KR101616187B1 - Method for manufacturing fermented beverage with Schisandra chinensis using GABA producing Lactobacillus - Google Patents

Abstract

The present invention relates to a process for producing fermented Omija beverage using lactic acid bacteria producing gamma-aminobutyric acid (GABA). More particularly, the present invention relates to a method for producing fermented Omija beverage using Lactobacillus plantarum plantarum ) EJ2014 (KCCM11545P). The present invention relates to a method for producing fermented Omija beverage containing high-concentration GABA. The present invention provides a method for producing GABA-enhanced Omija lactic acid fermentation product using Omiza fruit and GABA-producing lactic acid bacteria. The fermentation of GABA-enhanced Schizandra chinensis by lactic acid fermentation in a short period of time is considered to be an excellent fermentation product in terms of economic, nutritional and functional properties. This fermentation technology is considered as a fermentation method that can improve the manufacturing method of enzyme food by conventional sugar - picking.

Description

Technical Field [0001] The present invention relates to a method for manufacturing fermented beverages of Schizandra chinensis using GABA producing Lactobacillus,

The present invention relates to a method for producing a fermented drink of Omija using GABA-producing lactic acid bacteria, and more particularly to a method for producing GABA fermented beverage by using Lactobacillus plantarum plantarum ) EJ2014 (KCCM11545P). The present invention relates to a method for producing fermented Omija beverage containing high concentration GABA.

It has been found that plants present in the natural environment contain a large amount of functional components having biological control functions such as prevention of disease and suppression of aging, and studies on natural food materials have been actively conducted.

Schisandra chinensis ) is an evergreen vine plant belonging to the Magnoliaceae family. It has a distinctive flavor that is a mixture of sweetness, sourness, bitter taste, spicy taste and salty taste, but also a bright red color by anthocyanin . Carbohydrates, crude protein, crude fat, K, Mn, and Ca are abundant in Omija, and fruits are reported to have excellent alcohol detoxification and antioxidant activity. Such Omiza is rich in anthocyanins as well as flavonoids and organic acids, and has been used as a genome, a nourishing agent, a soy sauce, and the like in herbal compositions since ancient times. In addition, Omija is a herbal medicine known to have central inhibitory action, hypotensive action and alcohol detoxifying action, and various physiological functions such as cancer prevention, aging suppression and immunoregulatory action have been reported. On the other hand, the red color of Omiza extract is attributed to anthocyanin and imparts a natural red color to processed products such as tea and liquor.

In the recent global beverage market, beverages account for an increasing proportion of beverages using health functional extracts. Due to these characteristics, omija has attracted attention as a high-commodity raw material.

Gamma amino butyric acid (GABA) is a 4-aminobutyric acid, which is produced by a decarboxylation reaction of L-glutamate substrate. Glutamate decarboxylase (glutamate decarboxylase) ; GAD) and is synthesized as a pyridoxid-5'-phosphate dependent pathway. GABA is a nonprotein amino acid that is not found in proteins. It is a neurotransmitter in the brain or spinal cord. It is known as a brain nutrient that promotes blood flow through the brain and promotes brain metabolism and brain memory by increasing oxygen supply to the brain. GABA is known to inhibit neuronal activity as opposed to glutamate activating neurons, and this function has a profound effect on the function and information processing of neurons, especially in sensory brains such as direction sensitivity, angle sensitivity It is also known to coordinate and elaborate exercise functions. GABA stimulates the cerebral blood flow and increases oxygen supply, promotes the metabolism of brain cells, so that the after effects of stroke and cerebral atherosclerosis is improving and is being used as a medicine. As the function of GABA is known, many studies have been made to use it as a functional food material as well as pharmaceuticals. These studies have focused on increasing the content of GABA, which is naturally contained in brown rice, green tea, malt and cabbage have. However, since there is a limit to the production of GABA using natural plant materials, studies using microorganisms are required for mass production.

Korean Patent Publication No. 10-2013-0084887 (published on July 26, 2013)

It is an object of the present invention to provide a method for producing an omega-lactic acid fermented product in which gamma-aminobutyric acid (GABA) is enhanced by using Omiza fruit and GABA-producing lactic acid bacteria.

