JP2843963B2 - Method for improving survival of bifidobacteria - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for improving the survival of bifidobacteria and to a food and drink containing bifidobacteria. In particular,
It is an invention suitable for fermented milk products.

Here, a liquid fermented milk product will be mainly described as an example. However, the present invention is not limited to the fermented milk product, and the form of food and drink includes liquid, paste, solid, and the like. In addition,
Bifidobacterium refers to Bifidobacterium
erium) is a generic name for bacteria classified as:

[0003]

2. Description of the Related Art Bifidobacteria are bacteria that grow in the human intestine and exert physiologically advantageous effects such as pathogen control and intestinal regulation. They play an important role in human health. It is very useful to use it. In particular, fermented milk products containing milk as a main component of the medium and containing a culture of bifidobacteria are popular because of their high nutritional value.

[0004] However, bifidobacteria have different bacteriological properties from dairy lactic acid bacteria conventionally used for fermented milk production, and are obligate anaerobic bacteria that cannot grow in the presence of oxygen as a growth environment. The requirement is complicated and strict, it does not grow in pure milk medium containing no growth promoting substances such as yeast extract, and it has low acid resistance, so it is difficult to survive for a long time in a low pH region such as fermented milk,
However, a rapid decrease in the viable count of bifidobacteria in fermented milk is observed. For this reason, it is not expected that consumers will actually consume a large amount of bifidobacteria.

[0005] Therefore, as a method for improving the survival of bifidobacteria by generating carbon dioxide in the liquid of fermented milk product and increasing the anaerobicity in the product, a method of containing Streptococcus diacetylactis has been proposed. However, there is a possibility that diacetyl and acetoin which cause an unpleasant odor will be produced together with carbon dioxide when it becomes excessive, thereby impairing the flavor of the fermented milk product. For this reason, it has been proposed to suppress the production of diacetyl and acetoin in a container having a high gas barrier property. However, it is difficult to completely prevent the flavor from being impaired, and furthermore, the gas barrier property In such a case, the container must be filled in a state without a high space and a head space, so that the cost of the container increases and the manufacturing equipment needs to be improved.

[0006] Therefore, Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium spp., Which are a kind of intestinal bacteria in fermented milk and dairy products and used for the purpose of health, are used. Functions of Fermented Milk "
(1988), Food Materials Research Association, edited by Yuji Nakazawa and Akiyoshi Hosono: p1
Focusing on 93), the flavor is excellent among the bacterial species, fermented milk,
There has been proposed a method for improving the viability of bifidobacteria by including bacteria having excellent viability in bifidobacteria in dairy products in fermented milk.

There is a method for producing a fermented milk containing Bifidobacteria in which the survival rate of Bifidobacterium longum is increased by coexisting Lactobacillus casei and Bifidobacterium longum (Japanese Patent Application Laid-Open No. 63-209542). ).

Further, the present inventors have previously proposed a method for producing a culture having a high viable count of bifidobacteria by mixing and cultivating Bifidobacterium breve and Lactobacillus casei (Japanese Patent Laid-Open No. Hei 5 (1993) -105). 227946).

[0009]

However, all of them specify the species of bifidobacteria, and there are strains having a physiological effect that is useful for humans among human common resident bifidobacteria. Even can not be used.

[0010] Further, from the viewpoint of increasing the survival rate of bifidobacteria in the fermented milk of bifidobacteria, as a container for fermented milk,
Since it is necessary to use an aluminum-coated paper container, a multi-layer plastic container, a glass container and the like having a high gas barrier property, the cost is generally higher than that of a polystyrene container and the like used for fermented milk.

In addition, it is often necessary to improve the filling apparatus in order to fill these containers in such a way that the head space is minimized.

In view of the above, an object of the present invention is to provide a method capable of improving the survival of a general resident bifidobacterium species without using a container having a high gas barrier property.

[0013]

According to the present invention, fermented milk is prepared by adding malic acid or a salt containing malic acid, and a fruit juice, using a common resident bifidobacterium species and Lactobacillus casei. The present inventors have found that the survival of Bifidobacterium is significantly improved, and conceived a method for improving the survival of Bifidobacterium of the present invention.

