KR101501827B1 - Method for manufacturing base of watery radish kimchi using GABA producing Lactobacillus and Opuntia humifusa - Google Patents

Abstract

More particularly, the present invention relates to a method for producing a dongchimi base using GABA-producing lactic acid bacteria and a milky basil, and more particularly, to a method for producing a dongchimi base using a lactic acid bacteria, The present invention also provides a method for producing a donchimi base to which a liquid and a foil are mixed and added with milled water in a paste state without addition of water. The dongchimi base prepared by the above method was tried to give the color and functionality by attaching the dongchunsik to the dongchimi base. Thus, a dongchimi base without added water can be manufactured to contribute to industrial development.

Description

TECHNICAL FIELD The present invention relates to a method for producing a dongchimi base using lactic acid bacteria and a ginseng seed,

More particularly, the present invention relates to a method for producing a Dongchimi base using GABA-producing lactic acid bacteria and T. kansenensis. More particularly, the present invention relates to a method for producing a Dongchimi base using GABA- The present invention also relates to a method for producing a Dongchimi base to which a millet is added in a paste state without adding water.

Dongchimi is a type of kimchi, which is an old kind of Korean kimchi. It is a kind of kimchi which is largely or largely sun-dried and served in a soup. In particular, it is known that Dongchimi is a food which is fermented by adding radish and a small amount of raw material to a proper concentration of salt water, becomes a supply source of NaCl and minerals, and has alkaline properties in controlling the acidity balance of body fluids. Because Dongchimi does not contain a lot of seasonings and uses a lot of water, it is a soup kimchi that I have been enjoying since the past because of the unique taste of lactic acid, various organic acids and carbon dioxide produced in broth, the freshness of carbonated fish, Lactic acid fermented food.

Gamma-aminobutyric acid (GABA), which is produced as a metabolite of lactic acid bacteria, is a kind of nonprotein amino acid distributed in the natural environment. As a inhibitory neurotransmitter, blood pressure It is a substance known to suppress the rise, to work on improving the visual acuity, and to calm anxiety and nervousness. In animals, it is distributed in the brain, heart, lung, kidney, etc. In the case of plants, it is mainly found in germinated brown rice and green tea. Hypnotists, examinees, and alcohol-consuming sufferers suffering from stress are reported to have low levels of GABA in the brain and blood, and a severe shortage of GABA in the body is known to cause seizures, convulsions, and epilepsy. The function of GABA is to improve the blood flow of the brain, thereby increasing the oxygen supply to the brain, which is used as a therapeutic agent for improving the metabolism of the brain and lowering the motivation. It is also known as brain food. In addition, it acts on the blood vessel center of the soft tissue, Various physiological actions such as anti-hypertensive effect and diuretic effect which suppress the secretion and dilate the blood vessel and lower the blood vessel have been reported.

Fermented soybean fermented food by the Bacillus strain is decomposed by the action of the enzyme produced by the strain, and a sticky viscous substance is produced to give its distinctive taste and flavor. (Γ-PGA) produced by soybean fermented food is an amino acid-based polymer produced by a microorganism as a γ-polypeptide in which the α-amino group of glutamine and the γ-carboxyl group of glutamic acid are amide bonded. Gamma-polyglutamic acid (γ-PGA) has been reported to produce several strains, but γ-PGA produced by Bacillus subtilis , the soybean fermentation microorganism, is the most common. The reported efficacy of γ-PGA includes immunosuppressive effect, inhibition effect on intracellular bacterial and viral infection, cancer cell killing effect, cell wall protection and antioxidant effect, calcium absorption promotion and excellent moisturizing effect. Is a functional material that is well-received.

And Opuntia humifusa is a Korean native cactus belonging to the palm cactus and is a perennial plant capable of cultivating for several to several decades since it can survive the cold at 20 ℃. The fruit and stem contain 163.8 and 71 mg% of vitamin C, respectively, and it is reported that the total polyphenol compound content is high. In the study of functionalities of the viscous substances and pigments extracted from fruits, the extracts isolated from cactus fruit contain polysaccharides which are stable to heat and maintain the stability and physical properties of red pigment even under acidic conditions, Which is a useful food material that can be used to enhance nutritional value and promote nutritional value.

