JP5980785B2 - New bifidobacteria and their use - Google Patents

Description

  The present invention relates to a novel Bifidobacterium bacterium that remains in the intestine and can grow in the intestine, and uses thereof.

  Bifidobacterium is a major bacterium in the intestinal flora of humans and has beneficial effects on human health such as bowel regulation such as constipation and diarrhea improvement, serum cholesterol elevation suppression, and immunostimulation. Many products are sold in the form of various fermented foods and beverages, viable bacteria preparations, and the like.

  Bifidobacterium is considered to exert the above-mentioned effects mainly in the state of viable bacteria, increasing the survival of Bifidobacterium in fermented foods and beverages and viable preparations, and increasing the number of living organisms. Various devices for ingesting bacteria have been made. However, it has also been pointed out that the above effect may not be fully exerted by simply passing the Bifidobacterium bacterium through the intestinal tract, and the Bifidobacterium bacterium remains in the intestine and proliferates in the intestine. It is considered important to do.

  In order to remain in the intestinal tract, it is necessary to have the ability to adhere to the intestinal tract. Lactobacillus GG (ATCC 53103), Lactobacillus paracasei KW3110, etc. (Patent Documents 1 and 2) have been reported as lactic acid bacteria as bacteria having the ability to adhere to the intestinal tract. Moreover, although there are few report examples, Bifidobacterium bifidum WB1003 (patent document 3) is reported as a Bifidobacterium genus bacterium.

Japanese National Patent Publication No. 11-507934 JP 2005-139160 A WO99 / 64023 international publication pamphlet

  However, the aforementioned lactic acid bacteria and Bifidobacterium bacteria adhere to the intestinal tract, but they cannot fully exert their effects unless they grow in the intestinal tract.

  Accordingly, an object of the present invention is to provide a bacterium belonging to the genus Bifidobacterium that can not only stay in the intestine but also grow in the intestine.

  Therefore, in order to solve the above-mentioned problems, the present inventors have used a number of Bifidobacterium bacteria as an index of having an ability to assimilate mucin, which is a main component of the intestinal mucosa, in addition to the ability to adhere to intestinal cells. As a result, the present inventors have found a novel Bifidobacterium genus that has an ability to adhere to intestinal cells and an ability to utilize mucin and can not only stay in the intestinal tract but also grow in the intestinal tract, thereby completing the present invention.

  That is, the present invention provides a Bifidobacterium bacterium characterized by having the ability to adhere to intestinal cells and the ability to assimilate mucin.

Moreover, this invention provides the food / beverage products characterized by containing the said Bifidobacterium genus bacteria, especially fermented milk food / beverage products.
Furthermore, the present invention relates to a Bifidobacterium genus that remains in the intestinal tract and is capable of growing in the intestinal tract, wherein the Bifidobacterium genus bacterium is selected using as an index the ability to adhere to intestinal cells and the ability to assimilate mucin. The screening method is provided.

  Since the Bifidobacterium of the present invention has the ability to adhere to intestinal cells and the ability to assimilate mucin, it can stay in the intestine and can grow in the intestine, and the Bifidobacterium in the intestine Can exert the physiological effect of the bacterium effectively and continuously. Moreover, it can utilize suitably for manufacture of the food / beverage products which have these physiological effects, especially fermented milk food / beverage products.

B. Adhesive ability to HT-29 cells. It is a figure which shows the adhesion image of bifidum YIT 11926 (In the figure, the place which has developed the yellow fluorescence shows YIT 11926 adhere | attached on HT-29 cell). B. Adhesive ability to HT-29 cells. It is a figure which shows the adhesion | attachment image of bifidum YIT 11939 (In the figure, the part which has developed the yellow fluorescence shows YIT 11939 adhere | attached on HT-29 cell). B. Does not have the ability to adhere to HT-29 cells. It is a figure which shows the adhesion image of bifidum Y11077 (In the figure, the location which has developed the yellow fluorescence shows Y11077 adhere | attached on HT-29 cell). B. Does not have the ability to adhere to HT-29 cells. It is a figure which shows the adhesion | attachment image of bifidum YIT 11082 (In the figure, the part which has developed the yellow fluorescence shows YIT 11082 adhere | attached on HT-29 cell).