That is, Lactobacillus casei has
Considering that the metabolic action of generating CO ₂ gas from malic acid is used to increase anaerobicity in product liquids and culture liquids, L-malic acid and DL-malic acid were used, and the results were as follows. The present inventors have found that there is an effect of improving the growth and survival of the bacterium (especially remarkable with L-malic acid), and completed the present invention having the following constitution.

[0015] A method for improving the survival of bifidobacteria in food and drink, characterized in that the food and drink contain Lactobacillus casei and malic acid.

[0016]

[Detailed description of means]

(1) The bifidobacteria suitable for the present invention include, specifically, known strains of bifidobacteria, such as Bifidobacterium longum ATCC 15707, Bifidobacterium
Adressentis ATCC 15703, Bifidobacterium breve ATCC 15700, and newly isolated Bifidobacterium breve SBR3212 (FERMP-11915). Bifidobacterium breve is recommended for infants and infants, and Bifidobacterium longum is recommended for infants and adults.

(2) Lactobacillus casei suitable for the present invention includes Lactobacillus casei AST-8 and IF0353.
3, the same IF012004, the same ATCC4352, and the like.

(3) As the malic acid, L-malic acid, D
L-malic acid, D-malic acid, salts thereof, and apple and grape juices containing malic acid can be used. In particular, L-malic acid has a large effect and is available. It is desirable because it is easy to do.

The amount of these malic acids added is, for example,
The amount is usually 0.0001-0.05 mol, preferably 0.0003-0.04 mol, per liter of fermented milk product. If the amount is less than 0.0001 mol, the effect of improving the survival of bifidobacteria is hardly expected. If the amount exceeds 0.05 mol, the effect of improving the survival of bifidobacteria reaches a saturation value, and further improvement cannot be expected.

The malic acid may be contained in the fermented milk at the time when the fermented milk of Bifidobacterium is filled in a container and moved to the distribution / consumption stage, and the timing of addition in the production process is arbitrary. That is, it may be during the manufacturing process or after the manufacturing process is completed.

(4) In the production of foods and beverages containing bifidobacteria, growth promoting substances, sugars, fruits and the like are appropriately added to whole milk, skim milk, or reduced milk from these milk powders which are generally used for the production of fermented milk. A medium containing the above is used as a medium, but a semi-synthetic or synthetic medium containing no milk can also be used. Using this medium, Bifidobacteria and Lactobacillus casei and, if necessary, dairy lactic acid bacteria are separately cultured. If possible, the respective bacteria may be mixed and cultured.

Next, the obtained culture may be edible as a food containing Bifidobacterium as it is, or a fermented dairy lactic acid lactic acid bacterium product may be added with sweeteners, fruit juice, water, flavors and the like as appropriate. The same processing as described above may be performed to obtain food and drink.

[0023]

The method for improving the survival of bifidobacteria according to the present invention is directed to a method for improving the viability of bifidobacteria in foods and beverages by including Lactobacillus casei and malic acid or a salt thereof in the foods and beverages. The persistence can be improved.

In the method of the present invention, a container having no oxygen permeability such as a glass bottle or a container having a low oxygen permeability is naturally effective, but the cost tends to be relatively high. The method for improving the viability of bifidobacteria of the present invention has an advantage that a container for ordinary use (polystyrene or the like) having a relatively high oxygen permeability can be used for the container and the cost can be reduced.

The fermented milk containing bifidobacteria produced by the method of the present invention uses polystyrene containers which are usually used for low-cost fermented milk, and can be cultivated with conventional human resident bifidobacteria species using conventional equipment. Bifidobacterium-containing fermented milk using Bacillus casei can be produced, and a high Bifidobacterium count can be maintained until consumers eat and drink.

Prior to the present invention, the inventors have determined that Lactobacillus casei contains malic acid with or without oxygen.
It was confirmed that O ₂ gas was produced (in particular, the action of L-malic acid was large).

In the flavor evaluation, it was confirmed that there was no change in the flavor regardless of the presence or absence of oxygen.

This metabolic mechanism, Lactobacillus casei, is an enzyme that decomposes L-malic acid.
Having J. LONDON, EYMEYER, SRKULCZYK (197
1): Journal of Bacteriology, Vol 1.108, 196-201, and this enzyme is known to produce CO ₂ gas from L-malic acid. (J.LONDON, EYMEYER, (1969):
"Journal of Bacteriology Vo1.98" 705-711p) the future, Lactobacillus casei is, L- malic acid and it can be seen that to produce a CO ₂ gas enzyme from the have L- malic acid decomposes.

