JP3315826B2 - Bifidobacterium growth promotion composition - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a composition for promoting the growth of bifidobacteria comprising a naphthoquinone derivative and a naphthalene derivative having an activity of promoting the growth of bifidobacteria at an extremely low concentration as an active ingredient. It is used as a selective medium for improving intestinal flora by adding it to various foods and beverages and for counting the number of bifidobacteria.

[0002] Comparative studies on the intestinal flora of breastfeeding infants and artificially fed infants have suggested that bifidobacteria are useful for human health. At present, it is effective to significantly reduce bifidobacteria with various digestive tract diseases and aging, and to promote the growth of intestinal bifidobacteria, to suppress carcinogenesis, inhibit intestinal rot, and prevent infectious diseases. That has been confirmed. Therefore, it can be said that the selective growth of bifidobacteria in the intestine is extremely important from the viewpoint of maintaining health and preventing and treating various adult diseases.

In view of the above, the number of foods (fermented milk, yogurt, etc.) to which bifidobacteria have been added in recent years has increased. From the viewpoint of production and quality control of these foods, the number of bifidobacteria in these foods has been increasing. It has become necessary to develop a selective medium that can specifically measure the concentration of the selective medium.

[0004]

2. Description of the Related Art As substances useful for promoting the growth of useful bifidobacteria, so-called bifidobacteria, several substances have been studied and reported. For example, N-acetylglucosamine contained in breast milk (Proc.
Soc. Exp. Biol. Med. , 90 , 219
(1955)), peptide-related substances (Am. J. Cli)
n. Nutr. , 32 , 1428 (1974); Agr
ic. Biol. Chem. , 48 , 2159 (198
4)), ginseng extract (Nichino Kagaku Magazine, 55 , 499 (198)
1); Chem. Pharm. Bull. , (Tokyo
o) 14 , 1191 (1966)) and sugar-related substances (Bulletin of the Tohoku Fukushi University, 10 , 313 (1986)). However, the preparation of these Bifidobacterium growth-promoting substances is complicated, and the effect of selectively growing only Bifidobacteria is not always sufficient.

[0005]

DISCLOSURE OF THE INVENTION The present invention relates to a composition for promoting the growth of bifidobacteria comprising a compound capable of selectively and rapidly growing only bifidobacteria, and a system for measuring the number of bifidobacteria using these compounds. The purpose is to provide.

[0006]

DISCLOSURE OF THE INVENTION The present invention has been made to achieve the above-mentioned object, and the present inventors have intensively studied various compounds which selectively promote the growth of Bifidobacteria. As a result, certain naphthoquinone derivatives and naphthalene derivatives were converted to various Bifidobacteria (Bifi
dobacterium longum, B.C. brev
e, B. adolescentis, B.A. bifidu
m, B. infantis, B .; animalis,
B. Pseudolongum, etc.). The present invention has been made based on this new finding.

That is, according to the present invention, a hydrogen atom, a chlorine atom, and / or a functional group (a hydroxyl group, a carboxyl group, an amino group, an aldehyde group, and / or an alkyl group or other various functional groups) are added to 1,4-naphthoquinone or naphthalene. The present invention relates to a composition for promoting the growth of bifidobacteria, which comprises a compound to which the group (1) is added as an active ingredient.

Examples of naphthoquinone derivatives and naphthalene derivatives having a growth promoting action specific to Bifidobacteria include the following. 1) α-naphthoquinone, 2) 2-hydroxy-1,4-
Naphthoquinone, 3) 2,3-dichloro-1,4-naphthoquinone, 4) 5-hydroxy-1,4-naphthoquinone,
5) filoquinone, 6) menaquinone-n, 7) menadione, 8) 4-amino-2-methyl-1-naphthol,
9) 1-naphthoic acid, 10) 2-naphthoic acid, 11)
1,4-dihydroxy-2-naphthoic acid, 12) 1-hydroxy-2-naphthoic acid, 13) 1,4-naphthalenedicarboxylic acid, 14) 1-naphthylacetic acid, 15) α-naphthylamine, 16) α-naphthyl Aldehyde, 17)
β-naphthylaldehyde.