Conventional dongchimi were prepared by dipping various materials into salt water and matureting them at low temperature. However, there was no study on the Dongchimi base which was fermented by inoculating lactic acid bacteria starter by mixing both the extract and the pak which were all juices of the ingredients without adding water.

Korean Patent Application No. 10-2010-0046387 is directed to a method for commercial production of Dongchimi by a short-term property fermentation method after adding a fermentation strain to an uncongested product concentrated at a low temperature below 60 ° C . However, there is no mention of a method of producing a Dongchimi base which is prepared by inoculating a two-stage fermentation starter using the Bacillus subtilis and lactic acid bacteria of the present invention, .

Therefore, the inventors of the present invention have found that a starch is inoculated through a two-stage fermentation using Bacillus subtilis and lactic acid bacteria, and all of the materials used in the dongchimi are mixed with the extracted extract and the foil, And completed the present invention.

It is an object of the present invention to provide a method for manufacturing a donancho base additionally made by a two-stage fermentation using Bacillus subtilis and lactic acid bacteria and inoculating it with a starter.

Another object of the present invention is to provide a Dongchimi base manufactured by the above method.

In order to achieve the above object, the present inventors have conducted the preparation of a Dongchimi base to which Ganoderma lucidum Bacillus was produced using GABA-producing lactic acid bacteria and various lactic acid bacteria. First, NB (nutrient broth) liquid medium was added to Bacillus subtilis Subtilis HA was cultured at 42 ℃ for 24 hours at 160 rpm and centrifuged at 8,000 rpm for 20 minutes to remove the supernatant. The cells were diluted in sterilized water. The mixture was inoculated 5% in Bacillus subtilis culture medium and subjected to 1-stage fermentation at 42 ° C for 72 hours at 160 rpm. And four kinds of lactic acid bacteria [ Lactobacillus plantarum) K154 (KACC91727P), Lactobacillus brevis (Lactobacillus brevis) YC10 (KCCM43012), Lactobacillus tareom Plan (Lactobacillus plantarum ) YC11 (KCCM43013), Lactobacillus fermentum ) YC12 (KCCM43014) was cultured in MRS liquid medium at 30 ℃ for 48 hours, centrifuged at 15,000 rpm for 10 minutes, and the supernatant was removed from the supernatant to dilute it in sterilized water to obtain a lactic acid bacteria starter. In order to prepare this as a lactic acid fermentation starter of the second stage, first stage fermentation product and 10% defatted milk solution were mixed at a ratio of 1: 1, 1% of each lactic acid bacterial starter was inoculated and cultured at 30 ° C for 72 hours, I made a fermentation starter. In order to prepare the dongchimi base, the first part of the perilla seeds which had been removed from the seeds was heat treated at 85 ° C for 30 minutes. Then, 5% of the heat-treated milky goat was mixed with the paste mixed with the extract and the pak which were all juice of the Dongchimi base materials. A 10% fermented starter (2.5% each) was inoculated therein, fermented at 30 ° C for 1 day, and then fermented at 12 ° C for 6 days to prepare a Dongchimi base.

The present invention relates to (1) a primary fermentation step of preparing, inoculating and cultivating a Bacillus subtilis starter for primary fermentation; (2) mixing the primary fermentation product with 10% skim milk powder; (3) a secondary fermentation step of preparing a lactic acid bacteria starter for secondary fermentation, inoculating and culturing the mixture in step (2); (4) juicing a Dongchwami material consisting of radish, pear, persimmon, apple, ginger, garlic, sugar and salt; (5) Inoculating the secondary fermentation product of step (3) into the juice and juice mixture juiced in step (4); And (6) fermenting the mixture of step (5) at 25 to 30 ° C for 1 day, followed by fermentation at 8 to 12 ° C for 4 to 6 days.