In order to achieve the above object, the present invention provides a method for preparing an Omija diluted solution, comprising the steps of: (1) preparing an Omija diluted solution by sifting and diluting Omija; (2) adding 1 to 5 parts by weight of monosodium glutamate (MSG) and 0.1 to 1 part by weight of yeast extract to 100 parts by weight of the total of the omiza diluted solution; (3) a step of inoculating a lactic acid bacteria starter into the mixture, followed by culturing and fermenting the same, wherein the gamma-aminobutyric acid (GABA) is promoted.

The present invention also provides a GABA-enhanced Omija fermented beverage comprising the fermented product of Omija lactic acid produced by the above method.

The present invention relates to a method for producing fermented Omija beverage using lactic acid bacteria producing GABA, and aims at the development of a functional food containing GABA by lactic acid fermentation using a lactic acid bacterium for the fruit having excellent pharmacological effect. We tried to develop a functional fermented product containing lactic acid bacteria enhanced functional substance GABA while preserving color, taste and aroma.

FIG. 1 shows the pH and acidity of the fermentation broth of Schizosaccharomyces lactis according to the dilution factor.
FIG. 2 shows changes in the content of GABA in the fermentation broth of Omija lactic acid according to the dilution ratio.
Fig. 3 shows changes in pH and acidity of the fermentation broth of Omija lactic acid without heat treatment.
FIG. 4 shows changes in pH and acidity of the lactic acid fermentation solution according to yeast extract (YE *) concentration.
FIG. 5 shows changes in the content of GABA in the fermentation broth of Omija lactic acid according to yeast extract (YE) concentration.
FIG. 6 shows the color change of the fermentation broth of Omija lactic acid according to yeast extract (YE) concentration (A: YE 0%, B: YE 0.25%, C: YE 0.5%, D: YE 1%).
Fig. 7 shows changes in pH and acidity of the filtered Schizosaccharomyces cerevisiae.
Fig. 8 shows the change in the GABA content of the filtrated Schizandra chinensis (M + Y = MSG 2%, YE 0.5%).
Fig. 9 shows the color change of the filtered Schizosaccharum fermentation broth (A: control group, B: M + Y).
Fig. 10 shows the process of fermentation of omiga lactic acid by addition of MSG and YE.
Fig. 11 shows pH and acidity changes of the Schizandra chinensis solution according to MSG and YE addition conditions.
Fig. 12 shows changes in the GABA content of the Schizosaccharomyces cerevisiae according to MSG and YE addition conditions (M + Y = MSG 2%, YE 0.5%).
13 shows the color change of the fermentation broth of MSM and YE added with fermentation time according to the fermentation time (A ¹⁾ : MSG 0%, YE 0%, B ²⁾ : MSG 2%, C ^{3 ) 4 )} : MSG 2%, YE 0.5%).
Fig. 14 shows the characteristics of the final product after stepwise mixing the Omija extract with the fermentation broth of Omija.

The present invention relates to (1) a step of preparing an Omija diluted solution by juicing and diluting Omija; (2) adding 1 to 5 parts by weight of monosodium glutamate (MSG) and 0.1 to 1 part by weight of yeast extract to 100 parts by weight of the total of the omiza diluted solution; (3) a step of inoculating a lactic acid bacteria starter into the mixture, followed by culturing and fermenting the same, wherein the gamma-aminobutyric acid (GABA) is promoted.

Preferably, the dilution ratio of the diluted crude acidic milk of step (1) may be 1 to 5 times, but is not limited thereto.

Specifically, the lactic acid bacteria is Lactobacillus plan tareom (Lactobacillus plantarum ). The Lactobacillus plantarum used in the examples of the present invention plantarum ) strain EJ2014 (KCCM11545P).

In detail, the fermentation in the step (3) may be performed at 25 to 30 DEG C for 1 to 120 hours, but is not limited thereto.

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 GABA-enhanced Omija fermented beverage comprising the fermented product of Omija lactic acid produced by the above method.

The Omija fermented beverage can be prepared by mixing the Omija lactic acid fermented product and the raw stock solution of Omija at a ratio of 1-100: 1, but is not limited thereto.

