KR102104114B1 - Preparing method for lactic acid bacteria culture - Google Patents

Abstract

The present invention relates to a production method of a composition for culturing lactic acid bacteria and, more specifically, to a production method of a composition for culturing lactic acid bacteria, which is produced by using: water; powder of Artemisia annua L., Erigeron annuus, mulberry Leaves and Helianthus tuberosus; and a reed root extract. The composition for culturing lactic acid bacteria produced by the production method according to the present invention can control the growth rate of lactic acid bacteria by controlling the particle size of wild grass, such as Artemisia annua L., Erigeron annuus, mulberry Leaves and Helianthus tuberosus, which are added to the composition, thereby being usefully used for culturing lactic acid bacteria.

Description

Production method of composition for lactic acid bacteria culture {PREPARING METHOD FOR LACTIC ACID BACTERIA CULTURE}

The present invention relates to a method for preparing a composition for lactic acid bacteria culture, specifically water; Firefly mugwort, gaetweed, mulberry leaves and pork potato powder; And it relates to a method for producing a composition for lactic acid bacteria culture using reed root extract.

Lactobacillus is one of microorganisms that have been used since ancient times in human diet, and is mainly used in fermented foods such as kimchi in Korea and in dairy foods in the West. Recently, research on lactic acid bacteria has been actively conducted, and its application range has been broadened, such as being used as a health functional food or medicine as well as general food.

Lactobacillus exhibits various physiological activities as well as nutritional value. For example, the lactic acid produced in the manufacturing process of dairy products increases the shelf life of the product, exhibits a unique refreshing acidity, and inhibits the growth of harmful microorganisms. On the other hand, lactic acid bacteria increase the digestibility of milk proteins and promote absorption of calcium, iron and phosphorus. In addition, the health effects of lactic acid bacteria identified so far include prevention of diarrhea and constipation by intestinal action, anti-cancer and anti-aging activity by suppression of harmful bacteria, promotion of development by vitamin production, prevention of adult diseases by cholesterol control, and enhancement of immunity, etc. There is this.

Therefore, in order to commercialize such beneficial lactic acid bacteria, the use of the culture medium thereof is gradually increasing, and the development of a culture medium using natural products rather than a synthetic medium is required. In this regard, Korean Patent Publication No. 10-2015-0144394 discloses that the medium composition for lactic acid bacteria culture including frost extract, sesame leaf extract, buttercup extract, and lettuce extract is inexpensive, but excellent in lactic acid bacteria growth.

Republic of Korea Patent Publication No. 10-2015-0144394

An object of the present invention is to provide a method for producing a composition for lactic acid bacteria culture.

In order to achieve the above object, the present invention is a step of preparing a powder by washing, drying and pulverizing thrush, mango, mulberry leaves and pork potatoes; 2) washing and drying the roots of the reed; 3) adding an extraction solvent to the dried reed root and preparing a reed root extract; 4) provides a method for preparing a composition for lactic acid bacteria culture, comprising the step of mixing and maturing the step 1) 's thrush, primrose, mulberry leaf and pork potato powder, and step 3) reed root extract with water.

The composition for lactic acid bacteria cultivation prepared by the production method according to the present invention can control the growth rate of lactic acid bacteria by controlling the particle size of wild grass, such as thrush, manganese grass, mulberry leaves, and pork potato, which are added to the composition. It can be useful.

1 is prepared in an embodiment of the present invention, the fine powder (A) crushed to a diameter of 12 µm at a temperature of 30 ° C. and the coarse powder (B) crushed to a diameter of 50 µm at a temperature of 65 ° C. It is a result of observing the powder of the forelimbs under an electron microscope (SEM).
Figure 2 is a graph confirming the change in pH shown in the process of fermenting cabbage using the composition for lactic acid bacteria culture prepared in an embodiment of the present invention.
Figure 3 is a photograph of the result of observing the microorganisms present in the fermented product using a composition for lactic acid bacteria culture prepared using a fine powder (A) or crude powder (B) prepared in an embodiment of the present invention.
Figure 4 is a graph showing the results of confirming the pH change in the process of fermenting cabbage using a composition for lactic acid bacteria culture prepared from a fine powder, coarse powder, or a mixture of fine powder and coarse powder in one embodiment of the present invention.

