Beverage with suppressed culture medium odor of lactic acid bacteria

Abstract

The present invention addresses the problem of providing: a beverage which contain dead cells of lactic acid bacteria and in which a culture medium odor of lactic acid bacteria is suppressed; a method for producing the same; and the like.　The problem of the present invention can be solved by a beverage in which the concentration of dead cells of one or more types of lactic acid bacteria is at least 1.3 billion / L and the concentration of hesperidin is 10-1,000 mg / L.

Description

Beverage with reduced lactic acid bacteria culture odor

[0001] The present invention relates to a beverage and a method for producing the same, and more particularly to a beverage containing killed lactic acid bacteria cells and hesperidin at a predetermined concentration and having a suppressed culture medium odor of lactic acid bacteria, and a method for producing the same.

[0002] In response to the increasing health consciousness of people, many types of beverages containing killed lactic acid bacteria have been put on the market. However, in beverages containing killed lactic acid bacteria, the odor of the culture medium of the lactic acid bacteria can sometimes impair the flavor of the beverage.

[0003] As a technology for suppressing the culture medium odor of lactic acid bacteria in beverages, for example, Patent Document 1 discloses blending at least one selected from the group consisting of less than 20,000 ppb of nootkatone, less than 50,000 ppb of linalool, and less than 50,000 ppb of valencene to a beverage containing lactic acid bacteria powder and having a pH of over 4.6. However, it has not been known until now that the culture medium odor of lactic acid bacteria in beverages can be suppressed by adding hesperidin to the beverage at a predetermined concentration.

[0004] Japanese Patent Application Laid-Open No. 2019-103408

[0005] Beverages containing killed lactic acid bacteria cells have the problem that the flavor of the beverage may be impaired by the odor of the culture medium of the lactic acid bacteria. An object of the present invention is to provide a beverage containing killed lactic acid bacteria cells in which the odor of the culture medium of the lactic acid bacteria is suppressed, and a method for producing the same.

[0006] The present inventors have intensively investigated various methods to solve the above-mentioned problems, and have found that the culture medium odor of lactic acid bacteria can be suppressed by adjusting the hesperidin concentration to 10 to 1000 mg / L in a beverage containing one or more species of killed lactic acid bacteria at a concentration of 1.3 billion cells / L or more, thereby completing the present invention.

[0007] That is, the present invention provides the following inventions. (1) A beverage containing one or more species of killed lactic acid bacteria at a concentration of 1.3 billion cells / L or more and a hesperidin concentration of 10 to 1,000 mg / L. (2) The beverage according to (1) above, having a pH of 2.5 or more and 8.0 or less. (3) The beverage according to (1) or (2) above, wherein the lactic acid bacteria are one or more species selected from the group consisting of bacteria of the genus Lactococcus. (4) A method for producing the beverage, characterized in that the beverage contains killed lactic acid bacteria of one or more species of lactic acid bacteria at a concentration of 1.3 billion cells / L or more and hesperidin at a concentration of 10 to 1,000 mg / L. (5) A method for suppressing the odor of lactic acid bacteria culture media in a beverage, the method comprising the steps of: adding killed cells of one or more types of lactic acid bacteria to the beverage at a concentration of 1.3 billion cells / L or more; and adding hesperidin to the beverage at a concentration of 10 to 1,000 mg / L.

[0008] According to the present invention, it is possible to provide a beverage containing killed lactic acid bacteria cells in which the odor of the culture medium of lactic acid bacteria is suppressed, and a method for producing the same.

[0009] FIG. 1 is a diagram showing the relationship between Lactococcus lactis subsp. lactis JCM5805 strain and strains equivalent to said strain (strains derived from said strain and strains from which said strain is derived).

[0010] The present invention includes: [1] a beverage containing one or more species of killed lactic acid bacteria at a concentration of 1.3 billion cells / L or more and a hesperidin concentration of 10 to 1,000 mg / L (hereinafter also referred to as "the beverage of the present invention"); [2] a method for producing the beverage, characterized in that the beverage contains killed lactic acid bacteria of one or more species at a concentration of 1.3 billion cells / L or more and hesperidin at a concentration of 10 to 1,000 mg / L (hereinafter also referred to as "the production method of the present invention"); [3] a method for suppressing the culture medium odor of lactic acid bacteria in the beverage, characterized in that the beverage contains killed lactic acid bacteria of one or more species at a concentration of 1.3 billion cells / L or more and hesperidin at a concentration of 10 to 1,000 mg / L (hereinafter also referred to as "the suppression method of the present invention"); and the like.

[0011] In this specification, unless otherwise specified, a numerical range expressed with "to" naturally includes the numerical values ​​on both ends of "to." Furthermore, although "containing" and "added" are not synonymous, in all descriptions explaining the present invention in this specification, inventions in which "containing" is replaced with "added" are also described, and for example, inventions in which the word "containing" is replaced with "added" or "containing" is replaced with "added" are also described.

[0012] (Killed Cells of Lactic Acid Bacteria) In the present invention, killed cells of one or more types of lactic acid bacteria are used.

