JP2577692B2 - 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 viability of bifidobacteria and a food or beverage containing bifidobacteria, and is particularly 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 grow in the human large intestine,
It is a bacterium that exerts physiologically beneficial effects such as pathogen suppression and intestinal regulation, and plays an important role in human health. In particular, fermented milk products (such as lactic acid bacteria drinks) 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 the production of fermented milk, and (1) obligate anaerobic bacteria that cannot grow in the presence of oxygen as a growth environment. (2) nutrient requirements are complex and strict, do not grow in pure milk medium containing no growth promoting substances such as yeast extract, (3) because of low acid resistance,
It is difficult to survive for a long period of time in a low pH region such as fermented milk, and the like, and the viable count of bifidobacteria in fermented milk is rapidly reduced.

[0005] For this reason, it has been proposed to add, for example, 0.2 to 1.0 mol per liter of fermented dairy product to a medium or a culture of the main culture as an agent for improving the survival of bifidobacteria. . (See Japanese Patent Publication No. 57-4291)

[0006]

Recently, from the viewpoint of preventing obesity, fermented milk products containing bifidobacteria have, as with other foods and drinks, a certain level of sweetness and the lowest possible calories. Is increasing.

[0007] However, in the case of the method for improving the survival of bifidobacteria, there is almost no problem in improving the survival, but it is becoming difficult to respond to the increase in the required characteristics.

In view of the above, the present invention relates to a method for improving the survival of bifidobacteria, which is satisfactorily capable of responding to an increase in the required characteristics of a food and drink containing bifidobacteria having a certain sweetness and being as low in calories as possible. The purpose is to provide.

[0009]

Means for Solving the Problems In order to solve the above-mentioned problems, the inventors of the present invention have made intensive efforts to develop a non-caloric sweetener having a higher sweetness than sorbitol (sorbitol is a low-calorie sweetener). It has been found that erythritol, which is classified as a sweetener, has approximately the same viability-improving effect as sorbitol on bifidobacteria, and has completed the present invention having the following constitution.

[0010] A method for improving the survival of bifidobacteria, which comprises adding erythritol to the culture or culture of bifidobacteria in order to improve the survival of bifidobacteria in foods and drinks.

[0011]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The configuration of the present invention will be described below in detail.

(1) The bifidobacteria suitable for the present invention include, specifically, known Bifidobacterium strains such as Bifidobacterium breve ATCC 15700, Bifidobacterium longum ATCC 15707 and Separated Bifidobacterium breve SBR3212 (Microbial Bacteria No. 11915) can be mentioned. Bifidobacterium breve is recommended for infants and infants, and Bifidobacterium longum is recommended for infants and adults.

(2) As erythritol (tetravalent sugar alcohol) suitable for the present invention, meso form, L form, D form, L form
Any of the racemic form, which is an equivalent mixture of the isomer and the D-form, may be used, but the meso-form is preferred because of its easy synthesis and availability.

[0014] Erythritol is not included in natural foods such as lichens, mushrooms and fruits, and is not an artificial sweetener because its material is a natural product. In foods, fermented foods such as soy sauce, soy sauce, wine, and sake are particularly contained. The production method of erythritol can be divided into a chemical synthesis method and a fermentation method. In many cases, glucose is produced by fermentation using yeast as a raw material.
(See “Food Industry: Volume 4,” p. 73, published by Korinsha, 1989.) The amount of erythritol added is, for example, 1 fermented milk product.
Based on L, it is usually 0.05 to 2.0 mol, preferably 0.15 to 1.5 mol. If the amount is less than 0.05 mol, the effect of improving the survival of bifidobacteria is hardly expected, and if it exceeds 1.5 mol, the effect of improving the survival of bifidobacteria reaches a substantially saturated value.

(3) As a medium for culturing bifidobacteria, any of the following “milk medium” and “synthetic medium” may be used.
When producing fermented milk products, a "milk medium" is usually used.

"Milk medium": animal milk, vegetable milk, skim milk concentrated milk or reduced milk from powdered milk or concentrated milk generally used for the production of fermented milk, or these milks as appropriate.
Those containing growth promoting substances.

