WO2018034203A1 - Novel lactic acid bacteria, natural immunoactivator containing novel lactic acid bacteria as active ingredient, and food or beverage containing novel lactic acid bacteria - Google Patents

Abstract

This lactic acid bacteria is a lactic acid bacteria that belongs to the genus Leuconostoc and that has the accession number NITE BP-02307, a lactic acid bacteria that belongs to the genus Leuconostoc and that has the accession number NITE BP-02306, or a lactic acid bacteria that belongs to the genus Leuconostoc and that has the accession number NITE BP-02308 in the Patent Microorganism Depositary (NPMD) of the National Institute of Technology and Evaluation (NITE). The lactic acid bacteria has a high natural immunity activation effect.

Description

Novel lactic acid bacteria, natural immunity activator containing novel lactic acid bacteria as active ingredients, and food and drink containing new lactic acid bacteria

The present invention relates to a novel lactic acid bacterium, an innate immunity activator containing the novel lactic acid bacterium as an active ingredient, and a food or drink containing the novel lactic acid bacterium.

Lactic acid bacteria have been used for fermented foods for a long time, and have been used for the production of food and drinks, pharmaceuticals, probiotics and the like. Lactic acid bacteria are characterized by Gram-positive, catalase-negative, no endospore formation, no motility, etc.

Also, lactic acid bacteria are most utilized as probiotics. Establishment of a method for efficiently separating lactic acid bacteria with high functionality and establishment of a method for culturing these in foods are useful for the development of foods using functional lactic acid bacteria.

One of the functions attracting attention in probiotics is “innate immunity promoting activity”. In mammals, innate immunity is the forefront of biodefense and triggers subsequent immune responses, including antibody production. In natural immunity in mammals, cytokine secretion by immune cells such as macrophages is promoted by various stimuli, and invading pathogens are eliminated and information is transmitted to other immune cells.

On the other hand, invertebrates such as insects do not have acquired immunity, and pathogens are eliminated only by natural immunity. It is known that the innate immune mechanism has a lot in common between insects and mammals. For example, insect blood cells called hemocytes engulf foreign invaders like macrophages in mammals. In addition, it is known that Toll-like Receptor involved in innate immune response in mammals has high homology with Toll Receptor involved in innate immune response in Drosophila.

So far, the present inventors have developed an evaluation method (screening method) that can easily measure an innate immune activation reaction in silkworms having only an innate immune mechanism (Patent Document 1, etc., Non-Patent Document 1, etc.). ). Further, it has been confirmed that the method can be used to evaluate an innate immune activator (screening method) having an innate immune activation effect on vertebrates such as humans (Patent Document 1, etc.).
Further, it has been confirmed by the present inventors that silkworms are useful as model animals for evaluating resistance to microbial infection (Patent Documents 2, 3, etc.).
In addition, the present inventors have confirmed that a fraction purified from green tea using silkworm muscle contraction activity as an index has the ability to activate macrophages of mammalian cells (Non-patent Document 2).

Abnormal immune mechanism causes various diseases. Therefore, it is desired to develop excellent innate immunity activators and foods and drinks that activate innate immunity that can regulate such an immune mechanism as desired.

International Publication No. 2008/126905 JP 2007-327964 A JP 2012-006917 A

Ishii K, Hamamoto H, Kamimura M, Sekimizu K. Activation of the silkworm cytokine by bacterial and fungal cell wall components via a reactive oxygen species-triggered mechanism. J Biol Chem. 2831 Dhital, S. Purification of innate immunostimulant from green tea using a silkworm muscle contraction assay. Drug Discov. Ther. 5, 18-25 (2011).

An object of the present invention is to provide a novel lactic acid bacterium having high innate immunity activation ability, and further, an innate immunity activator comprising the lactic acid bacterium or killed bacteria or processed product of the lactic acid bacterium as an active ingredient, An object of the present invention is to provide a food or drink containing a lactic acid bacterium or an innate immune activator derived from the lactic acid bacterium.

As a result of intensive studies to solve the above problems, the present inventors have isolated a new lactic acid bacterium from kimchi and straw. And as a result of examining using the method disclosed in Patent Document 1 that can easily measure the innate immunity activation reaction, it has higher innate immunity activation ability than lactic acid bacteria known so far Was confirmed.

Furthermore, the above-described lactic acid bacteria having the ability to activate innate immunity were all found to be novel lactic acid strains belonging to the genus Leuconostoc, as a result of analysis of their properties and analysis of the 16S DNA sequence.

Further, the present invention was completed by finding that a highly useful food can be provided by subjecting the lactic acid bacteria to lactic acid fermentation with milk, vegetable juice and fruit juice, which are difficult to purely culture.

That is, the present invention provides a lactic acid bacterium belonging to the genus Leuconostoc whose accession number is NITE BP-02307 at the Patent Microorganism Deposit Center (NPMD) of the National Institute of Technology and Evaluation (NITE). is there.

The present invention also provides a lactic acid bacterium belonging to the genus Leuconostoc whose accession number is NITE BP-02306 in the Patent Microorganism Deposit Center (NPMD) of the National Institute for Product Evaluation and Technology (NITE). is there.

The present invention also provides a lactic acid bacterium belonging to the genus Leuconostoc whose accession number is NITE BP-02308 in the Patent Microorganism Deposit Center (NPMD) of the National Institute of Technology and Evaluation (NITE). is there.

Further, the present invention is an innate immunity activator comprising as an active ingredient the above-mentioned lactic acid bacteria, killed bacteria of the lactic acid bacteria, or processed products of the lactic acid bacteria,
The processed product of lactic acid bacteria is a culture of lactic acid bacteria; concentrate; pasted product; spray-dried product, freeze-dried product, vacuum-dried product, drum-dried product, etc .; liquefied product; diluted product; And an innate immunity activation agent characterized by being at least one processed product selected from the group consisting of extracts from the culture.

Moreover, this invention provides the food / beverage products containing the said lactic acid bacteria or the said innate immunity activator, or the food / beverage products manufactured using the process fermented using the said lactic acid bacteria.

According to the present invention, a novel lactic acid bacterium having a higher innate immunity activation ability than lactic acid bacteria known so far can be provided.
Furthermore, the innate immunity activator comprising the lactic acid bacterium as an active ingredient, the food and drink containing the lactic acid bacterium or the innate immunity activator, and the food and drink produced using the step of fermentation using the lactic acid bacterium Goods can be provided.

In addition, the lactic acid bacteria of the present invention can be used not only for the production of general foods and drinks, health foods, drugs, fermented foods and drinks, probiotics, etc., but also for the prevention and treatment of diseases due to innate immune activation. . In addition, since it is resistant to acids, it is easy to reach the small intestine without being decomposed in the stomach.

Hereinafter, the present invention will be described, but the present invention is not limited to the following specific embodiments, and can be arbitrarily modified within the scope of the technical idea.

