KR101952758B1 - Manufacturing Method of Liquid Culture - Google Patents

Abstract

The present invention relates to a method for producing a liquid culture medium, which comprises a yeast extract; Malt extract; Tryptone; Preparing a mixed solution by mixing any one of dextrose, fructose, and galactose saccharide and purified water; Preparing a culture solution by inoculating saccharomyces boulardii into the mixed solution; And a step of fermenting the culture liquid to produce a liquid culture medium. By significantly increasing the viable cell count, it is possible to improve various symptoms such as constipation of the livestock, dilute stool, northern bloating and intestinal fermentation by preventing diarrhea, improving digestibility, Improving the quality of the fattening, improving the anti-infectivity and immunity.

Description

[0002] Manufacturing Method of Liquid Culture [0003]

The present invention relates to a method for producing a liquid culture medium, and more particularly, to a method for producing a liquid culture medium in which the number of viable bacteria is greatly increased by optimizing a culture medium.

In general, the feed used for raising livestock, etc., is mainly used as a natural formulated feed mixed with various cereals.

However, the protein, fat, and carbohydrate of the grains contained in these feeds formed a strong binding force so that the digestion and absorption rate of the livestock was very low, so that the livestock could not absorb enough nutrients.

In addition, since the toxin called phytin, which is a storage form of phosphorus, exists in the grains of the compound feed, phytin is decomposed into phytic acid by the action of gastric acid to form essential essential minerals and phytate salts in the small intestine.

It is also known that when the cereals are fed to the livestock, the livestock can be digested by using microorganisms and then decomposed and supplied. However, this is merely a function of enhancing the physiological activity of livestock, promoting growth, and improving feed efficiency It is difficult to expect effects such as intestinal digestion function and immunity enhancement of livestock.

In order to solve this problem, Korean Patent Laid-Open Publication No. 10-2009-0070631 discloses a process for producing a mixture of ginseng, rice bran, safflower, Fermented feedstuffs prepared by mixing fermented feed additives prepared by fermenting, aging, drying and pulverizing with a mixture of common feed and proper ratio are disclosed, but the cost is high and the cost of livestock Diarrhea, constipation, abnormal fermentation, and intestinal abnormality resulted in poor quality of the fattening due to weakening of the facial weakness.

In order to solve the problems of the prior art, the applicant of the present invention has proposed a method for preventing diarrhea, improving digestibility, improving appetite, improving feed efficiency, improving quality of flesh, A method for producing a fermented feed which can improve the fermented feed is disclosed.

However, the number of viable cells obtained by the above-described method of the prior art is 2.0 x 10 ⁹ CFU / ml, which means that the viable cell count is significantly increased as compared with the commercial medium.

On the other hand, in Chinese Patent Laid-Open No. 10-2224885, in the production of liquid food stimulants for pigs, saccharomyces cerevisiae boulardii ) medium was prepared by using an aqueous solution of 1 wt% of yeast, 2 wt% of tryptone, 2 wt% of glucose and 2 wt% of agar powder.

In this way, the feeding stimulant for pigs is produced and significant results are obtained in terms of weight gain and feeding rate, which is predicted to be due to the increase in viable cell count through the medium.

Korean Patent Publication No. 2009-0070631 Korean Patent Publication No. 10-1705320 Chinese Patent Publication No. 10-2224885

SUMMARY OF THE INVENTION The present invention has been made in view of the above-described prior arts, and it is an object of the present invention to improve various symptoms such as constipation of livestock, dilute stool, northern bloating and intestinal fermentation to prevent diarrhea, improve digestibility, improve appetite, It is an object of the present invention to provide a method for producing a liquid culture medium in which the number of viable cells and the number of viable cells are increased.

It is also an object of the present invention to provide a production method capable of mass production of a liquid culture medium through a production method which is remarkably improved as compared with the prior art Korean Patent Publication No. 10-1705320.

In order to accomplish the above object, the present invention provides a method for preparing a liquid culture medium comprising: a yeast extract; Malt extract; Tryptone; Preparing a mixed solution by mixing any one of dextrose, fructose, and galactose saccharide and purified water; To the mixed solution was added saccharomyces < RTI ID = 0.0 > boulardii ) to prepare a culture solution; And fermenting the culture solution to prepare a liquid culture medium.

