Lactic acid bacteria and method for obtaining same, and food and beverage containing lactic acid bacteria

Abstract

The present invention provides a lactic acid bacterium that grows well in a low pH and high sucrose environment. A lactic acid bacterium, which is a species belonging to the genus Lactobacillus (Lactobacillus sp.) and is preserved under the preservation number: NITE BP-03116 (identification number: WR16-4).

Description

Technical Field

[0001] The present invention relates to novel lactic acid bacteria and a method for obtaining the same, and lactic acid bacteria-containing food and beverages containing the lactic acid bacteria. Background Art

[0002] It is said that the optimal growth pH range of general lactic acid bacteria is pH 6 to 7, the growth limit in the acidic region is pH 4, and the growth limit in the alkaline region is pH 8. However, some lactic acid bacteria have excellent acid resistance and can grow even in harsh environments (around pH 3) where general lactic acid bacteria have difficulty growing (for example, Patent Documents 1 and 2).

[0003] There are also lactic acid bacteria that are able to grow even in an environment with a high salt concentration such as miso or soy sauce and have excellent salt resistance (for example, Patent Documents 3 and 4).

[0004] As described above, there are many lactic acid bacteria that can grow even in harsh environments. However, to the best of the applicant's knowledge, there have been no reports on the existence of lactic acid bacteria that can grow well in harsh environments of low pH (high acidity) and high sucrose concentration.

[0005] Therefore, the applicant believes that through the presence of such lactic acid bacteria, for example, liquids obtained by squeezing a mixture of plants and sugar or beverages based on such liquids, or materials such as jams and fruit sauces that have high acidity and sucrose concentrations and are difficult to ferment using ordinary lactic acid bacteria can be easily fermented, thereby providing new fermented foods or fermented beverages; research has been conducted on lactic acid bacteria that can grow well in a low pH environment and a high sucrose environment as described above, and lactic acid bacteria that can grow well in the harsh environment of a low pH and a high sucrose concentration have finally been successfully obtained.

[0006] Prior art literature

[0007] Patent Literature

[0008] Patent Document 1: Japanese Patent Application Publication No. 2009-225792

[0009] Patent Document 2: WO2013 / 001862

[0010] Patent Document 3: Japanese Patent Application Publication No. 2007-236344

[0011] Patent Document 4: Japanese Patent Application Publication No. 2018-42557 Summary of the invention

[0012] Problems to be solved by the invention

[0013] An object of the present invention is to provide a lactic acid bacterium that can grow well in a low pH and high sucrose environment.

[0014] Means for solving problems

[0015] The gist of the present invention will be described with reference to the accompanying drawings.

[0016] The present invention relates to a lactic acid bacterium, characterized in that it is a species belonging to the genus Lactobacillus (Lactobacillus sp.) and is preserved under the preservation number: NITE BP-03116 (identification number: WR16-4).

[0017] The present invention also relates to the lactic acid bacteria according to claim 1, characterized in that the lactic acid bacteria have the base sequence of the 16S rDNA region shown in SEQ ID NO: 1.

[0018] In addition, the present invention relates to the lactic acid bacteria according to claim 1, characterized in that the lactic acid bacteria are obtained from a plant fermentation extract.

[0019] In addition, the present invention relates to the lactic acid bacteria according to claim 2, characterized in that the lactic acid bacteria are obtained from a plant fermentation extract.

[0020] In addition, the present invention relates to the lactic acid bacteria according to claim 3, characterized in that the plant fermentation extract is a radish fermentation extract.

[0021] In addition, the present invention relates to the lactic acid bacteria according to claim 4, characterized in that the plant fermentation extract is a radish fermentation extract.

[0022] The present invention also relates to the lactic acid bacteria according to claim 5, wherein the radish fermentation extract is obtained by mixing radish, granulated sugar and a bacterial solution and fermenting the mixture.

[0023] The present invention also relates to the lactic acid bacteria according to claim 6, wherein the radish fermentation extract is obtained by mixing radish, granulated sugar and a bacterial solution and fermenting the mixture.