On the other hand, the method further comprises adding 1 to 5% by weight of natural herbs to the whole Dongchimi material after the step (4), wherein 1 to 10% by weight of garlic is added to the whole Dongchimi material after the step (6) Wherein the primary fermentation product and the 10% non-fat milk powder are mixed in a weight ratio of 1: 1 in the step (2).

Specifically, the Bacillus subtilis is Bacillus subtilis HA (Bacillus subtilis And HA) (KCCM 10775P), the lactic acid bacteria is Lactobacillus plan tareom (Lactobacillus plantarum) K154 (KACC91727P) , Lactobacillus brevis (Lactobacillus brevis) YC10 (KCCM43012), Lactobacillus tareom Plan (Lactobacillus plantarum ) YC11 (KCCM43013), Lactobacillus fermentum YC12 (KCCM43014).

The Bacillus subtilis HA (Bacillus used in the present invention subtilis HA) (KCCM 10775P) is a strain isolated and identified in a conventional Chungkukjang, and it is disclosed in Korean Patent No. 10-0864850 (registered on October 16, 2008). In addition, the Lactobacillus plantarum used in the present invention plantarum ) K154 (KACC91727P) was purchased from Korea Food Research Institute.

Specifically, the primary fermentation step is characterized by fermentation at 37 to 42 DEG C for 72 hours, and the secondary fermentation step is characterized by fermentation at 25 to 30 DEG C for 48 to 72 hours.

In the present invention, the "dongchimi base" is an extract fermented on the basis of a mixture of a juice of Dongchimi material and an extract of fruit juice. The extract may be diluted with purified water to a desired concentration when dongchimi is prepared.

In the present invention, the term "starter" refers to a culture medium of microorganisms used for producing a fermented product. Therefore, the types of starter microorganisms determine the characteristics of the product and have an important influence on the quality of the product. Bacteria, fungi, yeast, etc., which are used as starters in microorganisms, can be used alone or in combination.

The present invention also provides a Dongchimi base manufactured by the above method.

The present invention relates to a method for producing a dongchimi base using GABA-producing lactic acid bacteria and T. acutissimae, which comprises mixing all the materials used in the production of Dongchimi with juice extract and foil, The starter containing γ-PGA and GABA is inoculated through fermentation. In addition, we tried to give color and functionality by attaching to the base of Dongchimi. Thus, a dongchimi base without added water can be manufactured to contribute to industrial development.

Figure 1 schematically illustrates the preparation and fermentation process of a Dongchimi base.
FIG. 2 is a graph showing the change in GABA production of isolated microorganisms by TLC.
3 is a graph showing changes in pH and acidity of a two-stage fermented lactic acid bacteria starter.
4 is a graph showing the change in GABA production of the two-stage fermented lactic acid bacteria starter by TLC.
5 is a graph showing changes in pH and acidity of a Dongchimi base using a single strain and a mixed strain. Single strains: Lactobacillus tareom Plan (Lactobacillus plantarum) K154, mixed strains: Lactobacillus plan tareom (Lactobacillus plantarum) K154 / Lactobacillus brevis (Lactobacillus brevis ) YC10 / Lactobacillus plantarum ( Lactobacillus plantarum YC11 / Lactobacillus fermentum YC12 /
FIG. 6 is a graph showing the change in GABA production on a Dachimi-base based on TLC using a single strain and a mixed strain.
7 is a graph showing changes in pH and acidity of the Dongchimi bases according to the addition of salt.
FIG. 8 is a graph showing the change in GABA production on a donchomy base according to the addition of salt by TLC.
FIG. 9 is a graph showing changes in pH and acidity of the Dongchimi bases according to addition of tsunami seeds.
FIG. 10 is a graph showing the change in GABA production on a donchimi base by TLC in accordance with the addition of tannins.
11 is a graph showing changes in pH and acidity of the Danchemi base added with the garlic ratio.
FIG. 12 is a graph showing the change in GABA production by TLC according to garlic ratio.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the present invention is not limited by these examples.

