Bottled unsweetened beverages

Abstract

The present invention addresses the problem of providing a packaged sugar-free beverage which suppresses an "in-mouth astringency" unique to sugar-free beverages containing a prescribed concentration of magnesium, a method for producing the same, and the like. Provided as a solution is a packaged sugar-free beverage comprising one or more types of lactic acid bacteria and / or dead bacteria of the present invention 1 in the amount of not less than 500,000,000 / L (for example, not less than 20,000,000,000 / L), wherein the magnesium concentration is not less than 1.2 mg / L.

Description

[Technical Field]

[0001] The present invention relates to a packaged sugar-free beverage containing magnesium, and dead cells of "lactic acid bacteria" and / or "bacteria capable of activating plasmacytoid dendritic cells," and to a method for producing the same. More specifically, in packaged sugar-free beverages containing a predetermined concentration of magnesium, a characteristic "mouth squeaking" (i.e., "a squeaking sensation as if the tongue or throat is being pulled," more specifically, "a feeling of fatigue as if the tongue or throat is being pulled as if it is cramping") occurs when drinking. The present invention relates to a packaged sugar-free beverage in which such characteristic "mouth squeaking" is suppressed, and to a method for producing the same. [Background technology]

[0002] The innate immune system is primarily responsible for the primary response to bacterial or viral infections, and dendritic cells are among its most powerful and important constituent cells. Furthermore, plasmacytoid dendritic cells (pDCs), in particular, are major producers of various interferons (IFNs), including type I interferons, which exhibit inhibitory activity against viral replication, and thus play an extremely important role in antiviral defense (Patent Document 1). On the other hand, magnesium is one of the essential minerals, present in bones and bodily fluids, and plays an important role in various enzymatic processes. Magnesium, along with calcium, forms bones and plays crucial roles in muscle and nerve function. It is said that many people are deficient in magnesium due to factors such as the Westernization of their diet.

[0003] Sports drinks are known as beverages for replenishing fluids, magnesium, and other nutrients. Magnesium is known to have a bitter taste, and in beverages containing relatively high amounts of magnesium, sweeteners are added to mitigate this bitterness (background art of Patent Document 2). Patent Document 2 discloses a method for suppressing bitterness caused by magnesium and other factors by combining rebaudioside A and mogroside V, which are natural high-intensity sweeteners.

[0004] However, it was previously unknown that a novel problem arises with magnesium-containing sugar-free beverages: a unique "mouth squeaking" sensation (i.e., a "pushing sensation around the tongue and throat," or more specifically, a "fatiguing feeling of being pulled and strained around the tongue and throat"), and that this unique "mouth squeaking" sensation can be suppressed by including dead bacteria of "lactic acid bacteria" and / or "bacteria capable of activating plasmacytoid dendritic cells" in such sugar-free beverages at a predetermined concentration. [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] International Release 2012 / 091081 Pamphlet [Patent Document 2] Japanese Patent Publication No. 2019-129769 [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] The inventors of this invention have discovered that a novel problem, a unique "gritty feeling in the mouth," arises in sugar-free beverages containing magnesium. The object of the present invention is to provide a packaged sugar-free beverage in which the "gritty feeling in the mouth" characteristic of sugar-free beverages containing a predetermined concentration of magnesium is suppressed, as well as a method for producing the same. [Means for solving the problem]

[0007] The inventors of the present invention, after diligent research to solve the problems of the present invention, found that by including 500 million or more dead cells (for example, 20 billion or more) of "lactic acid bacteria" and / or "bacteria capable of activating plasmacytoid dendritic cells" in a sugar-free beverage containing a predetermined concentration of magnesium, the aforementioned characteristic "squeaky feeling in the mouth" can be suppressed, and thus the present invention was completed.

[0008] In other words, the present invention provides the following inventions, etc. (1) A bottled, sugar-free beverage containing 500 million or more dead lactic acid bacteria of one or more types (e.g., 20 billion or more) and having a magnesium concentration of 1.2 mg / L or more; (2) A carbonated beverage with a gas pressure of 0.1 to 0.3 MPa, as described in (1) above; (3) A packaged sugar-free beverage as described in (1) or (2) above, further containing one or more organic acids, wherein the total concentration of the organic acids is 0.005 to 0.5% by weight; (4) A packaged sugar-free beverage containing 500 million or more dead cells (for example, 20 billion or more) of one or more types of lactic acid bacteria, and having a magnesium concentration of 1.2 mg / L or more, wherein the lactic acid bacteria include bacteria of the genus Lactobacillus, Streptococcus, Lactococcus, Leuconostoc, and Pediococcus. The packaged sugar-free beverage contains one or more bacteria selected from the group consisting of bacteria of the genus *Ccus*, *Enterococcus*, *Oenococcus*, *Bifidobacterium*, *Lentilactobacillus*, *Weissella*, and *Tetragenococcus*; (5) The unsweetened beverage in a container as described in (4) above, characterized in that the lactic acid bacteria is Lactobacillus rhamnosus; (6) A bottled sugar-free beverage containing 500 million or more dead bacteria (e.g., 20 billion or more) of one or more species capable of activating plasmacytoid dendritic cells, and having a magnesium concentration of 1.2 mg / L or more; (7) A method for producing a packaged sugar-free beverage having a magnesium concentration of 1.2 mg / L or more, characterized in that the beverage contains 500 million or more dead cells (for example, 20 billion or more) of one or more types of bacteria capable of activating plasmacytoid dendritic cells; (8) A method for producing a packaged unsweetened beverage having a magnesium concentration of 1.2 mg / L or more, characterized in that the beverage contains 500 million or more dead cells (for example, 20 billion or more) of one or more types of lactic acid bacteria; (9) A method for suppressing a squeaky feeling in the mouth when drinking a packaged sugar-free beverage having a magnesium concentration of 1.2 mg / L or more, characterized by containing 500 million or more (e.g., 20 billion or more) dead bacteria of one or more types of bacteria capable of activating plasmacytoid dendritic cells in the beverage; (10) A method for suppressing a squeaky feeling in the mouth when drinking a packaged sugar-free beverage having a magnesium concentration of 1.2 mg / L or more, characterized by containing 500 million or more dead cells (for example, 20 billion or more) of one or more types of lactic acid bacteria in the beverage; The present invention also provides the following inventions, etc. [1] A bottled, sugar-free beverage containing 20 billion or more dead lactic acid bacteria of one or more types, and having a magnesium concentration of 1.2 mg / L or more; [2] A carbonated beverage with a gas pressure of 0.1 to 0.3 MPa, as described in [1] above; [3] A bottled sugar-free beverage according to [1] or [2] above, further containing one or more organic acids, wherein the total concentration of the organic acids is 0.005 to 0.5% by weight; [4] A method for producing a packaged sugar-free beverage having a magnesium concentration of 1.2 mg / L or more, the method comprising causing the beverage to contain 20 billion or more dead cells of one or more lactic acid bacteria per liter; [5] A method for suppressing astringent roughness in the mouth when drinking a packaged sugar-free beverage having a magnesium concentration of 1.2 mg / L or more, the method comprising causing the beverage to contain 20 billion or more dead cells of one or more lactic acid bacteria per liter; Effects of the Invention

[0009] According to the present invention, there can be provided a packaged sugar-free beverage in which "astringent roughness in the mouth" (hereinafter sometimes simply referred to as "astringent roughness in the mouth" in the present specification) characteristic to sugar-free beverages containing a predetermined concentration of magnesium is suppressed, a method for producing the same, and the like. Brief Description of Drawings

[0010] [Figure 1] Fig. 1 is a diagram showing the relationship between Lactococcus lactis subsp. lactis strain JCM5805 and strains equivalent to said strain (strains derived from said strain and strains from which said strain is derived). Mode for Carrying Out the Invention

[0011] The present invention provides: [1] A packaged sugar-free beverage containing 500 million or more (for example, 20 billion or more) dead cells of one or more bacteria capable of activating plasmacytoid dendritic cells per liter, and having a magnesium concentration of 1.2 mg / L or more (hereinafter, also referred to as "the beverage of Invention 1"); [2] A packaged sugar-free beverage containing 500 million or more (for example, 20 billion or more) dead cells of one or more lactic acid bacteria per liter, and having a magnesium concentration of 1.2 mg / L or more (hereinafter, also referred to as "the beverage of Invention 2"); [3] In the production of a packaged sugar-free beverage having a magnesium concentration of 1.2 mg / L or more, the method for producing the packaged sugar-free beverage, which is characterized in that the beverage contains 500 million cells / L or more (for example, 20 billion cells / L or more) of dead cells of one or more bacteria capable of activating plasmacytoid dendritic cells (hereinafter also referred to as "the production method of the present invention 1"); [4] In the production of a packaged sugar-free beverage having a magnesium concentration of 1.2 mg / L or more, the method for producing the packaged sugar-free beverage, which is characterized in that the beverage contains 500 million cells / L or more (for example, 20 billion cells / L or more) of dead cells of one or more lactic acid bacteria (hereinafter also referred to as "the production method of the present invention 2"); [5] In the production of a packaged sugar-free beverage having a magnesium concentration of 1.2 mg / L or more, the method for suppressing astringency in the mouth when drinking the packaged sugar-free beverage, which is characterized in that the beverage contains 500 million cells / L or more (for example, 20 billion cells / L or more) of dead cells of one or more bacteria capable of activating plasmacytoid dendritic cells (hereinafter also referred to as "the suppression method of the present invention 1"); [6] In the production of a packaged sugar-free beverage having a magnesium concentration of 1.2 mg / L or more, the method for suppressing astringency in the mouth when drinking the packaged sugar-free beverage, which is characterized in that the beverage contains 500 million cells / L or more (for example, 20 billion cells / L or more) of dead cells of one or more lactic acid bacteria (hereinafter also referred to as "the suppression method of the present invention 2"); The present invention includes embodiments such as those described above.

