JPS6152253A - Method of stabilizing bifidobacterium longum atcc 15707 in food - Google Patents

Abstract

PURPOSE:To prevent reduction in number of molds of Bifidobacterium longum ATCC 15707 during preservation, and to stabilize the quality of the food, by adding a cell-free extracted solution of hygienically innoxious catalase-positive microorganism to Bifidobacterium longum ATCC 15707-containing liquid or pasty food such as fermented milk, etc. CONSTITUTION:A hygienically innoxious catalase-positive miroorganism [preferably bread yeast (Saccharomyces cerevisiae), etc.] is treated with ultrasonic wave, 0.1-5wt% of the prepared cell-free extracted solution is added to Bifidobacterium longum ATCC 15707-containing liquid or pasty food, and H2O2 formed by the mold is converted into H2O to stabilize the food.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for stabilizing bifidobacteria added to foods.

More specifically, the present invention relates to a method of adding a cell-free extract of a catalase-positive microorganism as a catalase source to quickly convert H2O2 produced by bifidobacteria in foods to H2O, thereby preventing the death of bifidobacteria.

In general, improving the number of bacteria in the human intestines to one dominated by bifidobacteria is thought to be useful for the prevention and treatment of various diseases. Many of them are now commercially available.

However, Bifidobacterium is an anaerobic bacterium that easily dies in the presence of oxygen, and in acidic foods such as fermented milk and lactic acid bacteria drinks, the number of bacteria rapidly decreases due to acid. Therefore, simply adding a bifidobacterium culture to liquid or pasty foods such as milk drinks, fermented milk, and lactic acid bacteria drinks does not produce high-quality products in which the bifidobacteria do not die during storage. For these products, which require the presence of a large amount of live bifidobacteria for health purposes, it is necessary to prevent the number of bifidobacteria from decreasing due to oxygen and acid during storage to stabilize the quality. .

However, as a result of intensive research by the inventors on the behavior of bifidobacteria in liquid or pasty foods containing bifidobacteria, such as milk drinks, fermented milk, and lactic acid bacteria drinks, they found that NADH, -oxidase possessed by bifidobacteria. It was confirmed that the bifidobacterium itself generates hydrogen peroxide using hydrogen peroxide-producing enzymes such as Bifidobacteria in food, and that the hydrogen peroxide accelerates the death of Bifidobacterium itself in foods.

So, Bifidobacterium lon
Figure 1 shows the results of measuring the amount of H2O2 produced when a Bifidobacterium culture solution prepared by neutralizing gom ATCC15707 under anaerobic conditions was stored in the air.

Furthermore, Table 2 shows the results of measuring the bacterial survival rate when catalase was added to the bifidobacteria culture solution.

Table 2: Viability of seedlings when catalase is added to bifidobacteria culture solution Catalase: Commercially available product used: 700 units added per bacterial solution As a result of intensive research in search of a suitable source of catalase, we collected cell-free extracts of catalase-producing microorganisms that are hygienically harmless, and added this to foods together with bifidobacteria culture solution to stabilize bifidobacteria. The inventors have discovered that this can be done, and have completed the present invention.

The present invention involves adding a cell-free extract of catalase-positive microorganisms that is hygienically harmless in order to prevent the number of bifidobacteria added to liquid or pasty foods from decreasing during storage and to stabilize the quality. This is a method for stabilizing bifidobacteria in food, which is characterized by:

The catalase-producing microorganisms that are hygienically harmless to be used in the present invention include S# bacteria for food and brewing, or Bacillus natto (
Bacillus natto), etc., but baker's yeast and Bacillus natto, which can be easily obtained commercially, are preferred.

Baker's yeast, Bacillus natto, etc. can be cultured as is or once cultured to collect bacterial cells, treated with ultrasonic waves to completely destroy the bacterial cells, and subjected to sterile p-filtration to obtain a cell-free extract.

The obtained cell-free extract is added to a liquid food or pasty food together with a bifidobacteria culture solution.

The amount of cell-free extract added to food varies depending on the catalase production ability of each microorganism, and must also be varied depending on the amount of bifidobacteria added at the same time, but as a rough guide, approximately 0. It is about 1 to 5 inches.

Examples of the liquid or pasty foods of the present invention include various milk drinks, yogurt, yogurt drinks, lactic acid bacteria drinks, etc.
About 0.01 to 3 inches of concentrated liquid is added, and about 0.1 to 5 inches of a cell-free extract of catalase-positive microorganisms, which is hygienically harmless, is added. As a result, the survival rate of bifidobacteria in food is extremely high, and viable bifidobacteria are stabilized in food for a long period of time.