[0009] In addition to the above compounds, compounds obtained by adding some functional groups such as amino group, hydroxyl group, carboxyl group, aldehyde group and alkyl group to α-naphthoquinone or naphthalene also exhibit the activity of promoting the growth of bifidobacteria. It is expected that these compounds can also be used as the active ingredient compound of the composition according to the present invention.

Furthermore, in addition to these known compounds, a new bifidobacterium growth-promoting substance has been sought, and as a result of intensive studies on metabolites of microorganisms, Propionobacterium spp. It was found to produce a highly active bifidobacterial growth-promoting substance.

The physicochemical properties of this substance were further studied in detail. As a result, it was confirmed that the substance was a novel substance which had not been known before, the structure was successfully determined, and an industrial production method was established. The invention has been completed.

The novel Bifidobacterium growth promoting substance (hereinafter, also referred to as a bifidobacterium factor) according to the present invention, which has been found in this way, has a conventionally unknown novel physicochemical property shown in (1) or (2) below. Substance.

[0013]

[Outside 1]

[0014]

[Outside 2]

The bifidogenic factor derived from this microorganism shows the properties of a low-molecular-weight quinone compound in view of the above-mentioned physicochemical properties, and succeeded in the determination of its structure. ) Was obtained, and a previously unknown novel compound, 2-amino-3, was obtained.
-Carboxy-1,4-naphthoquinone.

[0016]

Embedded image

The novel substance 2-amino-3-carboxy-1,4-naphthoquinone according to the present invention is 2-amino-3-
It can be obtained by culturing a strain having the ability to produce carboxy-1,4-naphthoquinone and collecting 2-amino-3-carboxy-1,4-naphthoquinone from the culture. Further, it can also be produced by a chemical synthesis method.

2-amino-3-carboxy-1,4-naphthoquinone (hereinafter sometimes abbreviated as the substance of the present invention)
Examples of the genus of genus producing naphthoquinone derivatives such as Propionibacteria (Propionibacterium)
acterium), Enterobacter
bacter) and Bacillus. For example, Propionibacterium freudenreich fr
eudenreichii) ATCC 6207 strain
It is suitable as a bacterium producing the substance of the present invention.

In order to produce 2-amino-3-carboxy-1,4-naphthoquinone according to the present invention, a strain having the ability to produce the substance of the present invention is first prepared by including a nutrient source capable of growing ordinary microorganisms. Culture aerobically or anaerobically in a medium. Known nutrients that have been conventionally used for culturing microorganisms can be used. In particular, a medium consisting of trypticase, phyton, yeast extract and glucose is preferably used.

As the culturing method, various known aerobic and anaerobic culturing methods can be used, but the aerobic or anaerobic culturing method using a liquid medium is most preferable from the viewpoint of mass production.
The cultivation temperature is about 20 to 40 ° C., and the pH of the medium is cultivated under neutral to slightly acidic conditions. In liquid culture, about 1
After 3 days, the substance of the present invention is accumulated in the medium and the cells. The culture is stopped, the culture supernatant is separated from the cells, and the cells are subjected to separation and purification of the substance of the present invention.

The method for separating and purifying the substance of the present invention is described below. First, cells are separated from the culture solution by a usual centrifugation method. Chloroform: methanol =
A 2: 1 solution is added and the active substance is extracted with stirring. Then
The extract fraction is concentrated under reduced pressure and then applied to a silica gel column. Thereafter, when elution is performed while increasing the concentration of ethyl acetate in n-hexane, the activity of promoting the growth of bifidobacteria is observed in the eluted fraction having an ethyl acetate concentration of 45 to 60% in n-hexane. After concentrating this eluted fraction under reduced pressure,
Gel filtration chromatography on an ephadex LH-20 column. When eluted with methanol, the growth promoting activity of Bifidobacterium is observed at the relatively low molecular weight elution position.

After the eluted fraction is concentrated under reduced pressure, it is further subjected to high performance liquid chromatography using a reversed phase column.
Acetonitrile: methanol: water: acetic acid = 250: 10
When eluted with 0: 900: 0.6 (pH 5.6), the growth promoting activity of Bifidobacterium is eluted as a single peak. The eluted fraction is concentrated under reduced pressure to obtain a yellow powdered substance of the present invention.