[0013] "Lactic acid bacteria" is a general term for all those taxonomically recognized as lactic acid bacteria, and is not limited by genus, species, strain, etc. Such "lactic acid bacteria" include bacteria that lactic acid ferment sugar to produce a large amount of lactic acid (preferably 50% or more of the lactic acid consumed), such as bacteria of the genus Lactococcus, Lactobacillus, Streptococcus, Leuconostoc, Pediococcus, Enterococcus, Oenococcus, Bifidobacterium, Weissella, and Tetragenococcus.

[0014] 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.

[0015] The genus or species of lactic acid bacteria used in the present invention is not particularly limited, and examples thereof include one or more bacteria selected from the group consisting of bacteria of the genus Lactococcus, Lactobacillus, Streptococcus, Leuconostoc, Pediococcus, Enterococcus, Oenococcus, Bifidobacterium, Weissella, and Tetragenococcus, and preferably one or more bacteria selected from the group consisting of bacteria of the genus Lactococcus, Lactobacillus, and Pediococcus. Examples of bacteria include the above bacteria, more preferably one or more bacteria selected from the group consisting of bacteria of the genus Lactococcus and bacteria of the genus Lactobacillus, even more preferably one or more bacteria selected from the group consisting of bacteria of the genus Lactococcus, even 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 subsp. lactis.

[0016] More specific preferred embodiments of the lactic acid bacteria of the present invention include Lactococcus lactis subsp. lactis (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 biovar diacetylactis), Lactococcus lactis subsp. cremoris (Lactococcus lactis subsp. cremoris JCM16167, Lactococcus lactis subsp. cremoris NBRC100676, etc.), Lactococcus raffinolactis, Lactococcus piscium, Lactococcus plantarum (Lactococcus plantarum JCM11056, etc.), Lactococcus garvieae (Lactococcus garvieae NBRC100934, etc.), Lactococcus lactis subsp. hordniae (Lactococcus lactis subsp. hordniae JCM1180, Lactococcus lactis subsp. hordniae JCM11040, etc.), Lactobacillus acidophilus (Lactobacillus acidophilus L-92, etc.),Lactobacillus delbrueckii subsp. bulgaricus (e.g., Lactobacillus bulgaricus OLL1073R-1), Lactobacillus delbrueckii subsp. delbrueckii, Lactobacillus delbrueckii subsp. lactis, Lactobacillus casei, Lactobacillus paracasei (e.g., Lactobacillus paracasei KW3110 and Lactobacillus paracasei MCC1849), Lactobacillus gasseri, Lactobacillus Lactobacillus gasseri (Lactobacillus gasseri SBT2055, etc.), Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus parakefiri (Lactobacillus parakefiri JCM8573, etc.), Lactobacillus plantarum (Lactobacillus plantarum L-137, etc.), Lactobacillus brevis, Lactobacillus rhamnosus (Lactobacillus rhamnosus GG, Lactobacillus rhamnosus CRL1505, etc.), Lactobacillus pentosus pentosus (Lactobacillus pentosus ONRICb0240, etc.), Lactobacillus fermentum, Lactobacillus fructivorans, Lactobacillus hilgardii,Streptococcus salivarius subsp. thermophilus, Leuconostoc mesenteroides subsp. cremoris, Leuconostoc lactis (Leuconostoc lactis NBRC12455, etc.), Leuconostoc carnosum (Leuconostoc carnosum JCM9695, etc.), Pediococcus damnosus (Pediococcus damnosus JCM5886, etc.), Pediococcus pentosaceus, Pediococcus acidilactici (Pediococcus Pediococcus acidilactici) (Pediococcus acidilactici JCM8797 and Pediococcus acidilactici K15, etc.), Pediococcus cellicola, Pediococcus claussenii, Pediococcus ethanolidurans, Pediococcus inopinatus, Pediococcus parvulus, Pediococcus stilesii, Enterococcus faecalis, Enterococcus faecium, Enterococcus alcedinis alcedinis), Oenococcus oeni (Oenococcus oeni JCM6125, etc.), Bifidobacterium animalis subsp. lactis (Bifidobacterium animalis subsp. lactis JCM10602, etc.),infantis (Bifidobacterium longum subsp. infantis JCM1222, etc.), Weissella paramesenteroides (Weissella paramesenteroides JCM9890, etc.), Weissella viridescens (Weissella viridescens JCM1174, etc.), and Tetragenococcus halophilus halophilus) (e.g., Tetragenococcus halophilus NRIC0098), and preferably, Lactococcus lactis subsp. lactis, Lactococcus lactis biovariant diacetylactis, Lactococcus lactis subsp. cremoris, Lactococcus raffinolactis, Lactococcus pythium, Lactococcus plantarum, Lactococcus garvieae, Lactococcus lactis subsp. holdoniae, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. delbrueckii, Lactobacillus delbrueckii Lactobacillus subsp. lactis, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus parakephili, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus rhamnosus, Lactobacillus pentosus, Lactobacillus fermentum, Lactobacillus fructivorans, Lactobacillus hilgardii, Pediococcus damnosus, Pediococcus pentosaceus, Pediococcus acidilactici, Pediococcus coericola, Pediococcus claussenii, Pediococcus ethanolidurans, Pediococcus inopinatus, Pediococcus parvulus, andExamples of the bacteria include one or more bacteria selected from the group consisting of Lactococcus lactis subsp. lactis, Lactococcus lactis biovariant diacetylactis, Lactococcus lactis subsp. cremoris, Lactococcus raffinolactis, Lactococcus pythium, Lactococcus plantarum, Lactococcus garvieae, Lactococcus lactis subsp. holdoniae, and Lactobacillus acidophilus. philus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. delbrueckii, Lactobacillus delbrueckii subsp. lactis, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus parakeophili, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus rhamnosus (Lactobacillus rhamnosus GG, and and Lactobacillus rhamnosus CRL1505), Lactobacillus pentosus, Lactobacillus fermentum, Lactobacillus fructivorans, and Lactobacillus hilgardii, and more preferably, one or two or more bacteria selected from the group consisting of Lactococcus lactis subsp. lactis, and more preferably, one or two or more bacteria selected from the group consisting of Lactococcus lactis subsp. lactis JCM5805, Lactococcus lactis sa lactis JCM20101, Lactococcus lactis subsp. lactis NBRC12007, Lactococcus lactis subsp. lactis NRIC1150, Lactococcus lactis subsp. lactis JCM7638, and Lactococcus lactis subsp. lactis ATCC11454, with Lactococcus lactis subsp. lactis JCM5805 being particularly preferred.