[0024] Furthermore, the present invention relates to the lactic acid bacteria according to claim 7, wherein the bacterial solution is composed of yeast.

[0025] Furthermore, the present invention relates to the lactic acid bacteria according to claim 8, wherein the bacterial solution is composed of yeast.

[0026] The present invention also relates to the lactic acid bacteria according to any one of claims 1 to 10, characterized in that the lactic acid bacteria proliferate in a low pH environment of pH 3.5 to 5.5 and also proliferate in a high sucrose concentration environment of 30 to 60 wt%.

[0027] The present invention also relates to the lactic acid bacteria according to any one of claims 1 to 10, characterized in that the lactic acid bacteria proliferate in a low pH environment of pH 3.5 to 5.5 and also proliferate in a high fructose / glucose concentration environment of 10 to 30 wt% fructose and glucose concentrations, respectively.

[0028] In addition, the present invention relates to the lactic acid bacteria described in any one of claims 1 to 10, characterized in that the lactic acid bacteria proliferate in a low pH environment of pH 3.5 to 5.5, and also proliferate in a high concentration sucrose environment of 30 to 60 wt% of sucrose concentration, and further, proliferate in a low pH environment of pH 3.5 to 5.5, and also proliferate in a high concentration fructose / glucose environment of 10 to 30 wt% of fructose concentration and glucose concentration, respectively.

[0029] In addition, the present invention relates to a method for obtaining lactic acid bacteria, characterized in that a plant fermentation extract is added to a sterilized liquid culture medium to obtain a culture solution, then the culture solution is enriched and cultured to obtain an enriched culture solution, then the enriched culture solution and a sterilized separation medium are poured to prepare a plate culture medium, and then colonies appearing in the plate culture medium are fished out to obtain lactic acid bacteria that can proliferate in a low pH environment and a high concentration of sucrose environment.

[0030] The present invention also relates to a method for obtaining lactic acid bacteria according to claim 14, wherein the plant fermentation extract is a radish fermentation extract obtained by mixing radish, fine sugar and a bacterial solution and fermenting the mixture.

[0031] Furthermore, the present invention relates to the method for obtaining lactic acid bacteria according to claim 15, characterized in that the bacterial solution is composed of yeast.

[0032] In addition, the present invention relates to a method for obtaining lactic acid bacteria according to any one of claims 14 to 16, characterized in that the liquid culture medium is prepared by adding an antibiotic to a mixture of a mixed fermentation extract and a wild vegetable-fine sugar mixture, the mixed fermentation extract is obtained using either or both of a vegetable fermentation extract and a fruit fermentation extract, and the wild vegetable-fine sugar mixture is obtained by mixing a decoction of wild grasses with fine sugar.

[0033] Furthermore, the present invention relates to the method for obtaining lactic acid bacteria according to claim 17, characterized in that the antibiotic is cycloheximide.

[0034] The present invention also relates to a lactic acid bacteria-containing food or drink, characterized in that it contains the lactic acid bacteria according to any one of claims 1 to 10.

[0035] The present invention also relates to a lactic acid bacteria-containing food or drink, characterized in that it contains the lactic acid bacteria according to claim 11.

[0036] The present invention also relates to a lactic acid bacteria-containing food or drink, characterized in that it contains the lactic acid bacteria according to claim 12.

[0037] The present invention also relates to a lactic acid bacteria-containing food or drink, characterized in that it contains the lactic acid bacteria according to claim 13.

[0038] Furthermore, the present invention relates to the lactic acid bacteria-containing food and drink according to claim 19, characterized in that the lactic acid bacteria-containing food and drink is a fermented food and drink.

[0039] Furthermore, the present invention relates to the lactic acid bacteria-containing food and drink according to claim 20, characterized in that the lactic acid bacteria-containing food and drink is a fermented food and drink.