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> Fermentation of Omija lactic acid according to dilution factor

After isolating and isolating Schizandra chinensis, MSG was added 2%, sucrose 5%, yeast extract 0.5%, and diluted 2, 3, 4 times with distilled water. Lt; 0 &gt; C for 30 minutes. Lactobacillus Lactobacillus plantarum ( Lactobacillus plantarum ) EJ2014 strain was cultured in MRS broth for 1 day and 1% starter was added for 3 days. Table 1 shows the results of changes in the number of viable cells in the fermentation broth of Omija. In the two-fold diluted Omija liquid, 5.5 × 10 ⁷ CFU / mL was obtained on the third day and 1.2 × 10 ⁹ CFU / mL was diluted four times. Therefore, the condition of 4-fold dilution of Omija for bacterial growth is considered to be appropriate. Also, the pH and acidity of the fermented product tended to increase with increasing dilution and decrease in acidity (Fig. 1). As the fermentation progressed in 4 times diluted fermentation broth, the pH increased from 3.75 to 4.01 on the 1st day and decreased to 3.54 on the 3rd day. The acidity increased continuously from 1.91% before fermentation to the 3rd day and increased to 2.81% . &Lt; / RTI &gt; When TLC analysis was used to determine the GABA content of the fermented Omija, the GABA content did not differ according to the dilution ratio of the omija (Fig. 2).

Changes in the Number of Live Cells in the Lactic Acid Fermentation Liquor according to Dilution Factor (CFU / mL) Dilution (x) Fermentation time (days) 0 One 2 3 2 3.8.x10 ⁷ 3.7.x10 ⁷ 5.7 × 10 ⁷ 5.5 × 10 ⁷ 3 3.8.x10 ⁷ 5.5 × ^{10 8} 6.7 × ^{10 8} 9.3 × ^{10 8} 4 3.8.x10 ⁷ 8.5 × ^{10 8} 1.1 X ^{10 9} 1.2 × ^{10 9}

< Example  2> Lactic acid fermentation of the untreated fruit of Omiza

Based on the results of <Example 1>, the change in the number of viable cells and the change in sugar content when the omza fruit was diluted 4 times with no heat treatment under the same conditions are shown in Table 2. The number of viable cells was generally higher on the 3rd day than on the heat treatment of 2.1 × 10 ⁹ CFU / mL. The pH decreased from 4.26 to 3.59 at 3 days before fermentation, and the acidity increased from 1.8% to 3.8% at 3 days (Fig. 3). This suggests that the sugar content decreased from 9.40 ° Brix to 8.53 ° Brix on the third day before fermentation, so it seems to use the added sugar as a nutrient and does not seem to affect GABA production.

Number of viable cells, change in sugar content (CFU / mL) of fermented milk of Omija lactate without heat treatment Fermentation time (days) 0 One 2 3 Viable cell count (CFU / mL) 2.9 × 10 ⁷ 2.0 × ^{10 9} 1.1 X ^{10 10} 2.1.x10 ⁹ Soluble solid
(° Brix) 9.40 8.83 8.63 8.53

< Example  3> Sugar Do not add  Yeast extract of non-Omiza fruit yeast  Extraction of Lactic Acid

As shown in Table 3, when the yeast extract (YE) concentration was changed under the condition that no sugar was added, the viable cell counts were 10 ⁹ CFU / mL and maintained until the third day. The pH decreased from 4.14 at 0% to 3.93 at 3 days and slightly increased from 4.13 to 4.30 at 1% addition (Fig. 4). The sugar content decreased by about 1.03 ° Brix from the addition of 0.5% YE to 5.17 ° Brix at 6.20 ° Brix and decreased by 0.76 ° Brix from 6.53 ° Brix to 5.77 ° Brix under the condition of adding 1% YE. As shown in FIG. 5, the GABA content was found to increase in a yeast extract concentration-dependent manner. In the case of 0.5% addition, about 0.25% GABA was added on the third day and 0.25% Was produced. However, the survival rate of MSG remaining in Omija fermentation broth was not converted to GABA. L value was decreased to similar value with fermentation time under 4 conditions. The value of a decreased with increasing yeast extract concentration in concentration - dependent manner (Table 4). b values also decreased with fermentation time depending on YE concentration (Table 4). 6 showing changes in color depending on the fermentation time. As the YE concentration was increased, the original color of the omija was lost and the yellow color became darker and the red color appeared to be lost as the fermentation progressed.