[0012] (Bacteria capable of activating pDC) The "bacteria capable of activating pDC" in the present invention 1 (hereinafter also referred to as "the bacteria in the present invention 1") is not particularly limited, and refers to, for example, lactic acid bacteria, acetic acid bacteria, bacteria of the genus Escherichia, bacteria of the genus Bacillus, cyanobacteria and the like capable of activating pDC, and one or more bacteria selected from the group consisting of lactic acid bacteria, acetic acid bacteria, and bacteria of the genus Bacillus can be mentioned. The bacteria in the present invention 1 are dead cells.

[0013] The "bacteria capable of activating pDCs" in Invention 1 are capable of inducing the production of IFN. Preferably, the IFN is at least one of Type I IFN (Type I interferon), Type II IFN (Type II interferon), or Type III IFN (Type III interferon). Type I IFN refers to cytokines that are effective against viral infections, and includes, for example, IFN-α (including subtypes such as 1, 2, 4, 5, 6, 7, 8, 10, 13, 14, 16, 17, or 21) or IFN-β. Type II IFN includes IFN-γ, and Type III IFN includes IFN-λ. Preferably, the "bacteria capable of activating pDCs" in Invention 1 have the activity to induce the production of at least Type I IFN.

[0014] The IFNs that can be induced to be produced by the "bacteria capable of activating pDCs" in Invention 1 are not particularly limited as long as they belong to any of Type I IFN, Type II IFN, or Type III IFN, but it is preferable that they be one or more selected from the group consisting of IFN-α, IFN-β, and IFN-λ, more preferably that at least one of the IFNs is IFN-α, even more preferably that at least one of the IFNs is IFN-α and two or more selected from the group consisting of IFN-α, IFN-β, and IFN-λ, and it is particularly preferable that at least two of the two or more IFNs are IFN-α and IFN-β.

[0015] Whether bacteria can activate pDCs can be confirmed by whether the pDCs phagocytose the bacteria, whether cellular protrusions appear on the surface of the phagocytosed pDCs, or whether the phagocytosed pDCs produce IFNs (Type I IFNs and / or Type III IFNs, etc.) when the bacteria are presented to the pDCs.

[0016] Phagocytosis of pDCs or the appearance of cellular protrusions on the surface of pDCs can be confirmed, for example, by microscopic observation or flow cytometry. Preferably, this involves applying bacteria modified with a fluorescent dye to pDCs and then observing the pDCs.

[0017] The production of IFNs (Type I IFNs and / or Type III IFNs, etc.) can be confirmed, for example, by measuring the amount or concentration of IFNs such as IFN-α or IFN-β in the culture system when bacteria are cultured in the presence of pDCs derived from mammalian bone marrow cells such as those of mice.

[0018] The IFN concentration can be determined by measuring it using the following procedure (i) to (iv). (i) De-erythrocyte-removed mouse bone marrow cells are placed in RPMI medium prepared to the following composition in a 1 × 10⁶ state. 6 Suspend the cells to a concentration of cells / mL to prepare a cell suspension. <Composition of culture medium> • 10% FBS by volume • 100 U / mL Penicillin / Streptomycin 1 mM sodium pyruvate 2.5mM HEPES ·1 mass%MEM NEAA 50 μM β-mercaptoethanol • 100 ng / mL Flt-3L (ii) Seed 1 mL of the prepared cell suspension into each cell and culture in a CO2 incubator at 37°C and 5% CO2 for 1 week to induce pDCs. (iii) Bone marrow cells containing induced pDCs, 2 × 10 5 Suspend the bacteria to a concentration of cells / mL, seed 200 μL into each 96-well plate, and add 2 μL of bacterial suspension, such as lactic acid bacteria, adjusted to a concentration of 1 mg / mL in PBS, to each plate. (iv) After 24 hours, collect the culture supernatant and measure the IFN-α concentration by ELISA using an IFN-α measurement kit.

[0019] In Invention 1, the "bacteria capable of activating pDCs" refers to "the bacterium with a final concentration of 10 μg / mL" and "a bacterium containing pDCs, obtained by recovering it from mouse bone marrow and culturing it for 7 days in a cell culture medium containing 100 ng / mL Flt3-L, with a final concentration of 2 × 10⁻¹⁶." 5 This can be expressed as an index that produces 30 pg / mL (preferably 50 pg / mL, more preferably 60 pg / mL, more preferably 70 pg / mL, 80 pg / mL, 90 pg / mL, 100 pg / mL, 150 pg / mL, even more preferably 200 pg / mL, 210 pg / mL, 220 pg / mL, 230 pg / mL, 240 pg / mL, 250 pg / mL, even more preferably 300 pg / mL, 400 pg / mL, 500 pg / mL, 600 pg / mL, 700 pg / mL, particularly preferably 800 pg / mL) or more of IFN-α by co-culturing with bone marrow cells at a concentration of 1 / mL / mL for 24 hours.

[0020] In the "bacteria capable of activating pDCs" of the present invention 1, pDC activation can be achieved by promoting the expression of activation markers such as CD80, CD86, or MHC class II.

[0021] The above-mentioned "lactic acid bacteria capable of activating pDCs" are not particularly limited, but include, for example, bacteria of the genera Oenococcus, Bifidobacterium, Weissella, Tetragenococcus, Lactococcus, Leuconostoc, Pediococcus, Streptococcus, Enterococcus, and Examples include bacteria of the genus Lactobacillus, with Oenococcus, Bifidobacterium, Weissella, Tetragenococcus, Lactococcus, Leuconostoc, Pediococcus, Enterococcus, and Lactobacillus being preferred. Furthermore, the Lactobacillus bacteria in this invention include bacteria that were classified under the Lactobacillus genus before the reclassification of the Lactobacillus genus. For example, with the reclassification of the Lactobacillus genus, new genera have been added: Acetilactobacillus, Agrilactobacillus, Amylolactobacillus, Apilactobacillus, Bombilactobacillus, Companilactobacillus, Dellaglioa, Fructilactobacillus, Furfurilactobacillus, Holzapfelia, Lacticaseibacillus, Lactiplantibacillus, and Lapidilactobacillus. This includes bacteria classified under genera such as Lapidilactobacillus, Latilactobacillus, Lentilactobacillus, Levilactobacillus, Ligilactobacillus, Limosilactobacillus, Liquorilactobacillus, Loigolactobacillus, Paralactobacillus, Paucilactobacillus, Schleiferilactobacillus, and Secundilactobacillus.

[0022] Examples of the Oenococcus genus mentioned above include Oenococcus oeni. A specific example of an Oenococcus genus is Oenococcus oeni JCM6125.

[0023] Examples of Bifidobacterium species mentioned above include Bifidobacterium animalis subsp. lactis and Bifidobacterium longum subsp. infantis. Specific examples of Bifidobacterium species include Bifidobacterium animalis subsp. lactis JCM10602 and Bifidobacterium longum subsp. infantis JCM1222.

[0024] Examples of the Weissella genus mentioned above include Weissella paramesenteroides and Weissella viridescens. Specific examples of Weissella genus include Weissella paramesenteroides JCM9890 and Weissella viridescens JCM1174.

[0025] Examples of the Tetragenococcus genus mentioned above include Tetragenococcus halophilus. A specific example of Tetragenococcus genus is Tetragenococcus halophilus NRIC0098.

[0026] Examples of Lactococcus species mentioned above include Lactococcus lactis, Lactococcus lactis subsp. lactis, Lactococcus garvieae, Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. hordniae, and Lactococcus plantarum.

[0027] Specific examples of the above Lactococcus species include Lactococcus lactis subspecies lactis JCM5805, Lactococcus lactis subspecies lactis NBRC12007, Lactococcus lactis subspecies lactis NRIC1150, Lactococcus lactis subspecies lactis JCM20101, Lactococcus lactis subspecies lactis JCM7638, Lactococcus lactis subspecies Examples include Lactosus lactis ATCC11454, Lactococcus garvieae NBRC100934, Lactococcus lactis subspecies cremoris JCM16167, Lactococcus lactis subspecies cremoris NBRC100676, Lactococcus lactis subspecies heldniae JCM1180, Lactococcus lactis subspecies heldniae JCM11040, and Lactococcus plantarum JCM11056.

[0028] Examples of Leuconostoc species mentioned above include Leuconostoc carnosum and Leuconostoc lactis. Specific examples of Leuconostoc species include Leuconostoc carnosum JCM9695 and Leuconostoc lactis NBRC12455.