<Lactic acid bacteria # 7-2>
The lactic acid bacterium of the present invention is a lactic acid bacterium belonging to the genus Leuconostoc whose accession number is NITE BP-02307 at the Patent Microorganism Deposit Center (NPMD) of the National Institute for Product Evaluation and Technology (NITE) (hereinafter referred to as “Leuconostoc”) (hereinafter referred to as “Leuconostoc”). (May be abbreviated as “lactic acid bacteria # 7-2”).

Hereinafter, a novel lactic acid strain (lactic acid bacterium # 7-2) belonging to the genus Leuconostoc will be described in detail.
Lactic acid bacteria # 7-2 of the present invention was isolated for the first time using kimchi as a separation source.

Gram staining result: positive Cell shape: spherical Aerobic / anaerobic: anaerobic Lactic acid producing ability: Yes

Physiological properties: The physiological and chemical taxonomic properties of lactic acid bacteria # 7-2 of the present invention are as follows.
(1) Catalase:-
(2) Acid phosphatase: +
(3) Alkaline phosphatase: +
(4) Naphthol-AS-BI-phosphohydrolase: +
(5) Esterase (C4): +
(6) α-Galactosidase:-
(7) Esterase lipase (C8): +
(8) β-galactosidase:-
(9) Lipase (C14):-
(10) β-glucuronidase:-
(11) Leucine allylamidase: +
(12) α-Glucosidase: +
(13) Valine allylamidase:-
(14) β-glucosidase:-
(15) Cystine allylamidase:-
(16) N-acetyl-β-glucosaminidase:-
(17) Trypsin:-
(18) α-Mannosidase:-
(19) α-chymotrypsin:-
(20) α-fucosidase:-

(21) Acid and gas generating ability from the following saccharides etc. Glycerol:-
Erythritol:-
D-Arabinose:-
L-Arabinose:-
D-Ribose: +
D-Xylose:-
L-Xylose:-
D-Adonitol:-
β-Methyl-D-xylopyranoside:-
D-Galactose: +
D-Glucose: +
D-Fructose: +
D-Mannose: +
L-Sorbose:-
L-Rhamnose:-
Dulcitol:-
Inositol:-
D-Mannitol:-
D-Sorbitol:-
α-Methyl-D-mannopyranoside:-
α-Methyl-D-glucopyranoside: +
N-Acetyl glucosamine: +
Amygdalin: +
Arbutin: +
Esculin ferric citrate: +
Salicin: +
D-Cellobiose: +
D-Maltose: +
D-Lactose:-
D-Melibiose: +
D-Sucrose: +
D-Trehalose: +
Insulin:-
D-Melezitose: +
D-Raffinose: +
Starch: +
Glycogen:-
Xylitol:-
Gentiobiose: +
D-Turanose: +
D-Lyxose:-
D-Tagatose: +
D-Fucose:-
L-Fucose:-
D-Arabitol:-
L-Arabitol:-
Gluconate: +
2-Keto-gluconate:-
5-Keto-gluconate:-

Molecular biological analysis result: The analysis result of lactic acid bacteria # 7-2 on 16S rDNA used as an index of molecular biological systematic classification is as shown in SEQ ID NO: 1 in the attached sequence listing.
That is, from the genomic DNA of lactic acid bacteria # 7-2, the base sequence of the 16S rDNA region was amplified by PCR and analyzed by a sequencer. As a result, a base sequence corresponding to almost the entire length of 16S rDNA was found.
When this base sequence was subjected to homology search by BLAST analysis of NCBI, the base sequence of the 16S rDNA region of lactic acid bacteria # 7-2 was the base sequence of Leuconostoc carnosum JB16 strain (registration number: NR — 102781.1). And lactic acid bacteria # 7-2 belong to Leuconostoc carnosum.
However, even when only the 16S rDNA region is compared, the lactic acid bacteria # 7-2 of the present invention is a lactic acid strain different from the above strains because they do not completely match.

Physiological and chemical taxonomic properties of the lactic acid bacteria # 7-2 in the light of classification according to Bergey's Manual of Systematic Bacteriology (vol.3 1989) and other literature In addition, as a result of determination in consideration of the result of the 16S rDNA analysis, “lactic acid bacteria # 7-2” of the present invention is a novel microorganism belonging to the genus Leuconostoc.
In addition, there is no microorganism having a 16S rDNA region base sequence that matches the 16S rDNA region base sequence of lactic acid bacteria # 7-2, and it exhibits a higher innate immunity activity than known strains belonging to the genus Leuconostoc. As a result of comprehensive examination including the above, it was determined that lactic acid bacteria # 7-2 was an isolated novel microbial strain.

Lactic acid bacteria # 7-2 is a patented microorganism of the National Institute of Technology and Evaluation (hereinafter abbreviated as “NITE”), 2-5-8, 122, Kazusa Kamashi, Kisarazu City, Chiba Prefecture It is a microorganism deposited domestically at the Deposit Center (NPMD) and deposited under the deposit number: NITE P-02307 (deposit date: July 26, 2016).
Lactic acid bacteria # 7-2 then submitted the original deposit application form to the patent microorganisms deposit center (NPMD) of 2-5-8 Kazusa-Kamashita, Kisarazu, Chiba Prefecture, Room 122, NITE Submission and application for transfer from domestic deposit (original deposit date: July 26, 2016) to deposit under the Budapest Treaty (transfer date (international deposit date): July 25, 2017) As a result of receiving an application for transfer to a deposit based on the Budapest Treaty (international deposit), it has received the deposit number “NITE BP-02307”.

As a general property of bacteria, since its properties as a strain are easily mutated, lactic acid bacteria # 7-2 of the present invention may not be within the range of the physiological properties shown above. Needless to say, such “mutation” includes both natural and artificial mutations.

The culture method of lactic acid bacteria # 7-2 is described below. The culture method of lactic acid bacteria # 7-2 may be performed according to a general culture method performed for microorganisms belonging to the genus Leuconostoc.
The culture is preferably performed under anaerobic conditions. Examples of the carbon source in the medium include D-ribose, D-galactose, D-glucose, D-fructose, D-mannose, D-mannitol, N-acetylglucosamine, amygdalin, arbutin, esculin, salicin, D-cellobiose Organic carbon compounds such as D-maltose, sucrose, D-trehalose, gentiobiose, molasses, starch syrup, and fats and oils are used, and nitrogen sources include meat extract, casein, peptone, yeast extract, dry yeast, germ, soybean Organic / inorganic nitrogen compounds such as powder, urea, amino acids and ammonium salts can be used.
Further, as the salts, inorganic salts such as sodium salt, potassium salt, calcium salt, magnesium salt, phosphate, iron salt, copper salt, zinc salt, cobalt salt and the like are appropriately added as necessary. Furthermore, it is preferable to add a growth promoting substance such as biotin, vitamin B1, cystine, methyl oleate, lard oil or the like in terms of increasing the production amount of the target product.
Moreover, you may add antifoamers, such as a silicone oil and surfactant. As the prepared medium, for example, an MRS medium, a GAM medium or the like is preferably used.