The substance of the present invention separated and purified as described above exhibits the physicochemical properties as described above. The structure was also determined. As a result, the novel substance 2 represented by the formula (I) 2
-Amino-3-carboxy-1,4-naphthoquinone.

The substance of the present invention widely includes not only the naphthoquinone derivative represented by the formula (I) but also a salt thereof. Examples of the salt of the substance of the present invention with a base include an alkali metal salt such as a sodium salt and a potassium salt; an alkaline earth metal salt such as a calcium salt and a magnesium salt; and the salt with an acid include hydrochloride and sulfate. And the like.

As will be apparent from the description below, all of the active ingredient compounds of the composition of the present invention, including the substance of the present invention, have a very strong bifidobacterial growth promoting action. The composition containing the active ingredient compound of the present invention can be used as a bifidobacterium growth promoting composition in the form of a pharmaceutical type, a food and drink type, and / or an analytical agent type. By direct administration or by direct administration or ingestion as nutritional food or functional food such as food for specified health use, or by adding it to various foods (fermented milk, yogurt, etc.) and ingesting it In addition, the intestinal flora can be improved. Further, the composition of the present invention can also be used in an assay system such as a bifidobacteria counting method.

When the composition according to the present invention is used as a pharmaceutical composition, the substance of the present invention and / or the active ingredient compound of the present invention are administered in various forms. Examples of the administration form include oral administration by tablets, capsules, granules, powders, syrups and the like. These various preparations are commonly used in the pharmaceutical preparation technical field such as excipients, binders, disintegrants, lubricants, flavoring agents, solubilizing agents, suspending agents, coating agents, and the like, in accordance with the usual methods for the main drug. It can be formulated using known adjuvants.

When this preparation is applied to humans, oral administration is preferred. The therapeutically effective amount of the active ingredient varies depending on the age and conditions of each patient to be treated, but generally the active ingredient is administered in a daily dose of 0.01 to 1. 1 kg / kg human body weight.
0 mg is orally administered.

Since the various compounds used in the present invention can achieve the intended purpose by oral administration, the composition according to the present invention can be used as a food or drink type. To that end, the substance of the present invention and / or the active ingredient compound of the present invention are prepared using various auxiliaries and other foods and drinks, drinks, tablets, and other various food and drink types, or directly added to foods and drinks. Various methods can be used. Since the composition of the present invention in the form of a food and drink can be ingested for a long period of time, in addition to ordinary food and drink, it is marketed as a food for specified health use, a nutritional supplement, a health food and the like. be able to.

Each of the various compounds used in the present invention, including the substance of the present invention, has not only a very high ability to promote the growth of bifidobacteria but also a very high selectivity and specificity. In addition, according to the present invention, even if the growth of bifidobacteria is suppressed by the presence of an antibiotic or the like, the growth of the bifidobacterium can be increased by adding the growth promoting substance (the various active ingredient compounds used in the present invention, including the substance of the present invention). Restores ability. Therefore, in addition to the method of increasing the number of bifidobacteria in the digestive tract by directly administering the present growth-promoting composition to humans or animals, it is also used as a selective medium for analyzing bifidobacteria in foods, large intestine contents, fecal samples, etc. It is possible to

That is, as one of the systems for counting the number of bifidobacteria in a sample containing bifidobacteria, an agar medium containing a selective agent for suppressing the growth of contaminants other than bifidobacteria has been used. A method is known in which only Bifidobacterium is specifically or selectively grown by inoculating and incubating the Bifidobacterium, and the presence or absence of the Bifidobacterium and the number of viable bacteria are measured. However,
In the selection medium to which the selection agent is added, especially when a high concentration of the selection agent is used, the growth of contaminants other than Bifidobacterium is naturally suppressed, and the growth of Bifidobacterium itself may be suppressed, It was difficult to accurately determine the number of bifidobacteria.

On the other hand, when the composition for promoting the growth of Bifidobacterium according to the present invention is added to the selective medium, the growth of Bifidobacterium can be specifically promoted and colonies can be formed. You can. Moreover, this effect is achieved by using paromomycin sulfate,
In addition to antibiotics such as neomycin sulphate, propionate, lithium chloride and any other known selection agent can be used. It can be used effectively for applications.