"Synthetic medium": MRS medium, Brix
Essentially a combination of various types of nutrients and growth factors, such as Briggs liverbroth medium.

Erythritol may be added to the above-mentioned medium or culture at any time before or after culturing, but is usually added before culturing.

All cultures having a milk solids concentration of about 8 to 20% can be used, and the resulting culture may be used as it is as a food containing bifidobacteria.
In addition, a sweetener, fruit juice, water, a flavor, and the like may be appropriately added, and the same treatment as that for the fermented milk product of dairy lactic acid bacteria may be performed to obtain a beverage.

Erythritol is not a nutrient source of Bifidobacterium and its mechanism is not clear,
Just by coexisting with bifidobacteria, it exerts some protective action.

[0021]

According to the present invention, in producing a food and drink containing bifidobacteria, erythritol is added to a culture or culture of bifidobacteria in order to improve the survival of bifidobacteria in the food and drink. As a result, the following operations and effects can be obtained.

In order to obtain the same level of effect of improving the survival of bifidobacteria in foods and drinks containing bifidobacteria, it has a sweetness of a certain level or more and has as low a calorie as possible, as compared with a conventional example in which sorbitol is added. It is easy to respond to the increase in required characteristics.

[0023]

Test Examples Hereinafter, test examples performed to confirm the effect of erythritol used in the present invention to improve the survival of bifidobacteria will be described.

[0024] The "viable cell count" in each case 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.

<Test Example 1> 17% containing 0.2% yeast extract
200 ml of the reduced skim milk medium was dispensed into a 300 ml Erlenmeyer flask, covered with a cotton plug, and sterilized at 95 ° C. for 30 minutes. Thereafter, the mixture was cooled to 37 ° C., and 2% of a starter of Bifidobacterium breve ATCC 15700, SBR3212, or Bifidobacterium longum ATCC 15707 was inoculated singly, and each was allowed to stand still at 37 ° C. for 18 hours.

The obtained culture was mixed with a sugar solution containing various sugars at various sugar concentrations and the culture solution at a ratio of 1: 1.
And stored for 7 days, and the viable cell count was measured. The results are shown in Table 1 and in FIGS.

The sugars used in the experiments shown in each figure are sugar-free, erythritol, glucose, and sucrose. The pH of each sweetened culture immediately after mixing was 4.4, and the number of surviving bacteria was 4.7 × 10 ⁹ / ml for Bifidobacterium breve ATCC 15700, 4.5 × 10 ⁹ / ml for SBR3212, 3.7 × 10 Bifidobacterium longum ATCC 15707
⁹ / ml.

Bifidobacterium breve ATCC 15
700, 0.05 to 1.
It can be seen from the figure that the number of surviving bacteria tends to be higher than that of other saccharides at any of the concentrations of 5-mol sugar. As one of them, the number of surviving bacteria in each sweetened culture at 0.6 mol was 5.0 × 10 ⁴ / ml for erythritol, 7.2 × 10 ³ / ml for glucose, and 2.1 for sucrose. × 10 ⁴ / ml
Met. Also, Bifidobacterium breve SB
R3212, bifidobacterium longum ATCC15707
However, at any erythritol concentration of 0.05 to 1.5 mol, there is a clear effect of improving the number of surviving bacteria as compared with other saccharides. Bifidobacterium breve SBR3212 has a cell count of 3.2 × 10 ⁶ / ml for erythritol, 2.0 × 10 ⁵ / ml for glucose, and 1.6 for sucrose.
× 10 ⁵ / ml, Bifidobacterium longum ATCC 157
07, erythritol 2.5 × 10 ⁸ / ml, glucose 5.2 × 10 ⁷ / ml, sucrose is 6.2 × 10 ⁷ /
ml.

<Test Example 2> Bifidobacterium breve ATCC 15700 and SBR3212, and Bifidobacterium longum ATCC 15707 were cultured in a liquid culture medium for enrichment at 37 ° C for 20 hours, and then collected by centrifugation. After washing several times, a cell suspension (OD ₆₆₀ = 1.2) was prepared. Separately, 0.1 M acetic acid buffer solutions of various pHs containing 0.6 M of sugar were prepared, and 9 mL of the buffer was mixed with 1 mL of the above cell suspension, followed by storage at 5 ° C. On the third, seventh and tenth days of storage, the number of surviving bacteria was measured.