The culture condition may be performed according to the general culture conditions performed for microorganisms belonging to the genus Leuconostoc as described above. If it is a liquid culture method, stationary culture is desirable. If it is a small scale, you may use the stationary culture method by the glass bottle with a lid | cover.
The culture temperature is preferably maintained between 25 ° C. and 37 ° C., more preferably 30 ° C. to 37 ° C. The culture pH is preferably performed in the vicinity of 7. The culture period is a factor that varies depending on the composition of the medium used, the culture temperature, and the like. In the case of lactic acid bacteria # 7-2, a sufficient amount of the desired product is preferably obtained in 12 to 72 hours, more preferably in 24 to 48 hours. Can be secured.
It is also preferable to pick up a colony obtained by culturing and form a single colony on the medium again.

<Lactic acid bacteria # 4-2>
The lactic acid bacterium of the present invention is a lactic acid bacterium belonging to the genus Leuconostoc which is NITE BP-02306 in the Patent Microorganism Deposit Center (NPMD) of the National Institute of Technology and Evaluation (NITE) (hereinafter referred to as “lactic acid bacteria #”). 4-2 ”in some cases).

Hereinafter, a novel lactic acid strain (lactic acid bacteria # 4-2) belonging to the genus Leuconostoc will be described in detail.
Lactic acid bacteria # 4-2 of the present invention was isolated for the first time using sputum as a separation source.

Gram staining result: positive Cell shape: spherical Aerobic / anaerobic: anaerobic Lactic acid producing ability: Yes

Physiological properties: Physiological and chemical taxonomic properties of lactic acid bacteria # 4-2 of the present invention are as follows.
(1) Catalase:-
(2) Acid phosphatase: +
(3) Alkaline phosphatase:-
(4) Naphthol-AS-BI-phosphohydrolase: +
(5) Esterase (C4): +
(6) α-Galactosidase:-
(7) Esterase lipase (C8): +
(8) β-galactosidase:-
(9) Lipase (C14):-
(10) β-glucuronidase:-
(11) Leucine allylamidase: +
(12) α-Glucosidase: +
(13) Valine allylamidase:-
(14) β-glucosidase: +
(15) Cystine allylamidase:-
(16) N-acetyl-β-glucosaminidase:-
(17) Trypsin:-
(18) α-Mannosidase:-
(19) α-chymotrypsin: +
(20) α-fucosidase:-

(21) Acid and gas generating ability from the following saccharides etc. Glycerol:-
Erythritol:-
D-Arabinose:-
L-Arabinose: +
D-Ribose: +
D-Xylose: +
L-Xylose:-
D-Adonitol:-
β-Methyl-D-xylopyranoside:-
D-Galactose: +
D-Glucose: +
D-Fructose: +
D-Mannose: +
L-Sorbose:-
L-Rhamnose:-
Dulcitol:-
Inositol:-
D-Mannitol: +
D-Sorbitol:-
α-Methyl-D-mannopyranoside:-
α-Methyl-D-glucopyranoside: +
N-Acetyl glucosamine: +
Amygdalin: +
Arbutin: +
Esculin ferric citrate: +
Salicin: +
D-Cellobiose: +
D-Maltose: +
D-Lactose:-
D-Melibiose: +
D-Sucrose: +
D-Trehalose: +
Insulin:-
D-Melezitose: +
D-Raffinose:-
Starch: +
Glycogen:-
Xylitol:-
Gentiobiose: +
D-Turanose: +
D-Lyxose:-
D-Tagatose: +
D-Fucose:-
L-Fucose:-
D-Arabitol:-
L-Arabitol:-
Gluconate: +
2-Keto-gluconate: +
5-Keto-gluconate:-

Molecular biological analysis result: The analysis result of lactic acid bacteria # 4-2 on 16S rDNA used as an index of molecular biological systematic classification is as shown in SEQ ID NO: 2 in the attached sequence listing.
That is, from the genomic DNA of lactic acid bacteria # 4-2, the base sequence of the 16S rDNA region was amplified by PCR and analyzed by a sequencer. As a result, a base sequence corresponding to almost the entire length of 16S rDNA was found.
When this base sequence was subjected to homology search by BLAST analysis of NCBI, the base sequence of the 16S rDNA region of lactic acid bacteria # 4-2 was found to be the base sequence of Leuconostoc gelidum POUF4d strain (registration number: NR — 133769.1). And lactic acid bacteria # 4-2 belong to Leuconostoc gelidum.
However, even when only the 16S rDNA region is compared, the lactic acid bacteria # 4-2 of the present invention are lactic acid strains different from the above strains because they do not completely match.

The physiological and chemical taxonomic properties of lactic acid bacteria # 4-2 are classified according to the classification by Bergey's Manual of Systematic Bacteriology (vol.3 1989) and other literature. In addition, as a result of determination in consideration of the result of the 16S rDNA analysis, “lactic acid bacteria # 4-2” of the present invention is a novel microorganism belonging to the genus Leuconostoc.
In addition, there is no microorganism having a 16S rDNA region base sequence that matches the base sequence of the 16S rDNA region of lactic acid bacteria # 4-2, and it exhibits a higher innate immunity activity than known strains belonging to the genus Leuconostoc. As a result of comprehensive examination including the above, it was determined that lactic acid bacteria # 4-2 was an isolated novel microorganism strain.

Lactic acid bacteria # 4-2 is a patented microorganism manufactured by the National Institute of Technology and Evaluation (hereinafter referred to as “NITE”), Room 2-5-8, Kazusa Kamashichi, Kisarazu City, Chiba Prefecture. It is a microorganism deposited domestically at the Deposit Center (NPMD) and deposited under the deposit number: NITE P-02306 (deposit date: July 26, 2016).
Lactic acid bacteria # 4-2 will then submit an original deposit application form to the patent microorganisms deposit center (NPMD) of Kazusa-Kamashita 2-5-8 122, Kisarazu-shi, Chiba, and National Institute of Technology and Evaluation (NITE). Submission and application for transfer from domestic deposit (original deposit date: July 26, 2016) to deposit under the Budapest Treaty (transfer date (international deposit date): July 25, 2017) As a result of receiving an application for transfer to a deposit based on the Budapest Treaty (international deposit), it has received the deposit number “NITE BP-02306”.

As a general property of bacteria, since its properties as a strain are easily mutated, lactic acid bacteria # 4-2 of the present invention may not be within the range of the physiological properties shown above. Needless to say, such “mutation” includes both natural and artificial mutations.