[0029] Examples of the Pediococcus species mentioned above include Pediococcus acidilactici, Pediococcus pentosaceus, Pediococcus cellicola, Pediococcus claussenii, Pediococcus damnosus, Pediococcus ethanolidurans, Pediococcus inopinatus, Pediococcus parvulus, and Pediococcus stilesii. Specific examples of Pediococcus species include Pediococcus acidilactici JCM8797, Pediococcus acidilactici K15, and Pediococcus damnosus JCM5886.

[0030] Examples of the Streptococcus species mentioned above include Streptococcus thermophilus. A specific example of a Streptococcus species is Streptococcus thermophilus SBC8781.

[0031] Examples of Enterococcus species mentioned above include Enterococcus alcedinis.

[0032] Examples of Lactobacillus species include Lactobacillus paracasei, Lactobacillus delbrueckii, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus fructivorans, Lactobacillus hilgardii, Lactobacillus rhamnosus, Lactobacillus gasseri, Lactobacillus acidophilus, and Lactobacillus bulgaricus. Examples include Lactobacillus bulgaricus, Lactobacillus parakefiri, Lactobacillus plantarum, and Lactobacillus pentosus.

[0033] Specific examples of Lactobacillus species include Lactobacillus paracasei KW3110, Lactobacillus paracasei MCC1849, Lactobacillus rhamnosus GG, Lactobacillus rhamnosus GG, Lactobacillus rhamnosus CRL1505, Lactobacillus gasseri SBT2055, Lactobacillus acidophilus L-92, Lactobacillus bulgaricus OLL1073R-1, Lactobacillus parakephyli (Lentilactobacillus parakephyli in the new classification) JCM8573, Lactobacillus plantarum (Lactipruntilabacillus plantarum in the new classification) L-137, and Lactobacillus pentosus (Lactipruntilabacillus pentosus in the new classification) ONRICb0240.

[0034] The above-mentioned "acetic acid bacteria capable of activating pDCs" are not particularly limited, but examples include bacteria of the genus Gluconacetobacter, Acetobacter, and Gluconobacter, preferably Gluconacetobacter, more preferably Gluconacetobacter hanzenii, and even more preferably Gluconacetobacter hanzenii GK-1.

[0035] The above-mentioned "Bacillus species capable of activating pDCs" are not particularly limited, but examples include Bacillus coagulans. A specific example of a Bacillus species is Bacillus coagulans SANK70258.

[0036] (Dead lactic acid bacteria cells) In the present invention 2, dead cells of one or more types of lactic acid bacteria are used.

[0037] "Lactic acid bacteria" is a general term for all bacteria that are taxonomically recognized as lactic acid bacteria, and is not limited to genus, species, or strain. Examples of such "lactic acid bacteria" include bacteria that ferment sugars to produce a large amount of lactic acid (preferably 50% or more of the consumed sugar as lactic acid), and include bacteria of the genera Lactobacillus, Streptococcus, Lactococcus, Leuconostoc, Pediococcus, Enterococcus, Oenococcus, Bifidobacterium, Weissella, and Tetragenococcus.

[0038] The genus and species of dead lactic acid bacteria used in Invention 2 are not particularly limited, but examples include one or more bacteria selected from the group consisting of Lactobacillus, Streptococcus, Lactococcus, Leuconostoc, Pediococcus, Enterococcus, Oenococcus, Bifidobacterium, Weissella, and Tetragenococcus, and also Lactobacillus. Examples include one or more bacteria selected from the group consisting of Streptococcus, Lactococcus, Leuconostoc, Pediococcus, and Enterococcus, preferably one or more bacteria selected from the group consisting of Lactococcus, more preferably one or more bacteria selected from the group consisting of Lactococcus lactis, and even more preferably one or more bacteria selected from the group consisting of Lactococcus lactis subspecies lactis.

[0039] More specifically preferred embodiments of the dead bacterial cells in Invention 2 include Lactobacillus acidophilus (e.g., Lactobacillus acidophilus L-92), Lactobacillus delbrueckii subsp. bulgaricus (e.g., Lactobacillus bulgaricus OLL1073R-1), Lactobacillus delbrueckii subsp. delbrueckii, Lactobacillus delbrueckii subsp. lactis, Lactobacillus casei, and Lactobacillus paracasei. Lactobacillus paracasei (e.g., Lactobacillus paracasei KW3110 and Lactobacillus paracasei MCC1849), Lactobacillus gasseri (e.g., Lactobacillus gasseri SBT2055), Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus plantarum (Lactiplantibacillus plantarum in newer classifications) (e.g., Lactobacillus plantarum L-137), Lactobacillus brevis, Lactobacillus casei subspecies rhamnosus.Lactobacillus rhamnosus (GG, Lactobacillus rhamnosus CRL1505, etc.), Lactobacillus pentosus (Lactobacillus pentosus (Lactiplantibacillus pentosus in the new classification) ONRICb0240, etc.), Lactobacillus fermentum, Lactobacillus fructivorans, Lactobacillus hilgardii, Streptococcus salivarius subsp. thermophilus, Lactococcus lactis subsp. Lactococcus lactis subsp. lactis biovar diacetylactis, Lactococcus lactis subsp. lactis cremoris (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, etc.), Lactococcus lactis biovariant diacetylactis, Lactococcus lactis subsp. lactis cremorisLactococcus cremoris (Lactococcus lactis subspecies cremoris JCM16167, Lactococcus lactis subspecies cremoris NBRC100676, etc.), Lactococcus raffinolactis, Lactococcus piscium, Lactococcus plantarum (Lactococcus plantarum JCM11056, etc.), Lactococcus garvieae (Lactococcus garvieae NBRC100934, etc.), Lactococcus lactis subsp. Lactococcus lactis subspecies hordniae (JCM1180, Lactococcus lactis subspecies hordniae JCM11040, etc.), Leuconostoc mesenteroides subsp. cremoris.Lactococcus lactis subspecies cremoris (JCM16167, Lactococcus lactis subspecies cremoris NBRC100676, etc.), Leuconostoc lactis (Leuconostoc lactis NBRC12455, etc.), Leuconostoc carnosum (Leuconostoc carnosum JCM9695, etc.), Pediococcus damnosus (Pediococcus damnosus JCM5886, etc.), Pediococcus pentosaceus, Pediococcus acidilactici (e.g., Pediococcus acidilactici JCM8797 and Pediococcus acidilactici K15), Pediococcus cellicola, Pediococcus claussenii, Pediococcus ethanolidurans, Pediococcus inopinatus, Pediococcus parvulus, Pediococcus stilesii, Enterococcus faecalis, Enterococcus faecium faecium), Enterococcus alcedinis, Oenococcus oeni (e.g., Oenococcus oeni JCM6125), Bifidobacterium animalis subsp. lactis (e.g., Bifidobacterium animalis subsp. lactis JCM10602), Bifidobacterium longum subsp. infantisExamples include dead bacterial cells of one or more species of bacteria selected from the group consisting of Bifidobacterium longum subspecies infantis (e.g., JCM1222), Weissella paramesenteroides (e.g., JCM9890), Weissella viridescens (e.g., JCM1174), and Tetragenococcus halophilus (e.g., NRIC0098). Also included are dead bacterial cells of Lactobacillus acidophilus (e.g., Lactobacillus acidophilus L-92) and Lactobacillus delbrueckii subspecies bulgaricus. Lactobacillus bulgaricus (OLL1073R-1, etc.), Lactobacillus delbrueckii subsp. delbrueckii, Lactobacillus delbrueckii subsp. lactisLactobacillus lactis), Lactobacillus casei, Lactobacillus paracasei (Lactobacillus paracasei KW3110 and Lactobacillus paracasei MCC1849, etc.), Lactobacillus gasseri (Lactobacillus gasseri SBT2055), Lactobacillus shelveticus, Lactobacillus johnsonii, Lactobacillus parakefiri (Lentilactobacillus parakefiri in the new classification) (Lactobacillus parakefiri JCM8573, etc.), Lactobacillus plantarum (Lactiplantibacillus in the new classification) Lactobacillus plantarum (e.g., L-137), Lactobacillus brevis, Lactobacillus casei subsp. rhamnosus (e.g., Lactobacillus rhamnosus GG, Lactobacillus rhamnosus CRL1505), Lactobacillus pentosus (e.g., Lactobacillus pentosus ONRICb0240), Lactobacillus fermentum, Lactobacillus fructivorans, Lactobacillus hilgardii, Streptococcus salivarius subsp. thermophilus Salivarius subsp. thermophilus, Lactococcus lactis subsp.Lactococcus lactis subsp. lactis biovar diacetylactis, Lactococcus lactis subsp. lactis cremoris (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, etc.), Lactococcus lactis biovariant diacetylactis, Lactococcus lactis subsp. lactis cremoris Lactococcus cremoris (Lactococcus lactis subspecies cremoris JCM16167, Lactococcus lactis subspecies cremoris NBRC100676, etc.), Lactococcus raffinolactis, Lactococcus piscium, Lactococcus plantarum (Lactococcus plantarum JCM11056, etc.), Lactococcus garvieae (Lactococcus garvieae NBRC100934, etc.), Lactococcus lactis subsp. Lactococcus lactis subspecies hordniae (JCM1180, Lactococcus lactis subspecies hordniae JCM11040, etc.), Leuconostoc mesenteroides subsp. cremoris.Lactococcus lactis subspecies cremoris (JCM16167, Lactococcus lactis subspecies cremoris NBRC100676, etc.), Leuconostoc lactis (Leuconostoc lactis NBRC12455, etc.), Leuconostoc carnosum (Leuconostoc carnosum JCM9695, etc.), Pediococcus damnosus (Pediococcus damnosus JCM5886, etc.), Pediococcus pentosaceus, Pediococcus acidilactici (e.g., Pediococcus acidilactici JCM8797 and Pediococcus acidilactici K15), Pediococcus cellicola, Pediococcus claussenii, Pediococcus ethanolidurans, Pediococcus inopinatus, Pediococcus parvulus, Pediococcus stilesii, Enterococcus faecalis, and Enterococcus faecium Examples include dead bacterial cells of one or more bacteria selected from the group consisting of Lactobacillus faecium, preferably Lactobacillus acidophilus, Lactobacillus delbrueckii subspecies bulgaricus, Lactobacillus... Lactobacillus delbrookii subspecies lactis, Lactobacillus delbrookii subspecies delbrookii, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus parakefilii (Lactobacillus parakefilii JCM8573, etc.), Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus casei subspecies rhamnosus (Lactobacillus rhamnosus GG, Lactobacillus Lactobacillus rhamnosus CRL1505, etc., Lactobacillus pentosus, Lactobacillus fermentum, Lactobacillus fructivorance, Lactobacillus hilgardii, Streptococcus salivarius subspecies thermophilus, Lactococcus lactis subspecies lactis, Lactococcus lactis biovariant diacetylactis, Lactococcus lactis subspecies cremoris, Lactococcus raffinolactis, Lactococcus pythium, Lactococcus Examples include dead bacterial cells of one or more bacteria selected from the group consisting of *Cucus plantarum*, *Lactococcus garbieae*, *Lactococcus lactis subspecies holdniae*, *Leuconostoc mecentroides subspecies cremoris*, and *Leuconostoc lactis*, more preferably *Lactococcus lactis subspecies lactis*, *Lactococcus lactis biovariant diacetylactis*, *Lactococcus lactis subspecies cremoris*, and *Lactococcus... Examples include dead bacterial cells of one or more bacteria selected from the group consisting of Raffinolactis, Lactococcus pythium, Lactococcus plantarum, Lactococcus garbieae, and Lactococcus lactis subspecies heldniae, more preferably dead bacterial cells of Lactococcus lactis subspecies lactis, and more preferably Lactococcus lactis subspecies lactis JCM5805, Lactococcus lactis subspecies lactis JCM20101,Examples include dead bacterial cells of one or more bacteria selected from the group consisting of Lactococcus lactis subspecies lactis NBRC12007 and Lactococcus lactis subspecies lactis NRIC1150, with particular preference being dead bacterial cells of Lactococcus lactis subspecies lactis JCM5805.

