or more species of Lactobacillus rhamnosus) was diluted 35 times with water and then heated at 80°C for 1 hour to prepare a 10 billion / g killed rhamnosus bacteria aqueous solution. This was mixed with pyrazines and water to the types and concentrations listed in Table 4, filled into a can, and heated at 121°C for 5 minutes to prepare samples corresponding to test plots 13-21, H (roasted green tea) 3, and M (barley tea) 3. Pyrazines were not added to the sample corresponding to test plot 6. Multiple coffee extracts were added to the sample corresponding to test plot 21 so that the types and concentrations of pyrazines contained in the sample were as shown in Table 4. Furthermore, roasted green tea (H3) and barley tea (M3) were added to test plots H3 and M3, respectively, so that the total concentration of pyrazines was as shown in Table 4.

[0075] <Sensory Evaluation> For the beverages obtained in the above test groups, a sample without added pyrazines was given a fixed score of 0, and a sample with added pyrazines at 20,000 ppb was given a fixed score of 4. The interval from 0 to 4 was equally divided to set a standard for each score of "1 point." Based on this standard, the beverages were rated on a scale of 0.5 points from 0 to 4 points (the higher the score, the better). The average scores of the six panelists were calculated. The standard error of the average scores of all panelists was less than 0.2. The results are shown in Table 4.

[0076] (Test 4) Effect of pyrazine concentration on reducing the unpleasant odor of beverages with a dead beneficial bacteria concentration of 10 billion / L The following test was conducted to investigate the relationship between the pyrazine concentration and the unpleasant odor specific to beneficial bacteria in beverages with a dead beneficial bacteria concentration of 10 billion / L.

[0077] <Sample Preparation> 350 billion / g live rhamnosus bacteria powder (containing one or more species of Lactobacillus rhamnosus) was diluted 35 times with water and then heated at 80°C for 1 hour to prepare a 10 billion / g killed rhamnosus bacteria aqueous solution. This was mixed with pyrazines and water to the types and concentrations listed in Table 5, filled into a can, and heated at 121°C for 5 minutes to prepare samples corresponding to test plots 22-30, H4, and M4. No pyrazines were added to the sample corresponding to test plot 7. For the sample corresponding to test plot 30, multiple coffee extracts were added so that the types and concentrations of pyrazines contained in the sample were as shown in Table 5. Furthermore, for test plots H4 and M4, roasted green tea (H4) and barley tea (M4) were added, respectively, so that the total concentration of pyrazines was as shown in Table 5.

[0078] <Sensory Evaluation> The beverages obtained in the above test plots were subjected to a sensory evaluation in the same manner as in the sensory evaluation test in Test 3. The results are shown in Table 5.

[0079] (Test 5) Effect of pyrazine concentration on reducing the unpleasant odor of a beverage with a dead beneficial bacteria concentration of 100 billion / L The following test was conducted to investigate the relationship between the pyrazine concentration and the unpleasant odor specific to beneficial bacteria in a beverage with a dead beneficial bacteria concentration of 100 billion / L.

[0080] <Sample Preparation> 350 billion / g live rhamnosus bacteria powder (containing one or more species of Lactobacillus rhamnosus) was diluted 35 times with water and then heated at 80°C for 1 hour to prepare a 10 billion / g killed rhamnosus bacteria aqueous solution. This was mixed with pyrazines and water to the types and concentrations listed in Table 6, filled into a can, and heated at 121°C for 5 minutes to prepare samples corresponding to test plots 31-39, H5, and M5. No pyrazines were added to the sample corresponding to test plot 8. For test plot 39, multiple coffee extracts were added so that the types and concentrations of pyrazines contained in the sample were as shown in Table 6. Furthermore, for test plots H5 and M5, roasted green tea (H5) and barley tea (M5) were added, respectively, so that the total concentration of pyrazines was as shown in Table 6.

[0081] <Sensory Evaluation> The beverages obtained in the above test plots were subjected to a sensory evaluation using the same method as in the sensory evaluation test in Test 3. The results are shown in Table 6.