[0040] Furthermore, the present invention relates to the lactic acid bacteria-containing food and drink according to claim 21, characterized in that the lactic acid bacteria-containing food and drink is a fermented food and drink.

[0041] Furthermore, the present invention relates to the lactic acid bacteria-containing food and drink according to claim 22, characterized in that the lactic acid bacteria-containing food and drink is a fermented food and drink.

[0042] Effects of the Invention

[0043] The lactic acid bacteria of the present invention grow (proliferate) dominantly in a low pH and high sucrose environment, and therefore, for example, can easily ferment liquids obtained by squeezing a mixture of plants and sugar or beverages based on the liquids, or materials such as jams and fruit sauces that have high acidity and sucrose concentrations and are difficult to ferment using general lactic acid bacteria, thereby providing new fermented foods or fermented beverages. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 The Lactobacillus acidophilus JCM 1132 in this example is shown. T A graph of the pH tolerance test results.

[0045] Figure 2 This is a graph showing the results of a pH resistance test of Lactococcus lactis subsp. lactis NBRC12007 in this Example.

[0046] Figure 3The NBRC100495 of Leuconostoc mesenteroides subsp. dextranicum in this example is shown. T A graph of the pH tolerance test results.

[0047] Figure 4 This is a graph showing the results of the pH tolerance test of the WR16-4 strain in this Example.

[0048] Figure 5 The Lactobacillus acidophilus JCM 1132 in this example is shown T A graph showing the results of a sucrose tolerance test.

[0049] Figure 6 This is a graph showing the results of a sucrose tolerance test of Lactococcus lactis subsp. lactis NBRC12007 in this Example.

[0050] Figure 7 The figure shows the Leuconostoc mesenteroides subspecies glucan NBRC100495 in this example. T A graph showing the results of a sucrose tolerance test.

[0051] Figure 8 This is a diagram showing the results of a sucrose tolerance test of the WR16-4 strain in this Example.

[0052] Fig. 9 This is a table showing the composition of the MRS medium in the sucrose tolerance test of this example.

[0053] Fig.10 This is a table showing the composition of the MRS medium in the fructose / glucose tolerance test of this example.

[0054] Fig.11 This is a diagram showing the results of the fructose / glucose tolerance test of the WR16-4 strain in this Example.

[0055] Fig.12 This is a data list showing the homology search results in the gene database (NCBI) of the present invention. DETAILED DESCRIPTION

[0056] Embodiments of the present invention which are considered to be preferred will be briefly described with reference to the drawings to illustrate the effects of the present invention.

[0057] The lactic acid bacteria of the present invention belong to the genus Lactobacillus (Lactobacillus sp.), and are deposited under the accession number: NITE BP-03116 (identification number: WR16-4).

[0058] Based on the sequence data (16S rDNA sequence) obtained from the lactic acid bacteria of the present invention, a homology search was performed using the gene database of NCBI (National Center for Biotechnology Information, U.S. National Center for Biotechnology Information), and the result showed that the highest homology with the above sequence was 93% (refer to Fig.12 ), it can be said from this result that the lactic acid bacteria of the present invention are a new species of lactic acid bacteria (species belonging to the genus Lactobacillus) that have not been seen before.

[0059] The lactic acid bacteria of the present invention have the following characteristics: they are difficult to grow at pH 6 to 7, which is considered to be the best growth environment for general lactic acid bacteria, and grow well in an environment of pH lower than pH 6 to 7 (for example, pH 3.5 to 5.5).

[0060] Furthermore, the lactic acid bacteria of the present invention have the characteristics of growing well even in a low pH (eg, pH 5 or less) and high sucrose (eg, 30 wt % or more) environment where general lactic acid bacteria have difficulty growing.

[0061] Thus, since the lactic acid bacteria of the present invention have the characteristic of predominantly proliferating in a low pH and high sucrose environment, for example, liquids obtained by squeezing a mixture of plants and sugar or beverages based on the liquids, or materials such as jams and fruit sauces that have high acidity and sucrose concentrations and are difficult to ferment using general lactic acid bacteria can be easily fermented.