Next, test examples and examples of the present invention will be shown.

Test Example 1 liter of the commercially available baker's yeast cell-free extract obtained in Example 1 was added to a milk drink, and at the same time, 0.25% of the concentrated Bifidobacterium solution obtained in Example 1 was added, and the mixture was heated at 10°C for 7 hours. The samples were stored for several days and changes in the number of bifidobacteria were observed. The same amount of water was added to the cell-free extract (as a control) and changes in the number of Bifidobacteria were observed in the same manner.

The results are shown in Table 3.

Example 1 Commercially available protease 0.034 is added to skim milk and kept at 50°C for 6 hours to hydrolyze proteins. Add 0.5 g of yeast extract to this and sterilize at 120°C for 15 minutes.

This includes Bifidobacterium Iongum
ATCC15707 was inoculated and neutralized cultured at 37°C for 24 hours at pH 5.0 while replacing the gas phase with 002 gas. The culture solution is centrifuged aseptically to prepare a concentrated bifidobacteria solution.

Separately, commercially available baker's yeast (Saceharomyces ce
revisiae) in sterile water, the bacterial cells were destroyed by ultrasonication (25 kHz for 20 minutes), and ultracentrifuged (25,00 Or
, pm 50 minutes) to obtain a supernatant. This supernatant liquid was added to 0.
Filter through a 45 μm sterile filter to obtain a cell-free extract.

0.25 t of the bifidobacterium concentrate for the milk drink;
Add 1% cell-free extract to make Bifidus Painter 9 milk drink. This milk drink was in good condition with almost no decrease in Bifidobacterium during storage.

Example 2 0.051 of a commercially available protease is added to a 59I+casein solution, and the solution is maintained at 50° C. for 4 hours to hydrolyze casein.

Add 1 part yeast extract and 5% lactose to this, and sterilize at 120°C for 15 minutes. Bifidobacterium breve
ATCC15700 was inoculated, and neutralized culture was carried out at 67° C. for 20 hours at pH 6.0 while replacing the gas phase with N2 gas. The culture solution is centrifuged aseptically to prepare a concentrated bacterial solution of Bifidobacterium.

Separately, commercially available Bacillus natto was cultured at 37° C. for 12 hours and then centrifuged to obtain a concentrated bacterial solution. This is subjected to ultrasonication (25 kHz 1f: lI'8
'l) to disrupt the bacterial cells and perform ultracentrifugation (20.00 Or, p, m 40 minutes) at low temperature (2° to 3° C.) to obtain a supernatant. This supernatant was filtered through a 0.45 mm sterile filter to obtain a cell-free extract. 4% of the Bifidus IO and 2% of the cell-free extract are added to the yogurt to prepare Bifidus Painter Schogeld. This yogurt was in good condition with a small decrease in bifidobacteria during storage. Example 6 0.1% of commercially available protease was added to skim milk and held at 45°C for 2 hours to hydrolyze proteins. This has 0 fish extract
．． Add 4% and sterilize at 120°C for 20 minutes.

In this, Bi fidobacterium Iongu
m ATCC15707 was inoculated, and neutralized culture was carried out for 37'c at pH 6.5 for 20 hours while replacing the gas phase with CO□ gas. The culture solution is centrifuged aseptically to prepare a concentrated bacterial solution of Bifidobacterium.

Separately, edible yeast (Candida pseudotor)
608C72 human CC8645) to obtain a concentrated bacterial solution. This is treated with ultrasonic waves (20 kHz for 15 minutes) to reveal bacterial cells. fj!
dl181 was disrupted and subjected to ultracentrifugation (3 cycles) at low temperature (about 5°C).
0.00 Or, p, m for 50 minutes) to obtain a supernatant. This supernatant is filtered through a 0.45 μm sterile filter to obtain a cell-free extract. To a lactic acid bacteria drink, 1% of the Bifidobacterium concentrate and 2 parts of the cell-free extract are added to obtain a Bifidobacterium lactic acid bacteria drink. This lactic acid bacteria drink showed a small decrease in bifidobacteria during storage and was in good condition.

Claims (1)

[Claims] It is characterized by adding a cell-free extract of catalase-positive microorganisms, which is harmless from a sanitary point of view, in order to prevent the number of Bifidobacterium added to liquid or pasty foods from decreasing during storage and to stabilize the quality. Method for stabilizing Bifidobacterium in food.