[0017] Further, in still another preferred embodiment of the genus or species of killed lactic acid bacteria used in the present invention, one or more species selected from the group consisting of bacteria of the genus Lactococcus can be mentioned, and preferred are Lactococcus lactis subsp. lactis JCM5805, Lactococcus lactis subsp. lactis NBRC12007, Lactococcus lactis subsp. lactis NRIC1150, Lactococcus lactis subsp. lactis JCM20101, and Lactococcus lactis subsp. lactis 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, and Lactococcus lactis subsp. holdoniae JCM11040.

[0018] Furthermore, preferred examples of the lactic acid bacteria of the present invention include lactic acid bacteria other than Lactobacillus paracasei MCC1849, Lactobacillus bacteria other than Lactobacillus paracasei MCC1849, lactic acid bacteria other than Lactobacillus paracasei, Lactobacillus bacteria other than Lactobacillus paracasei, and lactic acid bacteria other than Lactobacillus bacteria. Preferred examples of Lactobacillus bacteria other than Lactobacillus paracasei include Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. delbrueckii, Lactobacillus delbrueckii subsp. lactis, Lactobacillus casei, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus parakephili, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus rhamnosus, Lactobacillus pentosus, Lactobacillus fermentum, Lactobacillus fructivorans, and Lactobacillus hilgardii.

[0019] In the present invention, the lactic acid bacteria strains listed in this specification also include strains equivalent to the strains listed above, as long as they can suppress the culture medium odor of lactic acid bacteria in a specified beverage when the beverage contains killed lactic acid bacteria cells and hesperidin at a specified concentration. Here, "equivalent strains" refers to strains derived from the above strains, strains from which the above strains are derived, or descendant strains of those strains. Equivalent strains may also be preserved in other strain collection institutions. Figure 1 shows strains derived from Lactococcus lactis subsp. lactis JCM5805 and the strain from which Lactococcus lactis subsp. lactis JCM5805 is derived. Strains equivalent to Lactococcus lactis subsp. lactis JCM5805 shown in Figure 1 can also be used as the lactic acid bacteria of the present invention, as long as they can suppress the culture medium odor of lactic acid bacteria in a beverage containing killed lactic acid bacteria cells and hesperidin at a predetermined concentration. In this specification, the term "Lactococcus lactis subsp. lactis JCM5805" also includes these equivalent strains. Furthermore, the term "Lactobacillus rhamnosus CRL1505" also includes these equivalent strains.

[0020] Among the above lactic acid bacteria strains, the JCM strain can be obtained from the Microbial Materials Development Laboratory, RIKEN BioResource Center (3-1-1 Takanodai, Tsukuba, Ibaraki Prefecture), the NBRC strain from the Biological Genetic Resources Division, National Institute of Technology and Evaluation (2-5-8 Kazusa Kamatari, Kisarazu, Chiba Prefecture), the NRIC strain from the Strain Collection, Tokyo University of Agriculture (1-1-1 Sakuragaoka, Setagaya-ku, Tokyo), and the ATCC strain from the American Type Culture Collection (USA). Lactococcus lactis subsp. lactis JCM5805 can be obtained from the Microbial Materials Development Laboratory, RIKEN BioResource Center, as described above, but in the present invention, the same strain of JCM5805 stored in a collection institution other than the Microbial Materials Development Laboratory, RIKEN BioResource Center can also be used. Specifically, the same strain as JCM5805 can be obtained from the Biological Genetic Resources Division of the National Institute of Technology and Evaluation (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture), the Strain Collection of Tokyo University of Agriculture (1-1-1 Sakuragaoka, Setagaya-ku, Tokyo), the American Type Culture Collection (USA), etc.

[0021] The term "lactic acid bacteria" as used herein refers to killed cells of lactic acid bacteria. The killed cells of lactic acid bacteria used in the present invention may be dried or non-dried, but are preferably dried from the viewpoint of storage stability of the killed cells of lactic acid bacteria, and a suitable example is a dry powder.