[0062] Example

[0063] Specific embodiments of the present invention will be described based on the drawings.

[0064] The lactic acid bacteria of this example have the base sequence of the 16S rDNA region shown in SEQ ID NO: 1, and are lactic acid bacteria belonging to the species (Lactobacillus sp.) of the genus Lactobacillus deposited under the accession number: NITE BP-03116 (identification number: WR16-4).

[0065] The cell morphology of the lactic acid bacteria of this example is short rod-shaped, and in the form of colonies, it is white and produces extracellular polysaccharides (viscous substances).

[0066] Hereinafter, the lactic acid bacteria of this example (hereinafter referred to as WR16-4 strain) will be described in detail.

[0067] <About the strain of WR16-4>

[0068] Microbial identification was performed based on the 16S rDNA data of the WR16-4 strain, and the strain of WR16-4 was investigated. It should be noted that the microbial identification was entrusted to FASMAC Co., Ltd.

[0069] The results of the microbial identification test showed that the strain with the highest homology value was Lactobacillus kefiri (ATCC = 35441) (homology value 93.14%).

[0070] <Regarding the novelty of WR16-4 strain>

[0071] Based on the 16S rDNA data of the WR16-4 strain, a homology search was conducted using the gene database of NCBI to investigate the novelty of the WR16-4 strain.

[0072] Fig.12 This is a list of data that showed high homology with the sequences from the sample in this homology search. As shown in this Fig.12 As shown, the highest homology with this WR16-4 strain was about 93%. Generally, when there is no species with a homology value of more than 98.7%, it is judged as a new species. Therefore, it can be said that this WR16-4 strain is a new (new species) lactic acid bacterium.

[0073] <Isolation source of WR16-4 strain>

[0074] The WR16-4 strain was isolated from a plant fermentation extract, specifically from a radish fermentation extract.

[0075] The radish fermentation extract used as the isolation source was obtained by mixing radish, fine granulated sugar, and a bacterial solution and allowing it to ferment. Specifically, cut radish was mixed with the same amount of fine granulated sugar as the radish, fermented using yeast, then pressed, and the resulting liquid component was further fermented and ripened. It should be noted that the plant used as the isolation source can also be vegetables other than radish.

[0076] <Isolation method of WR16-4 strain>

[0077] When isolating the WR16-4 strain from the above radish fermentation extract, it cannot be isolated using the general isolation method for lactic acid bacteria. It can be isolated using the following isolation method discovered by the applicant.

[0078] (1) Add 80 μl of the above radish fermentation extract to 10 ml of a liquid medium sterilized wet (121 °C, 15 minutes), and incubate at 25 °C for 6 days to obtain a culture solution.

[0079] (2) Add 80 μl of the obtained culture solution to 10 ml of the above liquid medium, and incubate again at 25 °C for 3 - 4 days to obtain a culture solution.

[0080] (3) Perform enrichment culture by repeating the treatment in step (2) multiple times (8 times in this example) to obtain an enriched culture solution.

[0081] (4) 1 ml of the obtained enriched culture solution and 15 ml of a wet-sterilized (121° C., 15 minutes) separation medium were poured, and the mixture was cooled and solidified to prepare a plate.

[0082] (5) The plate was incubated at 25°C for 10 days.

[0083] (6) The colonies appearing on the plate were collected to obtain the WR16-4 strain.

[0084] It should be noted that the above-mentioned liquid culture medium is prepared by adding antibiotics to a mixture of a mixed fermentation extract and a weed-fine sugar mixture, the above-mentioned mixed fermentation extract is obtained using either or both of a vegetable fermentation extract and a fruit fermentation extract, and the above-mentioned weed-fine sugar mixture is obtained by mixing a decoction of weeds with fine sugar, specifically, a mixture of the above-mentioned mixed fermentation extract, a weed-fine sugar mixture obtained by mixing fine sugar in a ratio of 1:1 with decoctions of multiple weeds, and a 1g / L cycloheximide solution in a ratio of 10:90:1.