  The Bifidobacterium of the present invention has the ability to adhere to intestinal cells and the ability to assimilate mucin, and can remain in the intestine by having the ability to adhere to intestinal cells and has the ability to assimilate mucin Can grow in the intestine.

  In the present invention, “having the ability to adhere to intestinal cells” means having the ability to adhere to intestinal cells to the extent that the target Bifidobacterium genus bacteria can remain in the intestinal tract. There are no restrictions on the type of adhesion or the like. Specifically, when a sufficient amount (50 or more per HT-29 cell) of Bifidobacterium is allowed to act on HT-29 cells, which are human colon cancer-derived cells, for 2 hours, HT -Refers to the attachment of one or more Bifidobacterium per 29 cells. It is preferable that 2 or more bacteria adhere to one HT-29 cell, more preferably 3 or more bacteria adhere, and further preferably 3 to 10 bacteria adhere. The larger the number of bacteria that adhere, the more efficiently it can stay in the intestinal tract, and therefore it can be suitably used as the Bifidobacterium bacterium of the present invention.

  In addition, “having mucin utilization ability” means turbidity (Klett value, Klett-Summerson colorimetry) when the target Bifidobacterium bacterium is cultured in a medium containing only mucin as a sugar source for 48 hours. Total, No. 66 filter) is 50 or more. The kret value is preferably 75 or more, more preferably 100 or more, and still more preferably 100 to 200. The higher the Klett value, the higher the ability to grow in the intestinal tract, so that it can be suitably used as the Bifidobacterium bacterium of the present invention.

  The culture medium used for the mucin utilization test may be a medium obtained by adding mucin to a medium excluding the sugar source, and the composition of the medium excluding the sugar source is Rogosa medium, M-ILS medium, PY. The composition can be a medium, a GAM medium, a TOS propionic acid medium, or the like. The mucin to be added is preferably human intestinal mucin, but can be substituted with porcine stomach mucin. The amount of mucin added is preferably 0.1 to 10% by mass, more preferably 0.5 to 5% by mass, and still more preferably 0.5 to 2% by mass in the medium. In addition, since there are rare bacteria that can grow even if there is no sugar source, culture the target Bifidobacterium bacterium in a medium excluding the sugar source as a control and confirm the cret value. Is desirable. If the kret value in the medium excluding the sugar source is 10 or more, the klet value in the medium excluding the sugar source is subtracted from the klet value in the medium using only mucin as the sugar source, and the above criteria are applied. Good.

The type of Bifidobacterium genus subject to the present invention is not particularly limited. For example, Bifidobacterium breve ( Bifidobacterium breve ), Bifidobacterium longum ( B. longum ), Bifidobacterium bifidum (B.bifidum), Bifidobacterium Animarisu (B.animalis), Bifidobacterium Zuisu (B.suis), Bifidobacterium infantis (B.infantis), Bifidobacterium address Sen Eustis (B.adolescentis), Bifidobacterium Katenuratamu (B.catenulatum), Bifidobacterium shoe Doka Tenu error Tam (B.pseudocatenulat m), Bifidobacterium lactis (B.lactis), Bifidobacterium Gurobosamu (B.globosum), Bifidobacterium angular Tam (B.angulatum), Bifidobacterium Dentiumu (B. dentium ) and the like. Among them, Bifidobacterium breve, Bifidobacterium longum, and Bifidobacterium bifidum are highly safe species that have been used in dairy products for a long time, and various physiological effects have been reported. In particular, Bifidobacterium bifidum is preferred.