[0022] The method for preparing killed lactic acid bacteria cells is not particularly limited, and examples thereof include a method in which a medium in which the lactic acid bacteria have been cultured is sterilized and then the cells are collected by filtration, centrifugation, etc., or a method in which the cells are collected from a medium in which the lactic acid bacteria have been cultured by filtration, centrifugation, etc. and then sterilized, and further drying or crushing treatments can be performed as necessary. The means of sterilization is not particularly limited, and conventional means for killing bacteria, such as heating, ultraviolet light, or gamma-ray irradiation, can be used. Alternatively, killed lactic acid bacteria cells can be obtained by blending live lactic acid bacteria cells and killing the lactic acid bacteria in the beverage during sterilization treatment during beverage preparation.

[0023] In the present invention, the concentration of killed cells of one or more types of lactic acid bacteria in a beverage is not particularly limited as long as it is 1.3 billion cells / L or more. However, since the culture medium odor of lactic acid bacteria, which is the subject of the present invention, increases and the significance of the present invention becomes greater, examples of the concentration include 5 billion cells / L or more, 10 billion cells / L or more, 30 billion cells / L or more, preferably 50 billion cells / L or more, 80 billion cells / L or more, 100 billion cells / L or more, and more preferably 200 billion cells / L or more, 300 billion cells / L or more, 600 billion cells / L or more. Furthermore, the upper limit of the concentration of dead cells of one or more types of lactic acid bacteria in a beverage is not particularly limited, but examples thereof include, from the viewpoint of keeping the level of culture medium odor of lactic acid bacteria within a certain range, a total of 6 trillion cells / L or less, 3 trillion cells / L or less, 2 trillion cells / L or less, 1 trillion cells / L or less, 800 billion cells / L or less, 600 billion cells / L or less, 300 billion cells / L or less, 200 billion cells / L or less, 100 billion cells / L or less, 80 billion cells / L or less, 50 billion cells / L or less, 30 billion cells / L or less, 10 billion cells / L or less, etc. These lower and upper limits can be combined as desired.

[0024] The concentration of killed lactic acid bacteria cells in a beverage can be adjusted, for example, by adjusting the amount of killed lactic acid bacteria cells blended into the beverage. The method for measuring the number of killed lactic acid bacteria cells in a beverage includes, without particular limitation, known methods for measuring the number of lactic acid bacteria cells, such as direct microscopy, particle electrochemical detection zone assay, PCR, and flow cytometry, with flow cytometry being preferred.

[0025] (Hesperidin) Hesperidin is a type of polyphenol found in the peel, veins, sacs, pulp, etc. of fruits such as citrus fruits. The hesperidin used in the present invention is not particularly limited, and hesperidin extracted and / or purified from citrus fruits, hesperidin as a food additive, fruit juice or processed fruit juice containing hesperidin from citrus fruits, etc., can be used alone or in combination as appropriate. The type of citrus fruit is not particularly limited, and examples include lemon, lime, orange, mandarin orange, grapefruit, yuzu, iyokan, natsumikan, hassaku orange, ponkan, shiikuwasha, and kabosu, with orange and mandarin orange being preferred. Furthermore, in the present invention, one or more of hesperidin and hesperidin derivatives may be used as hesperidin. The hesperidin derivative is not particularly limited, and examples thereof include methyl hesperidin, hesperidin glycoside (transglycosylated hesperidin), etc., which have higher solubility in water and allow a beverage to contain a larger amount of hesperidin. An example of a hesperidin glycoside is glucosyl hesperidin (more preferably monoglucosyl hesperidin).

[0026] In the present invention, the hesperidin concentration (concentration in the beverage or concentration added to the beverage) is not particularly limited as long as it is 10 to 1,000 mg / L relative to the total volume of the beverage, but from the viewpoint of further suppressing the odor of the culture medium of lactic acid bacteria, which is the issue at hand, the lower limit can be 10 mg / L or more, 12 mg / L or more, preferably 15 mg / L or more, 20 mg / L or more, 30 mg / L or more, 40 mg / L or more, more preferably 50 mg / L or more, 60 mg / L or more, 70 mg / L or more, 80 mg / L or more, 90 mg / L or more, and even more preferably 100 mg / L or more. Furthermore, from the viewpoint of preventing the unpleasant odor of hesperidin itself from becoming a problem, the upper limit can be less than 1,500 mg / L, preferably 1,000 mg / L or less. Other embodiments of the upper limit of hesperidin include 900 mg / L or less, 800 mg / L or less, 700 mg / L or less, 600 mg / L or less, 500 mg / L or less, 400 mg / L or less, 300 mg / L or less, 200 mg / L or less, and 100 mg / L or less. These lower and upper limits can be combined arbitrarily. Therefore, taking into consideration the balance between them, examples include 10 to 1000 mg / L, 15 to 1000 mg / L, preferably 50 to 1000 mg / L, 50 to 800 mg / L, and 50 to 500 mg / L. Note that the hesperidin concentration in the present invention refers to the concentration relative to the total amount of all hesperidin contained in the beverage and the amount of hesperidin derivatives converted to hesperidin, relative to the total amount of the beverage.

[0027] One preferred embodiment of the beverage of the present invention is a beverage to which hesperidin-containing fruit juice or processed fruit juice, etc., from citrus fruits or the like is added, thereby resulting in hesperidin being contained in the beverage at the predetermined concentration specified in the present invention. Another preferred embodiment of the beverage of the present invention is a beverage to which hesperidin-containing fruit juice or processed fruit juice, etc., from citrus fruits or the like is added, and then further hesperidin is added, thereby resulting in hesperidin being contained in the beverage at the predetermined concentration specified in the present invention.