The culture method for lactic acid bacteria # 4-2 is described below. The culture method of lactic acid bacteria # 4-2 may be performed according to a general culture method performed for microorganisms belonging to the genus Leuconostoc.
The culture is preferably performed under anaerobic conditions. Examples of the carbon source in the medium include D-ribose, D-galactose, D-glucose, D-fructose, D-mannose, D-mannitol, N-acetylglucosamine, amygdalin, arbutin, esculin, salicin, D-cellobiose Organic carbon compounds such as D-maltose, sucrose, D-trehalose, gentiobiose, molasses, starch syrup, and fats and oils are used, and nitrogen sources include meat extract, casein, peptone, yeast extract, dry yeast, germ, soybean Organic / inorganic nitrogen compounds such as powder, urea, amino acids and ammonium salts can be used.
Further, as the salts, inorganic salts such as sodium salt, potassium salt, calcium salt, magnesium salt, phosphate, iron salt, copper salt, zinc salt, cobalt salt and the like are appropriately added as necessary. Furthermore, it is preferable to add a growth promoting substance such as biotin, vitamin B1, cystine, methyl oleate, lard oil or the like in terms of increasing the production amount of the target product.
Moreover, you may add antifoamers, such as a silicone oil and surfactant. As the prepared medium, for example, an MRS medium, a GAM medium or the like is preferably used.

The culture condition may be performed according to the general culture conditions performed for microorganisms belonging to the genus Leuconostoc as described above. If it is a liquid culture method, stationary culture is desirable. If it is a small scale, you may use the stationary culture method by the glass bottle with a lid | cover.
The culture temperature is preferably maintained between 25 ° C. and 37 ° C., more preferably 30 ° C. to 37 ° C. The culture pH is preferably performed in the vicinity of 7. The culture period is a factor that varies depending on the composition of the medium used, the culture temperature, etc. In the case of lactic acid bacteria # 4-2, a sufficient amount of the desired product is preferably obtained in 12 to 72 hours, more preferably in 24 to 48 hours. Can be secured.
It is also preferable to pick up a colony obtained by culturing and form a single colony on the medium again.

<Lactic acid bacteria 8 / 11-3>
The lactic acid bacterium of the present invention is a lactic acid bacterium belonging to the genus Leuconostoc whose accession number is NITE BP-02308 in the Patent Microorganism Deposit Center (NPMD) of the National Institute of Technology and Evaluation (NITE) (hereinafter referred to as “Leuconostoc”). (May be abbreviated as “lactic acid bacteria 8 / 11-3”).

Hereinafter, a novel lactic acid strain (Lactic acid bacterium 8 / 11-3) belonging to the genus Leuconostoc will be described in detail.
The lactic acid bacteria 8 / 11-3 of the present invention were isolated for the first time using kimchi as a separation source.

Gram staining result: positive Cell shape: spherical Aerobic / anaerobic: anaerobic Lactic acid producing ability: Yes

Physiological properties: The physiological and chemical taxonomic properties of the lactic acid bacteria 8 / 11-3 of the present invention are as follows.
(1) Catalase:-
(2) Acid phosphatase: +
(3) Alkaline phosphatase: +
(4) Naphthol-AS-BI-phosphohydrolase: +
(5) Esterase (C4): +
(6) α-galactosidase: +
(7) Esterase lipase (C8): +
(8) β-galactosidase: +
(9) Lipase (C14):-
(10) β-glucuronidase:-
(11) Leucine allylamidase: +
(12) α-Glucosidase: +
(13) Valine allylamidase:-
(14) β-glucosidase: +
(15) Cystine allylamidase:-
(16) N-acetyl-β-glucosaminidase:-
(17) Trypsin:-
(18) α-Mannosidase:-
(19) α-chymotrypsin:-
(20) α-fucosidase:-

(21) Acid and gas generating ability from the following saccharides etc. Glycerol:-
Erythritol:-
D-Arabinose:-
L-Arabinose: +
D-Ribose: +
D-Xylose: +
L-Xylose:-
D-Adonitol:-
β-Methyl-D-xylopyranoside:-
D-Galactose: +
D-Glucose: +
D-Fructose: +
D-Mannose: +
L-Sorbose:-
L-Rhamnose:-
Dulcitol:-
Inositol:-
D-Mannitol: +
D-Sorbitol: +
α-Methyl-D-mannopyranoside:-
α-Methyl-D-glucopyranoside: +
N-Acetyl glucosamine: +
Amygdalin: +
Arbutin: +
Esculin ferric citrate: +
Salicin: +
D-Cellobiose: +
D-Maltose: +
D-Lactose: +
D-Melibiose: +
D-Sucrose: +
D-Trehalose: +
Insulin:-
D-Melezitose: +
D-Raffinose: +
Starch:-
Glycogen:-
Xylitol:-
Gentiobiose: +
D-Turanose: +
D-Lyxose:-
D-Tagatose: +
D-Fucose:-
L-Fucose:-
D-Arabitol:-
L-Arabitol:-
Gluconate: +
2-Keto-gluconate:-
5-Keto-gluconate:-

Molecular biological analysis result: The analysis result of lactic acid bacteria 8 / 11-3 regarding 16SrDNA used as an index of molecular biological systematic classification is as shown in SEQ ID NO: 3 in the attached sequence listing.
That is, from the genomic DNA of lactic acid bacteria 8 / 11-3, the base sequence of the 16S rDNA region was amplified by PCR and analyzed by a sequencer. As a result, a base sequence corresponding to almost the entire length of 16S rDNA was found.
When this base sequence was subjected to homology search by NCBI BLAST analysis, the base sequence of the 16S rDNA region of Lactobacillus 8 / 11-3 was the base sequence of Leuconostoc mesenteroides ATCC8293 strain (registration number: NR_074957. Since it showed 99% homology with 1), lactic acid bacteria 8 / 11-3 belong to Leuconostoc mesenteroides.
However, even when only the 16S rDNA region is compared, the lactic acid bacteria 8 / 11-3 of the present invention are lactic acid strains different from the above strains because they do not completely match.

Physiological and chemical taxonomic properties of the lactic acid bacteria 8 / 11-3 are classified according to Bergey's Manual of Systematic Bacteriology (vol. 3 1989) and other publications. In addition, as a result of determination in consideration of the result of the 16S rDNA analysis, “lactic acid bacteria 8 / 11-3” of the present invention is a novel microorganism belonging to the genus Leuconostoc.
In addition, there is no microorganism having a base sequence of 16S rDNA region that matches the base sequence of 16S rDNA region of lactic acid bacteria 8 / 11-3, and it exhibits a high innate immunity activity compared to known strains belonging to the genus Leuconostoc As a result of comprehensive examination including the above, it was determined that lactic acid bacteria 8 / 11-3 was an isolated novel microbial strain.