[0085] The above-mentioned separation medium was prepared by wet-sterilizing and mixing the medium A and medium B shown in the following table.

[0086] Table 1

[0087]

[0088] In order to confirm the characteristics of the WR16-4 strain obtained by the above-mentioned isolation method, a pH tolerance test and a sucrose tolerance test were performed using common lactic acid bacteria and comparing with these.

[0089] Specifically, as a comparative lactic acid bacteria for the WR16-4 strain, Lactobacillus acidophilus JCM 1132 belonging to a species of the genus widely known among lactic acid bacteria and having an optimal growth pH range of pH 6 to 7 like general lactic acid bacteria was used. T , Lactococcus lactis subsp. lactis NBRC12007 of the genus Lactococcus, Leuconostoc mesenteroides subsp. dextranicum NBRC100495 of the genus Leuconostoc T These three types of lactic acid bacteria.

[0090] <Pre-culture of each lactic acid bacteria>

[0091] When conducting the above two tolerance tests, the following pre-cultures were carried out for each lactic acid bacterium.

[0092] (1) Adjust with 2-fold concentration Fig. 9 The MRS medium shown. At this time, 4N sodium hydroxide solution or 4N hydrochloric acid was added to adjust the medium for strain WR16-4 to pH 4.5 and the medium for the comparison object to pH 6.5.

[0093] (2) Prepare a 4 wt% sucrose solution or a 4 wt% glucose solution.

[0094] (3) For the above (1) and (2), autoclaving for moist heat sterilization (121 °C, 20 minutes) was carried out respectively.

[0095] (4) After cooling the solutions of (1) and (2) after moist heat sterilization, they were mixed in a 1:1 (v / v) manner in a clean bench to prepare an MRS medium containing 2 wt% sucrose (pH 4.5) or an MRS medium containing 2 wt% glucose (pH 6.5).

[0096] (5) Strain WR16-4 was cultured using the MRS medium containing 2 wt% sucrose (pH 4.5), and the comparison object was cultured using the MRS medium containing 2 wt% glucose (pH 6.5) under anaerobic conditions at 30 °C for 24 - 48 hours.

[0097] <pH Tolerance Test>

[0098] The pH tolerance test of each pre-cultured lactic acid bacterium was carried out as follows.

[0099] (1) Adjust with 2-fold concentration Fig. 9 The MRS medium shown. At this time, 4N sodium hydroxide solution or 4N hydrochloric acid was added to adjust to pH 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5.

[0100] (2) Prepare a 4 wt% sucrose solution.

[0101] (3) For the above (1) and (2), autoclaving for moist heat sterilization (121 °C, 20 minutes) was carried out respectively.

[0102] (4) After cooling the solutions of (1) and (2) after moist heat sterilization, they were mixed in a 1:1 (v / v) manner in a clean bench to prepare an MRS medium containing 2 wt% sucrose (pH 3 - 6.5).

[0103] (5) Each pre-cultured lactic acid bacterial strain was centrifuged (15,000×g, 10 minutes), and after removing the supernatant, at an initial concentration of A660nm The mixture was resuspended in the culture medium prepared in (3) at a concentration of ≒0.05 to 0.10.

[0104] (6) In a 96-well microplate, 200 μl / well was dispensed into 3 wells (n=3), and then cultured at 30°C under anaerobic conditions.

[0105] (7) Turbidimetry was performed over time using an ELISA reader.

[0106] Figures 1 to 3 The comparison object (Lactobacillus acidophilus JCM 1132 T ( Figure 1 ), Lactococcus lactis subsp. lactis NBRC12007( Figure 2 ), Leuconostoc mesenteroides subsp. glucan NBRC100495 T ( Figure 3 )) is a graph showing the results of turbidity measurement of lactic acid bacteria, Figure 4 It is a graph showing the turbidity measurement results of the WR16-4 strain.

