JP3261571B2 - Method for improving survival of bifidobacteria - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a method for improving the survival of bifidobacteria and a food and beverage containing the bifidobacteria, and is particularly suitable for fermented milk products.

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

[0003]

2. Description of the Related Art Bifidobacteria are not limited to infants,
Its effects on health, from adults to the elderly, are highly valued.

[0004] For example, with regard to the physiological significance, the intestinal p-type associated with the production of lactose-derived organic acids such as lactic acid, acetic acid, and formic acid.
Numerous studies have been clarified, such as an inhibitory action on pathogens due to a decrease in H and an intestinal action. As described above, since bifidobacteria are expected to have health effects, they are widely used in medicines and foods.

[0005] In particular, various products containing bifidobacteria are widely used in dairy products such as fermented milk (yogurt), and have formed a market. The used bacterial species to such as fermented milk, bi- to infants Bifidobacterium-blade base, is for adult from infant has been recommended Bifidobacterium longum ( "chemical and biological" vol.24 No. See paragraphs 18 and 9).

[0006]

However, bifidobacteria differ in mycological properties from dairy lactic acid bacteria conventionally used for the production of fermented milk and have the following problems.

[0007] The growth environment is an obligate anaerobic bacterium that grows slowly in the presence of oxygen.

[0008] The nutritional requirements are complex and strict and do not grow on a pure milk medium containing no growth promoting substances such as yeast extract.

Due to low acid resistance, low p like fermented milk
It is difficult to survive for a long time in the H region.

[0010] For this reason, the sharp decline in the number of viable bacteria of bifidobacteria in fermented dairy products in is recognized, it becomes difficult to achieve the original purpose of trying to use alive bifidobacteria.

From such a viewpoint, many studies have been made on methods for improving the survival of bifidobacteria in fermented milk products, and various proposals have been made.

For example, a method for improving the survival of bifidobacteria using sucrose (sucrose) or sorbitol (D-glucitol) (Japanese Patent Publication No. 57-4291) and a method for improving the survival of bifidobacteria using erythritol (Japanese Patent No. 2577692) No.).

However, sucrose is difficult to respond to the recent demand for low cariogenicity (hard caries) and low calories, and sorbitol is hard to meet the demand for low calories although it has hard caries. Although erythritol can meet the above-mentioned requirements, erythritol has a much less sweet aftertaste than sugar, making it difficult to obtain a sweetness similar to that of sugar. In other words, foods and drinks that require a sweetness similar to sugar have to be used in combination with a sweetener having sweetness characteristics similar to sugar.

In view of the above, the present invention can improve the survival of bifidobacteria which is not inferior to the prior art, can meet the demand for low calorie and low caries, and has the same effect as sugar. An object of the present invention is to provide a method for improving the survival of bifidobacteria, which can easily obtain a sweet taste.

[0015]

SUMMARY OF THE INVENTION In order to improve the survival of bifidobacteria in foods and drinks, the method for improving the survival of bifidobacteria of the present invention comprises the steps of: The problem is solved by a configuration added to the material.

Further, the food and drink containing bifidobacteria of the present invention solves the above-mentioned problems by providing a food and drink containing bifidobacteria together with a survival improver, wherein the survival improver comprises lactitol as an essential component. Is what you do.

Further, the method for producing a bifidobacterium-containing fermented milk product of the present invention is a method for producing a fermented milk product containing bifidobacteria, which is obtained by culturing the bifidobacterium under aerobic conditions as it is or as appropriate. Product 1 when manufacturing
The object is achieved by a configuration in which lactitol is added to and mixed with a culture medium or a culture for main culture so as to have a concentration of 0.01 to 2 mol per L.

[0018]

Description of the Structure of the Invention The structure of the present invention will be described in detail below. In the following description, “%” and the ratio are by weight unless otherwise specified.