Further, the method may further include a drying step of adding and drying a solution of the above-mentioned liquid culture broth.

The mixed solution is a mixed solution obtained by mixing 0.1 to 0.5% by weight of yeast extract, 0.1 to 0.5% by weight of malt extract, 0.1 to 1% by weight of tryptone, 0.5 to 1.5% by weight of saccharides and the balance of purified water .

In addition, the yeast extract and malt extract may be prepared by extracting yeast or malt with purified water at 40 to 60 ° C.

The method for producing a liquid culture liquid according to the present invention can provide a production method capable of mass production of a liquid culture medium as a production method for obtaining a remarkably improved viable cell count as compared with the prior art Korean Patent Publication No. 10-1705320.

In addition, the liquid culture solution produced by the production method of the present invention contains a large amount of saccharomyces boulardii to improve various symptoms such as constipation of livestock, dilute stool, northern bloating, intestinal fermentation, Improvement of appetite, improvement of feed efficiency, improvement of quality of fattening, improvement of anti-illness and immunity.

1 is a flowchart showing a method of manufacturing a fermented feed using a liquid culture liquid according to the present invention.

Hereinafter, the present invention will be described in more detail. The terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary terms and the inventor may appropriately define the concept of the term in order to best describe its invention It should be construed as meaning and concept consistent with the technical idea of the present invention.

As shown in FIG. 1, the method for producing a liquid culture medium according to the present invention comprises: a yeast extract; Malt extract; Tryptone; Preparing a mixed solution by mixing any one of dextrose, fructose, and galactose saccharide and purified water; To the mixed solution was added saccharomyces < RTI ID = 0.0 > boulardii ) to prepare a culture solution; And fermenting the culture solution to prepare a liquid culture medium.

In addition, in order to accelerate the drying of the liquid culture medium, it is possible to further include a drying step in which the starch is added and dried.

In the present invention, tryptone and dextrose are respectively applied as a protein source and a carbon source of a culture medium, and yeast extract and malt extract are applied as a fermentation source for fermentation.

The yeast extract and malt extract may be prepared by extracting yeast or malt with purified water at 40 to 60 ° C, but may be extracted using functional water at 40 to 60 ° C.

When functional water is used, the first functional water and purified water prepared by dissolving sodium metasilicate pentahydrate, sodium borate decahydrate, sodium chloride or potassium carbonate in purified water, Zinc sulphate heptahydrate or sodium selenite to prepare a mixed functional water and mixing the yeast or malt with a mixed functional water at 40 to 60 ° C . ≪ / RTI >

The first functional water is a functional water containing sodium and potassium ions, and the second functional water is functional water containing zinc and selenium ions, which is disclosed in Korean Patent Publication No. 10-1705320.

However, in the present invention, even when the extract is prepared using ordinary purified water without using such functional water, the number of viable cells in the liquid culture liquid to be produced is greatly increased, so that the process and the effect are improved as compared with the prior art. However, when functional water is used, nutrients such as sodium, potassium, zinc, and selenium are contained in the liquid culture liquid, so that it can exhibit a better effect on the growth of livestock. Therefore, have.

When the yeast and malt are extracted using the functional water, the extract is subjected to a filtration process. At this time, it is preferable to perform filtration using a filter having a size of 1 to 2 mm. If the size of the filtration hole is less than 1 mm, there may be a problem that the filtration of the extract is blocked due to the clogging of the fine particulate matter in the filter. Conversely, if the size of the filtration hole is more than 2 mm, the filtrate is filtered by the fine particulate matter, Can occur.

In addition, the yeast extract and malt extract are mixed with sugar to prepare a culture medium, sterilized, and then the extract is treated with tryptone as a protein source, dextrose, fructose, galactose, galactose, and purified water are mixed to prepare a mixed solution. Saccharomyces cerevisiae is inoculated thereto to prepare a culture solution.

The tryptone is a component used as a protein source, and it can be shown that the use of protein instead of the protein used in Korean Patent Registration No. 10-1705320 can greatly increase the viable cell count.

In addition, the saccharides are used as a carbon source. Among the various saccharides, fructose, galactose, and dextrose showed the best effects. Especially, when fructose was used, the highest number of viable cells was obtained.