< Example  1> Isolation, Growth and Identification of Microorganisms GABA  exam

Lactobacillus plantarum with excellent GABA productivity ( Lactobacillus plantarum ) strain K154 was purchased from Korea Food Science Research Institute. In order to isolate lactic acid bacteria from Dongchimi, 20 μl of sample was inoculated in MRS agar medium (Table 1) at 1,000 times dilution and cultured at 30 ° C for 48 hours. Each isolate was then purified by MRS agar medium and identified as a potential lactic acid bacterium. The isolated lactic acid bacteria were identified using the 16S rRNA gene sequence (Gene Sequence). Then, all of the purely isolated lactic acid bacteria were used for Dongchimi fermentation. In addition, Bacillus subtilis NB medium (Table 2) was used for the growth of subtilis HA.

Composition of MRS medium  ingredient Component Ratio (grams / liter)  Proteose Peptone # 3  10.0 Beef extract  10.0 Yeast extract 5.0 Dextrose 20.0 Polysorbate 80 1.0 Ammonium citrate 2.0 Sodium acetate 5.0 Magnesium sulfate 0.1 Manganese sulfate  0.05 Dipotassium phosphate 2.0

Composition of NB medium  ingredient Component Ratio (grams / liter)   peptone  3.0 Beef extract  5.0

Lactobacillus isolated from Dongchimi was identified by using 16S rRNA gene sequence (Gene Sequence). Lactobacillus brevis brevis , Lactobacillus &lt; RTI ID = 0.0 &gt; plantarum , and Lactobacillus fermentum . The present inventors have found that Lactobacillus brevis (lactic acid bacteria Lactobacillus each brevis) YC10, Lactobacillus tareom Plan (Lactobacillus plantarum ) YC11, Lactobacillus fermentum YC12, and deposited with the Korean Microorganism Conservation Center (KCCM) on Feb. 5, 2013 under the accession numbers KCCM43012, KCCM43013 and KCCM43014.

In order to confirm the GABA production ability of lactic acid bacteria, one platinum was inoculated into the MRS medium and the MRS medium containing 0.5% MSG, respectively, and the culture was conducted for 48 hours. TLC (Thin Layer Chromatography) was used to confirm GABA production . Of three strains isolated from Dongchimi, GABA was produced by Lactobacillus brevis YC10. The other two kinds of lactic acid bacteria did not produce GABA in the MRS medium (Fig. 2).

< Example  2> Bacillus first stage Fermentation product  Manufacturing and content testing

Table 3 shows the composition of the restriction media required for Bacillus subtilis fermentation. The ingredients were each prepared as a solution, sterilized at 121 ° C for 15 minutes, and mixed immediately before fermentation.

ingredient unit(%) Monosodium glutamate 3 Glucose 3 Ammonium sulfate 0.5 Monopotassium phosphate 0.1 Sodium phosphate die basic 12-water 0.1 Magnesium sulfate 7-water 0.05 Calcium chloride 0.04 Biotin 0.005

The pH was measured with a pH meter and the acidity was calculated as the amount of 0.1N-NaOH consumed at that time by dropping 0.1N NaOH into an appropriate ending point at pH 8.3. As for the content of the viscous substance, the first step fermented product was centrifuged at 8,000 rpm for 20 minutes, and the supernatant was mixed with 3 times volume of 95% ethanol, and the precipitated solid was separated, dried at 50 ° C for 48 hours, .

Fermentation time (days) pH Number of living cells (CFU / mL) Viscosity (%) One 5.64 ± 0.19 1.24X10 ⁹ 0.49 + 0.27 2 5.94 + - 0.27 9.20X10 ⁸ 0.98 + - 0.34 3 6.36 ± 0.22 8.50X10 ⁸ 1.94 + - 0.28

As shown in Table 4, the pH gradually increased to 6.36 ± 0.22 on the third day of fermentation. The number of viable cells was the highest at 1.24 × ^{10 9} CFU / mL on the first day of fermentation, but tended to decrease gradually with increasing fermentation time. The content of viscous substance increased from 0.49 ± 0.27% on the first day of fermentation to 1.94 ± 0.28% on the third day of fermentation.