Further, from these documents, in addition to Lactobacillus casei, Streptococcus faecalis also has an enzyme that decomposes L-malic acid. Therefore, the effect of the present invention can be expected in the same manner as Lactobacillus casei.

[0030]

EXAMPLES Hereinafter, the present invention will be described in detail with reference to examples and experimental examples, and the effects of the present invention will be confirmed. In the following description, a blending unit is a weight unit unless otherwise specified.

The "viable cell count" in each example is based on Mitsuoka's anaerobic diluent (Mitsuoka: Clinical Laboratory, Vol. 18, p. 1163,
1974), blood liver agar (Bl
oodLiver Ager, BL agar)
Anaerobic culture was carried out at 7 ° C for 72 hours by the steel wool method, and the number of colonies that appeared was counted, and the value per 1 ml of the sample was shown. The “acidity” was represented by% lactic acid per 1 g of the sample, based on the number of ml of 0.1N sodium hydroxide solution required to neutralize 9 g of the sample.

The Lactobacillus casei AST-8 used in the experimental examples was deposited with the Institute of Biotechnology, National Institute of Advanced Industrial Science and Technology as FERM P-12704.

<Experimental Example 1> (1) Bifidobacterium longum ATCC1 was added to a reduced skim milk medium containing 0.5% yeast extract and 16% non-fat milk solids.
3% of 5707 starter was inoculated and cultured at 37 ° C. for 24 hours to obtain a culture having a pH of 4.4. The resulting culture had a milk solid content of 8%, sucrose 8%, and a viable cell count of Lactobacillus casei ATCC 5432 of 2.3 × 10 ⁸ CF.
U / ml, DL-Na malate 0, 0.01, 0.1,
A 10 ° C. storage test was performed on the product liquid to which 0.5% was added.

(2) From Table 1 showing the results of the storage test,
Bifidobacterium in proportion to the concentration of malic acid
It was found that the survival of longum could be improved. In addition, it can be seen that the survival of the lactic acid bacteria is hardly related to the presence or absence of the addition of DL-malic acid.

<Experimental Example 2> (1) After Bifidobacterium adrecentis ATCC 15703 was cultured in Briggs River Broth, the culture was collected by a centrifuge to prepare a concentrated bacterial solution.
Also, 3% Lactobacillus casei IFO3533 starter was added to a 16% reduced skim milk medium supplemented with 0.2% yeast extract.
Inoculate and incubate at 37 ° C. for 40 hours.
A culture of 52%, pH 4.2, with a viable cell count of 3.1 × 10 ⁹ CFU / ml was obtained. Using the concentrated bacterial body fluid of Bifidobacterium adrecentis and a culture of Lactobacillus casei obtained here, the milk solids concentration is 4%, sucrose 10%,
A liquid to which 0, 0.05, and 0.3% of L-malic acid was added was prepared. Each prepared solution was subjected to a 10 ° C. storage test.
In addition, the lactic acidity of each preparation was adjusted with citric acid.

(2) From Table 2 showing the results of the storage test, it was found that the survival of Bifidobacterium longum could be improved in proportion to the concentration of L-malic acid added. In addition, it can be seen that the survival of the lactic acid bacteria hardly depends on the presence or absence of L-malic acid.

Example 1 (1) A medium consisting of 17.1 kg of skim milk powder, 0.2 kg of yeast extract and 82.7 L of water was sterilized at 95 ° C. for 30 minutes, and 3% of a Bifidobacterium breve ATCC 15700 starter was added. The cells were inoculated and cultured at 37 ° C. for 18 hours. After sterilizing a medium consisting of 17.1 kg of skim milk powder and 82.9 L of water at 95 ° C. for 30 minutes, Lactobacillus casei A
ST-8 starter was inoculated at 1% and cultured at 37 ° C for 40 hours. 15 L of Bifidobacterium breve culture obtained here, 33 L of Lactobacillus casei culture
And sucrose 8Kg, DL-malate 50g, water 43.9
After mixing 5 L of sterilizing solution, homogenization was performed to obtain 90 L of fermented milk. This was filled in a 160 ml polystyrene container, and an aluminum seal was attached to obtain about 540 fermented milks.
Stored at 10 ° C.