[0028] The hesperidin concentration can be calculated and / or measured by methods known to those skilled in the art. For example, an analytical method using HPLC (high performance liquid chromatography) can be used. A more specific example of a method for measuring hesperidin concentration is described below. 3 mL of water and 30 mL of methanol are added to 4 g of the sample to be measured, and the mixture is extracted by ultrasonic irradiation for 5 minutes. The volume is then adjusted to 50 mL. This can be appropriately diluted to prepare a sample solution. High performance liquid chromatography can be performed under the following conditions. [High performance liquid chromatography operating conditions] Detector: UV-visible absorption spectrophotometer Column: XBridge C18, φ4.6 mm × 150 mm, particle size 3.5 μm Column temperature: 40°C Mobile phase: Mixture of water, acetonitrile, and phosphoric acid (850:150:1) Flow rate: 1.0 mL / min Measurement wavelength: 280 nm

[0029] (pH) The pH of the beverage of the present invention is not particularly limited, and examples thereof include 2.5 or more, 2.8 or more, 3.0 or more, 3.2 or more, 3.5 or more, 3.8 or more, and 4.0 or more, and 8.0 or less, 7.0 or less, 6.0 or less, 5.0 or less, 4.6 or less, 4.3 or less, 4.0 or less, 3.8 or less, 3.5 or less, 3.2 or less, and 3.0 or less, and more specifically, 2.5 to 8.0, 2.5 to 7.0, and 2.5 to 4.6. These pH values ​​are measured at 20°C. The pH of the beverage of the present invention can be adjusted by conventional methods, such as by adding an acidulant or a pH adjuster. Examples of such pH adjusters include substances that exhibit alkaline or acidic properties when dissolved in water, such as baking soda (sodium bicarbonate), sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, trisodium phosphate, tripotassium phosphate, sodium citrate, and carbon dioxide. The pH of the beverage of the present invention can be measured by a conventional method using a pH meter or the like.

[0030] (Beverage of the Present Invention) The beverage of the present invention is not particularly limited as long as it contains one or more species of killed lactic acid bacteria at a concentration of 1.3 billion cells / L or more and a hesperidin concentration of 10 to 1000 mg / L.

[0031] The beverage of the present invention is not particularly different from ordinary "beverages" in terms of the raw materials, production method, and production conditions used, except that the beverage of the present invention contains one or more species of killed lactic acid bacteria at a concentration of 1.3 billion cells / L or more and a hesperidin concentration of 10 to 1,000 mg / L.

[0032] The beverage of the present invention contains, as essential ingredients, 1.3 billion or more killed cells of one or more species of lactic acid bacteria per liter and 10 to 1,000 mg / liter of hesperidin. The beverage of the present invention may contain optional ingredients to the extent that the effects of the present invention are not impaired. Such optional ingredients may or may not include one or more selected from the group consisting of acidulants, sweeteners, bittering agents, flavorings, colorants, fruit juice or processed fruit juices, vegetable juice or processed vegetable juices, antioxidants, preservatives, stabilizers (thickening stabilizers, etc.), milk components, soybean components, lactic acid fermentation products, emulsifiers, pH adjusters, dietary fiber, vitamins, and minerals.

[0033] Examples of acidulants include phosphoric acid, lactic acid, tartaric acid, citric acid, malic acid, adipic acid, etc. Examples of sweeteners include sugars such as sucrose, fructose, glucose, maltose, starch syrup, reduced starch syrup, dextrin, cyclodextrin, trehalose, brown sugar, honey, etc.; sugar alcohols such as sorbitol, erythritol, xylitol, mannitol, etc.; and high-intensity sweeteners such as sucralose, stevia, acesulfame potassium, saccharin sodium, aspartame, glycyrrhizin, dipotassium glycyrrhizinate, thaumatin, neotame, etc.

[0034] In this specification, "fruit juice" means juice squeezed from fruit. Fruit juice can be produced by a method commonly used for producing fruit juice as a raw material for fruit drinks, such as a method of squeezing whole fruit (the entire fruit including the skin, etc.) using an in-line juicer or the like to separate the juice from a residue including the skin, and then sterilizing and cooling the squeezed juice, or a method of separating the whole fruit into the skin and pulp, or halving the fruit, squeezing only the pulp, and then sterilizing and cooling it. Fruit juice includes concentrated fruit juice obtained by concentrating squeezed fruit juice, and reconstituted fruit juice obtained by diluting concentrated fruit juice. Fruit juice refers to a puree or the like obtained by crushing fruit and squeezing or straining it to remove the skin, seeds, etc.

[0035] The fruit juice may be cloudy or clear, and may be clarified by a method such as enzyme treatment, microfiltration, or ultrafiltration.

[0036] The fruit juice may be commercially available juice, concentrated juice, paste, etc. Specific examples include juices and concentrated juices specified by the JAS standard (Japanese Agricultural Standards for Fruit Drinks), and one or more of these may be used to prepare the beverage of the present invention.

[0037] In this specification, the term "processed fruit juice" refers to processed fruit juice products, including, for example, fruit juice extracts, fruit juice flavors, and the like.