Lactic acid bacteria 8 / 11-3 is a patent of Room 2-5-8, Kazusa Kamashichi, Kisarazu City, Chiba Prefecture, National Institute of Technology and Evaluation (hereinafter abbreviated as “NITE”). It is a microorganism deposited domestically at the Microorganism Deposit Center (NPMD) and deposited under the deposit number: NITE P-02308 (deposit date: July 26, 2016).
The lactic acid bacteria 8 / 11-3 was then submitted to the patent microorganisms deposit center (NPMD) of 2-5-8 Kazusa-Kamashita, Kisarazu City, Chiba Prefecture, and the National Institute for Product Evaluation and Technology (NITE). Apply for transfer from domestic deposit (original deposit date: July 26, 2016) to deposit under the Budapest Treaty (transfer date (international deposit date): May 22, 2017) and survive As a result of receiving an application for transfer to a deposit under the Budapest Treaty (international deposit), it has received the deposit number “NITE BP-02308”.

As a general characteristic of bacteria, since its properties as a strain are easily mutated, the lactic acid bacteria 8 / 11-3 of the present invention may not be within the range of the physiological characteristics shown above. . Needless to say, such “mutation” includes both natural and artificial mutations.

Hereinafter, a culture method of lactic acid bacteria 8 / 11-3 will be described. The culture method of lactic acid bacteria 8 / 11-3 may be performed according to a general culture method performed for microorganisms belonging to the genus Leuconostoc.
The culture is preferably performed under anaerobic conditions. Examples of the carbon source in the medium include D-ribose, D-galactose, D-glucose, D-fructose, D-mannose, D-mannitol, N-acetylglucosamine, amygdalin, arbutin, esculin, salicin, D-cellobiose Organic carbon compounds such as D-maltose, sucrose, D-trehalose, gentiobiose, molasses, starch syrup, and fats and oils are used, and nitrogen sources include meat extract, casein, peptone, yeast extract, dry yeast, germ, soybean Organic / inorganic nitrogen compounds such as powder, urea, amino acids and ammonium salts can be used.
Further, as the salts, inorganic salts such as sodium salt, potassium salt, calcium salt, magnesium salt, phosphate, iron salt, copper salt, zinc salt, cobalt salt and the like are appropriately added as necessary. Furthermore, it is preferable to add a growth promoting substance such as biotin, vitamin B1, cystine, methyl oleate, lard oil or the like in terms of increasing the production amount of the target product.
Moreover, you may add antifoamers, such as a silicone oil and surfactant. As the prepared medium, for example, an MRS medium, a GAM medium or the like is preferably used.

The culture condition may be performed according to the general culture conditions performed for microorganisms belonging to the genus Leuconostoc as described above. If it is a liquid culture method, stationary culture is desirable. If it is a small scale, you may use the stationary culture method by the glass bottle with a lid | cover.
The culture temperature is preferably maintained between 25 ° C. and 37 ° C., more preferably 30 ° C. to 37 ° C. The culture pH is preferably performed in the vicinity of 7. The culture period is a factor that varies depending on the composition of the medium used, the culture temperature, etc. In the case of lactic acid bacteria 8 / 11-3, it is preferably 12 to 72 hours, more preferably 24 to 48 hours. Can be secured.
It is also preferable to pick up a colony obtained by culturing and form a single colony on the medium again.

<Natural immune activator>
The innate immunity activator of the present invention is an innate immunity activator comprising as an active ingredient the above lactic acid bacteria, killed bacteria of the lactic acid bacteria, or processed products of the lactic acid bacteria,
The treated product of lactic acid bacteria is at least selected from the group consisting of cultures, concentrates, pasted products, dried products, liquefied products, diluted products, crushed products, sterilized processed products, and extracts from cultured products. It is one processed product.

The innate immunity activator of the present invention can contain the lactic acid bacterium of the present invention, a killed bacterium of the lactic acid bacterium, or a processed product of the lactic acid bacterium in various states. For example, the state containing a suspension, lactic acid bacteria, a culture supernatant, a culture medium component, etc. are mentioned.

The innate immunity activator of the present invention may contain the lactic acid bacterium as it is, or may contain a lactic acid bacterium-treated product obtained by subjecting the lactic acid bacterium to some kind of treatment.
Examples of the processed product of lactic acid bacteria used for the innate immunity activator include, for example, culture of lactic acid bacteria; concentrate; pasted product; Diluted product; crushed product; sterilized processed product; extract from the culture;

As the lactic acid bacteria, viable cells, wet bacteria, dry bacteria and the like can be used as appropriate. Moreover, the killed bacteria which performed sterilization, ie, a heat sterilization process, a radiation sterilization process, a crushing process, etc. may be sufficient.

The content of the lactic acid bacterium, the killed lactic acid bacterium, and the processed product of the lactic acid bacterium, which is an active ingredient in the innate immunity activator of the present invention, with respect to the entire innate immunity activator is not particularly limited, depending on the purpose. The total amount of lactic acid bacteria, dead bacteria of the lactic acid bacteria, and processed product of the lactic acid bacteria can be 0.001 to 100 parts by mass when the innate immunity activator is 100 parts by mass. It is preferably contained in an amount of 0.01 to 99 parts by mass, particularly preferably 0.1 to 95 parts by mass, and still more preferably 1 to 90 parts by mass.

In addition, any one of the above active ingredients may be used alone, or two or more may be used in combination. The content ratio of each active ingredient in the innate immunity activator when two or more are used in combination is not particularly limited and can be appropriately selected depending on the purpose.

The innate immunity activator of the present invention contains the lactic acid bacterium, the killed bacterium of the lactic acid bacterium, or the processed product of the lactic acid bacterium as an active ingredient, and can contain “other ingredients” in addition to these active ingredients. .
The “other components” in the innate immunity activator are not particularly limited and can be appropriately selected according to the purpose within a range not impairing the effects of the present invention. For example, pharmaceutically acceptable And a carrier that can be used.
There is no restriction | limiting in particular as this support | carrier, For example, it selects suitably according to the dosage form etc. which are mentioned later. In addition, the content of “other components” in the innate immunity activator is not particularly limited and can be appropriately selected depending on the purpose.

<Pharmaceuticals; Food and drink; Health foods>
The lactic acid bacterium of the present invention and the innate immunity activator of the present invention derived from the lactic acid bacterium include pharmaceuticals (drugs), quasi-drugs, general foods and drinks, health foods, fermented foods and drinks, food and drinks having standards such as powdered milk, etc. And can be applied to various pharmaceuticals, foods and drinks, etc. regardless of their form. It can also be used for probiotic production.
Among them, food and drink manufactured using the above-described process of fermenting using the lactic acid bacteria of the present invention, and among them, fermented milk is more likely to exhibit the normal effects of lactic acid bacteria and the effects specific to the present invention. preferable.