  In this invention, as shown in the below-mentioned Example, the Bifidobacterium genus bacteria with 1 or more adhesion | attachment bacteria counts per HT-29 cell and a kret value of 50 or more were selected. In general Bifidobacterium, the number of adherent cells per HT-29 cell is 0.1 to 0.6 and the kret value is about 10 to 40. The Bifidobacterium of the present invention has the ability to adhere to intestinal cells and the ability to assimilate mucin, so that it remains in the intestine and can grow in the intestine, and the physiological effects of Bifidobacterium Can be effectively and continuously exhibited. The Bifidobacterium of the present invention grows even in a bile acid-containing medium, has excellent bile acid resistance, and can reach the intestine in an orally alive state.

  The new strains thus selected were designated as Bifidobacterium bifidum YIT 11926 (FERM BP-11504) and Bifidobacterium bifidum YIT 11939 (FERM BP-11505) on September 6, 2011. Deposited at the Patent Biological Depositary Center of the National Institute for Product Evaluation Technology (1st, 1st East, 1st Street, Tsukuba City, Ibaraki Prefecture, 6th). These strains have the following mycological properties.

(1) Bifidobacterium bifidum YIT 11926
The bacterium is isolated from healthy adults and has scientific properties of Gram-positive polymorphic gonococci, obligate anaerobes, and optimal growth temperature of 37 ° C. The 16S rDNA base sequence obtained from YIT 11926 was searched from a database to identify the bacterial species. The 16S rDNA base sequence of Bifidobacterium bifidum YIT 11926 showed 99.5% homology with Bifidobacterium bifidum DSM20456 (accession No. M38018) and was identified as Bifidobacterium bifidum.

(2) Bifidobacterium bifidum YIT 11939
The bacterium is isolated from healthy adults and has scientific properties of Gram-positive polymorphic gonococci, obligate anaerobes, and optimal growth temperature of 37 ° C. The 16S rDNA base sequence obtained from YIT 11939 was searched from a database to identify the bacterial species. The 16S rDNA base sequence of Bifidobacterium bifidum YIT 11939 showed 99.6% homology with Bifidobacterium bifidum DSM20456 (accession No. M38018) and was identified as Bifidobacterium bifidum.

  The utilization form of the bacterium belonging to the genus Bifidobacterium of the present invention is not particularly limited, and may be lyophilized, or may be used as a culture containing these bacteria. The bacterium is preferably in a live state.

  The Bifidobacterium of the present invention can be mixed with a solid or liquid non-toxic pharmaceutical carrier and used in the form of a conventional pharmaceutical preparation. Examples of such preparations include solid preparations such as tablets, granules, powders and capsules, liquid preparations such as solutions, suspensions and emulsions, and freeze-dried preparations. These preparations can be prepared by conventional means on the preparation. Examples of the non-toxic pharmaceutical carrier include glucose, lactose, sucrose, starch, mannitol, dextrin, fatty acid glyceride, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid ester, amino acid, gelatin, albumin , Water, physiological saline and the like. In addition, conventional additives such as stabilizers, wetting agents, emulsifiers, binders, isotonic agents, excipients and the like can be appropriately added as necessary.

  The Bifidobacterium genus bacterium of the present invention can be used not only as a preparation as described above but also in a food or drink. What is necessary is just to make it contain with a various nutrient component as it is, when mix | blending with food-drinks. Specifically, when the Bifidobacterium genus bacterium of the present invention is blended in a food or drink, an additive that can be used as a food or drink is appropriately used, and a form suitable for food using conventional means, that is, a granular form , Granulated, tablets, capsules, pastes, etc., added to various foods such as processed meats such as ham and sausage, fishery processed foods such as kamaboko and chikuwa, bread, confectionery, butter and milk powder Or may be used by adding to beverages such as water, fruit juice, milk, soft drinks and tea drinks. The food and drink includes animal feed.

  The Bifidobacterium genus bacterium of the present invention can effectively and continuously exert physiological effects such as the intestinal regulating action of the Bifidobacterium genus bacteria. In particular, because it stays in the intestinal tract and can grow in the intestinal tract, even if it does not continuously ingest Bifidobacterium spp., For example, when the intake interval is opened, such as every other month, every other week, every other day Since the physiological effect can be expected even if the period is short, the economic merit is also great.