[0040] Furthermore, other preferred embodiments of the genus and species of dead lactic acid bacteria used in Invention 2 include one or more bacteria selected from the group consisting of Lactococcus and Lactobacillus species, preferably one or more bacteria selected from the group consisting of Lactococcus lactis and Lactobacillus rhamnosus, and more preferably one or more bacteria selected from the group consisting of Lactococcus lactis subspecies lactis and Lactobacillus rhamnosus.

[0041] More specific, other preferred embodiments of the dead bacterial cells in Invention 2 include Lactococcus lactis subspecies lactis, Lactococcus lactis biovariant diacetylactis, Lactococcus lactis subspecies cremoris, Lactococcus raffinolactis, Lactococcus pythium, Lactococcus plantarum, Lactococcus garbieae, Lactococcus lactis subspecies heldniae, Lactobacillus acidophilus, Lactobacillus delbrookii Lactobacillus subspecies bulgaricus, Lactobacillus delbrookii subspecies lactis, Lactobacillus delbrookii subspecies delbrookii, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus parakefilii (Lactobacillus parakefilii JCM8573, etc.), Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus casei subspecies rhamnosus (Lactobacillus Examples include dead bacterial cells of one or more bacteria selected from the group consisting of Lactobacillus rhamnosus GG, Lactobacillus rhamnosus CRL1505, etc., Lactobacillus pentosus, Lactobacillus fermentum, Lactobacillus fructivorans, and Lactobacillus hilgardii, preferably dead bacterial cells of one or more bacteria selected from the group consisting of Lactococcus lactis subspecies lactis and Lactobacillus rhamnosus, more preferably Lactococcus lactis subspecies lactis Examples include dead bacterial cells of one or more bacteria selected from the group consisting of Lactococcus lactis subspecies JCM5805, Lactococcus lactis subspecies JCM20101, Lactococcus lactis subspecies NBRC12007, Lactococcus lactis subspecies NRIC1150, Lactobacillus rhamnosus GG, and Lactobacillus rhamnosus CRL1505, with particular preference being Lactococcus lactis subspecies JCM5805 and,Examples include dead bacterial cells of one or two bacteria selected from Lactobacillus rhamnosus CRL1505, more preferably dead bacterial cells of Lactococcus lactis subspecies lactis JCM5805, or dead bacterial cells of Lactobacillus rhamnosus CRL1505.

[0042] Another preferred embodiment of the dead bacterial cells in Invention 2 includes one or more species selected from the group consisting of bacteria of the genus Lactococcus, preferably Lactococcus lactis subspecies lactis JCM5805, Lactococcus lactis subspecies lactis NBRC12007, Lactococcus lactis subspecies lactis NRIC1150, Lactococcus lactis subspecies lactis JCM20101, and Lactococcus lactis subspecies lactis JCM76. 38. One or more species selected from the group consisting of Lactococcus lactis subspecies lactis ATCC11454, Lactococcus garvieae NBRC100934, Lactococcus lactis subspecies cremoris JCM16167, Lactococcus lactis subspecies cremoris NBRC100676, Lactococcus lactis subspecies heldniae JCM1180, and Lactococcus lactis subspecies heldniae JCM11040.

[0043] With regard to the dead bacterial strains listed herein, in the present invention (i.e., Invention 1 and / or Invention 2), strains equivalent to the above strains are included as strains, provided that they exert an inhibitory effect on mouth irritation when dead bacterial cells are included at a predetermined concentration in a sugar-free beverage containing a predetermined concentration of magnesium. Here, equivalent strains refer to strains derived from the above strains, strains from which the above strains originate, or progeny strains of those strains. Equivalent strains may also be stored in other strain preservation institutions. Figure 1 shows strains derived from Lactococcus lactis subspecies lactis JCM5805, and strains from which Lactococcus lactis subspecies lactis JCM5805 originates. Equivalent strains of Lactococcus lactis subspecies lactis JCM5805 shown in Figure 1 can also be used as dead bacterial cells of the present invention, provided that they exert an inhibitory effect on the squeaky feeling in the mouth when included at a predetermined concentration in a sugar-free beverage containing a predetermined concentration of magnesium. In this specification, when referring to Lactococcus lactis subspecies lactis JCM5805 (Lactococcus lactis JCM5805), these equivalent strains are also included. Furthermore, in this specification, when referring to Lactobacillus rhamnosus CRL1505, these equivalent strains are also included. Furthermore, of the above-mentioned lactic acid bacteria strains, the JCM strain can be obtained from the BioResource Center, Microbial Materials Development Laboratory (3-1-1 Takanodai, Tsukuba City, Ibaraki Prefecture), the NBRC strain from the National Institute of Technology and Evaluation, Biological Genetic Resources Division (2-5-8 Kazusa-Kamatari, Kisarazu City, Chiba Prefecture), the NRIC strain from the Strain Preservation Room, Tokyo University of Agriculture (1-1-1 Sakuragaoka, Setagaya-ku, Tokyo), and the ATCC strain from the American Type Culture Collection (USA). As mentioned above, the Lactococcus lactis subspecies lactis JCM5805 strain can be obtained from the Microbial Materials Development Laboratory at the RIKEN BioResource Center. However, in this invention, the same JCM5805 strain can be used from other storage institutions. Specifically, the same JCM5805 strain can be obtained from the National Institute of Technology and Evaluation (NITE) Biological Genetics Division (2-5-8 Kazusa-Kamatari, Kisarazu City, Chiba Prefecture), the Strain Preservation Room of Tokyo University of Agriculture (1-1-1 Sakuragaoka, Setagaya-ku, Tokyo), and the American Type Culture Collection (USA), among others.

[0044] In this specification, "dead cells of lactic acid bacteria and / or bacteria in Invention 1" are not particularly limited as long as they are dead cells of lactic acid bacteria and / or bacteria in Invention 1, and may be dried or not. However, from the viewpoint of storage stability of dead cells of lactic acid bacteria and / or bacteria in Invention 1, they are preferably dried, and dried powder is a suitable example.