[0107] In the above-mentioned pH tolerance test using 2 wt% sucrose as a carbon source, as shown in the turbidity measurement results of each lactic acid bacteria, it was confirmed that the lactic acid bacteria used as the comparison object grew well in the acidic region close to neutrality of pH 5 to 6.5. In contrast, the WR16-4 strain grew in a region closer to the acidic side than the region where these lactic acid bacteria grew well, specifically in the low pH region of pH 3.5 to 5, and hardly grew in the region close to neutrality with a higher pH than that.

[0108] <Sucrose tolerance test>

[0109] The sucrose tolerance test of each pre-cultured lactic acid bacteria was performed as follows.

[0110] (1) Add sucrose Fig. 9 The MRS medium shown was adjusted to a sucrose concentration of 5wt%, 10wt%, 20wt%, 30wt%, 40wt%, 50wt%, 60wt%, and 70wt%, and 4N hydrochloric acid was added thereto to adjust the pH to 4.5.

[0111] (2) The above (1) was sterilized with moist heat (121° C., 20 minutes) using an autoclave.

[0112] (3) Each lactic acid strain pre-cultured was centrifuged (15,000 × g, 10 minutes), and after removing the supernatant, the initial concentration was A. 660nm The mixture was resuspended in the culture medium prepared in (2) at a concentration of ≒0.05 to 0.10.

[0113] (4) In a 96-well microplate, 200 μl / well was dispensed into 3 wells (n=3), and then cultured at 30°C under anaerobic conditions.

[0114] (5) Turbidimetry was performed over time using an enzyme reader.

[0115] Figures 5 to 8 The comparison object (Lactobacillus acidophilus JCM 1132 T ( Figure 5 ), Lactococcus lactis subsp. lactis NBRC12007( Figure 6 ), Leuconostoc mesenteroides subsp. glucan NBRC100495 T ( Figure 7 )) is a graph showing the results of turbidity measurement of lactic acid bacteria, Figure 8 It is a graph showing the turbidity measurement results of the WR16-4 strain.

[0116] In the sucrose tolerance test under the above-mentioned initial acidic conditions of pH 4.5, as shown in the turbidity measurement results of each lactic acid bacteria, the results of the lactic acid bacteria used as the comparison object showed that although growth was confirmed at a sucrose concentration of less than 20wt%, its growth state (proliferation rate) was low, and it was almost impossible to grow in a high sucrose environment of more than 30wt%. In contrast, it was confirmed that the WR16-4 strain showed a good growth state up to a sucrose concentration of about 40wt%, and grew even in a high sucrose environment of 60wt%.

[0117] <Fructose and glucose tolerance test>

[0118] Sucrose is a disaccharide formed by the combination of fructose, which is a monosaccharide, and glucose. As mentioned above, the WR16-4 strain was confirmed to be able to grow in a high-sucrose environment. Therefore, it is believed that it may be able to grow in a high-fructose / glucose concentration environment where fructose and glucose exist in the form of monosaccharides, just like sucrose. Similar to the above-mentioned sucrose tolerance test, a fructose / glucose tolerance test was conducted as follows.

[0119] (1) Add sucrose, fructose, and glucose to the mixture Fig.10 The MRS medium shown was adjusted to have a sugar concentration of 20 wt%, and 4 N hydrochloric acid was added thereto to adjust the pH to 4.5.

[0120] (2) In addition, fructose and glucose were added Fig.10 The MRS medium shown was adjusted to have a sugar concentration of 10 wt% for each of fructose and glucose (20 wt% in total), and then 4 N hydrochloric acid was added thereto to adjust the pH to 4.5.

[0121] (3) The above (1) and (2) were sterilized by moist heat (121° C., 20 minutes) using an autoclave.

[0122] (4) Centrifuge the pre-cultured WR16-4 strain (15,000×g, 10 minutes), remove the supernatant, and resuspend it in the adjusted medium at an initial concentration of A 660nm ≒0.05 - 0.10.