It is preferable that the mixed solution contains 0.1 to 0.5% by weight of yeast extract, 0.1 to 0.5% by weight of malt extract, 0.1 to 1% by weight of tryptone, 0.5 to 1.5% by weight of saccharides and the remaining purified water. When the content of each component was excessively small or large, the number of living cells tended to decrease. Although the exact cause is not known, it has been experimentally confirmed that, when at least the above-mentioned components and contents are prepared, it is possible to obtain an optimized result for the cultivation of Saccharomyces cerevisiae.

The sterilization treatment is performed at 85 ° C. First, steam is put into the tank jacket to raise the temperature to 100 ° C or higher, and the inside temperature of the culture liquid is set at 85 ° C for 20 minutes or more to sterilize microorganisms, minimizing nutrient destruction.

The liquid culture of the present invention can be prepared by fermenting the culture solution and drying it.

At this time, the drying step is carried out by adding starch. The starch is added in order to accelerate the drying of the culture by activating the fermentation of the culture, removing moisture, and preventing invasion or propagation of other harmful bacteria other than the saccharomyces cerevisiae. .

If the drying time is longer than 5 days, there may be a problem that a long time is consumed. On the other hand, if the drying time is more than 45 캜, There may be a problem of perishability.

The liquid culture liquid prepared by the production method of the present invention is prepared as an additive to be added to the feed and fed to the livestock. The feed includes cereals (for example, crushed or crushed wheat, oats, barley, corn, rice) Protein feeds (for example, feeds based on rapeseed, soybeans, and sunflower), animal protein feeds (e.g., blood, meat, bone meal and fish meal), sugar and dairy products (e.g., various powdered milk and whey powders And the like), and may further include nutritional supplements, digestion and absorption enhancers, growth promoters, and the like.

The feed containing the liquid culture liquid may be administered alone to animals or in combination with other feeds in an edible carrier, and the feed may be used as a top dressing or directly mixed with an animal feed, Can be easily administered to animals in oral formulations and, when administered separately from animal feed, can be prepared in immediate release or sustained release formulations in combination with a pharmaceutically acceptable excipient as is well known in the art .

Such edible carriers may be solid or liquid such as corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol, and when a solid carrier is used, the feed may be in the form of tablets, capsules, A topical dressing in the form of a troche or hamburger or a finely divided acid, and where a liquid carrier is used, the feed can be a gelatin soft capsule, or a syrup or suspension, emulsion, or solution formulation.

The feed may comprise any protein-containing organic fructose commonly used to meet an animal's dietary needs. These protein-containing flours usually consist mainly of corn, soy flour, or corn / soy flour mix.

The feed may also contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, solution promoters and the like. The feed can be used by adding to the animal's feed by pouring, spraying or mixing.

The liquid culture medium of the present invention can be applied to a large number of animal diets including mammals, poultry and fish by being included in the feed, and can be applied to pigs, cows, horses, sheep, goats, laboratory animals, It can be used for pets such as dogs and cats. The poultry can be used for chicken, turkey, duck, goose, pheasant, quail and the like. It is not.

In addition, the liquid culture liquid can be used in animal medicine, and can be administered in combination with improving intestinal function (rectal function), diarrhea treatment, and antibiotic administration, and is applicable to a large number of animal diets including mammals, poultry and fish .

Hereinafter, the present invention will be described more specifically by way of Experimental Examples and Examples, but the present invention is not limited to these Examples.

[Example]

Yeast and malt extracted with purified water at 40 캜 were extracted to prepare yeast extract and malt extract. 50 g of the yeast extract, 60 g of malt extract, and 10 kg of sugar were mixed and sterilized at 80 ° C for 30 minutes using steam, and the internal temperature of the culture was 80 ± 1 ° C.

(1.0 × 10 ⁷ CFU / g) of Saccharomyces cerevisiae, malt extract, tryptone and dextrose were prepared in the same amounts as shown in Table 1, and the remaining purified water was mixed to prepare a mixed solution. After the inoculation, the total amount was 1,000 g, followed by incubation at 25 ° C for 48 hours. After the incubation, 1 ml of each solution was diluted with the decimal system, and the solution was cultured in an incubator at 25 ° C for 48 hours (Example 1).