There is no restriction | limiting in particular as a dosage form of the innate immunity activation agent of this invention, For example, it can select suitably according to the desired administration method as mentioned later.
Specifically, for example, oral solid preparations (tablets, coated tablets, granules, powders, hard capsules, soft capsules, etc.), oral liquids (internal solutions, syrups, elixirs, etc.), injections (solvents, suspensions) Agents), ointments, patches, gels, creams, powders for external use, sprays, inhalation sprays and the like.

Examples of the oral solid preparation include, for example, excipients, and further additives such as binders, disintegrants, lubricants, coloring agents, flavoring and flavoring agents, and the like, in addition to the above-mentioned active ingredients. Can be manufactured.

Examples of the excipient include lactose, sucrose, sodium chloride, glucose, starch, calcium carbonate, kaolin, microcrystalline cellulose, silicic acid and the like.
Examples of the binder include water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, hydroxypropylcellulose, hydroxypropyl starch, methylcellulose, ethylcellulose, shellac, calcium phosphate, polyvinylpyrrolidone and the like. It is done.
Examples of the disintegrant include dry starch, sodium alginate, agar powder, sodium hydrogen carbonate, calcium carbonate, sodium lauryl sulfate, stearic acid monoglyceride, and lactose.
Examples of the lubricant include purified talc, stearate, borax, polyethylene glycol and the like.
Examples of the colorant include titanium oxide and iron oxide.
Examples of the flavoring / flavoring agent include sucrose, orange peel, citric acid, tartaric acid and the like.

The oral liquid preparation can be produced by a conventional method, for example, by adding additives such as a flavoring / flavoring agent, a buffer, a stabilizer, an edible (processed) oil, an animal and vegetable oil to the active ingredient.

Examples of the flavoring / flavoring agent include sucrose, orange peel, citric acid, tartaric acid and the like. Examples of the buffer include sodium citrate. Examples of the stabilizer include tragacanth, gum arabic, and gelatin.

As the injection, for example, a pH adjuster, a buffer, a stabilizer, a tonicity agent, a local anesthetic, etc. are added to the above active ingredient, and subcutaneous, intramuscular, intravenous use are performed by a conventional method. Etc. can be manufactured.
Examples of the pH adjuster and the buffer include sodium citrate, sodium acetate, sodium phosphate and the like. Examples of the stabilizer include sodium pyrosulfite, EDTA, thioglycolic acid, thiolactic acid, and the like. Examples of the isotonic agent include sodium chloride and glucose. Examples of the local anesthetic include procaine hydrochloride and lidocaine hydrochloride.

The ointment can be produced, for example, by mixing a known base, stabilizer, wetting agent, preservative and the like with the above active ingredients and mixing them by a conventional method.
Examples of the base include liquid paraffin, white petrolatum, white beeswax, octyldodecyl alcohol, paraffin and the like. Examples of the preservative include methyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, and the like.

As the patch, for example, a cream, gel, paste or the like as the ointment can be applied to a known support by a conventional method. Examples of the support include woven fabric, nonwoven fabric, soft vinyl chloride, polyethylene, polypropylene, polyurethane and other films made of cotton, suf, and chemical fibers, and foam sheets.

The innate immunity activator of the present invention can be suitably used for, for example, individuals, bacteria, and the like that require activation of the innate immune mechanism.
Specifically, for example, by administering to an individual who needs to maintain health or recover from fatigue; an individual who needs prevention or treatment of cancer or lifestyle-related diseases; an individual infected with bacteria, fungi, viruses, etc. Can be used.

The animal to be administered with the innate immunity activator of the present invention is not particularly limited. For example, humans; laboratory animals such as mice and rats; monkeys; horses; livestock such as cows, pigs, goats and chickens; And pets such as dogs.

Moreover, there is no restriction | limiting in particular as the administration method of the said innate immunity activator, For example, according to the above-mentioned dosage form etc., it can select suitably, Oral administration, intraperitoneal administration, injection in the blood, intestine Injection into the inside. Especially, oral administration is preferable from the point which is simple and exhibits the said effect, and oral administration as food / beverage products, such as general food / beverage products, health food, and fermented food / beverage products, is especially preferable.

The dose of the innate immunity activator is not particularly limited and can be appropriately selected according to the age, weight, desired degree of effect, etc. of the individual to be administered. The daily dose is preferably from 1 mg to 30 g, more preferably from 10 mg to 10 g, particularly preferably from 100 mg to 3 g as the amount of the active ingredient.
Moreover, there is no restriction | limiting in particular also as an administration time, According to the objective, it can select suitably, For example, you may administer prophylactically and may administer therapeutically.

"The above-mentioned lactic acid bacteria of the present invention, killed or treated products of the lactic acid bacteria, foods and drinks containing an innate immunity activator, and foods and drinks manufactured using the process of fermenting with the lactic acid bacteria" (hereinafter, in parentheses) Are simply abbreviated as “the food and drink of the present invention”), the content of the lactic acid bacteria and the innate immunity activator is not particularly limited, and is appropriately determined according to the purpose and the form (type) of the food and drink. The total amount is preferably 0.001 to 100 parts by mass, more preferably 0.01 to 99 parts by mass, when the total amount of food and drink is 100 parts by mass. Particularly preferred is a content of 0.1 to 95 parts by mass.

Any one of the above may be used alone, or two or more may be used in combination. There is no restriction | limiting in particular in the content ratio of each substance in the said food / beverage products in the case of using 2 or more types together, According to the objective, it can select suitably.

The food / beverage products of this invention have innate immunity activation ability and / or infectious disease prevention-treatment ability.
The food and drink of the present invention can further contain “other components” in addition to the above-described innate immunity activator and infectious disease preventive and therapeutic agent of the present invention.

The “other components” are not particularly limited and may be appropriately selected depending on the purpose within a range not impairing the effects of the present invention. Examples thereof include various food materials. Further, the content of “other components” is not particularly limited and can be appropriately selected depending on the purpose.

The lactic acid bacteria of the present invention can be used for production of general foods and drinks, health foods, drugs, fermented foods and drinks, probiotics, and the like. As fermented foods and drinks, it is particularly suitable for use as fermented milk, lactic acid bacteria beverages, yogurt, pickles, lactic acid bacteria starters for the production of pickles.

There is no restriction | limiting in particular as a kind of said foodstuff, It can select suitably according to the objective, For example, confectionery, such as jelly, a candy, chocolate, a biscuit, a gummy; Preference, such as green tea, tea, coffee, a soft drink Beverages; Dairy products such as fermented milk, yogurt, ice cream and lacto ice; Vegetables and fruit processed products such as vegetable drinks, fruit drinks and jams; Liquid foods such as soups; Grain processed products such as breads and noodles; The charge; etc. are mentioned. Of these, dairy products such as yogurt and fermented milk are preferred.
There is no restriction | limiting in particular as a manufacturing method of these foodstuffs, For example, according to the manufacturing method of various normal foodstuffs, it can manufacture suitably.