[0123] (5) In a 96-well microplate, dispense 200 μl / well into each of 3 wells (n = 3), and then culture under anaerobic conditions at 30°C.

[0124] (6) Use a microplate reader to measure the turbidity over time.

[0125] The results of the above turbidity measurement in the fructose·glucose tolerance test under the acidic condition of the initial pH 4.5 are shown in Fig.11 .

[0126] As shown in this Fig.11 , when the WR16-4 strain is in the presence of either fructose or glucose alone, the proliferation rate is low (low growth state), but when both fructose and glucose are present, the proliferation rate increases (becomes a high growth state), similar to the case of sucrose. From this result, it is confirmed that the WR16-4 strain not only has sucrose tolerance but also has fructose·glucose tolerance.

[0127] In addition, as described above, sucrose is a disaccharide composed of fructose and glucose. Therefore, it is considered that the WR16-4 strain can grow sufficiently in a high sugar concentration environment containing, for example, 30 wt% fructose and glucose (total sugar concentration is 60 wt%) separately, similar to the case of sucrose.

[0128] <Use of WR16-4 strain>

[0129] The WR16-4 strain can be used in food and beverages containing lactic acid bacteria, specifically in fermented food and beverages.

[0130] As described above, the WR16-4 strain is a lactic acid bacterium characterized by preferentially proliferating in a low pH and high sucrose environment. Therefore, for example, it can easily ferment liquids obtained by mixing and pressing plants with sugar or beverages based on such liquids, or materials with high acid values and sucrose concentrations such as jams and fruit sauces, which are difficult to ferment with general lactic acid bacteria (acidic and high sugar concentration extracts), thereby enabling the above extracts to contain organic acids and obtaining effects such as imparting sourness (reducing sweetness or imparting a refreshing feeling) or防腐 (no need for preservatives as additives).

[0131] In addition, the WR16-4 strain is a lactic acid bacterium that has the characteristic of predominantly proliferating even in a low pH and high fructose / glucose environment. Therefore, in addition to the above, it can also be used in food and beverages using sugars containing both fructose and glucose (sugars mixed with isomerized sugars, refined sugars of glucose and fructose, etc.).

[0132] In addition, an example of a method for obtaining the above-mentioned acidic and high-sugar concentration extract is shown below. It should be noted that the method for obtaining the extract is not limited to this.

[0133] (1) Mix chopped vegetables or fruits with fine sugar in the same proportion, add bacterial liquid and ferment.

[0134] (2) After fermentation, the liquid component is recovered by squeezing, and the recovered liquid is further fermented and aged to obtain fermentation liquid A.

[0135] (3) Fine sugar is added to the decoction of wild grass, and fermentation liquid A is added to the resultant, and the mixture is fermented and matured to obtain fermentation liquid B.

[0136] The fermentation liquid A or the fermentation liquid B becomes an acidic extract with a high sugar concentration. It should be noted that the fermentation liquid A and the fermentation liquid B are produced into beverages, and by fermenting them with the WR16-4 strain, a sour taste can be imparted and an antiseptic effect can be obtained without adding a preservative, thereby providing an unprecedented beverage.

[0137] In addition, as shown in the above test results, the WR16-4 strain has the characteristic of hardly growing in the optimum pH environment of general lactic acid bacteria, i.e., pH 6 to 7. Therefore, for example, when it is desired to ferment a material having a high sucrose concentration and a pH near neutral using the WR16-4 strain and other microorganisms (e.g., general lactic acid bacteria or yeast, etc.), since the WR16-4 strain does not grow before reaching the optimum pH, it will not hinder the growth of other microorganisms. Therefore, it is useful in the case of using multiple microorganisms for composite fermentation.

[0138] In addition, due to the above characteristics, the WR16-4 strain can stop fermentation without heating by adjusting the pH of the raw material to near neutrality, and is also useful for use in fermented products that cannot be heated.