A liquid culture (Example 2) prepared by the same method as Example 1 but using fructose instead of dextrose (Example 2) and a liquid culture (Example 3) prepared using galactose were prepared.

For comparative experiments, media were prepared as shown in Table 1. In the comparative example, the total amount of the medium is 1,000 g. In Table 1, the content units are all g.

ingredient Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Yeast extract 3 3 3 3 3 3 10 Malt extract 3 3 3 3 3 3 Tryptone 5 5 5 20 peptone 5 Protein extract 5 Beef extract 5 Agar powder 20 Dextrose 10 10 10 10 Fructose 10 Galactose 10 L-glucose 20

Table 2 shows the results of measurement of viable cell counts for Examples and Comparative Examples. In Table 2, the number of live cells is CUF / ml.

collection Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Primary 2.4 × 10 ¹¹ 2.0 × 10 ¹¹ 1.6 × 10 ¹¹ 1.4 × 10 ¹¹ 8.8 × 10 ¹⁰ 1.8 × 10 ¹¹ 1.5 × 10 ¹¹ Secondary 2.2 × 10 ¹¹ 2.0 × 10 ¹¹ 1.8 × 10 ¹¹ 1.0 × 10 ¹¹ 8.5 × 10 ¹⁰ 1.7 × 10 ¹¹ 1.5 × 10 ¹¹ Third 2.2 × 10 ¹¹ 1.9 × 10 ¹¹ 1.8 × 10 ¹¹ 1.4 × 10 ¹¹ 8.7 × 10 ¹⁰ 2.0 × 10 ¹¹ 1.4 × 10 ¹¹ Average 2.3 × 10 ¹¹ 2.0 × 10 ¹¹ 1.7 × 10 ¹¹ 1.3 × 10 ¹¹ 8.7 × 10 ¹⁰ 1.8 × 10 ¹¹ 1.5 × 10 ¹¹

As shown in Table 2, the number of viable cells was significantly increased as compared to the peptone-and-peptides-treated Comparative Examples 1 and 2, in which the tryptone was applied as a protein source. The number of viable cells was increased in Comparative Example 3 in which BEEF extract was applied. However, when the number of experiments was repeated 10 times, the minimum value was 1.3 × 10 ¹¹ CUF / ml and the maximum value was 2.0 × 10 ¹¹ CUF / And it was found that it is difficult to cultivate a reliable mass.

In the case of the carbon source, the number of viable cells was also significantly increased in Example 1 to which dextrose was applied, and in Examples 2 and 3 in which fructose and galactose were applied. In particular, in comparison with Comparative Example 4 and Examples prepared according to the prior art Chinese Patent Laid-Open No. 10-2224885, optimization of the protein source and carbon source and optimization of the amount of use can provide a better effect on the viable cell count .

In addition, in Comparative Example 4, the content was different from that required in the present invention, which was also found to be related to an increase in viable count.

Therefore, it was confirmed that the culture of Saccharomyces cerevisiae could be greatly improved by optimizing the composition and composition of the mixed solution in the present invention.

In addition, the number of viable cells increased by 100 times or more as compared with the viable cell count of the prior art Korean Patent Registration No. 10-1705320, and it was confirmed that the number of viable cells was increased more than the conventional commercial medium.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments and that various changes and modifications may be made therein without departing from the spirit and scope of the invention. Change is possible. Such variations and modifications are to be considered as falling within the scope of the invention and the appended claims.

Claims (5)

A method for producing a liquid culture medium to be added to a feed for livestock,
0.1 to 0.5% by weight of yeast extract prepared by extracting yeast with purified water at 40 to 60 캜, 0.1 to 0.5% by weight of malt extract prepared by extracting malt with purified water at 40 to 60 캜, 0.1 to 1% by weight of tryptone, 0.5 to 1.5% by weight of a saccharide selected from dextrose, fructose and galactose and the balance of purified water to prepare a mixed solution ;
Preparing a culture solution by inoculating saccharomyces boulardii into the mixed solution;
Fermenting the culture solution to prepare a liquid culture medium;
≪ / RTI >
The method according to claim 1,
Further comprising a drying step of adding and drying the liquid culture broth to the liquid culture broth.
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