Hereinafter, the present invention will be described in detail.

The present invention comprises the steps of: 1) washing, drying and pulverizing dog wormwood, manganese leaves, mulberry leaves, and pork potatoes; 2) washing and drying the roots of the reed; 3) adding an extraction solvent to the dried reed root and preparing a reed root extract; 4) provides a method for preparing a composition for lactic acid bacteria culture, comprising the step of mixing and maturing the step 1) 's thrush, primrose, mulberry leaf and pork potato powder, and step 3) reed root extract with water.

In one aspect, the method for preparing a composition for lactic acid bacteria culture according to the present invention provides a step of washing, drying, and grinding potato wormwood, manganese grass, mulberry leaves, and pork potatoes to prepare a powder.

The term " Artemisia annua L. ", as used herein, refers to the annual grass of the dicotyledonous campanula. The wormwood mugwort is also referred to as a mugwort mugwort or dog ground stalk, and is distributed in Korea, Japan, China and Taiwan, and is known for its antioxidant activity and immune-enhancing effect, and thus can be used for treatment and prevention of various diseases.

The term " erigeron annuus (L.) Pers. ", As used herein, refers to a perennial plant belonging to the family Asteraceae. The forget-me-not is distributed all over the world, grows to a height of 30 to 100 cm, branches are split from the top, and has thick hairs throughout. In addition, it may have activity such as gastroenteritis, indigestion, abdominal pain and diarrhea treatment due to enteritis, hypoglycemia, hematuria treatment, antimalarial, and anti-inflammatory.

As used herein, the term "mulberry leaves" refers to the leaves of a mulberry tree. The mulberry tree may be a deciduous tree or shrub belonging to the dicotyledon nettle mulberry family Mulberry, and its origin is temperate and subtropical. Leaves can be used for sericulture, bark for medicinal or papermaking, and fruit for edible and medicinal purposes.

The term " Helianthus tuberosus ", as used herein, means a plant of the asteraceae that blooms yellow. The pig potato grows to a height of 1.5 to 3 m, and the tip of the subterranean stem becomes thicker, and the tuber develops, thus having hair along with leaves. Pork potato extract contains a large amount of phenolic compounds, which can have a strong antioxidant effect.

The wormwood, manganese, mulberry leaves, and pork potatoes can be used for all purchased, cultivated or harvested. The thrush, mango, mulberry leaves and pork potatoes can be washed and dried before use. The washing and drying can be performed by any method known in the art. Specifically, the drying may be performed at a temperature of 60 ° C or less, 55 ° C or less, 50 ° C or less, or 47 ° C or less.

Dried thrush, mango, mulberry leaves, and pork potatoes can be pulverized in a conventional manner, and the particle size of the powder after grinding can be appropriately selected by a person skilled in the art. Specifically, the powder is 5 to 60 ㎛, 7 to 60 ㎛, 9 to 60 ㎛, 5 to 57 ㎛, 7 to 57 ㎛, 9 to 57 ㎛, 5 to 54 ㎛, 7 to 54 ㎛, 9 to 54 ㎛ , 5 to 51 μm, 7 to 51 μm or 9 to 51 μm in diameter.

The pulverization may be performed by any method known in the art, and for example, the pulverization may be low temperature pulverization. The crushing process may be performed under a temperature that does not denature the powder of dog wormwood, manganese grass, mulberry leaves, and pork potatoes. Specifically, the temperature may be 50 ° C or less, 48 ° C or less, 46 ° C or less, 44 ° C or less.