[0139] It should be noted that the present invention is not limited to this embodiment, and the specific structure of each component can be designed appropriately.

[0140] Accession number

[0141] NITE BP-03116

[0142] Sequence Listing <110> National Institute of Technology MIYATOU YASOU KENKYUJO CO., LTD <120> Lactic acid bacteria and method for obtaining same, and food or drink containing lactic acid bacteria (Lactic acid bacterium (LAB), method for separating LAB and food or drink using LAB) <130> TY0304-PCT <150> JP2020-171989 <151> 2020-10-12 <150> JP2020-214170 <151> 2020-12-23 <160> 1 <170> PatentIn version 3.5 <210> 1 <211> 1494 <212> DNA <213> Lactobacillus sp. <400> 1 cgctggcggc atgcctaata catgcaagtc gaacgaggtt tccttaatga tagttggtgc 60 ttgcatcaat ttgacttaag atctaaccga gtggcgaact ggtgagtaac acgtgggtaa 120 cctgccctga agtagggggat aacatttgga aacaaatgct aataccgtat aacaacataa 180 accacatggt ttatgtttga aagatggctc tgctatcgct tttggatgga cccgcggcgt 240 attagctagt tggtgagata atggctcacc aaggcgatga tacgtagccg acctgagagg 300 gtaatcggcc acattgaac tgagacacgg tccagactcc tacgggaggc agcagtaggg 360 aatctttcac aatggacgaa agtctgatga agcaatgccg cgtgagtgat gaagggtttc 420 ggctcgtaaa gctctgttgt tagagaa caggtgtaag agtaactgtt tacatctcga 480 cggtatctaa ccagaaagtc acggctaact acgtgccagc agccgcggta atacgtaggt 540 ggcaagcgtt gtccggattt attgggcgta aagtgagcgc aggcggtttt ttaagtctaa 600 tgtgaaagcc ttcggcttaa ccgaagaagt gcattggaaa ctaagaaact tgagtgcaga 660 aaaggatagt ggaactccat gtgtagcggt gaatgcgta gatatatgga agaacaccag 720 tggcgaaggc gggctatctgg tctgttactg acgttgaggc tcgaagcat gggtagcgaa 780 caggattaga taccctggta gtccatgccg taaacgatga atgctaggtg ttagaggatt 840 tccgtccttt agtgccgcag ctaacgcatt aagcattccg cctggggagt acgaccgcaa 900 ggttgaaact caaggaatt gacggggggcc cgcacaagtg gtggagcatg tggtttaatt 960 cgatgctacg cgagaacct taccaggctct tgacatcttt ctgctagccct agagattag 1020 gtgttccctt cggggacaga ttgacaggtg gtgcatggtt gtcgtcagct cgtgtcgtga 1080 gatgttgggt taagtcccgc aacgagcgca acccttatta tcagttgcca gcattcagtt 1140 gggcactctg gtgagactgc cggtgataaa ccggaggaag gcggggatga cgtctaatca 1200 tcatgcccct tacgacctgg gctacacacg tgctacaatg gacagtacaa agagttgcaa 1260 aaccgcgagg tcaagcaaat cccataaagc tgttctcagt tcggattgca ggctgcaact 1320 cgcctgcatg aagttggaat cactagtaat cgtggatcag aatgccacgg tgaatacgtt 1380 ctcaggcctt gtacataccg cccgtcacac catgagagtt tgtaacaccc aaagccggtg 1440 atgtaaccct tcggggaact agccgtctaa ggtgggacag atgattaggg tgaa 1494

Claims

1. A lactic acid bacterium, characterized in that Its accession number is: The species Lactobacillus sp. deposited in NITE BP-03116 belongs to the genus Lactobacillus and its identification number is WR16-4.

2. A food or beverage containing lactic acid bacteria, characterized in that: It contains the lactic acid bacteria according to claim 1.

3. The lactic acid bacteria-containing food and drink according to claim 2, wherein The food and drink containing lactic acid bacteria is a fermented food and drink.

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