[0045] The method for preparing dead lactic acid bacteria and / or bacteria according to Invention 1 is not particularly limited. For example, methods include sterilizing the culture medium on which lactic acid bacteria and / or bacteria according to Invention 1 are cultured, and then collecting the cells by filtration, centrifugation, etc., or collecting the cells from the culture medium on which lactic acid bacteria and / or bacteria according to Invention 1 are cultured by filtration, centrifugation, etc., and then sterilizing the culture medium. If necessary, further drying or crushing treatments may be performed. Furthermore, there are no particular restrictions on the sterilization methods; conventional methods for killing bacteria, such as heating, ultraviolet light, or gamma ray irradiation, can be used. In addition, lactic acid bacteria and / or bacteria in the beverage may be killed during the sterilization process in beverage preparation.

[0046] In the present invention, the bacterial cell concentration of one or more types of lactic acid bacteria and / or bacteria in the present invention 1 in the packaged sugar-free beverage is not particularly limited, but the total number of dead lactic acid bacteria and / or bacteria in the present invention 1 can be, for example, 500 million cells / L or more, 1 billion cells / L or more, 1.3 billion cells / L or more, 5 billion cells / L or more, 15 billion cells / L or more, or 20 billion cells / L or more. From the viewpoint of obtaining a greater inhibitory effect on the squeaky feeling in the mouth, it is preferably 25 billion cells / L or more, more preferably 50 billion cells / L or more, 100 billion cells / L or more, 200 billion cells / L or more, or 400 billion cells / L or more. Furthermore, there are no particular limitations on the upper limit of the bacterial cell concentration of one or more types of lactic acid bacteria and / or bacteria in Invention 1 in a bottled sugar-free beverage. However, examples of total dead bacterial cells of lactic acid bacteria and / or bacteria in Invention 1 include 8 trillion cells / L or less, 6 trillion cells / L or less, 4 trillion cells / L or less, 2 trillion cells / L or less, 1 trillion cells / L or less, 500 billion cells / L or less, 350 billion cells / L or less, 200 billion cells / L or less, 175 billion cells / L or less, and 150 billion cells / L or less. These lower and upper limits can be arbitrarily combined within the range of combinations that can be set as lower and upper limits. The bacterial concentration of lactic acid bacteria and / or bacteria according to Invention 1 in a packaged sugar-free beverage can be adjusted by adjusting the amount of dead lactic acid bacteria and / or bacteria according to Invention 1 that are added to the beverage. Furthermore, as a method for measuring the number of dead lactic acid bacteria and / or bacteria in the present invention 1 in a packaged sugar-free beverage, any known method for measuring the number of lactic acid bacteria and / or bacteria in the present invention 1 can be mentioned without particular limitation, such as direct microscopy, particle electrical detection zone method, PCR method, or flow cytometry method, with flow cytometry method being preferred.

[0047] (magnesium) The magnesium concentration of the bottled sugar-free beverage in this invention is 1.2 mg / L or higher. In this specification, "magnesium concentration" refers to the concentration of magnesium dissolved in the water in the bottled sugar-free beverage. Therefore, for example, magnesium contained in solid lactic acid bacteria and / or dead bacterial cells in this invention 1 is not considered in the magnesium concentration as defined herein.

[0048] The magnesium concentration of the bottled sugar-free beverage in the present invention is not particularly limited as long as it is 1.2 mg / L or higher. However, from the viewpoint of achieving a stronger mouth-cleaning sensation and enjoying the full benefits of the present invention, preferred magnesium concentrations are 2.4 mg / L or higher, more preferably 4.9 mg / L or higher, even more preferably 7.3 mg / L or higher, even more preferably 9.7 mg / L or higher, even more preferably 12.1 mg / L or higher, even more preferably 24.3 mg / L or higher, even more preferably 77.7 mg / L or higher, even more preferably 109.3 mg / L or higher, and even more preferably 145.7 mg / L or higher. Furthermore, there are no particular upper limits on the magnesium concentration in bottled sugar-free beverages, but examples include 150 mg / L or less, 200 mg / L or less, 250 mg / L or less, and 500 mg / L or less. These lower and upper limits can be combined arbitrarily. The magnesium concentration in bottled sugar-free beverages can be adjusted by adjusting the amount of one or more substances selected from the group consisting of magnesium, magnesium compounds, and magnesium-containing compositions that are added to the beverage. Furthermore, known measurement methods can be used to measure the magnesium concentration in bottled sugar-free beverages without any particular limitations, such as inductively coupled plasma atomic emission spectroscopy (ICP-OES).

[0049] The ratio of bacterial cell concentration to magnesium concentration in this invention, i.e., (content of dead lactic acid bacteria and / or bacteria in Invention 1 [billion cells / L]) / (magnesium concentration [mg / L]), is not particularly limited. However, from the viewpoint of obtaining a greater inhibitory effect on oral irritation, examples of lower limits include 0.4 or more, 1 or more, 80 or more, 200 or more, or 400 or more, and examples of upper limits include 60,000 or less, 30,000 or less, or 8,000 or less. These lower and upper limits can be combined arbitrarily.

[0050] Preferably, the magnesium compound is a water-soluble magnesium compound, of which magnesium chloride and magnesium sulfate are preferred, and of which magnesium chloride is more preferred. The magnesium-containing composition is a composition containing the aforementioned magnesium compound. Commercially available magnesium compounds and compositions can be used.

[0051] (sugar-free drink) In this specification, "sugar-free beverage" means a beverage containing less than 0.5 g of sugars per 100 mL of beverage. The term "sugars" above means one or more selected from the group consisting of crystalline sugars and amorphous sugars. Examples of "crystalline sugars" above include monosaccharides such as fructose, glucose, tagatose, and arabinose, and disaccharides such as lactose, trehalose, maltose, and sucrose. Examples of "amorphous sugars" above include corn syrup and isomerized liquid sugar (e.g., high fructose corn syrup).

[0052] In the present invention, the sugar concentration of the packaged sugar-free beverage is not particularly limited as long as it is less than 0.5 g of sugar per 100 mL of beverage. However, preferred sugar concentrations are 0.25 g or less, 0.2 g or less, 0.15 g or less, 0.1 g or less, 0.5 g or less, 0.2 g or less, 0.1 g or less, 0.05 g or less, and 0.01 g or less per 100 mL of beverage, with beverages that do not contain or do not contain sugars being more preferred.

[0053] (Carbon dioxide) The packaged sugar-free beverage of the present invention does not have to be a carbonated beverage, but it is preferable that it be a carbonated beverage from the viewpoint of suppressing the grittiness in the mouth to a greater extent. In this specification, "carbonated beverage" refers to a carbonated beverage having a gas pressure of 0.05 to 0.5 MPa, preferably 0.1 to 0.3 MPa, 0.15 to 0.3 MPa, or 0.15 to 0.25 MPa.

[0054] In this specification, gas pressure refers to the internal gas pressure of a bottled unsweetened beverage at 1 atmosphere and 20°C. This pressure can be measured by first bringing the sample to 20°C, attaching a gas pressure gauge, opening a stopcock to release the gas (snifting), immediately closing the stopcock, shaking vigorously, and converting the value obtained when the pressure becomes constant to MPa.

[0055] (optional ingredient) The packaged sugar-free beverage of the present invention may or may not contain one or more of the following: organic acids, colorants, flavorings, acidulants (except organic acids), sugar alcohols, high-intensity sweeteners, antioxidants, preservatives, thickening stabilizers, emulsifiers, and pH adjusters. In one aspect of the present invention, the packaged sugar-free beverage of the present invention may contain either a sugar alcohol, a high-intensity sweetener, or both, but may not contain either a sugar alcohol or a high-intensity sweetener. Examples of the "sugar alcohol" mentioned above include maltitol, lactitol, sorbitol, mannitol, xylitol, and erythritol. Examples of the "high-intensity sweetener" mentioned above include acesulfame potassium (acesulfame K), sucralose, stevia, licorice extract, thaumatin, glycyrrhizin, saccharin, and aspartame.

[0056] (organic acid) The bottled sugar-free beverage of the present invention does not necessarily have to contain organic acids, but it is preferable that it further contains organic acids from the viewpoint of further suppressing the feeling of dryness in the mouth. Examples of such organic acids include one or more selected from the group consisting of citric acid, malic acid, tartaric acid, lactic acid, gluconic acid, succinic acid, pyruvic acid, formic acid, phytic acid, and acetic acid, or one or more selected from the group consisting of citric acid, malic acid, tartaric acid, lactic acid, gluconic acid, and succinic acid, with one or more selected from the group consisting of citric acid, malic acid, tartaric acid, and lactic acid being preferred, and one or more selected from the group consisting of citric acid, malic acid, and tartaric acid being more preferred from the viewpoint of further suppressing the feeling of dryness in the mouth. When manufacturing the beverage of the present invention that contains organic acids, organic acids may be used, or salts of organic acids or organic acid-containing compositions may be used. The salts are not particularly limited, and examples include sodium salts, potassium salts, calcium salts, etc.

[0057] When the beverage of the present invention contains organic acids, the total content of organic acids in the beverage (for example, one or two organic acids selected from any of the aforementioned groups of organic acids) is not particularly limited. However, from the viewpoint of suppressing mouth irritation to a greater extent, it is preferably 0.005% by weight or more, more preferably 0.01% by weight or more, even more preferably 0.05% by weight or more, even more preferably 0.12% by weight or more, and even more preferably 0.4% by weight or more or 0.5% by weight or more, relative to the total amount of the beverage. Upper limits include, for example, less than 0.7% by weight, 0.65% by weight or less, 0.6% by weight or less, and 0.5% by weight or less. When an organic acid salt is used, the organic acid content (by weight) is calculated by converting the organic acid salt to organic acid.