From the results shown in Table 1, it can be seen that erythritol has the highest survival cell count at each pH. In particular, it is remarkable at pH 4.5, and the results are shown in FIGS. In addition, the sugar used in the experiment of each figure is sugar-free, sucrose, and erythritol.

As can be seen from the figure, Bifidobacterium breve ATCC 15700 and SBR3212 during storage.
The number of surviving bacteria of Bifidobacterium longum ATCC 1570 was sharply reduced from the third day of storage when no sugar was added, and the number of surviving bacteria was three days after storage when sucrose was added. The decline was slow. When the pH was low, it decreased significantly. In contrast, when erythritol was added, the number of surviving bacteria was improved as compared with other sugars.

Test Example 3 Bifidobacterium longum ATCC 15707 was inoculated 3% into 400 ml of a 12% reduced skim milk medium containing 0.5% yeast extract, and cultured at 37 ° C. for 18 hours to prepare a starter. . Immediately after this starter, 17% reduced skim milk medium 1 containing 0.5% of yeast extract was used.
0L was inoculated with 3% and cultured at 37 ° C for 18 hours. For each of erythritol, fructose, and glucose,
The sugar solution and culture solution contained at each sugar concentration shown in Table 2 were 1: 1.
, And homogenized with a homogenizer to obtain a fermented milk beverage 10
L was prepared, stored at 5 ° C. for 7 days, and the number of surviving bacteria was measured.

From the results shown in Table 2, it can be seen that the addition of erythritol improves the retention of the number of surviving bacteria in the stored product.

<Test Example 4> 17 containing 0.2% yeast extract
100 L of skim milk reduced at 95 ° C for 30 minutes,
Cooled to 37 ° C. Bifidobacterium breve
A 3% starter of SBR3212 was inoculated and cultured at 37 ° C. for 18 hours. Erythritol was mixed with a sugar solution containing 0.2 mol of sugar in the final product and a culture solution at a ratio of 1: 1 to prepare 150 L of a fermented milk beverage. 3. Immediately after product production.
It contained 5 × 10 ⁹ / ml bifidobacteria, pH 4.7, and lactic acidity 0.7%. After storage at 5 ° C. for 7 days, the number of surviving bacteria was 2.4 × 10 ⁷ / ml.

[0035]

[Table 1]

[0036]

[Table 2]

[Brief description of the drawings]

[Fig. 1] Bifidobacterium breve ATCC 15700
When various saccharides were added to the culture of
FIG. 3 is a graph showing the relationship with the number of surviving bacteria after storage on a day.

FIG. 2 is a graph showing the relationship between each sugar concentration and the number of surviving bacteria after storage for 7 days when various sugars are added to a culture of Bifidobacterium breve SBR3212.

FIG. 3 is a graph showing the relationship between the concentration of each saccharide and the number of surviving bacteria after storage for 7 days when various saccharides are added to a culture of Bifidobacterium longum ATCC 15707.

Fig. 4 Bifidobacterium breve ATCC 15700
FIG. 7 is a graph showing the number of surviving Bifidobacterium in a 0.1 molar acetate buffer when various sugars were added to the culture of Example 1.

FIG. 5 is a graph showing the number of surviving Bifidobacterium in a 0.1 M acetate buffer when various saccharides are added to a culture of Bifidobacterium breve SBR 3212.

FIG. 6: Bifidobacterium breve ATCC 15707
FIG. 7 is a graph showing the number of surviving Bifidobacterium in a 0.1 molar acetate buffer when various sugars were added to the culture of Example 1.

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Claims (3)

(57) [Claims] 1. A method for improving the survival of bifidobacteria, which comprises adding erythritol to a culture or culture of bifidobacteria in order to improve the survival of bifidobacteria in food and drink. 2. A food and drink containing bifidobacteria together with a survival improver, wherein the survival improver comprises erythritol as an essential component. 3. The food and drink containing bifidobacteria according to claim 2, wherein the food and drink are fermented milk products, and the concentration of the erythritol is 0.05 to 1.5 molar.