Changes in the Number of Live Cells of Lactic Acid Lactic Acid Fermented with Yeast Extract (CFU / mL) YE
concentration (%) Fermentation time (days) 0 One 2 3 0 3.4 X 10 ⁷ 4.6 × ^{10 8} 3.8 × ^{10 8} 3.4 X ^{10 8} 0.25 3.4 X 10 ⁷ 1.6 X ^{10 9} 1.5 X ^{10 9} 1.5 X ^{10 9} 0.5 3.4 X 10 ⁷ 1.5 X ^{10 9} 1.5 X ^{10 9} 1.3 X ^{10 9} One 3.4 X 10 ⁷ 1.5 X ^{10 9} 1.5 X ^{10 9} 1.4 X ^{10 9}

Chromatographic Changes in the Lactic Acid Lactic Acid Fermentation Solution by the Yeast Extract Concentration (CFU / mL) YE
concentration (%) Color
value Fermentation time (days) 0 One 2 3 0 L ^{1 )} 29.68 28.01 27.67 27.94 a ^{2 )} 17.96 14.99 13.41 10.95 b ^{3 )} 6.53 5.39 5.49 5.55 0.1 L 29.60 25.03 26.32 28.39 a 17.22 11.92 9.22 6.24 b 7.52 5.04 4.86 4.66 0.5 L 27.97 24.34 25.59 25.96 a 16.20 10.55 7.80 6.22 b 9.82 5.58 5.35 5.75 One L 26.48 22.88 17.92 24.83 a 15.27 10.10 7.15 5.91 b 12.63 6.70 6.16 6.96

L ^{1 )} : Lightness, a ^{2 )} : redness, b ^{3 )} yellowness

< Example  4> GABA  Filtration for production optimization

In order to increase the GABA content, the omelet was crushed without using crushed seeds except for the seeds, and then diluted with boiled distilled water. After the fermentation, only juice was collected. When 0.5% YE and 2% MSG were added and fermented for 3 days, the number of viable cells increased to 10 ⁸ on day 1 in the control (control) and maintained until 3 days. In MSG and YE addition conditions of GABA production, it can be seen that maintain increased to 10 ⁹ to day 3 (Table 5). Figure 7 shows the change in pH and acidity, with the control increasing from 3.23 to 3.29 and the acidity decreasing from 1.58% to 1.48% overall. On the other hand, pH of lactic acid fermented with MSG and YE increased from 4.21 to 4.92 and acidity decreased from 1.65% to 0.87%. When GABA content was measured, it was found that GABA was not present in the control (control). In MSG and YE addition conditions, it increased to the size of 0.5% of GABA standard on the 2nd day of fermentation and increased to about 1% (Fig. 8). As shown in FIG. 9, in the control (control), the color did not change and lactic acid fermentation showed that the color changed to yellow as the pH value increased.

Changes in viable cell counts of filtrate and omija fermentation broth (CFU / mL) Fermentation time (days) 0 One 2 3 Control 3.4 X 10 ⁷ 4.6 × ^{10 8} 3.8 × ^{10 8} 3.4 X ^{10 8} M + Y 3.4 X 10 ⁷ 1.5 X ^{10 9} 1.5 X ^{10 9} 1.4 X ^{10 9}