[0032]

Embodiments of the present invention will be described below.

[0033]

Example 1: Activity (1) 1/2 concentration TPYG medium supplemented with 1.5% agar (Trypticase (BBL))
4 g, phyton peptone (BBL) 1.5 g, yeast extract 2.5 g, L-cysteine hydrochloride 0.25 g, glucose 20 g, K ₂ HPO ₄ 1 g, KH ₂ PO ₄ 1.5
g, MgCl ₂ .6H ₂ O 0.25 g, FeSO ₄ .7
5 mg of H ₂ O, 1000 ml of purified water, pH 7.5) were dissolved by heating, and the mixture was dissolved in Bifidobacterium at 50 ° C.
m longumATCC 15707 to 10 ⁶ cfu
/ Ml, and 10 ml was dispensed into a sterilized petri dish (8.5 cm in diameter) to prepare an agar plate. On this plate, 1 mg / m
A growth promoting circle observed when a paper disc (diameter 6 mm, thickness 0.7 mm) impregnated with ethanol solution serially diluted to 1 to 100 ng / ml and dried is placed at 37 ° C. for 15 hours under anaerobic culture. The diameter was measured.
The results are shown in Table 1 below.

[0034]

[Table 1]

(A) α-naphthoquinone (b) 2-hydroxy-1,4-naphthoquinone (c) 2,3-dichloro-1,4-naphthoquinone (d) 5-hydroxy-1,4-naphthoquinone (e) phylloquinone (vitamin K ₁₎ (f) menaquinone (vitamin K ₂₎ (g) menadione (vitamin K ₃₎ (h) 4- amino-2-methyl-1-naphthol (vitamin K ₅₎ (i) 1- naphthoic acid (J) 2-naphthoic acid (k) 1,4-dihydroxy-2-naphthoic acid (l) 1-hydroxy-2-naphthoic acid (m) 1,4-naphthalenedicarboxylic acid (n) 1-naphthylacetic acid (o ) Α-Naphthylamine (p) α-naphthylaldehyde (q) β-naphthylaldehyde

As shown in Table 1, all of the present growth-promoting substances showed growth-promoting activity against bifidobacteria.
Among them, in particular, 1,4-dihydroxy-2-naphthoic acid and 1-hydroxy-2-naphthoic acid showed strong activity.

[0037]

Example 2: Degree of activity (2) The substance of the present invention (2-amino-3-carboxy-1,4-naphthoquinone) obtained in Examples described later was adjusted to a concentration of 1 to 0.01 ng / ml with T.
PYG medium (8 g of trypticase (BBL), 3 g of phyton peptone (BBL), 5 g of yeast extract, 0.5 g of L-cysteine hydrochloride, 20 g of glucose, K ₂ HPO ₄
2 g, KH ₂ PO ₄ 3 g, MgCl ₂ .6H ₂ O 0.5
_{g, FeSO 4 · 7H 2 O} 10mg, purified water 1000
ml, pH 6.5), followed by Bifidobacterium longum A.
TCC 15707, Bif. breve ATCC
15700, Bif. bifidum ATCC 11
146, Bif. adolescentis ATCC
15703, Bif. infantis ATCC
15697).
After anaerobic culturing for hours, the OD ₅₈₀ (absorbance at a wavelength of 580 nm) of each culture solution was measured. The results are shown in Table 2 below. As a control, a TPYG medium (pH 6.5) in which each of the test bacteria was inoculated without adding the substance of the present invention was used.

[0038]

[Table 2]

As shown in Table 2, the substance of the present invention contained 0.
It was confirmed that a very low concentration of 01 to 0.1 ng / ml exhibited a growth promoting effect on various bifidobacteria.