(1) The medium of the fermented milk product of the present invention includes:
Usually, whole milk used in the production of fermented milk, skimmed milk, or reduced milk from these milk powders and the like containing a growth promoting substance or the like as appropriate, but a semi-synthetic or synthetic medium containing no milk Can also be used. In addition, all of the medium can be used at a milk solids concentration of about 8 to 20%, and the obtained culture may be used as it is as a food containing bifidobacteria, and may be used as a sweetener, fruit juice, water, or the like. , A fragrance and the like may be added as appropriate, and the same treatment as that for fermented milk products of dairy lactic acid bacteria may be performed to obtain a beverage.

The above-mentioned "synthetic medium" refers to a medium formed essentially by a combination of various types of nutrients and growth factors, such as an MRS medium and a Briggsliver broth medium.

The above culture is usually performed under aerobic conditions. The aerobic condition refers to a culture condition in a dairy lactic acid bacterium beverage production condition.

(2) The bifidobacteria used for producing the fermented milk product of the present invention is a known strain.
Bifidobacterium breve ATCC 1570
0, Bifidobacterium longum ATCC 1570
7 and Bifidobacterium breve SBR3212
(FARM P-11915).

(3) Lactitol is a sugar alcohol in which the glucose group of lactose has been reduced to a sorbitol group, and is produced by adding (reducing) hydrogen to lactose. The method of adding hydrogen has been applied to hardening of fats and oils for a long time, and can be performed by a conventional method well-known as a production method such as margarine or shortening.

Lactitol has a low sweetness (30 to 40% of sugar), but has no habit of sweetness similar to sugar (sucrose: sucrose). In Europe, it is widely used for sugarless confectionery. ing. In addition, it is easily dissolved in water, easily crystallized like sugar, is crystalline powder, has no hygroscopicity, and can be treated in the same manner as sugar in production. Also, like other sugar alcohols, it has the following features (confectionery synthesis technology center, “Appropriate utilization technology series of new materials for confectionery N
o. 16 ", March 1989).

It is hardly decomposed by digestive enzymes and is hardly absorbed in the small intestine (the energy value is 2 kcal / g and half that of lactose 4 kcal / g).

The blood sugar level hardly rises.

It is hardly carious.

Further, lactitol, relatively well-utilized in Bifidobacterium and Lactobacillus is a lactate-producing bacteria, hard to be utilized bacter oi des or C l ostr i dium.
Therefore, an intestinal bacterial layer improving effect equivalent to that of lactose has been reported (“Bifidus Vol. 9, No. 1, p. 19-26”).
1995).

[0029] However, lactitol is not intended to be a nutrient source for all bifidobacteria, but are assimilated with respect to a test strain Bifidobacterium blade base, capital for Bifidobacterium longum Is not converted. For this reason, it is not clear what mechanism is used by lactitol, but it exhibits some protective effect only by coexisting with bifidobacteria.

Therefore, lactitol may be present in foods and drinks containing bifidobacteria. Therefore, lactitol can be added at any time before or after culturing, but after culturing, it is desirable to add lactitol as early as possible in order to reliably obtain the effects of the present invention.

The amount of lactitol added in the present invention varies depending on the type of food or drink, that is, the acidity and sweetness. For example, in the case of a fermented milk product, the amount is 0.01 to 2 mol, preferably 0.015 to 1.7 mol, more preferably 0.015 to 0.8 mol per liter of the product. If the amount is less than 0.01 mol, it is difficult to obtain a survival improving effect, and if it exceeds 2 mol, the survival improving effect is reduced.

[0032]

Action / Effect of the Invention The method for improving the survival of bifidobacteria of the present invention has the following action / effect by adding lactitol to a medium or a culture.

The viable cell count in the food and drink (fermented dairy product) obtained by using them, as shown in the Examples below, shows a small decrease in the viable cell count even after storage for 10 days.

That is, the number of surviving bacteria is stable to the extent that it can be sufficiently commercialized, the original purpose of utilizing bifidobacteria can be achieved, and the sweetness similar to sugar is obtained. The feeling can be easily obtained.

[0035]

[Test Example & applications The following describes the present invention by the test Rei及 beauty applications went to confirm the viability improving effects of bifidobacteria lactitol used in the present invention.