< Example  3> Two-stage fermentation of lactic acid bacteria starter pH , Acidity, Viable cell count  And GABA  Generation test

The first stage fermentation product and 10% nonfat milk solution were mixed at a ratio of 1: 1. Each of the lactic acid bacteria cultured in MRS for 24 hours was centrifuged at 8,000 rpm for 10 minutes, and the supernatant-removed cells were mixed with 1 mL of distilled water, inoculated, and fermented at 30 ° C for 3 days. The pH was measured with a pH meter and the acidity was calculated as the amount of 0.1N-NaOH consumed at that time by dropping 0.1N NaOH into an appropriate ending point at pH 8.3.

As shown in FIG. 3, the pH of the two-stage fermented lactic acid bacteria starter decreased with the fermentation time, and the acidity thereof was increased. Especially, the fastest fermentation is Lactobacillus plantarum plantarum K154 strain, pH 4.23 and acidity 1.13% on the third day of fermentation.

The number of viable cells was determined by using a 10-step dilution method on each MRS agar plate medium cultured for 72 hours at intervals of 24 hours.

Number of living cells (CFU / mL) 0 days 1 day 2 days 3 days L. plantarum K154 0 1.65X10 ⁹ 2.25X10 ⁹ 1.10X10 ⁹ L. brevis YC10 0 7.75X10 ⁸ 9.75X10 ⁸ 6.00X10 ⁸ L. plantarum YC11 0 1.17X10 ⁹ 1.84X10 ⁹ 9.75X10 ⁸ L. fermentum YC12 0 6.25X10 ⁸ 9.25X10 ⁸ 6.90X10 ⁸

As shown in Table 5, the viable cell counts of Lactobacillus plantarum ) strain K154 was the highest at 1.10 × ^{10 9} CFU / mL. The viable cell counts were maintained above 10 ⁸ CFU / mL in all conditions during 3 days of fermentation and were used as a dongchimi starter.

In order to confirm the GABA production ability in the two-stage fermented lactic acid bacteria starter, samples were collected at intervals of 24 hours after fermentation by the above method, and TAB (Thin Layer Chromatography) was used to confirm whether or not GABA was produced.

As shown in FIG. 4, all of the lactic acid bacteria were found to produce GABA during the incubation period of 3 days. In particular, the highest GABA production was found in Lactobacillus plantarum was plantarum) K154 strain, Lactobacillus brevis (Lactobacillus brevis) YC10 strain also showed that the content of the third day the culture rapidly increased GABA.

< Example  4> Single strain Mixed strains  Using dongchimi base pH , Acidity, Viable cell count  And GABA  exam

The juice of Dongchimi was mixed with the juice. A dongchimi was prepared in the ratio of the materials shown in Table 6 to 10% of the total weight of the two-stage lactic acid fermentation starter prepared by the method of Example 3 (2.5% starter for each strain). Dongchimi was fermented at 30 ℃ for 1 day and aged at 12 ℃ for 6 days. The pH was measured with a pH meter and the acidity was calculated as the amount of 0.1N-NaOH consumed at that time by dropping 0.1N NaOH into an appropriate ending point at pH 8.3.

material  Unit (Gram) radish 150 ship 6 wave 5 Apple 5 ginger 2 garlic 20 Sugar 10 Salt 2 Millennium (added separately) (10) Sum 200 (210 at the beginning of the year)

In order to add millenniaceps, seeds were removed by heat treatment at 85 ℃ for 30 min and cooled at room temperature. Then, in order to ferment Dongchimi, 5% of Dongchimi was added to the fermented starch before inoculation of the lactic fermentation starter.

As shown in FIG. 5, the fermentation speed was faster than that of the single strain when fermented using a mixed strain. As a result of fermentation with mixed strains for 7 days, pH was 4.05 and acidity was 1.03%.