[0058] If the beverage of the present invention contains organic acids, another aspect of the total content of organic acids in the beverage (for example, one or two organic acids selected from any of the aforementioned groups of organic acids) is preferably 1 × 10⁻¹⁶. -4mol / L or more, or 1×10 -3 mol / L or more, more preferably 3×10 -3 mol / L or more, or 5×10 -3 mol / L or more, 2.6×10 -2 mol / L or more. As the upper limit, for example, 3.6×10 -2 mol / L or less, 3.4×10 -2 mol / L or less, 3.1×10 -2 mol / L or less. When a salt of an organic acid is used, the content (mol / L) of the organic acid is calculated by converting the organic acid salt to the organic acid.

[0059] The total content of organic acids in the beverage of the present invention can be adjusted by adjusting the amount of the organic acid or a salt thereof, or the organic acid-containing composition to be used.

[0060] The content of an organic acid in the beverage of the present invention can be calculated by measuring the target organic acid by HPLC or the like.

[0061] The ratio of the total amount of organic acids to the magnesium concentration of the present invention, that is, (total amount of organic acids [% by weight]) / (magnesium concentration [mg / L]) is not particularly limited, but from the viewpoint of obtaining a greater inhibitory effect against astringency in the mouth, examples of the lower limit include 0.00001 or more, 0.00003 or more, 0.0001 or more, 0.001 or more, or 0.01 or more, and examples of the upper limit include 0.5 or less, 0.1 or less, 0.05 or less, and these lower limits and upper limits can be arbitrarily combined with each other.

[0062] (Beverage of the Present Invention) The beverage of the present invention is not particularly limited as long as it is a packaged sugar-free beverage containing 500 million or more (for example, 20 billion or more) dead cells of one or more lactic acid bacteria and / or the bacteria according to the first aspect of the present invention, and having a magnesium concentration of 1.2 mg / L or more.

[0063] The beverage of the present invention is a bottled sugar-free beverage that contains 500 million or more dead cells (for example, 20 billion or more) of one or more types of lactic acid bacteria and / or bacteria as described in Invention 1, and has a magnesium concentration of 1.2 mg / L or more. Other than these differences in the raw materials used, manufacturing method, and manufacturing conditions, it is no different from a normal "bottled sugar-free beverage".

[0064] The type of beverage of the present invention is not particularly limited as long as it is a bottled sugar-free beverage, but a bottled sugar-free carbonated beverage is preferred. Furthermore, the beverage of the present invention may be a chilled beverage (i.e., a beverage that is managed in a refrigerated state of 0°C to 10°C from the time of manufacture to distribution and sale), but it may also be a beverage that is not a chilled beverage.

[0065] The beverage of the present invention can be produced by adjusting the magnesium concentration to 1.2 mg / L or more in any stage of a general manufacturing method for "bottled sugar-free beverages," by adding 500 million or more dead lactic acid bacteria and / or bacteria according to Invention 1 (for example, 20 billion or more), and adjusting the magnesium concentration to 1.2 mg / L or more.

[0066] The beverage of the present invention is a packaged beverage. Examples of such containers include resin bottles such as PET bottles, polypropylene bottles, and polyvinyl chloride bottles; glass bottles; cans; and other containers.

[0067] The beverage of the present invention does not need to be heat-sterilized, but it may be heat-sterilized from the viewpoint of improving its shelf life. As for the heat-sterilization method and conditions, conventional methods and conditions used for beverages such as bottled beverages can be used.

[0068] (Manufacturing method of the present invention) The manufacturing method of the present invention is not particularly limited as long as it is a method for manufacturing a packaged unsweetened beverage having a magnesium concentration of 1.2 mg / L or more, characterized in that the beverage contains 500 million or more (for example, 20 billion or more) dead cells of one or more types of lactic acid bacteria and / or bacteria according to Invention 1.

[0069] The beverage of the present invention can be manufactured according to conventional known manufacturing methods for bottled unsweetened beverages, except that it contains 500 million or more dead cells (for example, 20 billion or more) of one or more types of lactic acid bacteria and / or bacteria in the present invention 1, and has a magnesium concentration of 1.2 mg / L or more.

[0070] More specifically, a method for incorporating 500 million or more (e.g., 20 billion or more) dead cells of one or more types of lactic acid bacteria and / or bacteria according to Invention 1 into a beverage is to incorporate 500 million or more (e.g., 20 billion or more) dead cells of one or more types of lactic acid bacteria and / or bacteria according to Invention 1 into the raw materials for the beverage (e.g., "water", "water containing 1.2 mg / L or more of magnesium", or "water further containing some or all of an arbitrary component") when manufacturing a bottled sugar-free beverage. Alternatively, a method is to incorporate "one or more types of lactic acid bacteria and / or dead cells of bacteria according to Invention 1" and "one or more selected from the group consisting of magnesium, magnesium compounds, and magnesium-containing compositions" into water, or to incorporate some or all of an arbitrary component into water simultaneously.

[0071] A more specific method (preferably a method of adjusting) to achieve a magnesium concentration of 1.2 mg / L or higher in a beverage is to add "one or more selected from the group consisting of magnesium, magnesium compounds, and magnesium-containing compositions" to the raw materials for the beverage during the production of a bottled sugar-free beverage (for example, "water," "water containing 500 million or more dead bacteria of one or more types of lactic acid bacteria and / or bacteria according to Invention 1 (e.g., 20 billion or more)," or "water further containing some or all of the optional components") to achieve a magnesium concentration of 1.2 mg / L or higher in the beverage. Alternatively, methods include adding "one or more types of lactic acid bacteria and / or dead bacteria according to Invention 1" and "one or more selected from the group consisting of magnesium, magnesium compounds, and magnesium-containing compositions" to water, or adding some or all of the optional components to water simultaneously.

[0072] In the manufacturing method of the present invention, lactic acid bacteria and / or dead bacterial cells of the bacteria described in Invention 1, along with magnesium, are included in the beverage as essential components. In the manufacturing method of the present invention, one or more of the following components may be included in the beverage: organic acids, colorants, flavorings, acidulants (except organic acids), antioxidants, preservatives, thickeners / stabilizers, emulsifiers, and pH adjusters.

[0073] In the manufacturing method of the present invention, there are no particular restrictions on the order in which the raw materials are added, as long as the beverage of the present invention can be produced. After preparing a liquid in which the raw materials are mixed, the liquid can be filled into a container and sealed to obtain the beverage of the present invention.

[0074] In the manufacturing method of the present invention, heat sterilization treatment is not required, but it may be performed from the viewpoint of improving shelf life. The method of heat sterilization treatment is not particularly limited, and examples include high-temperature short-time sterilization (HTST), pasteurization, ultra-high temperature heat treatment (UHT), and retort sterilization.

[0075] (Inventive suppression method of the present invention) The present invention provides a method for suppressing a squeaky feeling in the mouth when drinking a packaged unsweetened beverage, characterized by containing 500 million or more (for example, 20 billion or more) dead lactic acid bacteria and / or bacteria according to Invention 1 in the beverage, in the production of a packaged unsweetened beverage with a magnesium concentration of 1.2 mg / L or more.

[0076] Methods for incorporating 500 million or more dead cells (e.g., 20 billion or more) of one or more types of lactic acid bacteria and / or bacteria according to Invention 1 into a beverage, or for setting the magnesium concentration in the beverage to 1.2 mg / L or more, can be the same as those described above in (Production Method of the Invention).

[0077] (Suppression of the characteristic squeaky feeling in the mouth) The present invention provides a beverage in which the characteristic grittiness in the mouth of bottled unsweetened beverages with a magnesium concentration of 1.2 mg / L or higher is suppressed. In this specification, the term "mouth squeaking" means "a squeaking sensation as if the tongue or throat area is being pulled," and more specifically, "a feeling of fatigue as if the tongue or throat area is being pulled and cramped."

[0078] In the present invention, a beverage with "suppressed oral grittiness" means a beverage in which oral grittiness during consumption is suppressed compared to a beverage manufactured using the same raw materials and the same manufacturing method to the same final concentration, except that it does not contain one or more types of lactic acid bacteria and / or dead bacterial cells of the bacteria in Invention 1 (hereinafter also referred to as "control beverage").