  Furthermore, as food and drink, fermented milk food and drink, fermented soy milk, fermented fruit juice, fermented vegetable juice and the like containing the Bifidobacterium bacterium of the present invention in the state of viable bacteria are preferably used, particularly fermented. Use of dairy foods and drinks is preferred. Fermented milk foods and drinks may be produced in accordance with conventional methods. For example, when producing fermented milk, the sterilized milk medium is inoculated and cultured with the Bifidobacterium of the present invention alone or simultaneously with other microorganisms, This is homogenized to obtain a fermented milk base. Next, a separately prepared syrup solution may be added and mixed, homogenized with a homogenizer or the like, and then a flavor may be added to finish the final product. The fermented milk food and drink obtained in this way can be made into a product of any form such as a plain type, a soft type, a fruit flavor type, a solid form, and a liquid form that does not contain a syrup (sweetener).

  Such fermented milk foods and drinks can contain optional ingredients such as sweeteners such as syrup, emulsifiers, thickeners (stabilizers), and various vitamins. Syrup includes glucose, sucrose, fructose, fructose glucose liquid sugar, glucose fructose liquid sugar, palatinose, trehalose, lactose, xylose, maltose, honey, molasses and other sugars, sorbitol, xylitol, erythritol, lactitol, palatinit, reduced starch syrup High-sweetness sweeteners such as sugar alcohols such as reduced maltose starch syrup, aspartame, thaumatin, sucralose, acesulfame K, stevia. Fermented milk foods and drinks include sucrose fatty acid esters, glycerin fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, lecithin and other emulsifiers, agar, gelatin, carrageenan, guar gum, xanthan gum, pectin, locust bean gum, gellan gum, A thickening (stabilizing) agent such as carboxymethyl cellulose, soybean polysaccharide, propylene glycol alginate and the like may be blended. In addition, vitamins such as vitamin A, vitamin B, vitamin C and vitamin E, minerals such as calcium, magnesium, zinc, iron and manganese, citric acid, lactic acid, acetic acid, malic acid, tartaric acid, glucone Acid flavors such as acids, milk fat such as cream, butter, sour cream, yogurt, berry, orange, quince, perilla, citrus, apple, mint, grape, apricot, pair, custard cream Flavors such as peach, melon, banana, tropical, herbal, tea, and coffee, herb extract, brown sugar extract, and the like can also be blended.

For the production of fermented milk foods and drinks, microorganisms other than the Bifidobacterium of the present invention can be used in combination. Examples of such microorganisms include Bifidobacterium breve ( Bifidobacterium breve ), Bifidobacterium longum ( B. longum ), Bifidobacterium bifidum ( B. bifidum ), Bifidobacterium animalis ( B. animalis ), Bifidobacterium zuis ( B. suis ), Bifidobacterium infantis ( B. infantis ), Bifidobacterium adrecentis ( B. adolescentens ), Bifidobacterium catenuratam (B.catenulatum), Bifidobacterium shoe Doka Tenu error Tam (B.pseudocatenulatum), Bifidobacterium lactis (B.l CTIS), Bifidobacterium Gurobosamu (B.globosum), Bifidobacterium angular Tam (B.angulatum), bifidobacteria of the present invention, such as Bifidobacterium Dentiumu (B.dentium) Bifidobacteria other than bacteria, Lactobacillus casei , Lactobacillus acidophilus, L. plantarum, L. plantarum, L. buchneri L. gallinarum , L. amylovorus , Lactobacillus brevis ( L. brevis) ), Lactobacillus rhamnosus ( L. rhamnosus ), Lactobacillus kefir ( L. kefir), Lactobacillus paracasei ( L. paracasei), Lactobacillus cripatus ( L. cripatus ), Lactobacillus z. ), Lactobacillus helveticus ( L. helveticus), Lactobacillus salivarius ( L. salivarius ), Lactobacillus gasseri ( L. gasseri), Lactobacillus fermentum ( L. fermentum ), Lactobacillus reuteri ( L. fermentum ) reuteri ), Lactobacillus delbrukki Subspecies. Bulgaricus ( L. delbrueckii subsp. Bulgaricus ), Lactobacillus delbrukki subspecies. Deruburukkyi (L.delbrueckii subsp. Delbrueckii), Lactobacillus John Sony (L.johnsonii) Lactobacillus bacteria such as, Streptococcus thermophilus (Streptococcus thermophilus) Streptococcus bacteria such as, Lactococcus lactis subsp. Lactococcus lactis subsp. Lactis , Lactococcus lactis subspecies. Cremoris (Lactococcus lactis subsp. Cremoris) Lactococcus bacteria such as, Enterococcus faecalis (Enterococcus faecalis), Enterococcus faecium (E.faecium) Enterococcus bacteria such as Bacillus subtilis (Bacillus subtilis) bacteria belonging to the genus Bacillus such as, Saccharomyces cerevisiae ( Saccharomyces cerevisiae ), Torulaspora delbruecki ( Tollaspora delbrueckii ), Candida kefir ( Candida kefyr ), etc. When fermented milk foods and drinks are produced by using one or more selected from Lactobacillus bacteria, Streptococcus bacteria, and Lactococcus bacteria together with the Bifidobacterium of the present invention, high palatability is obtained and food and drink are easy This is preferable.