The viable counts of fermented Dongchimi samples were measured by using 10-step dilution method on MRS agar plate medium.

　
　 Number of living cells (CFU / mL) 0 days 1 day 3 days 5 days 7 days A single strain 0 2.75X10 ⁷ 1.10X10 ⁸ 6.80X10 ⁷ 6.50X10 ⁷ Mixed strain 0 1.80X10 ⁸ 2.00X10 ⁸ 1.42X10 ⁸ 1.42X10 ⁸

Single strains: Lactobacillus tareom Plan (Lactobacillus plantarum ) K154,

Mixed strains: Lactobacillus tareom Plan (Lactobacillus plantarum) K154 / Lactobacillus brevis (Lactobacillus brevis ) YC10 / Lactobacillus plantarum ( Lactobacillus plantarum YC11 / Lactobacillus fermentum YC12 /

As shown in Table 7, the number of viable cells of Dongchimi using mixed strains was high regardless of fermentation period. The number of viable cell counts was found to be 1.42 × ^{10 8} CFU / mL more than the viable cell count of 6.5 × 10 ⁷ CFU / mL in the case of mixed strains on the 7th day after fermentation. Therefore, mixed strains were used for the production of Dongchimi base.

Identification of GABA production ability of Dongchimi The samples were collected by dongchimi fermented by the above method and TLC (Thin Layer Chromatography) was used to confirm whether or not GABA was produced.

As shown in FIG. 6, there was no significant difference in the change of GABA according to the fermentation period of Dongchimi using a single strain and a mixed strain. The content of GABA produced at 30 ℃ for 24 hours was not significantly different from that of the fermented GABA on the 7th day of fermentation, but the content of GABA in Dongchimi was slightly higher than that of Dongchimi fermented as a single strain.

< Example  5> Effect of Salt Addition on Dongchimi Bases pH , Acidity, Viable cell count  And GABA  Generation test

The pH was measured with a pH meter and the acidity was calculated as the amount of 0.1N-NaOH consumed at that time by dropping 0.1N NaOH into an appropriate ending point at pH 8.3.

As shown in FIG. 7, the higher the salt addition ratio, the more inhibited the Dongchimi fermentation. In particular, it was found that fermentation hardly occurs at more than 3% of salt. There was little change in pH and acidity. However, the pH change was the highest at 1% salt condition, and pH fermentation rapidly proceeded at 7 days after the addition of 1% salt, pH of 3.93 and acidity of 1.26%.

The viable counts of fermented Dongchimi bases were measured by using 10-step dilution method on MRS agar plate medium.

Number of living cells (CFU / mL) 1 day 3 days 5 days 7 days Salt 1% 1.13X10 ⁸ 3.95X10 ⁸ 3.50X10 ⁸ 3.25X10 ⁸ Salt 2% 5.00X10 ⁷ 3.50X10 ⁸ 1.58X10 ⁸ 1.00X10 ⁸ Salt 3% 6.10X10 ⁷ 3.38X10 ⁷ 3.20X10 ⁷ 7.56X10 ⁶ Salt 4% 2.90X10 ⁷ 6.00X10 ⁶ 1.05X10 ⁴ 3.80X10 ³

As shown in Table 8, the higher the salt addition ratio, the more the viable cell count decreases. In the case of 4% salt, the viable cell count rapidly decreased after 5 days of fermentation and reached 3.80 × 10 ³ CFU / mL on the 7th day of fermentation. At the 1% salt level, the highest viable cell count was found at 3.25 × ^{10 8} CFU / mL on the 7th day of fermentation.

To confirm the GABA production ability of Dongchimi-based samples, samples were taken from fermented Dongchimi samples, and diluted 3 times with 95% ethanol. The precipitates were centrifuged at 15,000 rpm for 10 minutes, and the supernatant was removed by TLC (Thin Layer Chromatography) GABA production was confirmed.

As shown in FIG. 8, it can be seen that the generation of GABA is similarly generated without significantly affecting the salt addition ratio. In addition, the content of GABA produced on the first day of fermentation did not show a tendency to be continuously produced even when fermentation was prolonged.