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 performed by the steel wool method at 7 ° C. for 72 hours, and the number of colonies that appeared was counted, and the value per 1 mL of the sample was shown. The “acidity” is represented by the number of mL of 0.1 N sodium hydroxide solution required to neutralize 9 g of the sample.
The acidity per 1 g of the sample was indicated by% lactic acid.

<Test Example 1> 17 containing 0.2% yeast extract
200 mL of% reduced skim milk medium was dispensed into a 300 mL Erlenmeyer flask, covered with a cotton plug, and then sterilized at 95 ° C. for 30 minutes. Thereafter, the mixture was cooled to 37 ° C., and a 2% starter of Bifidobacterium breve ATCC15700, SBR3212, and Bifidobacterium longum ATCC15707 was alone inoculated, and each was allowed to stand at 37 ° C. for 18 hours. The obtained culture was mixed with a sugar solution containing various sugar concentrations and a culture solution at a ratio of 1: 1 and stored at 5 ° C. for 7 days.
The number of viable bacteria was measured.

The results are shown in Table 1 and FIGS. The sugars used in each experiment were sugar-free, lactitol, glucose, and sucrose. In addition, immediately after mixing,
The pH was 4.4 and the viable cell count was 4.7 × 10 ⁹ / mL for Bifidobacterium breve ATCC15700, 4.5 × 10 ⁹ / mL for SBR3212, and 3 for Bifidobacterium longum ATCC15707. It was 7 × 10 ⁹ / mL.

As shown in Table 1 and FIG. 1, in Bifidobacterium breve ATCC15700, lactitol was added to
At any of the 0.02-1.5 molar sugar addition concentrations,
The number of surviving bacteria tended to be higher than other sugars. For example, the number of surviving bacteria in each sweetened culture at 0.6 mole addition is
Lactitol 2.1 × 10 ⁶ / mL, glucose 1.4E × 10 ³ / mL, sucrose 6.6 × 10 ⁴
/ ML.

In addition, even in the case of SBR3212 and Bifidobacterium longum ATCC15707, the number of surviving bacteria tended to be clearly higher than that of other sugars at any of the concentrations of 0.02 to 1.5 molar sugar. there were. For example, the number of surviving bacteria in each sweetened culture at the addition of 0.6 mol is as follows: Bifidobacterium breve SBR3212 has 3.8 × 1 lactitol.
0 ⁸ / mL, glucose was 1.33 × 10 ⁶ / mL, and sucrose was 1.68 × 10 ⁸ / mL. In addition, Bifidobacterium longum ATCC15707 contains lactitol at 1.50 × 10 ⁸ / mL and glucose at 9.1 × 10 ⁸ / mL.
It was 10 ⁶ / mL and sucrose was 7.8 × 10 ⁷ / mL.

[0041]

[Table 1]

<Test Example 2> Bifidobacterium breve ATCC15700 and SBR3212 and Bifidobacterium longum ATCC15707 were added to a liquid culture medium for enrichment at 37 ° C. × 2.
After culturing for 0 hour, the cells were collected by centrifugation and washed several times to prepare a cell suspension (OD ₆₆₀ = 1.2). Separately, each of the pH ranges contains 0.6 moles of sugar.
A 1 M acetate buffer was prepared, 9 mL of the buffer was mixed with 1 mL of the cell suspension, and the mixture was stored at 5 ° C. On the third day of storage,
On days 7 and 10, the number of surviving bacteria was measured.

As is clear from Tables 2 and 3 showing the results,
Lactitol had the highest number of surviving bacteria at each pH for all bifidobacteria. In particular, pH4.
5 is remarkable, and the results are shown in FIGS. In addition,
The sugars used in the experiments in each figure are sugar-free, sucrose, and lactitol.

As is apparent from FIGS. 4 to 6, Bifidobacterium breve ATCC 15700 and SBR321 during storage.
2. The number of surviving bacteria of Bifidobacterium longum ATCC15707 is rapidly reduced from the third day of storage when no sugar is added.
From the day, the decrease in the number of surviving bacteria is more gradual than in the case without sugar. In the case of adding lactitol, the number of surviving bacteria was further improved as compared with the case of adding sucrose.