[0040]

Example 3: Growth-promoting action against various bifidobacteria
The growth promoting substances (a) to (q) shown in Example 1 were used in 10
A 濃度 concentration TPYG medium (pH
6.5), followed by Bifidobacterium ((A) B
ifidobacterium longum ATC
C 15707, (B) Bif. breve ATCC
15700, (E) Bif. bifidum ATC
C 29521, (C) Bif. adolescent
is ATCC 15703, (D) Bif. infa
ntis ATCC 15697), and after anaerobic cultivation at 37 ° C. for 15 hours by a gas pack method, the OD ₅₈₀ (absorbance at a wavelength of 580 nm) of each culture solution was measured. The results are shown in Table 3 below. As a control, a TPYG medium (pH 6.5) in which each of the test bacteria was inoculated was used without adding the growth promoting substance.

[0041]

[Table 3]

As shown in Table 3, 2-hydroxy-
All of the present growth-promoting substances except 1,4-natoquinone, 1,4-naphthalenedicarboxylic acid, α-naphthylamine and α-naphthylaldehyde show a growth-promoting action against various bifidobacteria at a concentration as low as 10 μg / ml. confirmed.

[0043]

Example 4: Bifidobacterium specificity of growth-promoting activity (1) The substance of the present invention obtained in Examples described later was 10 ng / day.
ml of TPYG medium (pH 6.5), followed by inoculation of various intestinal bacteria including bifidobacteria described below, followed by anaerobic cultivation at 37 ° C. for 20 hours by a gas pack method.

(A) Bifidobacterium longum ATCC 15707 (b) Bif. Breve ATCC 15700 (c) Bif. Adolescentis ATCC 15703 (d) Bif. Infantis ATCC 15697 (e) Bif. Bifidum ATCC 11146 (f) Clostridium perfringens ATCC 3626 (b) G) Cl. Butyricum ATCC 860 (h) Cl. Ramosum ATCC 13937 (h) Enterobacter cloacae ATCC 961 (h) Escherichia coli ATCC 26 (h) Fusobacterium varium ATCC 8501 (h) Bacteroides fragilis ATCC 23745 (h) Eubacterium aerofaciens ATCC 25986 (F) Enterococcus faecalis IFO 3971 (f) Staphylococcus aureus ATCC 4012

After completion of the culture, the OD _{580 of} each culture was
(Absorbance at a wavelength of 580 nm) was measured.
Was obtained. As a control, a TPYG medium to which each of the test bacteria was inoculated was used without adding the substance of the present invention.

[0046]

[Table 4]

As shown in Table 4 above, remarkable promotion of growth of various Bifidobacteria was observed by adding the substance of the present invention. On the other hand, against other intestinal bacteria, the growth promoting effect was extremely weak or absent, and the selectivity or specificity of the substance of the present invention was also demonstrated.

[0048]

Example 5: Bifidobacterium specificity of growth promoting action (2) Among the growth promoting substances, 1-hydroxy-2-
Naphthoic acid was added to a TPYG medium (pH 6.5) at a concentration of 100 ng / ml, and subsequently various intestinal bacteria including bifidobacteria described below were inoculated and anaerobically cultured at 37 ° C. for 15 hours by a gas pack method. did.

(A) Bifidobacterium longum ATCC 15707 (B) Bif. Breve ATCC 15700 (C) Bif. Adolescentis ATCC 15703 (D) Bif. Infantis ATCC 15697 (E) Bif. Bifidum ATCC 29521 (F) Clostridium perfringens ATCC 3626 (B) G) Lactobacillus acidophilus ATCC 4356 (H) Fusobacterium varium ATCC 8501 (I) Enterobacter cloacae ATCC 961 (J) Escherichia coli ATCC 26 (K) Peptostreptococcus products ATCC 27340 (L) Bacteroides vulgatus ATCC 8482 (M) Eubacterium aerofaciens ) Enterococcus faecalis IFO 3971 (O) Staphylococcus aureus ATCC 4012

After the completion of the culture, the CD _{580 of} each culture was used.
(Absorbance at a wavelength of 580 nm) was measured. The results are shown in Table 5 below. In addition, 1-hydroxy-
TPYG medium without adding 2-naphthoic acid (pH 6.
Each of the test bacteria inoculated in 5) was used.