In addition, the food may be produced as oral solid preparations such as tablets, granules and capsules, or oral liquid preparations such as internal use liquid preparations and syrup preparations. The method for producing the above oral solid preparation and oral liquid preparation is not particularly limited and can be appropriately selected depending on the purpose. For example, it can be produced in accordance with the above-described method for producing an oral solid preparation or oral liquid preparation. it can.

The food / beverage products of the present invention are particularly useful as functional foods, health foods, and the like for the purpose of activating the innate immune mechanism and imparting resistance to infectious diseases.
When the lactic acid bacteria of the present invention, the killed bacteria or the processed product are used for the production of food and drink, the production method can be carried out by methods well known to those skilled in the art. If it is an expert, the process which mixes the (dead) microbial cell or processed material of this invention with other components, a shaping | molding process, a sterilization process, a fermentation process, a baking process, a drying process, a cooling process, a granulation process, It is possible to combine the packaging process and the like as appropriate to make the desired food or drink.

Moreover, when using the lactic acid bacteria of this invention for manufacture of various fermented milk, it can manufacture using a method well-known to those skilled in the art. For example, food and drink manufactured using the process of adding the required amount of lactic acid bacteria of the present invention as dead bacteria to fermented milk and food and drink manufactured using the process of fermenting using the lactic acid bacteria of the present invention as a lactic acid bacteria starter Can be mentioned.
When performing fermentation using the lactic acid bacteria of this invention as a lactic acid bacteria starter, it can carry out on the conditions similar to the culture conditions of the lactic acid bacteria of this invention, etc.

Hereinafter, the present invention will be described in more detail based on examples and examination examples, but the present invention is not limited to the specific scope of the following examples and the like.

As described above, “Lactic acid bacteria # 7-2” and “Lactic acid bacteria 8 / 11-3” used in the examples were separated from Kimchi, and “Lactic acid bacteria # 4-2” was separated from straw.

Lactic acid bacteria # 7-2 belonging to Leuconostoc carnosum is located in the National Institute for Product Evaluation and Technology Patent Microorganism Depositary (NPMD) (Kazusa Kamashitsu, Chiba Prefecture 2-5-8 Room 122) It has been deposited (Accession Number: NITE P-02307, Deposit Date July 26, 2016).
Lactic acid bacteria # 7-2 then submitted the original deposit application form to the patent microorganisms deposit center (NPMD) of 2-5-8 Kazusa-Kamashita, Kisarazu, Chiba Prefecture, Room 122, NITE Submission and application for transfer from domestic deposit (original deposit date: July 26, 2016) to deposit under the Budapest Treaty (transfer date (international deposit date): July 25, 2017) As a result of receiving an application for transfer to a deposit based on the Budapest Treaty (international deposit), it has received the deposit number “NITE BP-02307”.

Lactic acid bacteria # 4-2 belonging to Leuconostoc gelidum has been established in the National Institute for Product Evaluation and Technology Patent Microbiology Depositary Center (NPMD) (Kazusa Kamashichi, Kisarazu City, Chiba Prefecture, Room No. 2-5-8 122) It has been deposited (Accession Number: NITE P-02306, Deposit Date July 26, 2016).
Similarly to lactic acid bacteria # 7-2, lactic acid bacteria # 4-2 also submitted an original deposit application to the Patent Microorganism Deposit Center (NPMD), from domestic deposit (original deposit date: July 26, 2016), An application for transfer to the deposit under the Budapest Treaty was made (transfer date (international deposit date): July 25, 2017), survival was proved, and an application for transfer to the deposit under the Budapest Treaty (international deposit) was received As a result, it has received the deposit number “NITE BP-02306”.

Lactic acid bacteria 8 / 11-3 belonging to Leuconostoc mesenteroides is the National Institute of Technology and Evaluation (NPMD), National Institute for Product Evaluation and Technology (NPMD) (Room 2-5-8 Kazusa Kamashichi, Kisarazu City, Chiba Prefecture) (Accession number: NITE P-02308, date of deposit July 26, 2016).
Lactic acid bacteria 8 / 11-3, as well as lactic acid bacteria # 7-2, submitted the original deposit application form to the Patent Microorganism Deposit Center (NPMD) and started domestic deposit (original deposit date: July 26, 2016). Made an application for transfer to the deposit under the Budapest Treaty (transfer date (international deposit date): May 22, 2017), and the survival was proved, and an application for transfer to the deposit under the Budapest Treaty (international deposit) was received As a result, it has received the deposit number “NITE BP-02308”.

<Materials and methods>
<< Measurement of innate immunity promoting activity of lactic acid bacteria using silkworm muscle specimens >>
The innate immunity promoting activity was measured using the method described in Patent Document 1, using silkworm muscle contraction activity as an index. Lactic acid bacteria were spread on 0.5% calcium carbonate MRS agar medium and anaerobically cultured at 30 ° C. to form colonies. Colonies were collected, inoculated into 14 mL of MRS medium added to a 15 mL tube, and statically cultured at 30 ° C. for 2 days. The whole culture solution was added to 100 mL of MRS liquid medium and cultured at 30 ° C. for 1 day. The culture was autoclaved at 121 ° C. for 20 minutes and centrifuged at 8000 rpm and 4 ° C. for 10 minutes. The precipitate was washed with 50 mL of 0.9% NaCl and then centrifuged again at 8000 rpm and 4 ° C. for 10 minutes, and the precipitate was suspended in 1 mL of 0.9% NaCl solution. 50 μL of this suspension was diluted and injected into a silkworm muscle specimen, and the change in the length of the silkworm specimen before and after the injection was measured. The sample amount by which the length of the silkworm muscle specimen was reduced by 15% was defined as 1 unit. On the other hand, 100 μL of the bacterial suspension was dried with a centrifugal evaporator, and the dry weight was measured.

<< Proliferation test of each lactic acid bacterium in milk, fruit and vegetable juice >>
Pre-culture of lactic acid bacteria was carried out by anaerobic culture at 30 ° C. using MRS liquid medium. A preculture solution was added to 50 mL of various vegetable / fruit juices, and static culture was performed at 30 ° C. Viable counts of lactic acid bacteria were measured by spreading the culture on an MRS agar medium and anaerobically culturing at 30 ° C., and counting the number of colonies that appeared. When it reached 1.0 × 10 ⁶ cells / mL or more, it was determined that the proliferation ability was “present”.