[0079] A trained panel can easily and clearly determine the degree of "mouth grittiness" in a particular beverage and how it compares to a control beverage (e.g., whether it is suppressed). General methods can be used for evaluation criteria and for summarizing evaluations among the panel members. In this invention, the number of panel members evaluating the sensory aspects may be one, but from the viewpoint of obtaining a more objective evaluation, the lower limit of the number of panel members can be, for example, two or more, preferably four or more. Also, from the viewpoint of conducting the evaluation test more simply, the upper limit of the number of panel members can be, for example, 20 or fewer, 10 or fewer, or 7 or fewer. When there are two or more panel members, the evaluation of mouth grittiness for each beverage may be, for example, by adopting the average of the evaluations of all panel members regarding the mouth grittiness of that beverage, or by adopting the lowest evaluation among the panel members. If evaluation points are assigned to each evaluation criterion, the average of the evaluation points of all panel members may be adopted as the evaluation of mouth grittiness for that beverage, or by adopting the lowest evaluation point among the panel members. As mentioned above, when using the average of the evaluation scores, the value obtained by rounding the average to the first or second decimal place (preferably the second decimal place) may be used. Alternatively, from among the points "1 point, 1.5 points, 2 points, 2.5 points, 3 points, 3.5 points, 4 points, 4.5 points, 5 points, 5.5 points, 6 points, 6.5 points, 7 points, 7.5 points, 8 points, 8.5 points, 9 points, 9.5 points, 10 points", the point closest to the average value of each panel may be used as the evaluation of the oral grittiness of the beverage. When there are two or more panelists, it is preferable to standardize the evaluation criteria so that the evaluation criteria of each panel are as consistent as possible before conducting the actual sensory evaluation test, in order to reduce the variability in the evaluations of each panel. One example of such standardization work is to have each panel evaluate the sensory perception of several standard beverages whose degree of oral grittiness is known in this invention, then compare the evaluation scores to confirm that there is no large discrepancy in the evaluation criteria of each panel. Furthermore, it is preferable to standardize the evaluation criteria in advance so that the standard deviation of the oral creaking evaluations by each panel is within 0.5.

[0080] The degree of mouth-grittiness in a given beverage according to the present invention can be evaluated using a method similar to the one described in Test 1 of the Examples below, which uses an evaluation criterion (10-point scale from 1 to 10), preferably the same method as the one described in Test 1 of the Examples below. More specifically, a sensory evaluation test can be conducted on multiple panels using a 10-point scale from 1 to 10, the average of these evaluation scores can be calculated, and the point closest to the average value of each panel among the 19 points (i.e., "1 point, 1.5 points, 2 points, 2.5 points, 3 points, 3.5 points, 4 points, 4.5 points, 5 points, 5.5 points, 6 points, 6.5 points, 7 points, 7.5 points, 8 points, 8.5 points, 9 points, 9.5 points, 10 points") can be used to evaluate the degree of mouth-grittiness of the beverage. When evaluating the squeaky feeling in the mouth using this method, beverages that receive a rating of one level (i.e., 0.5 points) or more lower than the control beverage can be listed as beverages with suppressed squeaky feeling in the mouth. Preferably, beverages that receive a rating of two levels (i.e., 1 point) or more lower, more preferably three levels (i.e., 1.5 points) or more lower, even more preferably four levels (i.e., 2 points) or more lower, more preferably five levels (i.e., 2.5 points) or more lower, even more preferably six levels (i.e., 3 points) or more lower, even more preferably seven levels (i.e., 3.5 points) or more lower, even more preferably eight levels (i.e., 4 points) or more higher, even more preferably nine levels (i.e., 4.5 points) or more higher, and even more preferably ten levels (i.e., 5 points) or more lower can be listed as beverages with suppressed squeaky feeling in the mouth.

[0081] The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples. [Examples]

[0082] [Test 1] Confirmation of the occurrence of "gritty feeling in the mouth" when drinking a sugar-free beverage containing magnesium. The following tests were conducted to investigate how adding magnesium to sugar-free beverages affects their flavor.

[0083] (1. Preparation of a sample of sugar-free beverage) Each of the sample beverages in Test Examples 1 to 11 was prepared by adding magnesium chloride to deionized water so that the magnesium concentration was as shown in Table 1.

[0084] (2. Sensory evaluation test) For the "gritty feeling in the mouth," the sensory evaluation focused on the "gritty feeling of the tongue and throat being pulled" when drinking sugar-free beverages, or more specifically, "a feeling of the tongue and throat being pulled and becoming tired as if they are cramping." The degree of "mouth grittiness" experienced when drinking the sample beverages from Test Examples 1 to 11 was evaluated by a panel of four trained experts using a 10-point scale (1, 2, 3, 4, 5, 6, 7, 8, 9, 10). The evaluation criteria used were as follows: the sample beverage from Test Example 1 (magnesium concentration 0.0 mg / L) was assigned a score of 1 ("no mouth grittiness"), and the sample beverage from Test Example 11 (magnesium concentration 145.7 mg / L) was assigned a score of 10 ("extremely strong mouth grittiness"). A higher score indicates a stronger mouth grittiness. For each test sample, the evaluation of oral irritation was based on a 19-point scale (1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10), with the score closest to the average of each panel's evaluation scores being used. Furthermore, the standard deviation of each panel's evaluation scores for each sample beverage was 0.43 or less for all sample beverages.

[0085] The results of the sensory evaluation test are shown in Table 1.

[0086] [Table 1]

[0087] As can be seen from the results in Table 1, a magnesium concentration of 1.2 mg / L or higher causes a characteristic "squeaky feeling in the mouth," and this squeaky feeling intensifies in a magnesium concentration-dependent manner.

[0088] [Test 2] Effect of containing dead lactic acid bacteria on mouth squeakiness The following tests were conducted to investigate how the inclusion of dead lactic acid bacteria affects the squeaky feeling in the mouth of magnesium-containing sugar-free beverages.

[0089] (1. Preparation of a sample of sugar-free beverage) Sample beverages for Test Examples 13-19 were prepared by adding magnesium chloride and dried dead cell powder of Lactococcus lactis subspecies Lactis JCM5805 strain to deionized water so that the magnesium concentration and the concentration of dead lactic acid bacteria were as shown in Table 2. Sample beverage for Test Example 12 was prepared using the same method, except that the dried dead cell powder of Lactococcus lactis subspecies Lactis JCM5805 strain was not used.

[0090] (2. Sensory evaluation test) The degree of "mouth grittiness" experienced when drinking the sample beverages from Test Examples 12-19 was evaluated by a panel of four trained experts using a 10-point scale (1, 2, 3, 4, 5, 6, 7, 8, 9, 10). The evaluation criteria used were as follows: the degree of "mouth grittiness" for the sample beverage in Test Example 1 (magnesium concentration 0.0 mg / L) in Table 1 was assigned a score of 1 ("no mouth grittiness"), and the degree of "mouth grittiness" for the sample beverage in Test Example 11 in Table 1 was assigned a score of 10 ("extremely strong mouth grittiness"). The evaluation criteria divided the degree of mouth grittiness into the aforementioned 10 levels. For each test sample, the evaluation of oral irritation was based on a 19-point scale (1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10), with the score closest to the average of each panel's evaluation scores being used. Furthermore, the standard deviation of each panel's evaluation scores for each sample beverage was 0.43 or less for all sample beverages.

[0091] The results of the sensory evaluation test are shown in Table 2.

[0092] [Table 2]

[0093] As can be seen from the results in Table 2, it was shown that adding more than 20 billion dead lactic acid bacteria per liter to a magnesium-containing sugar-free beverage has an inhibitory effect on the squeaky feeling in the mouth.

[0094] [Test 3] Effect of using dead lactic acid bacteria and carbon dioxide together on oral squeaking The following tests were conducted to investigate how adding carbon dioxide to dead lactic acid bacteria affects the squeaky feeling in the mouth of a magnesium-containing sugar-free beverage.

[0095] (1. Preparation of a sample of sugar-free beverage) Sample beverages for Test Example 21 were prepared by adding magnesium chloride and dried dead cell powder of Lactococcus lactis subspecies Lactis JCM5805 strain to deionized water containing carbon dioxide, so that the magnesium concentration and the concentration of dead lactic acid bacteria were as shown in Table 3. Sample beverages for Test Example 20 were prepared using the same method, except that dried dead cell powder of Lactococcus lactis subspecies Lactis JCM5805 strain was not used.

[0096] (2. Sensory evaluation test) The degree of "mouth grittiness" experienced when drinking the sample beverages from Test Examples 20 and 21 was evaluated by a panel of four trained experts using a 10-point scale (1, 2, 3, 4, 5, 6, 7, 8, 9, 10). The evaluation criteria used were as follows: the degree of "mouth grittiness" for the sample beverage in Test Example 1 (magnesium concentration 0.0 mg / L) in Table 1 was assigned a score of 1 ("no mouth grittiness"), and the degree of "mouth grittiness" for the sample beverage in Test Example 11 in Table 1 was assigned a score of 10 ("extremely strong mouth grittiness"). The evaluation criteria divided the degree of mouth grittiness into the aforementioned 10 levels. For each test sample, the evaluation of oral irritation was based on a 19-point scale (1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10), with the score closest to the average of each panel's evaluation scores being used. Furthermore, the standard deviation of each panel's evaluation scores for each sample beverage was 0.43 or less for all sample beverages.

[0097] The results of this sensory evaluation test are shown in Table 3.

[0098] [Table 3]

[0099] As can be seen from the results in Table 3, even when the magnesium-containing sugar-free beverage is a carbonated beverage, the inclusion of dead lactic acid bacteria has been shown to have an inhibitory effect on the squeaky feeling in the mouth. Furthermore, comparing Test Example 21 (Table 3), which is a carbonated beverage, with Test Example 19 (Table 2), which is not a carbonated beverage, it was shown that Test Example 21, being a carbonated beverage, showed a greater suppression of the squeaky feeling in the mouth.