[0051]

[Table 5]

As shown in Table 5, 1-hydroxy-2
-By adding naphthoic acid, remarkable growth promotion of various bifidobacteria was recognized. On the other hand, against other intestinal bacteria, the growth promoting effect was extremely weak or absent, and the selectivity or specificity was also demonstrated. Similar selectivity or specificity was confirmed for substances other than this substance.

[0053]

Example 6: Production of the substance of the present invention TPYG medium (pH
6.5) 10 L was dispensed into three 5 L Erlenmeyer flasks and sterilized in an autoclave at 121 ° C. for 15 minutes.
This is followed by Propionibacterium freudenreich (P
ropionobacterium freudenr
eichii) Activated culture of ATCC 6207 strain 1
The seed culture was prepared by inoculating 00 ml and statically culturing at 30 ° C. for 3 days.

In the main culture, 500 L of the above TPYG medium (pH 6.5) sterilized at 135 ° C. for 5 seconds was added to
10 L of the seed culture prepared as described above was inoculated and allowed to stand at 30 ° C. for 96 hours under nitrogen gas pressure (0.5 kg / cm ² ). The culture solution thus obtained was subjected to continuous centrifugation to separate the culture supernatant from the cells. The obtained cells were freeze-dried to obtain 1738 g of freeze-dried cells.

15 L of a 2: 1 chloroform: methanol solution was added to the dried cells obtained above, and the cells were added at room temperature for 30 minutes.
After stirring for minutes, the substance of the present invention was extracted. The obtained extract was subjected to suction filtration, and the filtrate was concentrated under reduced pressure to obtain 22.8 g of an extract. This was subjected to a silica gel column (column length 70 cm, column diameter 6.0 cm; Wako Gel C-300 manufactured by Wako Pure Chemical Industries, Ltd.).
Then, elution was carried out while increasing the ethyl acetate concentration in n-hexane, and the eluate fraction having an ethyl acetate concentration in n-hexane of 45 to 60% showed a bifidobacterium growth promoting activity.

The eluted fraction was concentrated to dryness under reduced pressure,
Dissolve in 8 ml of methanol and then Sephadex L
H-20 column (column length 70 cm, column diameter 2.6 c
m; manufactured by Pharmacia). Using methanol as eluent, flow rate 36 ml
/ Hr, the eluate volume was 497-539 ml
The fraction showed a bifidobacterium growth promotion activity. The eluted fraction was concentrated under reduced pressure.

Next, a reversed phase column (column length 250 mm,
Column diameter 20mm; Shiseido's CAPCELL PAK
High performance liquid chromatography using C ₁₈ ) was performed. Acetonitrile: methanol: water: acetic acid = 250:
100: 900: 0.6 (pH 5.6) as eluent,
The elution was performed at a flow rate of 10 ml / min.
The activity of promoting the growth of bifidobacteria was observed in the eluted fractions around the minute. The eluted fraction was concentrated under reduced pressure to obtain 7.1 mg of the substance of the present invention as a yellow powder. Its physicochemical properties were as previously described, and its chemical structure was determined as in formula (I).

[0058]

Example 7 50 g of granulated sugar, 100 g of an equal mixture of corn starch and lactose, and 100 mg of 1-hydroxy-2-naphthoic acid were added and mixed well. Mix 10
The mixture was divided into 0 equal portions and packed into bags, and 100 bags of 1.5 g stick-shaped bifidobacterium nutritional health food were produced.

[0059]

Example 8 The following formulations (1) to (4) were prepared. (1) 1 g of 1-hydroxy-2-naphthoic acid,
(2) lactose 140g, (3) corn starch 29g,
(4) 1 g of magnesium stearate.

First, (1), (2) and (3) (17
g) was mixed and granulated with the paste prepared from (3) (but 7 g). Add (3) (5
g) and (4) were added and mixed well, and the mixture was compressed using a compression tablet machine to produce 1000 tablets each containing 1 mg of 1-hydroxy-2-naphthoic acid.

[0061]

Example 9 50 g of granulated sugar, 100 g of an equal mixture of corn starch and lactose, substance 1 of the present invention obtained in Example 6
00 mg was added and mixed well. The mixture was divided into 100 equal parts and packed into bags to produce 100 bags of 1.5 g stick-shaped bifidobacterium nutritional health food.