[0045]

[Table 2]

[0046]

[Table 3]

<Application Example 1> Bifidobacterium longum ATCC15707 was inoculated at a concentration of 3% into 400 mL of a 12% reduced skim milk medium containing 0.5% of yeast extract, and cultured at 37 ° C. for 18 hours. Prepared. This starter was immediately inoculated into 10 L of a 17% reduced skim milk medium containing 0.5% of yeast extract, and cultured at 37 ° C. for 18 hours. For each of lactitol, fructose and glucose, a sugar solution and a culture solution containing at each sugar concentration shown in Table 4 were mixed at a ratio of 1: 1 and homogenized with a homogenizer to prepare a fermented milk beverage 10 L, Store at 5 ° C for 7 days,
The number of surviving bacteria was measured. As is apparent from Table 4 showing the results, the addition of lactitol improved the preservation of the surviving bacteria in the preserved product.

[0048]

[Table 4]

<Application Example 2> 17 containing 0.2% yeast extract
100 L of 100% reduced skim milk was sterilized at 95 ° C. for 30 minutes. Then, the mixture was cooled to 37 ° C, and 3% of a starter of Bifidobacterium breve SBR3212 was inoculated.
For 18 hours. Lactitol was mixed with a sugar solution containing 0.2 mol of sugar in the final product in a 1: 1 ratio of a culture solution to prepare 150 L of a fermented milk beverage. Immediately after preparation of the product, it contained 4.5 × 10 ⁹ / mL of bifidobacteria, pH was 4.7, and lactic acidity was 0.7%. After storage at 5 ° C. for 7 days, the number of surviving bacteria was 1.2 × 1
Was 0 ⁸ / mL.

[Brief description of the drawings]

FIG. 1 is a graph showing the relationship between the 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.

[Figure 2] Bifidobacterium breve ATCC1
FIG. 4 is a graph showing a similar relationship in 5700 cultures.

[Fig. 3] Bifidobacterium longum ATCC1
FIG. 7 is a graph showing a similar relationship in the 5707 culture.

FIG. 4 is a graph showing the relationship between the number of surviving bacteria of Bifidobacterium breve ATCC15700 and the number of storage days in a 0.1 M acetate buffer at pH 4.5 containing each sugar.

Fig. 5 Bifidobacterium breve SBR32
FIG. 13 is a graph showing a similar relationship in FIG.

FIG. 6: Bifidobacterium longum
FIG. 27 is a graph showing a similar relationship in ATCC15707.

──────────────────────────────────────────────────続 き Continuation of front page (72) Inventor Katsunobu Yamaguchi 3-45, Ogihigashi, Komaki-shi, Aichi Pref. Yukizai Raleigh Co., Ltd. Development Laboratory (56) References Bifidus, Vol. 9, No. 1, p. 19-26 (1995) Clinical Nutrition, Vol. 76, No. 5, p. 457 (1990) (58) Fields investigated (Int. Cl. ⁷ , DB name) A23C 9/12-9/13 C12N 1/20-1/38 BIOSIS (DIALOG) JICST file (JOIS) JAFIC file (JOIS)

Claims (4)

(57) [Claims] 1. A method for improving the survival of bifidobacteria, which comprises adding lactitol to a culture or culture of bifidobacteria in order to improve the survival of bifidobacteria in foods and drinks. 2. A food or drink containing bifidobacteria together with a survival improver, wherein the survival improver comprises lactitol as an essential component. 3. The food or drink is a fermented milk product, and the product 1
3. The food and drink containing bifidobacteria according to claim 2, which contains 0.01 to 2 mol of lactitol per L. 4. to leave or appropriate processing of that the culture obtained by culturing the Bifidobacterium under aerobic conditions, when producing a fermented milk product containing Bifidobacterium, 0.01 per product 1L A method for producing a fermented milk product containing bifidobacteria, wherein lactitol is added to and mixed with a medium or a culture for main culture so as to have a 2 molar concentration.