[0082] (Test 6) Effect of pyrazine concentration on reducing the unpleasant odor of a beverage with a dead beneficial bacteria concentration of 500 billion / L The following test was conducted to investigate the relationship between the pyrazine concentration and the unpleasant odor specific to beneficial bacteria in a beverage with a dead beneficial bacteria concentration of 500 billion / L.

[0083] <Sample Preparation> 350 billion / g live rhamnosus bacteria powder (containing one or more species of Lactobacillus rhamnosus) was diluted 35 times with water and then heated at 80°C for 1 hour to prepare a 10 billion / g killed rhamnosus bacteria aqueous solution. This was mixed with pyrazines and water to the types and concentrations listed in Table 7, filled into a can, and heated at 121°C for 5 minutes to prepare samples corresponding to test plots 40-48, H6, and M6. No pyrazines were added to the sample corresponding to test plot 10. For the sample corresponding to test plot 48, multiple coffee extracts were added so that the types and concentrations of pyrazines in the sample were as shown in Table 7. Furthermore, for test plots H6 and M6, roasted green tea (H6) and barley tea (M6) were added, respectively, so that the total concentration of pyrazines was as shown in Table 7.

[0084] <Sensory Evaluation> The beverages obtained in the above test plots were subjected to a sensory evaluation in the same manner as in the sensory evaluation test in Test 3. The results are shown in Table 7.

[0085] (Test 7) ​​Effect of pyrazine concentration on reducing the unpleasant odor of beverages with a dead beneficial bacteria concentration of 1,000 billion / L The following test was conducted to investigate the relationship between the pyrazine concentration and the unpleasant odor specific to beneficial bacteria in beverages with a dead beneficial bacteria concentration of 1,000 billion / L.

[0086] <Sample Preparation> 350 billion / g live rhamnosus bacteria powder (containing one or more species of Lactobacillus rhamnosus) was diluted 35 times with water and then heated at 80°C for 1 hour to prepare a 10 billion / g killed rhamnosus bacteria aqueous solution. This was mixed with pyrazines and water to the types and concentrations listed in Table 8, filled into a can, and heated at 121°C for 5 minutes to prepare samples corresponding to test plots 49-57, H7, and M7. No pyrazines were added to the sample corresponding to test plot 11. Multiple coffee extracts containing pyrazines of the types and concentrations listed in Table 8 were added to the sample corresponding to test plot 57. Furthermore, roasted green tea (H7) and barley tea (M7) were added to test plots H7 and M7, respectively, so that the total concentration of pyrazines was as shown in Table 8.

[0087] <Sensory Evaluation> The beverages obtained in the above test plots were subjected to a sensory evaluation using the same method as in the sensory evaluation test in Test 3. The results are shown in Table 8.

[0088] (Test 8) Effect of pyrazine concentration on reducing the unpleasant odor of beverages with a dead beneficial bacteria concentration of 2,000 billion / L The following test was conducted to investigate the relationship between the pyrazine concentration and the unpleasant odor specific to beneficial bacteria in beverages with a dead beneficial bacteria concentration of 2,000 billion / L.

[0089] <Sample Preparation> 350 billion / g live rhamnosus bacteria powder (containing one or more species of Lactobacillus rhamnosus) was diluted 35 times with water and then heated at 80°C for 1 hour to prepare a 10 billion / g killed rhamnosus bacteria aqueous solution. This was mixed with pyrazines and water to the types and concentrations listed in Table 9, filled into a can, and heated at 121°C for 5 minutes to prepare samples corresponding to test plots 58-66, H8, and M8. No pyrazines were added to the sample corresponding to test plot 12. For the sample corresponding to test plot 66, multiple coffee extracts were added so that the types and concentrations of pyrazines in the sample were as shown in Table 9. Furthermore, for test plots H8 and M8, roasted green tea (H8) and barley tea (M8) were added, respectively, so that the total concentration of pyrazines was as shown in Table 9.