[0100] [Test 4] Effect of using dead lactic acid bacteria and organic acids together on oral squeaking The following tests were conducted to investigate how the inclusion of organic acids, in addition to dead lactic acid bacteria, affects the mouth-clean feeling of magnesium-containing sugar-free beverages.

[0101] (1. Preparation of a sample of sugar-free beverage) Magnesium chloride and dried dead cell powder of Lactococcus lactis subspecies Lactis JCM5805 strain were added to ion-exchanged water containing carbon dioxide so that the magnesium concentration and the concentration of dead lactic acid bacteria were as shown in Table 4, and the acidulant listed in Table 4 was added in an amount of 6.3 × 10 -3 Test Examples 24-28 were prepared by adding the required amount to mol / L. A sample beverage for Test Example 23 was prepared using the same method, except that it did not contain an acidulant. Similarly, a sample beverage for Test Example 22 was prepared using the same method, except that it did not contain an acidulant or dried dead bacterial powder of Lactococcus lactis subspecies Lactis JCM5805 strain.

[0102] (2. Sensory evaluation test) The degree of "mouth grittiness" experienced when drinking the sample beverages from Test Examples 22-28 was evaluated by a panel of four trained experts using a 10-point scale (1, 2, 3, 4, 5, 6, 7, 8, 9, 10). The evaluation criteria used were as follows: the degree of "mouth grittiness" for the sample beverage in Test Example 1 (magnesium concentration 0.0 mg / L) in Table 1 was assigned a score of 1 ("no mouth grittiness"), and the degree of "mouth grittiness" for the sample beverage in Test Example 11 in Table 1 was assigned a score of 10 ("extremely strong mouth grittiness"). The evaluation criteria divided the degree of mouth grittiness into the aforementioned 10 levels. For each test sample, the evaluation of oral irritation was based on a 19-point scale (1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10), with the score closest to the average of each panel's evaluation scores being used. Furthermore, the standard deviation of each panel's evaluation scores for each sample beverage was 0.43 or less for all sample beverages.

[0103] The results of the sensory evaluation test are shown in Table 4.

[0104] [Table 4]

[0105] As can be seen from the results in Table 4, when phosphoric acid, an inorganic acid, was further added, there was no change in the squeaky feeling in the mouth (Test Example 28 compared with Test Example 23), but when organic acids such as citric acid, lactic acid, malic acid, or tartaric acid were further added, the squeaky feeling in the mouth was further suppressed (Test Examples 24-27 compared with Test Example 23).

[0106] [Test 5] Effect of citric acid concentration on oral squeaking when dead lactic acid bacteria and citric acid are used together. The following tests were conducted to investigate how the concentration of citric acid affects the feeling of dryness in the mouth when dead lactic acid bacteria and citric acid are used together.

[0107] (1. Preparation of a sample of sugar-free beverage) Test Examples 31-37 were prepared by adding magnesium chloride, dried dead cell powder of Lactococcus lactis subspecies Lactis JCM5805 strain, and citric acid to deionized water containing carbon dioxide, so that the magnesium concentration, the concentration of dead lactic acid bacteria, and the citric acid concentration were as shown in Table 5. A sample beverage for Test Example 30 was prepared using the same method, except that citric acid was not used. A sample beverage for Test Example 29 was prepared using the same method, except that citric acid and dried dead cell powder of Lactococcus lactis subspecies Lactis JCM5805 strain were not used.

[0108] (2. Sensory evaluation test) The degree of "mouth grittiness" experienced when drinking the sample beverages from Test Examples 29-37 was evaluated by a panel of four trained experts using a 10-point scale (1, 2, 3, 4, 5, 6, 7, 8, 9, 10). The evaluation criteria used were as follows: the degree of "mouth grittiness" for the sample beverage in Test Example 1 (magnesium concentration 0.0 mg / L) in Table 1 was assigned a score of 1 ("no mouth grittiness"), and the degree of "mouth grittiness" for the sample beverage in Test Example 11 in Table 1 was assigned a score of 10 ("extremely strong mouth grittiness"). The evaluation criteria divided the degree of mouth grittiness into the aforementioned 10 levels. For each test sample, the evaluation of oral irritation was based on a 19-point scale (1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10), with the score closest to the average of each panel's evaluation scores being used. Furthermore, the standard deviation of each panel's evaluation scores for each sample beverage was 0.43 or less for all sample beverages.

[0109] The results of this sensory evaluation test are shown in Table 5.

[0110] [Table 5]

[0111] The results in Table 5 show that adding 0.005% by weight or more of citric acid in addition to dead lactic acid bacteria can more effectively suppress the squeaky feeling in the mouth. In Test Example 37, which contained 0.7% by weight of citric acid, the squeaky feeling in the mouth was sufficiently suppressed, but the sourness derived from the citric acid was strong.

[0112] [Test 6] Example of a prescription using Lactobacillus rhamnosus CRL1505, a lactic acid bacterium other than Lactococcus lactis subspecies lactis JCM5805.

[0113] When magnesium-containing sugar-free beverages (containing 1.3 billion or 15 billion dead Lactobacillus rhamnosus CRL1505 cells / L, etc.) were prepared using dead Lactobacillus rhamnosus CRL1505 cells instead of dead Lactococcus lactis subspecies lactis JCM5805 cells, following the method of Test 2 above, it was confirmed that the use of dead Lactobacillus rhamnosus CRL1505 cells suppressed the squeaky feeling in the mouth. [Industrial applicability]

[0114] According to the present invention, it is possible to provide a packaged sugar-free beverage in which the "gritty feeling in the mouth" characteristic of sugar-free beverages containing a predetermined concentration of magnesium is suppressed, as well as a method for producing the same.

Claims

1. A bottled, sugar-free carbonated beverage containing 500 million or more dead lactic acid bacteria of one or more types per liter, a magnesium concentration of 1.2 mg / L or more, a total concentration of 0.01% by weight or more of one or more organic acids, and a gas pressure of 0.1 to 0.3 MPa.

2. The packaged sugar-free carbonated beverage according to claim 1, wherein the total concentration of one or more organic acids is 0.4% by weight or more.

3. A bottled, sugar-free carbonated beverage containing 500 million or more dead cells of one or more types of lactic acid bacteria per liter, with a magnesium concentration of 1.2 mg / L or more, a total concentration of one or more types of organic acids of 0.01% by weight or more, and a gas pressure of 0.1 to 0.3 MPa, wherein the lactic acid bacteria include bacteria of the genus Lactobacillus, Streptococcus, Lactococcus, and Leuconostoc. The packaged unsweetened carbonated beverage is one or more bacteria selected from the group consisting of bacteria, bacteria of the genus Pediococcus, bacteria of the genus Enterococcus, bacteria of the genus Oenococcus, bacteria of the genus Bifidobacterium, bacteria of the genus Lentilactobacillus, bacteria of the genus Weissella, and bacteria of the genus Tetragenococcus.

4. The bottled unsweetened carbonated beverage according to claim 3, characterized in that the lactic acid bacteria is Lactobacillus rhamnosus.

5. A bottled, sugar-free carbonated beverage containing 500 million or more dead bacteria of one or more species capable of activating plasmacytoid dendritic cells per liter, with a magnesium concentration of 1.2 mg / L or more, a total concentration of one or more organic acids of 0.01% by weight or more, and a gas pressure of 0.1 to 0.3 MPa.

6. In the production of a bottled unsweetened carbonated beverage with a magnesium concentration of 1.2 mg / L or higher, the beverage contains 500 million or more dead bacteria of one or more species capable of activating plasmacytoid dendritic cells. The product contains one or more organic acids such that the total concentration of one or more organic acids is 0.01% by weight or more. A method for producing the bottled unsweetened carbonated beverage, characterized by setting the gas pressure to 0.1 to 0.3 MPa.

7. In the manufacture of a bottled unsweetened carbonated beverage with a magnesium concentration of 1.2 mg / L or higher, the beverage contains 500 million or more dead lactic acid bacteria of one or more types. The product contains one or more organic acids such that the total concentration of one or more organic acids is 0.01% by weight or more. A method for producing the bottled unsweetened carbonated beverage, characterized by setting the gas pressure to 0.1 to 0.3 MPa.

8. In the production of a bottled unsweetened carbonated beverage with a magnesium concentration of 1.2 mg / L or higher, the beverage contains 500 million or more dead bacteria of one or more species capable of activating plasmacytoid dendritic cells. The product contains one or more organic acids such that the total concentration of one or more organic acids is 0.01% by weight or more. A method for suppressing the gritty feeling in the mouth when drinking the bottled unsweetened carbonated beverage, characterized by setting the gas pressure to 0.1 to 0.3 MPa.

9. In the manufacture of a bottled unsweetened carbonated beverage with a magnesium concentration of 1.2 mg / L or higher, the beverage contains 500 million or more dead lactic acid bacteria of one or more types. The product contains one or more organic acids such that the total concentration of one or more organic acids is 0.01% by weight or more. A method for suppressing the gritty feeling in the mouth when drinking the bottled unsweetened carbonated beverage, characterized by setting the gas pressure to 0.1 to 0.3 MPa.

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