[0062]

Example 10 The following formulations (1) to (4) were prepared. (1) 1 g of the substance of the present invention obtained in Example 6, (2) lactose 140 g, (3) corn starch 29 g, (4) magnesium stearate 1 g.

First, (1), (2) and (3) (17
g) was mixed and granulated with the paste prepared from (3) (but 7 g). Add (3) (5
g) and (4) were added and mixed well, and the mixture was compressed using a compression tablet machine to produce 1000 tablets each containing 1 mg of the substance of the present invention.

[0064]

Since the bifidobacterium comprising the compound according to the present invention is excellent in promoting the growth of bifidobacteria, it can be directly administered or ingested to humans or animals as a food or drink or an orally administered drug, and can be used in the digestive tract. Very effective in increasing bifidobacteria.

Further, the bifidobacterium according to the present invention comprises:
In addition to the excellent effect of promoting the growth of bifidobacteria, the selectivity or specificity is strong, and no growth effect is observed on other contaminating bacteria.Therefore, when the bifidobacterium is added to the bifidobacterium selection medium, A colony containing only bacteria is quickly formed, and can be advantageously used for measuring the viable cell count of bifidobacteria and other various assays.

[Brief description of the drawings]

FIG. 1 shows an ultraviolet absorption spectrum of 2-amino-3-carboxy-1,4-naphthoquinone dissolved in methanol to a concentration of 10 μg / ml.

Continued on the front page (51) Int.Cl. ⁷ Identification code FI A61K 31/19 A61K 31/19 31/70 31/70 A61P 1/12 A61P 1/12 C12N 1/38 C12N 1/38 C12Q 1/06 C12Q 1/06 // (C12N 1/38 (C12N 1/38 C12R 1:01) C12R 1:01) (C12Q 1/06 (C12Q 1/06 C12R 1:01) C12R 1:01) (72) Inventor Nao Taketomo 540 Narita, Odawara-shi, Kanagawa Prefecture Meiji Dairies Corporation Health Science Research Institute (72) Inventor Yoshiro Sato 540 Narita, Odawara-shi, Kanagawa Prefecture Meiji Dairies Corporation Health Science Research Institute (72) Inventor Tomotaka Kamiyama Odawara, Kanagawa Prefecture 540 Narita, Japan Meiji Dairy Products Co., Ltd. Health Science Laboratory (56) References JP-A-5-41995 (JP, A) JP-A 5-344882 (JP, A) (58) Fields investigated (Int. Cl. ⁷ , DB name) C12N 1/00-1/38 A23C 9/13 CA (STN) REGISTRY (STN) WPI (D IALOG) BIOSIS (DIALOG) JICST file (JOIS)

Claims (5)

(57) [Claims] 1. A composition for promoting the growth of bifidobacteria, comprising at least one of the following (a) to (q) as an active ingredient: (A) α-naphthoquinone (b) 2-hydroxy-1,4-naphthoquinone (c) 2,3-dichloro-1,4-naphthoquinone (d) 5-hydroxy-1,4-naphthoquinone (e) phylloquinone (vitamin K ₁₎ (f) menaquinone -n (vitamin _K 2) (g) menadione (vitamin _K 3) (h) 4- amino-2-methyl-1-naphthol (i) 1-naphthoic acid (j) 2-naphthoic Acid (k) 1,4-dihydroxy-2-naphthoic acid (l) 1-hydroxy-2-naphthoic acid (m) 1,4-naphthalenedicarboxylic acid (n) 1-naphthylacetic acid (o) α-naphthylamine (p ) Α-Naphthylaldehyde (q) β-naphthylaldehyde 2. The composition for promoting the growth of bifidobacteria according to claim 1.
2. The composition according to claim 1, which is a food or drink for promoting the growth of bifidobacteria, to which the composition according to claim 1 is added. 3. The composition according to claim 1, wherein the composition for promoting the growth of Bifidobacterium is an agent for promoting the growth of Bifidobacterium. 4. The composition according to claim 1, wherein the composition for promoting the growth of Bifidobacterium is a selective agent for Bifidobacterium. 5. A selective medium for analyzing bifidobacteria, comprising the composition according to claim 4 added thereto.