[0038] The beverage of the present invention can be produced by incorporating killed cells of one or more species of lactic acid bacteria at a concentration of 1.3 billion cells / L or more and hesperidin at a concentration of 10 to 1000 mg / L at any stage in a general method for producing a "beverage" that is well known in the art.

[0039] The beverage of the present invention does not have to be a bottled beverage, but is preferably a bottled beverage. Examples of such containers include resin bottle containers such as PET bottles, polypropylene bottles, and polyvinyl chloride bottles; glass bottles; and cans.

[0040] The beverage of the present invention may or may not be heat-sterilized. However, the culture medium odor of lactic acid bacteria in a beverage is further enhanced by heating a beverage containing a predetermined concentration of killed lactic acid bacteria cells (e.g., dried powder of lactic acid bacteria) at a predetermined high temperature (e.g., 65°C or higher, 70°C or higher, 75°C or higher, 80°C or higher, 85°C or higher, 100°C or higher, 120°C or higher, etc.) for a predetermined time (e.g., 4 minutes or more, 10 minutes or more, 20 minutes or more, 30 minutes or more) compared to when not heated. Therefore, since heat sterilization of a beverage increases the culture medium odor of lactic acid bacteria, which is the subject of the present invention, and thus further enhances the significance of the present invention, it is more preferable that the beverage of the present invention be a heat-sterilized beverage. Furthermore, from the viewpoint of improving shelf life, it is more preferable that the beverage of the present invention be a heat-sterilized beverage. The heat sterilization method and conditions can be the same as those typically used for bottled beverages and other beverages. Examples of heating temperature and time conditions for heat sterilization of the beverage of the present invention include 65°C for 10 minutes or more, 80°C for 10 minutes or more, 85°C for 10 minutes or more, 85°C for 30 minutes or more, 120°C for 4 minutes or more, or conditions that are equivalent to or more effective than these conditions.

[0041] The beverage of the present invention may be a beverage that is sold in a refrigerated state (0 to 10°C), at room temperature, or at a warm temperature (50 to 70°C), but may be a beverage other than a chilled beverage that is sold in a refrigerated state.

[0042] The type of beverage of the present invention is not particularly limited, but is preferably a fruit juice-containing beverage containing fruit juice or a processed fruit juice, more preferably a fruit juice-containing beverage containing citrus fruit juice or a processed fruit juice, and more preferably a fruit juice-containing beverage containing mandarin orange or orange juice or a processed fruit juice. In addition, the beverage of the present invention may be a beverage other than fermented milk or a lactic acid bacteria beverage.

[0043] (Production Method of the Present Invention) The beverage of the present invention can be produced according to a conventional method for producing a beverage, except that killed cells of one or more species of lactic acid bacteria are added to the beverage at a concentration of 1.3 billion cells / L or more and hesperidin is added at a concentration of 10 to 1000 mg / L.

[0044] The production method of the present invention is not particularly limited, as long as it is a method for producing a beverage, characterized in that killed cells of one or more types of lactic acid bacteria are contained in the beverage at a concentration of 1.3 billion cells / L or more and hesperidin is contained in a concentration of 10 to 1000 mg / L.

[0045] More specifically, a method for incorporating killed cells of one or more species of lactic acid bacteria at a concentration of 1.3 billion cells / L or more and hesperidin at a concentration of 10 to 1,000 mg / L in a beverage includes, during the production of a beverage, incorporating killed cells of one or more species of lactic acid bacteria and hesperidin into a raw material for the beverage (e.g., "water" or "water further containing some or all of the optional ingredients") (hereinafter collectively referred to as "water, etc.") so that the concentration of killed cells of one or more species of lactic acid bacteria is 1.3 billion cells / L or more (preferably adjusted to 1.3 billion cells / L or more) and the concentration of hesperidin is 10 to 1,000 mg / L (preferably adjusted to 10 to 1,000 mg / L). Alternatively, a method may be used in which killed cells of one or more species of lactic acid bacteria and hesperidin are incorporated into water, etc., at predetermined concentrations, simultaneously with some or all of the optional ingredients.

[0046] In the production method of the present invention, the beverage contains 1.3 billion killed cells / L of one or more species of lactic acid bacteria and 10 to 1,000 mg / L of hesperidin as essential ingredients. In the production method of the present invention, the beverage may or may not contain one or more optional ingredients selected from the group consisting of acidulants, sweeteners, bittering agents, flavorings, colorants, fruit juice or processed fruit juices, vegetable juice or processed vegetable juices, antioxidants, preservatives, stabilizers (thickening stabilizers, etc.), milk components, soybean components, lactic acid fermentation products, emulsifiers, pH adjusters, dietary fiber, vitamins, and minerals.

[0047] In the production method of the present invention, the order in which the ingredients are added is not particularly limited as long as the beverage of the present invention containing the ingredients to be used can be produced. When the beverage of the present invention is to be made into a bottled beverage, it can be produced by preparing a liquid containing the ingredients, filling it into a container, and sealing it.

[0048] In the production method of the present invention, heat sterilization of the beverage may or may not be performed. However, as described above, heat sterilization of the beverage increases the culture medium odor of lactic acid bacteria, which is the subject of the present invention, thereby further enhancing the significance of the present invention. Therefore, it is more preferable that the production method of the present invention includes heat sterilization of the beverage. Furthermore, from the viewpoint of improving shelf life, it is more preferable that the production method of the present invention includes heat sterilization of the beverage. The method of heat sterilization is not particularly limited, and examples thereof include high-temperature short-time sterilization (HTST method), pasteurizer sterilization, ultra-high-temperature heat treatment (UHT method), retort sterilization, etc.