< Example  6> Millennium  The dongchimi base pH , Acidity, Viable cell count , Chromaticity and GABA  exam

The dongchimi bass added with the millennium ginseng was added to each concentration by heat treatment and cooling for 30 minutes at 85 ℃ in which the seeds were removed before the second stage lactic acid fermentation starter was inoculated. Then, a second stage lactic acid fermentation starter (mixed strain) was inoculated to ferment a Dongchimi fermentation. The pH was measured with a pH meter and the acidity was calculated as the amount of 0.1N-NaOH consumed at that time by dropping 0.1N NaOH into an appropriate ending point at pH 8.3.

As shown in FIG. 9, even when the addition rate of the perennial plant was increased, the difference in pH was not significantly different. The acidity was maintained at the highest level after 3 days of fermentation with 5% addition of 5% TC, and 1.06% on the 7th day of fermentation.

The viable counts of fermented Dongchimi bases were measured by using 10-step dilution method on MRS agar plate medium.

　 Number of living cells (CFU / mL) 1 day 3 days 5 days 7 days 0% 2.18X10 ⁸ 4.50X10 ⁸ 9.25X10 ⁸ 6.25X10 ⁸ 1% 6.90X10 ⁸ 6.88X10 ⁸ 1.04X10 ⁹ 8.78X10 ⁸ 5% 4.38X10 ⁸ 7.63X10 ⁸ 1.80X10 ⁹ 9.25X10 ⁸

As shown in Table 9, the number of viable cells was increased as the addition rate of the tiller seeds was higher. The viable cell counts were higher in the 1% condition on the first day of fermentation, but higher in the 5% of the second day after fermentation. On the 7th day of fermentation, 6.90 × ^{10 8} CFU / mL showed the highest level of 5% per year.

The colorimetric test of the dongchimi base was carried out by measuring the L, a, and b values of the Hunter color system using a colorimeter. L value indicates brightness, a value indicates redness, and b value indicates yellowness.

　
　 　
　 0 One 3 5 7 L 0% 30.67 33.23 36.59 37.22 37.84 1% 29.60 31.81 35.81 36.21 36.81 5% 28.40 28.56 29.72 29.94 30.22 a 0% -1.64 -1.69 -2.07 -1.87 -1.95 1% -1.43 -1.39 -1.42 -1.24 -1.02 5% 2.06 2.36 3.37 5.67 6.32 b 0% 4.60 4.82 5.16 5.25 5.27 1% 4.97 4.87 4.89 4.39 4.59 5% 4.88 4.81 4.37 4.14 4.18

As shown in Table 10, the lightness increased gradually as the fermentation period was longer, and was highest at 0% at the beginning of the millennium and 37.84 at the 7th day of fermentation. The redness showed a trend of increasing redness at 5% of the millenniace at the early stage of fermentation from 2.06 to 6.32 at the 7th day of fermentation, but little change in other conditions. There was little change in yellowness regardless of the condition.

To confirm the GABA production ability of the dongchimi base, a sample was taken from the fermented Dongchimi bases, and diluted 3 times with 95% ethanol. The precipitate was centrifuged at 15,000 rpm for 10 minutes, and the supernatant was removed by TLC (thin layer chromatography) To confirm the presence of GABA production.

As shown in FIG. 10, it was found that the condition of addition of Ganoderma lucidum did not significantly affect the production of GABA. In addition, the content of GABA produced on the first day of fermentation did not show a tendency to increase continuously even when fermentation was prolonged.

< Example  7> According to garlic ratio Millennium  Additive Dongchimi base pH , Acidity, Viable cell count , Chromaticity and GABA  exam

Dongchimi base was prepared by varying the addition ratio of garlic from 0 to 10%. The pH was measured with a pH meter and the acidity was calculated as the amount of 0.1N-NaOH consumed at that time by dropping 0.1N NaOH into an appropriate ending point at pH 8.3.