According to an embodiment of the present invention, the larger the powder particle size of the thrush, manganese grass, mulberry leaves, and pork potato, the faster the growth rate of lactic acid bacteria, and the smaller the powder particle size, the slower the growth rate of lactic acid bacteria. In other words, the composition for lactic acid bacteria cultivation prepared using coarse powder of thrush, mango, mulberry leaf, and pork potato has a growth rate of lactic acid bacteria, lactic acid bacteria cultivation composition using pulverization Can be faster than

Specifically, the coarse powder is 40 to 60 μm, 43 to 60 μm, 48 to 60 μm, 40 to 56 μm, 43 to 56 μm, 48 to 56 μm, 40 to 52 μm, 43 to 52 μm, or 48 to 52 μm Can have a diameter of On the other hand, the fine powder is 5 to 22 ㎛, 7 to 22 ㎛, 10 to 22 ㎛, 5 to 18 ㎛, 7 to 18 ㎛, 10 to 18 ㎛, 5 to 14 ㎛, 7 to 14 ㎛ or 10 to 14 ㎛ It can have a diameter.

Therefore, the composition for lactic acid bacteria culture according to the present invention can control the growth rate of lactic acid bacteria by controlling the size of powder particles of thrush, manganese, mulberry leaves, and swine potato contained therein. In addition, powders of stoolworm wormwood, primrose, mulberry leaves, and pork potato having different particle sizes may be appropriately mixed as necessary.

In the above step, lotus leaf powder may be additionally used in addition to thrush, mango, mulberry leaves, and pork potatoes. The lotus leaf may be dried, washed and crushed in the same manner as described above.

In another aspect, the method for preparing a composition for lactic acid bacteria culture according to the present invention provides a step of washing and drying the root of a reed.

As used herein, the term “reed ( Phragmites communis Trin. )” Refers to a monocotyledonous plant that is classified as a family of flowering plants. The reeds are mainly distributed in temperate and handae areas and inhabit swamps, river banks, wetlands, and beach shores. The height is about 3 m, the stem is empty, and the leaves can have a size of 30-50 cm. In oriental medicine, dried reed roots are used as medicinal herbs, which can hemostasis, promote stomach movement, and diuretic.

The reed can be purchased, grown or harvested, which can be washed and dried before use. The washing and drying can be performed by any method known in the art. Specifically, the drying may be performed at a temperature of 70 ° C or lower, 65 ° C or lower, 60 ° C or lower, or 57 ° C or lower.

In another aspect, the method for preparing a composition for lactic acid bacteria culture according to the present invention provides a step of adding an extraction solvent to the roots of the dried reed and preparing a reed root extract.

The reed root extract can be prepared by a manufacturing method comprising the following steps:

a) preparing an extract by adding an extraction solvent to the reed root;

b) filtering the extract of step a); And

c) Concentrating the filtered filtrate of step b) under reduced pressure.

In addition, the extraction solvent may be water. The extraction solvent may be used in an appropriate amount by a person skilled in the art according to methods known in the art. Specifically, based on the weight of the reed root, water is 100 to 300 times, 130 to 300 times, 160 to 300 times, 190 to 300 times, 100 to 260 times, 130 to 260 times, 160 to 260 times, 190 to 260 Pear, 100 to 220 times, 130 to 220 times, 160 to 220 times, or may be added in an amount of 190 to 220 times.

The extraction may be performed using any commonly known extraction method, and specifically, it may be shaking extraction, Soxhlet extraction, or reflux extraction. On the other hand, the reduced pressure concentration of step c) may be used a vacuum decompressor or a vacuum rotary evaporator. In addition, the drying may be vacuum drying, vacuum drying, boiling drying, spray drying or freeze drying, and specifically, freeze drying.

The manufacturing method may further include the step of cooling the prepared reed root extract.

In another aspect, the method of preparing a composition for lactic acid bacteria culture according to the present invention provides a step of mixing and maturing the stoolwormwood wormwood, manganese grass, mulberry leaf and pork potato powder, and reed root extract with water.

In the above step, the thrush powder is 0.01 to 0.09% (w / v), 0.02 to 0.09% (w / v), 0.03 to 0.09% (w / v), and 0.01 to 0.07% based on the volume of thawed water. (w / v), 0.02 to 0.07% (w / v), 0.03 to 0.07% (w / v), 0.01 to 0.06% (w / v), 0.02 to 0.06% (w / v) or 0.03 to 0.06% (w / v).