(2) The fermented milk of this example had a lactic acidity of 0.76% immediately after production, a pH of 4.3, a Bifidobacterium breve count of 5.6 × 10 ⁸ CFU / ml, and a Lactobacillus casei. The bacterial count was 7.2 × 10 ⁸ CFU / ml.

On the other hand, after storage at 10 ° C. for 10 days,
The lactic acid acidity was 0.82%, pH 4.2, the number of Bifidobacterium breve bacteria was 9.3 × 10 ⁶ CFU / ml, and the number of Lactobacillus casei bacteria was 5.8 × 10 ⁸ CFU / ml.

<Example 2> A medium consisting of 17.1 kg of skim milk powder, 0.2 kg of yeast extract and 82.7 L of water was used in 95
After sterilization at 30 ° C. for 30 minutes, a 3% starter of Bifidobacterium longum ATCC 15707 was inoculated and cultured at 37 ° C. for 18 hours. In addition, 17.1 kg of skim milk powder, 8 water
After 2.9 L of the medium was sterilized at 95 ° C. for 30 minutes, 1% Lactobacillus casei AST-8 starter was inoculated and cultured at 37 ° C. for 40 hours. 10 L of the obtained Bifidobacterium longum culture, 15 L of Lactobacillus casei culture, 7 kg of sucrose, 5 apple juices
L, pectin 0.4 kg, fragrance 0.2 kg, water 62.4
After mixing the L germicidal solution, the mixture was homogenized to obtain 91 L of a dairy lactic acid bacteria beverage. This was filled in a 100 ml polystyrene container, and an aluminum seal was attached thereto to obtain about 880 dairy lactic acid bacteria drinks, which were stored at 10 ° C.

Immediately after production, this lactic acid lactic acid bacteria beverage had a lactic acidity of 0.54%, pH 4.2,
The number of Longum bacteria was 4.3 × 10 ⁸ CFU / ml, and the number of Lactobacillus casei bacteria was 3.7 × 10 ⁹ CFU / ml.

On the other hand, after storage at 10 ° C. for 10 days,
Lactic acidity was 0.67%, pH 4.1, the number of Bifidobacterium longum bacteria was 2.1 × 10 ⁷ CFU / ml, and the number of Lactobacillus casei was 1.9 × 10 ⁹ CFU / ml.

[0043]

[Table 1]

[0044]

[Table 2]

──────────────────────────────────────────────────続 き Continued on the front page (72) Katsunobu Yamaguchi 133-2 Fujishima-cho, Fujishima-cho, Komaki City, Aichi Prefecture (56) References JP-A-58-1111638 (JP, A) JP-A-60-164432 (JP, A JP-A-63-309138 (JP, A) JP-A-2-308754 (JP, A) JP-A-5-227946 (JP, A) JP-A-63-209542 (JP, A) (58) Field (Int.Cl. ⁶ , DB name) A23C 1/00-23/00

Claims (9)

(57) [Claims] 1. A method for improving the survival of bifidobacteria in foods and drinks, wherein said foods and drinks contain Lactobacillus casei and malic acid or a salt thereof. How to improve persistence. 2. The method according to claim 1, wherein the malic acid is L-
A method for improving the viability of bifidobacteria, which is malic acid or a salt thereof. 3. The method according to claim 1, wherein the malic acid is DL.
-A method for improving the survival of bifidobacteria, which is malic acid or a salt thereof. 4. The method for improving the survival of bifidobacteria according to claim 1, wherein malic acid or a salt thereof is contained at the end of the production process. 5. The method according to claim 1, wherein the number of Lactobacillus casei in food and drink is 1 × 10 ⁷ CFU / ml or more. 6. A food and drink containing bifidobacteria, which comprises Lactobacillus casei and malic acid or a salt thereof. 7. The method according to claim 6, wherein the malic acid is L-
A method for improving the viability of bifidobacteria, which is malic acid or a salt thereof. 8. The method according to claim 6, wherein the malic acid is DL.
-A method for improving the survival of bifidobacteria, which is malic acid or a salt thereof. 9. The food and drink containing bifidobacteria according to claim 6, wherein the number of bacteria of Lactobacillus casei in the food and drink is 1 × 10 ⁷ CFU / ml or more.