Although there is no strict limitation on the dosage when using the Bifidobacterium of the present invention, the suitable dosage is 10 ⁵ cfu to 10 ¹³ cfu per day as the number of viable bacteria, especially 10 ⁸ cfu. -10 ¹² cfu is preferred. Moreover, the physiological effect can be expected even when the administration interval is opened, such as every other month, every other week, every other day, or even in a short intake period, instead of continuous administration.

  By using the ability to adhere to intestinal cells and the ability to assimilate mucin as indicators, it is possible to screen Bifidobacterium bacteria that remain in the intestinal tract and can grow in the intestinal tract. That is, by selecting Bifidobacterium bacteria using the above-mentioned criteria for adhesion to intestinal cells and the criteria for mucin utilization, the ability to adhere to intestinal cells is selected. In addition, a Bifidobacterium genus having mucin utilization ability, staying in the intestinal tract, and capable of growing in the intestinal tract can be obtained.

  In addition, the above-mentioned criteria also apply to Bifidobacterium bacteria that have only the ability to adhere to intestinal cells and remain in the intestinal tract, and Bifidobacterium bacteria that have only mucin-assimilating ability and can grow in the intestinal tract. Can be used. In other words, if a strain having an ability to adhere to intestinal cells that exceeds the reference value and a mucin utilization ability that is less than the reference value is selected, a Bifidobacterium bacterium that has only the ability to adhere to intestinal cells and remains in the intestinal tract is obtained. Bifidobacterium genus that has only mucin-assimilating ability and can grow in the intestinal tract if a strain having a mucin-assimilating ability greater than the reference value and an ability to adhere to intestinal cells is less than the reference value is selected. Bacteria can be obtained.

  Furthermore, with respect to bacteria other than Bifidobacterium, it is possible to determine whether or not the bacteria have the ability to adhere to intestinal cells and the ability to assimilate mucin by using the above-mentioned criteria.

  Hereinafter, the content of the present invention will be described in more detail with reference to examples, but the present invention is not limited by these.

[Example 1] Selection of Bifidobacterium having an ability to adhere to intestinal cells and an ability to assimilate mucin (1) ability to assimilate mucin
B. adolescentis , B. et al . bifidum, B. breve , B.B. catenulatum, B. longum , B.B. pseudocateulatum , B.I. angulatum , B. et al . animalis, B. infantis, B. globosum, B. lactis , B. et al . dentium, L. Using the 13 species of test strains of rhamnosus.
Bifidobacteria were inoculated with 1% of a test bacterial solution cryopreserved in a GAM anaerobic liquid medium supplemented with 0.7% glucose and pre-cultured for 24 hours. Lactobacillus ( L. rhamnosus ) was inoculated with 1% of a test bacterial solution cryopreserved in MRS liquid medium and pre-cultured for 24 hours. Inoculate 1% of the pre-cultured culture medium (mucin addition amount is 1%) on the medium in which the lactose of M-ILS is replaced with mucin (Wako, manufactured by porcine stomach) (the initial bacterial count is 10 ⁷ / ml / medium) After 48 hours, the kret value was measured. Moreover, it culture | cultivated also for 48 hours also in the M-ILS culture medium which does not add carbohydrate, and measured the cret value.