On the other hand, forgotten powder is 0.05 to 0.25% (w / v), 0.07 to 0.25% (w / v), 0.09 to 0.25% (w / v), 0.05 to 0.23% (w /) based on the volume of thawed water. v), 0.07 to 0.23% (w / v), 0.09 to 0.23% (w / v), 0.05 to 0.21% (w / v), 0.07 to 0.21% (w / v) or 0.09 to 0.21% (w / v).

In addition, the mulberry leaf powder is 0.1 to 0.3% (w / v), 0.12 to 0.3% (w / v), 0.14 to 0.3% (w / v), and 0.1 to 0.28% (w /) based on the volume 1 of thawed water. v), 0.12 to 0.28% (w / v), 0.14 to 0.28% (w / v), 0.1 to 0.26% (w / v), 0.12 to 0.26% (w / v), 0.14 to 0.26% (w / v).

Furthermore, pork potato powder is 0.01 to 0.08% (w / v), 0.01 to 0.07% (w / v), 0.02 to 0.08% (w / v) or 0.02 to 0.07% (w) based on the volume of thawed water. / v).

In the above step, the reed root extract may be added to the mixture of thawed water, thrush, manganese, mulberry leaves, and pork potato powder. At this time, the reed root extract may be added in an appropriate amount by a person skilled in the art, specifically, the reed root extract is 1:50 to 250, 1:75 to 250, 1:90 to 250, 1 based on the mixture. It may be added in a volume ratio of: 50 to 230, 1:75 to 230, 1:90 to 230, 1:50 to 210, 1:75 to 210 or 1:90 to 210.

The water may include any water of any kind known in the art, and may specifically be water that does not contain a chlorine component. The water may be used after freezing and thawing, and may be thawed after freezing at a temperature of 0 ° C or lower, -4 ° C or lower, -10 ° C or lower, -25 ° C or lower, or -20 ° C or lower.

The thawing may be performed at a temperature suitable for the growth and activity of lactic acid bacteria. Specifically, the thawing is 5 to 50 ℃, 5 to 45 ℃, 5 to 40 ℃, 5 to 35 ℃, 5 to 30 ℃, 5 to 25 ℃, 7 to 50 ℃, 7 to 45 ℃, 7 to 40 ℃ , 7 to 35 ℃, 7 to 30 ℃, 7 to 25 ℃, 9 to 50 ℃, 9 to 45 ℃, 9 to 40 ℃, 9 to 35 ℃, 9 to 30 ℃ or 9 to 25 ℃ You can. In addition, the thawing for 1 to 60 hours, 1 to 55 hours, 1 to 50 hours, 3 to 60 hours, 3 to 55 hours, 3 to 50 hours, 5 to 60 hours, 5 to 55 hours or 5 to 50 hours Can be performed. The thawed water can be stored under appropriate temperature until further use.

The mixing can be carried out for a suitable time, which can be appropriately adjusted by the skilled person. In one example, the mixing may be performed for 1 to 50 minutes, 1 to 40 minutes, 1 to 45 minutes, 3 to 50 minutes, 3 to 40 minutes or 3 to 45 minutes.

The uniformly mixed mixture as described above can be aged under appropriate temperature and time conditions, which can be appropriately modified by those skilled in the art. Specifically, the aging is performed at a temperature of 8 to 30 ° C, 10 to 30 ° C, 8 to 27 ° C, 10 to 27 ° C, 8 to 25 ° C or 10 to 25 ° C for 12 hours or more, 20 hours or more, or 24 hours or more Can be.

When adding lotus leaf powder to the mixture, the lotus leaf powder may be added in an appropriate amount by a person skilled in the art, specifically 0.03 to 0.2% (w / v), 0.04 to 0.2% (w / v), 0.05 To 0.2% (w / v), 0.03 to 0.15% (w / v), 0.04 to 0.15% (w / v), and 0.05 to 0.15% (w / v).