Bitter gourd juice was prepared by adding bitter gourd to RO water and treating with juicer at 121 ° C. for 20 minutes.
Kiwi juice was prepared by adding 500 mL of RO water to 100 g of kiwi. The orange juice is “100% orange (manufactured by Tokyo Meiraku Co., Ltd.)”, the apple juice is “100% apple (manufactured by Tokyo Meiraku Co., Ltd.)”, and the vegetable mixed juice is “daily vegetables (manufactured by ITO EN Co., Ltd.)”. used.
Broccoli juice was prepared by incubating at 37 ° C. for 1 day after autoclaving.
The pH was adjusted by adding a 10N sodium hydroxide solution. Kiwi juice was adjusted to pH 7, and apple juice, orange juice and grapefruit juice were adjusted to pH 6.

Glycerol stock of three kinds of lactic acid bacteria was added to 50 mL of milk (manufactured by Meiji Co., Ltd., delicious milk), and anaerobic culture was performed at 30 ° C. for 1 day. The number of viable bacteria in the milk was calculated by spreading the milk dilution on the MRS agar medium and measuring the number of colonies that appeared after anaerobic culture at 30 ° C.
Glycerol stocks were spread on 0.5% calcium carbonate MRS agar medium and anaerobically cultured at 30 ° C. to form colonies. The colony was inoculated in 14 mL of MRS liquid medium and anaerobically cultured at 30 ° C. for 1 day, and then the precipitate centrifuged at 8000 rpm and 4 ° C. for 5 minutes was suspended in 2 mL of 0.9% NaCl, and the like. Mix well with 80% glycerol solution and store at -80 ° C.

<< Quantification of sugar concentration in vegetable and fruit juices >>
The total sugar concentration in the juice was quantified by the phenol sulfate method. The glucose concentration was quantified using Accu-Chek (Roche).
The phenol sulfuric acid method was performed as follows. The culture solution was centrifuged at 8000 rpm for 10 minutes, and 100 μL of the supernatant was recovered. Next, 100 μL of 5% phenol was added and stirred vigorously with a vortex mixer for 5 seconds, and 500 μL of sulfuric acid was added and stirred vigorously with a vortex mixer until heat was generated. After standing at room temperature for 20 minutes, OD490 was measured. Accu Check Aviva Trip Trip F was set in Accu Check Aviva, and the glucose concentration was quantified.

Example 1
<Measurement of innate immunity promoting activity of each lactic acid bacterium>
The separated various lactic acid bacteria were autoclaved and centrifuged, and then the bacterial cell component fraction was collected. The innate immunity promoting activity of each lactic acid bacterium was evaluated by injecting a bacterial suspension into a silkworm muscle specimen and measuring the contraction of the muscle. The results are shown in Table 1.
As a result, the specific activity of Leuconostoc carnosum # 7-2 (lactic acid bacterium # 7-2) was 460 units / mg, the specific activity of Leuconostoc gelidum # 4-2 (lactic acid bacterium # 4-2) was 250 units / mg, Leuconostoc mesenteroides 8/11 -3 (lactic acid bacteria 8 / 11-3) had a specific activity of 250 units / mg, and a high activity value was obtained for any lactic acid bacteria (Table 1).

Example 2
<Proliferation of lactic acid bacteria in fruit and vegetable juices and milk>
Each of the three strains of lactic acid bacteria was inoculated into fruit / vegetable juice and milk, and the presence or absence of bacterial growth was examined. The results are shown in Tables 2-4.

Only lactic acid bacteria 8 / 11-3 showed growth in non-neutralized fruit / vegetable juice (Table 2). On the other hand, lactic acid bacteria # 7-2 and lactic acid bacteria # 4-2 grew when the fruit juice was neutralized (Table 3).
Moreover, as a result of culturing each lactic acid bacterium in milk at 30 ° C. for 24 hours, proliferation was observed in all cases (Table 4).

Example 3
<Decrease in sugar content due to growth of each lactic acid bacterium in fruit and vegetable juice>
Each of the three strains of lactic acid bacteria was cultured in fruit juice, and the sugar concentration in the juice was quantified. The total sugar concentration was measured by the phenol sulfuric acid method. The glucose concentration was quantified with Accu-Chek (Roche). Table 5 shows the measurement results.

As a result of Table 5, orange grapefruit juice grown with lactic acid bacteria # 7-2, orange grapefruit juice grown with lactic acid bacteria # 4-2, kiwi grapefruit juice grown with lactic acid bacteria 8 / 11-3, Large reductions in sugar and glucose concentrations were observed.

The novel lactic acid bacteria and the processed product of the present invention have a high innate immunity activation ability, and further have an effect of preventing and treating infectious diseases. Therefore, it is possible to provide a drug or a food or drink containing an innate immunity activator that activates innate immunity or an infectious disease prevention / treatment agent using the lactic acid bacterium of the present invention, and widely used in the pharmaceutical industry, the food industry, etc. Is possible.

This application is based on Japanese Patent Application No. 2016-159557 filed on August 16, 2016 in Japanese Patent Application, the entire contents of which are cited herein as disclosure of the specification of the present invention. It is taken in.

NITE BP-02307
NITE BP-02306
NITE BP-02308

SEQ ID NO: 1 is a base sequence corresponding to almost the entire length of 16S rDNA of an unknown strain (lactic acid bacterium # 7-2) belonging to the genus Leuconostoc.
SEQ ID NO: 2 is a base sequence corresponding to almost the entire length of 16S rDNA of an unknown strain (lactic acid bacterium # 4-2) belonging to the genus Leuconostoc.
SEQ ID NO: 3 is a base sequence corresponding to almost the entire length of 16S rDNA of an unknown strain (lactic acid bacterium 8 / 11-3) belonging to the genus Leuconostoc.

Claims (7)

A lactic acid bacterium belonging to the genus Leuconostoc whose NITE BP-02307 has a deposit number at the Patent Microorganism Deposit Center (NPMD) of the National Institute of Technology and Evaluation (NITE). A lactic acid bacterium belonging to the genus Leuconostoc, whose deposit number is NITE BP-02306 at the Patent Microorganism Deposit Center (NPMD) of the National Institute of Technology and Evaluation (NITE). A lactic acid bacterium belonging to the genus Leuconostoc whose accession number is NITE BP-02308 at the Patent Microorganism Deposit Center (NPMD) of the National Institute of Technology and Evaluation (NITE). An innate immunity activator comprising as an active ingredient the lactic acid bacterium according to any one of claims 1 to 3, a killed bacterium of the lactic acid bacterium, or a processed product of the lactic acid bacterium,
The treated product of lactic acid bacteria is at least selected from the group consisting of cultures, concentrates, pasted products, dried products, liquefied products, diluted products, crushed products, sterilized processed products, and extracts from cultured products. An innate immunity activator characterized by being one processed product. A food or drink containing the lactic acid bacteria according to any one of claims 1 to 3. A food or drink containing the innate immunity activator according to claim 4. The food-drinks manufactured using the process fermented using the lactic acid bacteria of any one of Claims 1 thru | or 3.