(2) Adhesive ability to intestinal cells The same 13 strains of test strains as in (1) were used.
Human colon cancer-derived cells HT-29 were used after being cultured in RPMI-1640 medium supplemented with 10% FBS.
Bifidobacteria is a GAM anaerobic liquid medium supplemented with 0.7% glucose, and lactic acid bacteria ( L. rhamnosus ) are cultured for 24 hours in MRS liquid medium, washed twice with PBS, and then stained with DAPI in RPMI-1640 medium supplemented with 10% FBS. The suspension was suspended so that the number of bacteria by the method was 5 × 10 ⁸ / ml · medium.
After HT-29 was cultured on a glass slide with a chamber until it became confluent, 400 μl of a cell suspension suspended in RPMI-1640 supplemented with 10% FBS was added (the number of test strains per HT-29 cell was 300. Will be). After incubating at 37 ° C. under 5% CO ₂ for 2 hours, the plate was washed twice with RPMI-1640 medium supplemented with 10% FBS and once with PBS to remove non-adherent bacteria and added with 2% PFA at 4 ° C. Fixed overnight. After 96% ethanol treatment, Bifidobacterium-specific probe Bif153 labeled with the fluorescent dye Cy5 for Bifidobacteria (Intestinal Bacteriology Journal 18: 141-146, 2004), and for lactic acid bacteria with the fluorescent dye Cy5 Hybridization was performed using a labeled lactic acid bacteria-specific probe Lab158 (Microb. Ecol. Health Dis. 11: 3-12, 1999), and the mixture was sealed with a mounting medium to which a DAPI stain was added. The number of adherent bacteria of each test strain per HT-29 cell was quantified using image processing software.

(3) Results Tables 1 and 2 show the mucin utilization ability and adhesion ability of each test strain. FIGS. 1 and 2 show images of strains having the ability to adhere to HT-29 cells, and FIGS. 3 and 4 show images of strains having no ability to adhere.

Examination of mucin assimilation ability and adhesion ability to intestinal cells of 12 Bifidobacteria species, new selection that satisfies the selection criteria (the number of adherent bacteria per HT-29 cell is 1 or more and the kret value is 50 or more) 2 strains (YIT 11926, YIT 11939) were found.
In addition to the above two strains, there were 3 strains of bifidobacteria having the ability to assimilate mucin (Y11077, YIT11082, A-43). In addition to YIT 11926 and YIT 11939, YIT 10155, 31-2, YIT 4044, and YIT 4018 possessed the ability to adhere to intestinal cells, but these four strains all had low ability to assimilate mucin. As for the ability to assimilate mucin and the ability to adhere to intestinal cells, the variation among strains was very large even in the same species, and no particular tendency of the same species was observed.
L. Rhamnosus GG is known as a lactic acid bacterium having the ability to adhere to the intestinal tract (Patent Document 1), but this test system also showed the same ability to adhere to intestinal cells as YIT 11939.
On the other hand, regarding the mucin utilization ability of GG strain, since the Klett value in the medium without sugar was 76, it was judged that the GG strain could grow even without the sugar source. When the value (21) obtained by subtracting the above-mentioned value (76) from the klet value (97) was determined to have no ability to assimilate mucin.
In addition to the above, a number of bifidobacteria were used to examine the ability to assimilate mucin (total of 340 strains) and the ability to adhere to intestinal cells (total of 60 strains). Yet 11926, It was not obtained except YIT 11939.