[0049] (Suppression Method of the Present Invention) The suppression method of the present invention is not particularly limited as long as it is a method for suppressing the culture medium odor of lactic acid bacteria in a beverage, characterized in that, in the production of a beverage, killed cells of one or more types of lactic acid bacteria are added to the beverage at a concentration of 1.3 billion cells / L or more and hesperidin is added at a concentration of 10 to 1000 mg / L.

[0050] In producing a beverage, the same method as described above in (Production method of the present invention) can be used to contain killed cells of one or more types of lactic acid bacteria at a concentration of 1.3 billion cells / L or more and hesperidin at a concentration of 10 to 1000 mg / L in the beverage.

[0051] (Suppression of lactic acid bacteria culture medium odor in the present invention) The beverage of the present invention is a beverage in which the lactic acid bacteria culture medium odor is suppressed. In this specification, "culture medium odor" refers to the odor derived from the culture medium in which lactic acid bacteria are cultured, and is likely to become problematic when killed lactic acid bacteria cells (e.g., dried powder) cultured in a culture medium containing, for example, yeast extract, meat extract, peptone, etc. are blended. The lactic acid bacteria culture medium odor is accompanied by unpleasant flavors such as umami and animal odor, and is a unique flavor that is particularly undesirable in general soft drinks, etc. As mentioned above, heat sterilization of a beverage increases the lactic acid bacteria culture medium odor, which is the subject of the present invention, thereby further enhancing the significance of the present invention. Therefore, the suppression method of the present invention is more suitable for use with beverages that include heat sterilization of the beverage during production.

[0052] In the present invention, a beverage "in which the culture medium odor of lactic acid bacteria is suppressed" means a beverage containing killed cells of one or more types of lactic acid bacteria at a concentration of 1.3 billion cells / L or more, in which the culture medium odor of lactic acid bacteria is suppressed compared to a beverage (hereinafter also referred to as a "control beverage") produced by the same method using the same types of raw materials to the same final concentration, except that it does not contain hesperidin at a concentration of 10 to 1000 mg / L.

[0053] A trained panel can easily and clearly determine the degree of lactic acid bacteria culture medium odor in a particular beverage and how that degree compares with the control beverage of the present invention (for example, whether the lactic acid bacteria culture medium odor is suppressed and to what extent it is suppressed).

[0054] The present invention will be specifically described based on the following examples, but the present invention is not limited to these examples.

[0055] [Test 1] Effect of Inclusion of Hesperidin The following test was carried out to investigate the effect of including hesperidin in a beverage on the culture medium odor of lactic acid bacteria.

[0056] (1. Preparation of sample beverages) Sample beverages of Test Examples 1 to 12 were prepared by adding dried killed cell powder of Lactococcus lactis subsp. lactis JCM5805, hesperidin, and phosphoric acid to ion-exchanged water so as to achieve the Lactococcus lactis subsp. lactis JCM5805 concentration, hesperidin concentration, and pH shown in Table 2. Each sample beverage was filled into a can and heat sterilized at 80°C for 10 minutes.

[0057] (2. Sensory Evaluation Test) The degree of culture medium odor of the sample beverages obtained in Test Examples 1 to 12 was evaluated by four trained expert panelists using the evaluation criteria in Table 1 below. The differences between 1 point and 1.5 points, between 1.5 points and 2 points, between 2 points and 2.5 points, between 2.5 points and 3 points, between 3 points and 3.5 points, between 3.5 points and 4 points, between 4 points and 4.5 points, and between 4.5 points and 5 points were all considered to be approximately the same. Furthermore, the average of the evaluation results of the four expert panelists was calculated for each sample beverage, and the value obtained by rounding the average to one decimal place was used as the evaluation result for that sample beverage. All standard deviations were 0.5 or less. Beverages with an evaluation score that was 0.5 or more lower than that of Test Example 1 (control beverage) could be evaluated as beverages with suppressed culture medium odor.

[0058]

[0059] The results of the sensory evaluation test for the sample beverages of Test Examples 1 to 12 are shown in Table 2.

[0060]

[0061] As can be seen from the results in Table 2, all of Test Examples 3 to 12 demonstrated a greater suppression of culture medium odor than Test Example 1 (control beverage). These results demonstrate that adding hesperidin at a concentration of 10 mg / L or higher to a beverage containing killed lactic acid bacteria cells can suppress the culture medium odor of lactic acid bacteria. Furthermore, this effect was shown to increase depending on the concentration of hesperidin added. From the perspective of achieving a greater suppression effect on the culture medium odor of lactic acid bacteria, a hesperidin concentration of 15 mg / L or higher is preferred, with 50 mg / L or higher being more preferred. However, in Test Example 12, although the suppression effect on the culture medium odor of lactic acid bacteria was high, it was also evaluated that the hesperidin itself had a strong unpleasant odor. Therefore, the hesperidin concentration is preferably less than 1500 mg / L, and more preferably 1000 mg / L or lower.