As shown in FIG. 11, the pH was found to increase proportionally to the amount of garlic added. This is consistent with the report that fermentation is retarded by inhibiting the growth of bacteria due to the antimicrobial activity of garlic. The fermentation progression was slow at 7.5 and 10% of garlic until the third day of fermentation, but the difference in pH was not significant at 7th fermentation. The acidity was the highest in the group without added garlic.

The number of viable cell counts was measured by using a 10-step dilution method on MRS agar plate medium by collecting fermented Dongchimi base samples at intervals and shown in Table 11.

Number of living cells (CFU / mL) 1 day 3 days 5 days 7 days 0% of garlic 8.62X10 ⁸ 6.00X10 ⁸ 6.70X10 ⁸ 5.70X10 ⁸ Garlic 2.5% 4.20X10 ⁸ 4.50X10 ⁸ 6.00X10 ⁸ 5.80X10 ⁸ Garlic 5% 1.58X10 ⁸ 3.45X10 ⁸ 3.65X10 ⁸ 2.96X10 ⁸ 7.5% of garlic 1.18X10 ⁸ 8.80X10 ⁷ 3.25X10 ⁸ 3.35X10 ⁸ 10% of garlic 7.10X10 ⁷ 1.95X10 ⁸ 2.80X10 ⁸ 2.20X10 ⁸

As shown in Table 11, the number of viable cells decreased with increasing garlic addition ratio. There was no significant difference between garlic - added and non - garlic - added groups at 2.5% garlic during fermentation period.

To confirm the GABA production ability of the dongchimi base, a sample was taken from the fermented Dongchimi bases, and diluted 3 times with 95% ethanol. The precipitate was centrifuged at 15,000 rpm for 10 minutes, and the supernatant was removed by TLC (thin layer chromatography) To confirm the presence of GABA production.

As shown in FIG. 12, it was found that the production of GABA on the dongchimi based on the addition of garlic gradually increased with the fermentation period at 0% garlic condition. However, the production of GABA did not change with the 1st day of fermentation in garlic 2.5% or more. Therefore, it was shown that the addition of garlic after fermentation was suitable to increase the content of GABA by preparing Dongchimi base without adding garlic.

Korea Microorganism Conservation Center (overseas) KCCM43012 20130205 Korea Microorganism Conservation Center (overseas) KCCM43013 20130205 Korea Microorganism Conservation Center (overseas) KCCM43014 20130205

Claims (9)

(1) a primary fermentation step of preparing and inoculating and cultivating a yeast starter for primary fermentation;
(2) mixing the primary fermentation product with 10% skim milk powder;
(3) a secondary fermentation step in which a lactic acid bacteria starter capable of producing gamma-aminobutyric acid (GABA) for secondary fermentation is prepared, and inoculation and culture are carried out in the step (2);
(4) juicing dongchimi material consisting of radish, pear, wave, apple, ginger, sugar and salt;
(5) adding 1 to 5% by weight of the total dandruff material to the juice mixture and juice mixture to be juiced in step (4) and inoculating the second fermentation product of step (3);
(6) fermenting the mixture of step (5) for one day at 25 to 30 DEG C, and then fermenting for 4 to 6 days at 8 to 12 DEG C; And
(7) adding 1 to 10% by weight of garlic to the whole Dongchimi material after fermentation in step (6). delete delete The method according to claim 1, wherein the primary fermentation product and the 10% non-fat dry milk are mixed at a weight ratio of 1: 1 in the step (2). The method according to claim 1, wherein the Bacillus subtilis HA (KCCM 10775P) is Bacillus subtilis HA . The method according to claim 1, wherein the lactic acid bacteria are Lactobacillus plantarum K154 (KACC91727P) or Lactobacillus brevis YC10 (KCCM43012). The method of claim 1, wherein the primary fermentation step is performed at 37 to 42 DEG C for 72 hours. The method according to claim 1, wherein the secondary fermentation step fermentes at 25 to 30 DEG C for 48 to 72 hours. A dongchimi base containing GABA produced by the method of claim 1.

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