  In addition, some bifidobacteria were also examined for bile acid resistance. Specifically, 30 μl of the bacterial solution cultured anaerobically to the stationary phase in the GAM liquid medium was inoculated into 3 ml of the GAM liquid medium supplemented with 0.2% bile acid and cultured anaerobically at 37 ° C. After the time, microbial turbidity (Kret value) was measured with a Klett-Summerson colorimeter (No. 66 filter). A high Klett value indicating growth in a bile acid-containing medium indicates a high survival rate and growth rate in the gastrointestinal tract, and a Bifidobacterium having a Klett value of 100 or more in the bile acid-containing medium. Can reach the intestines with a large number of bacteria alive.

Klett value of YIT 11926 is 125, YIT 11939 is 135, B. YIT 4039, which is a type strain of bifidum , hardly grew on the BAM-added GAM medium. General bifidobacteria proliferating in a bile acid-containing medium has a Klett value of about 0 to several tens. Therefore, it was revealed that YIT 11926 and YIT 11939 have high bile acid resistance. .

[Example 2] Manufacture of fermented milk foods and beverages 15% skimmed milk was sterilized by adding 3% glucose, inoculated with 1% Bifidobacterium bifidum YIT 11939, cultured at 37 ° C for 24 hours, and yogurt base 210 g was obtained. On the other hand, 97 g of sugar and 0.2 g of emulsified iron were dissolved in water and sterilized with a total amount of 790 g to obtain a syrup. The yogurt base and syrup obtained as described above were mixed, 1 g of flavor was added, and then homogenized at 15 MPa and filled in a container to obtain a fermented milk beverage. The obtained fermented milk beverage had good appearance and flavor and good storage stability.

[Example 3] Manufacture of fermented milk food and drink 124 g of whole milk powder was dissolved in 506 g of water and sterilized at 135 ° C. for 3 seconds, and then Bifidobacterium breve was 0.5%, Bifidobacterium bifidum YIT 11926 1% and Lactobacillus acidophilus 0.001% were inoculated, cultured at 37 ° C. to pH 5.3, and homogenized at 15 MPa to obtain 630 g of a bacterial solution. On the other hand, 98 g of palatinose, 8 g of carrot juice, 2.5 g of DHA-containing oil, 7 g of emulsified calcilm, 0.1 g of lactoferrin, 0.02 g of vitamin D and 1 g of fragrance were dissolved in water, and water was added to make the total amount 370 g. Sterilized at 3 ° C. for 3 seconds to obtain a syrup solution. Bacteria solution and syrup solution are mixed and filled in a tetra brick container, pH: 5.6, acidity: 3.4, non-fat milk solid content: 8.7%, milk fat content: 3.2% fermentation I got milk. The obtained fermented milk beverage had good appearance and flavor and good storage stability.

[Example 4] Manufacture of tablets Various ingredients were mixed according to the following formulation, granulated, dried and sized, and then tableted to produce tablets.
(Prescription) (mg)
Dried product of bacterial cells of the present invention ¹⁾ 20
Microcrystalline cellulose 100
Lactose 80
Magnesium stearate 0.5
Methylcellulose 12
1) A living cell of Bifidobacterium bifidum YIT 11926 was produced by lyophilization.

[Example 5] Manufacture of soft drink According to the following prescription, each component was blended according to a conventional method and homogenized to obtain a soft drink. The obtained soft drink was filled in a brown bottle, sealed with an aluminum cap, and heat-treated. The resulting soft drink had good appearance and flavor and good storage stability.
(Prescription) (g)
Dried product of bacterial cells of the present invention ¹⁾ 5
Fragrance 0.8
Water 100
Reduced starch saccharified product 24
Fructose 18
1) Bifidobacterium bifidum YIT 11939 was produced by freeze-drying.

Claims (4)

  Bifidobacterium bifidum YIT11926 (FERM BP-11504) or Bifidobacterium bifidum YIT11939 (FERM BP-11505).   A food or drink comprising the Bifidobacterium bifidham according to claim 1.   The food or drink according to claim 2, which is a fermented milk food.   Furthermore, the food / beverage products of Claim 2 or 3 which contain a sweetener.