The composition for lactic acid bacteria culture prepared by the method according to the present invention can be used for culturing all kinds of lactic acid bacteria known as lactic acid bacteria in the ordinary technical field. More specifically, the lactic acid bacteria is Lactobacillus genus (Lactobacillus), Lactococcus genus (Lactococcus), Streptococcus genus (Streptococcus), Phedi O Rhodococcus genus (Pediococcus), flow Pocono stock in (Leuconostoc), Wei Cellar in (Weissella) and Bifidobacterium , and the like.

Hereinafter, the present invention will be described in detail by the following examples, provided that the following examples are only for illustrating the present invention, and the present invention is not limited by them. Any thing that has substantially the same configuration and the same operation and effect as the technical idea described in the claims of the present invention is included in the technical scope of the present invention.

Example 1. Preparation of fine powder of wild grass

First, as a wild grass, the wormwood ( Artemisia annua L. ), the primrose ( Erigeron annuus (L.) Pers. ), Mulberry leaves (mulberry leaves) and swine potato ( Helianthus tuberosus ), respectively, collected and washed, and the temperature below 45 ℃ It was dried in. The dried wormwood mugwort, Gaemancho, mulberry leaves and pork potatoes were prepared by pulverizing to a diameter of 12 μm at a temperature condition of 30 ° C., respectively.

Example 2. Preparation of crude powder of wild grass

First, as wild grass, dog wormwood, manganese, mulberry, and pork potatoes were collected and washed, respectively, and dried at a temperature of 45 ° C. or lower. The dried wormwood mugwort, gangwort, mulberry leaves, and pork potatoes were prepared by pulverizing to a diameter of 50 µm at a temperature of 65 ° C.

Example 3. Form confirmation of wild grass powder

The powders and powders according to the frictional heat and the size of the grinding were compared by pulverizing and pulverizing the wild powder prepared in Examples 1 and 2 with an electron microscope (SEM). At this time, the powder of Gaebangcho was observed as an object, and the result photograph is shown in FIG. 1.

As shown in Fig. 1, the powder of the forelimbs crushed in Example 1 had a uniform and soft particle size, whereas the powder of the forgets crushed in Example 2 had a non-uniform particle size and a angular shape.

Preparation Example 1. Preparation of composition for lactic acid bacteria culture- (1)

A composition for lactic acid bacteria culture capable of selectively activating lactic acid bacteria was prepared using the fine powder of wild grass prepared in Example 1.

First, water containing no chlorine component was frozen to a temperature of -18 ° C or lower, and then prepared by standing at a temperature of 10 ° C for 6 hours. On the other hand, the roots of the reed ( Phragmites communis Trin. ) Were collected and washed in a clean valley area, and dried at a temperature of 55 ° C. or lower. The roots of the dried reed were added to 200 times water based on the weight of the reed's roots, and heated to a low heat for 2 hours to prepare a reed root hot water extract.

After adding and mixing 20 g, 50 g, 75 g, and 15 g of manure wormwood powder, manganese powder, mulberry leaf powder, and pork potato powder prepared in Example 1 in water not containing 50 L of chlorine, Based on the volume of the mixture, 100 times the root hot water extract was added. This was stirred for 5 minutes to mix uniformly, and aged for 1 day or more in a sterile room at 12 ° C. As a result, a composition for lactic acid bacteria culture using a fine powder of wild grass was prepared.

Preparation Example 2. Preparation of composition for lactic acid bacteria culture- (2)

In the same conditions and method as in Preparation Example 1, except that the wild grass powder of Example 1 and the wild grass powder of Example 2 were added in a weight ratio of 75:25 instead of the wild grass powder of Example 1, and A composition for lactic acid bacteria culture using crude powder was prepared.

Preparation Example 3. Preparation of composition for lactic acid bacteria culture- (3)

In the same conditions and method as in Production Example 1, except that the wild grass powder of Example 1 and the wild grass powder of Example 2 were added in a weight ratio of 50:50 instead of the wild grass powder of Example 1, and A composition for lactic acid bacteria culture using crude powder was prepared.