[0062] [Test 2] Effect of Differences in Concentration of Killed Lactic Acid Bacteria Cells The following test was conducted to investigate how differences in concentration of killed lactic acid bacteria cells affect the inhibitory effect of hesperidin on the odor-inhibiting effect of lactic acid bacteria on culture media.

[0063] (1. Preparation of sample beverages) Sample beverages of Test Examples 13 to 24 were prepared and heat-sterilized in the same manner as in [Test 1] above, except that the concentrations of Lactococcus lactis subsp. lactis JCM5805 strain and hesperidin were adjusted to the values ​​shown in Table 3. Note that the sample beverages of Test Examples 1 and 6 were prepared and heat-sterilized in the same manner as in [Test 1] above.

[0064] (2. Sensory Evaluation Test) The sample beverages obtained in Test Examples 1, 6, and 13 to 24 were subjected to a sensory evaluation test using the same method as in Test 1 above. The results are shown in Table 3.

[0065]

[0066] As can be seen from the results in Table 3, when the concentration of killed lactic acid bacteria cells was 1.3 billion cells / L or more, a culture medium odor was generated, and the higher the concentration of killed lactic acid bacteria cells, the stronger the culture medium odor became. Furthermore, it was shown that adding hesperidin to a beverage had the effect of suppressing the culture medium odor, regardless of the concentration of killed lactic acid bacteria cells.

[0067] [Test 3] Effect of Differences in pH The following test was conducted to investigate how differences in the pH of a beverage affect the inhibitory effect of hesperidin on the culture medium odor of lactic acid bacteria.

[0068] (1. Preparation of sample beverages) Sample beverages of Test Examples 25 to 34 were prepared and heat sterilized in the same manner as in [Test 1] above, except that the pH was adjusted to the values ​​shown in Table 4. Note that sample beverages of Test Examples 1 and 6 were prepared and heat sterilized in the same manner as in [Test 1] above.

[0069] (2. Sensory Evaluation Test) The sample beverages obtained in Test Examples 1, 6, and 25 to 34 were subjected to a sensory evaluation test using the same method as in Test 1 above. The results are shown in Table 4.

[0070]

[0071] As can be seen from the results in Table 4, when hesperidin is added to sample beverages with a pH of 2.5 to 8, it is possible to obtain an inhibitory effect on culture medium odor.

[0072] [Test 4] Formulation example using citrus fruit juice The following test was carried out using citrus fruit juice containing hesperidin.

[0073] (1. Preparation of Sample Beverages) Sample beverages of Test Examples 35 to 42 were prepared and heat-sterilized in the same manner as in [Test 1] above, except that 2% orange juice containing hesperidin at a concentration of 355 mg / L was added to the beverages, resulting in 7.1 mg / L of hesperidin derived from orange juice, and further hesperidin was added as necessary to ultimately contain hesperidin at the concentrations shown in Table 5. The sample beverage of Test Example 1 was prepared and heat-sterilized in the same manner as in [Test 1] above.

[0074] The concentration of hesperidin contained in orange juice was measured by the following method. 3 mL of water and 30 mL of methanol were added to 4 g of the sample to be measured, and the mixture was extracted by ultrasonic irradiation for 5 minutes. The volume was then adjusted to 50 mL. This was then diluted appropriately to prepare a sample solution. High-performance liquid chromatography was performed under the following conditions. [High-performance liquid chromatography operating conditions] Detector: UV-visible absorption spectrophotometer Column: XBridge C18, φ4.6 mm × 150 mm, particle size 3.5 μm Column temperature: 40°C Mobile phase: mixture of water, acetonitrile, and phosphoric acid (850:150:1) Flow rate: 1.0 mL / min Measurement wavelength: 280 nm

[0075] (2. Sensory Evaluation Test) The obtained sample beverages of Test Example 1 and 35 to 42 were subjected to a sensory evaluation test in the same manner as in the above [Test 1]. The results are shown in Table 5.

[0076]

[0077] As can be seen from the results in Table 5, even when orange juice was used in part to achieve a predetermined concentration of hesperidin in the beverage, the inhibitory effect on the culture medium odor of lactic acid bacteria was achieved, similar to when no orange juice was used (Table 2).

[0078] According to the present invention, it is possible to provide a beverage containing killed lactic acid bacteria cells in which the odor of the culture medium of lactic acid bacteria is suppressed, and a method for producing the same.

Claims

1. A beverage containing one or more types of killed lactic acid bacteria at a concentration of 1.3 billion cells / L or more and a hesperidin concentration of 10 to 1,000 mg / L.

2. The beverage according to claim 1, having a pH of 2.5 or more and 8.0 or less.

3. The beverage according to claim 1 or 2, wherein the lactic acid bacteria is one or more species selected from the group consisting of bacteria of the genus Lactococcus.

4. A method for producing a beverage, comprising the steps of: containing killed cells of one or more types of lactic acid bacteria in the beverage at a concentration of 1.3 billion cells / L or more; and containing hesperidin in a concentration of 10 to 1,000 mg / L.

5. A method for suppressing the culture medium odor of lactic acid bacteria in a beverage, comprising the steps of: adding killed cells of one or more types of lactic acid bacteria to the beverage at a concentration of 1.3 billion cells / L or more; and adding hesperidin at a concentration of 10 to 1,000 mg / L.

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