[0090] <Sensory Evaluation> The beverages obtained in the above test plots were subjected to a sensory evaluation using the same method as in the sensory evaluation test in Test 3. The results are shown in Table 9.

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[0099] Table 2 shows that the higher the heating temperature for beverages containing beneficial bacteria, the stronger the off-flavor characteristic of the beneficial bacteria (in the above example, lactic acid bacteria). Furthermore, when heated at a constant temperature and time, specifically, when heated for 5 minutes at 121°C (see Table 3), the off-flavor characteristic of beneficial bacteria occurs in test plots 5-12, which contain beneficial bacteria at a concentration of 500 million / L or more. Tables 4 and 5 show that in test plots 13-30, H3-4, and M3-4, where the killed bacteria concentration of beneficial bacteria is 1 billion / L and 10 billion / L, respectively, the off-flavor reduction effect is observed when pyrazines are contained at 1 ppb or more. Furthermore, it can be seen that the off-flavor reduction effect is similar for coffee extracts, roasted green tea, and barley tea containing 2-methylpyrazine alone, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine alone, and mixtures of the above pyrazines. Tables 6 to 9 show that even in test plots 31 to 66, H5 to 8, and M5 to 8, where the concentration of dead bacteria of beneficial bacteria is in the range of 100 billion cells / L or more and 2,000 billion cells / L or less, the off-odor reduction effect is observed when pyrazines are contained in an amount of 1 ppb or more. It is also clear that the same level of off-odor reduction effect is achieved with coffee extracts, roasted green tea, and barley tea containing not only 2-methylpyrazine alone, but also 2,5-dimethylpyrazine and 2,6-dimethylpyrazine alone, as well as mixtures of the above pyrazines.

[0100] According to the present invention, a beverage can be provided in which the unpleasant odor specific to beneficial bacteria is masked.

Claims

1. A beverage containing dead beneficial bacteria, the concentration of the dead beneficial bacteria being 500 million / L or more, and containing pyrazines at a concentration of 1 ppb or more.

2. The beverage according to claim 1, wherein the concentration of dead beneficial bacteria is 2,000 billion / L or less.

3. The beverage according to claim 1, wherein the beneficial bacteria is one or more species selected from the group consisting of Lactobacillus and Lactococcus.

4. The beverage according to claim 1, wherein the beneficial bacteria is one or more species selected from the group consisting of Lactobacillus rhamnosus CRL1505, Lactococcus lactis subsp. lactis JCM5805, and Lactobacillus paracasei KW3110.

5. The beverage according to claim 1, wherein the concentration of pyrazines is 40,000 ppb or less.

6. The beverage according to claim 1, wherein the concentration of pyrazines and the concentration of dead beneficial bacteria satisfy the following relational expression (1): pyrazine concentration (ppb) ≧ 0.0009 × dead beneficial bacteria concentration (100 million / L) + 0.9 (1) 7. The beverage of claim 1, wherein the pyrazines include at least one of 2-methylpyrazine, 2,5-dimethylpyrazine, and 2,6-dimethylpyrazine.

8. The beverage according to any one of claims 1 to 7, which is a coffee beverage, a green tea beverage, a black tea beverage, or a cereal tea beverage.

9. The beverage according to any one of claims 1 to 7, which is a packaged beverage.

10. A method for producing a beverage containing dead useful bacteria, comprising a blending step of blending the dead useful bacteria and pyrazines so that the dead useful bacteria concentration is 500 million / L or more and the pyrazines concentration is 1 ppb or more.

11. A method for producing the beverage according to claim 10, further comprising a heat treatment step of heat treating the blend obtained in the blending step.

12. A method for masking an odor of a beverage containing dead useful bacteria, comprising a blending step of blending the dead useful bacteria and pyrazines so that the dead useful bacteria concentration is 500 million / L or more and the pyrazines concentration is 1 ppb or more.

13. The odor masking method according to claim 12, further comprising a heat treatment step of heat treating the mixture obtained in the blending step.

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