Preparation Example 4. Preparation of composition for lactic acid bacteria culture- (4)

In the same conditions and method as in Production Example 1, except that the wild grass powder of Example 1 and the wild grass powder of Example 2 were added in a weight ratio of 25:75, instead of the wild grass powder of Example 1, and A composition for lactic acid bacteria culture using crude powder was prepared.

Preparation Example 5. Preparation of composition for lactic acid bacteria culture- (5)

A composition for lactic acid bacteria cultivation using the crude powder of wild grass was prepared in the same conditions and method as in Preparation Example 1, except that the wild grass powder of Example 2 was used instead of the wild grass powder of Example 1.

Experimental Example 1. Confirmation of pH change according to particle size of wild grass powder

Using the composition for lactic acid bacteria culture prepared in Preparation Examples 1 and 5, the pH change during the fermentation process was confirmed by the following method.

Specifically, a composition for lactic acid bacteria culture prepared in Preparation Example 1 or 5 of 60 g was added to a mixture containing 1 kg of cabbage, 2 kg of water, and 60 g of oligosaccharide. This was placed in a fermentation container allowing internal air to move outward, and fermented for 5 days under a temperature of 28 ° C. On the other hand, Figure 3 shows a picture of lactic acid bacteria present in the fermentation product fermented in the above process under a microscope.

As a result, as shown in FIG. 2, when the composition for lactic acid bacteria culture of Example 1 was used, the pH rapidly dropped 2 days after the start of fermentation, and the active activity of lactic acid bacteria was shown. On the other hand, when the composition for lactic acid bacteria culture of Example 2 was used, fermentation was not good.

On the other hand, as shown in Figure 3, it was possible to confirm the lactic acid bacteria present in the fermentation product obtained in the process.

Experimental Example 2. Control of pH change rate using particle size of wild grass

Using the composition for lactic acid bacteria culture prepared in Preparation Examples 1 to 5 was confirmed the pH change during the fermentation process. Specifically, the experiment was conducted under the same method and conditions as described in Experimental Example 1, and the results are shown in FIG. 4.

As shown in Figure 4, when using a composition for lactic acid bacteria culture prepared using a fine powder of wild grass, the fermentation activity was the best, the composition for lactic acid bacteria culture using a wild powder of crude powder has a relatively fermentation rate late. On the other hand, when a mixture of fine powder and coarse powder was used, the fermentation rate tended to decrease as the content of coarse powder used increased.

Therefore, it was found from the above that the composition for lactic acid bacteria culture according to the present invention can control the fermentation rate using the particle size and content of the wild grass powder contained therein.

Claims (6)

1) Washing, drying and crushing the wormwood wormwood, manganese grass, mulberry leaves and pork potatoes to prepare a powder;
2) washing and drying the roots of the reed;
3) adding an extraction solvent to the dried reed root and preparing a reed root extract;
4) Method of producing a composition for lactic acid bacteria culture, comprising the step of mixing and maturing the root extract of step 1), wormwood, manganese mulberry leaf, and pork potato powder of step 1) and reed of step 3) with water.
The method of claim 1, wherein the powder of step 1) has a diameter of 5 to 60 μm, a method for preparing a composition for lactic acid bacteria culture.
The method of claim 1, wherein the water of step 4) does not contain a chlorine component.
The method of claim 1, wherein the extraction solvent of step 3) is water.
According to claim 1, In the step 4), the root extract of the reed is added in a volume ratio of 1:50 to 250 based on the mixture of the ground wormwood, manganese grass, mulberry leaves, and pork potatoes of step 1) mixed with water. , Method for producing a composition for lactic acid bacteria culture.
The method of claim 1, wherein the thrush in the step 4), wormwood, manganese leaves, mulberry leaves and pork potatoes are 0.01 to 0.09% (w / v), 0.05 to 0.25% (w / v) based on the volume 1 of thawed water. , 0.1 to 0.3% (w / v), and 0.01 to 0.08% (w / v), respectively, a method for preparing a composition for lactic acid bacteria culture.

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