Control: MSG, yeast extract 0%, M + Y: MSG 2%, yeast extract 0.5%

< Example  5> MSG Wow YE  Lactic Acid Fermentation by Addition of Omija

The addition of MSG and YE resulted in the fermentation of succinic acid lactic acid (Fig. 10). After the addition of MSG and YE after the fermentation of Omija, the color change of the fermentation broth of Omija was examined. The same conditions as the previous one were used to dilute only boiled distilled water using only Omiza fruit except for seeds, and only juice was collected after fermentation. The number of viable cell counts increased by more than 10 ⁸ CFU / mL before fermentation at 3.0 × 10 ⁷ CFU / mL before addition of nitrogen source. On the third day of fermentation under conditions using MSG and YE, the number of viable cells was the highest at 1.4 × ^{10 9} CFU / mL (Table 6). YE addition was good for microbial growth and nutrition. As shown in Table 7 showing the sugar content, it decreased to 1.2 ° Brix in the case of control and MSG addition, and decreased to 1.7 ° Brix in the case of addition of YE and MSG and YE together. When pH and acidity were measured (FIG. 11), the pH increased from 3.14 to 4.16 even after addition of MSG before fermentation. As shown in FIG. 13, when MSG was added, red color disappeared and YE Is not likely to have a significant effect. It is thought that the addition of MSG increases the pH value and affects the color. As the fermentation progresses, the pH of the control increases from 3.14 to 3.30 and the acidity decreases from 1.94% to 1.19%. When MSG and YE were added together, the pH increased from 4.19 to 5.03, and acidity changed from 1.74% to 3 days to 0.56%.

In FIG. 13, when the change of color was observed, it was found that red color was maintained under the condition of adding YE, and that yellowness increased on the third day of fermentation. GABA production conditions, MSG and YE, were added together and the red color appeared to be yellow from day 1. When the color was measured, no significant change was observed in the control (control) until the third day, and the a value decreased significantly from 18.24 to 8.61 as the fermentation proceeded under MSG addition conditions. 4.04, and the value of a decreased from 15.74 to 5.39 and the value of b decreased from 7.85 to 6.28 under the condition of addition of MSG and YE (Table 8). When GABA content was measured, MSG was not converted to GABA at all when control and MSG were added, and it was found that GABA was converted into GABA under the condition of adding only YE. MSG, and YE were converted to GABA by exhausting MSG from day 1, and MSG 2% was almost exhausted on day 3, and it was converted to a concentration similar to 1% of GABA standard 12). It was found that glutamic acid decreased from 1.4% before fermentation to 0.4% after fermentation, and GABA increased from 0.02% to 0.92 by about 1% when the free amino acid was quantitatively analyzed (Table 9) .

Changes in the number of viable cells according to MSG and YE addition conditions (unit: CFU / mL) Fermentation time (days) 0 One 2 3 Control 3.0 X 10 ⁷ 2.8 × 10 ⁷ 4.0.x10 ⁷ 8.1.x10 ⁷ MSG 3.0 X 10 ⁷ 4.4.x10 ⁸ 8.3.x10 ⁸ 6.4 × ^{10 8} YE 3.0 X 10 ⁷ 2.0 × 10 ⁷ 5.9 × ^{10 8} 4.2.x10 ⁸ M + Y 3.0 X 10 ⁷ 1.5 X ^{10 9} 2.2 X ^{10 9} 1.4 X ^{10 9}

Change in sugar content according to MSG and YE addition conditions (unit: ° Brix) Fermentation time (days) 0 One 2 3 Control 2.5 1.7 1.5 1.3 MSG 4.6 3.5 3.5 3.7 YE 3.2 2.1 1.9 1.5 M + Y 5.5 4.7 4.0 3.8

Change of chromaticity of Schizandra chinensis according to MSG and YE addition conditions Fermentation time (days) 0 One 2 3 Control L ^{1 )} 22.24 26.71 26.13 25.86 a ^{2 )} 22.38 23.48 23.96 22.89 b ^{3 )} 12.47 13.48 14.40 13.26 MSG L 27.11 32.25 32.05 27.81 a 18.24 10.16 13.09 8.61 b 6.15 3.15 3.70 4.04 YE L 23.18 24.62 23.80 23.75 a 23.05 20.33 17.73 10.54 b 13.66 12.51 9.97 4.53 M + Y ^{4 )} L 24.96 26.26 27.31 26.71 a 15.74 7.92 5.66 5.39 b 7.85 6.23 6.38 6.28

L ^{1 )} : Lightness, a ^{2 )} : redness, b ^{3 )} yellowness, M + Y ⁴⁾ : MSG 2%, YE 0.5%

Free amino acid changes before fermentation and after fermentation (unit: ㎍ / mL) Fermentation time (days) 0 3 Cys 0.00 0.00 ASP 82.67 182.69 GLU 14267.42 4362.14 ASN 48.97 50.88 SER 81.33 4.09 GLN 62.09 91.44 GLY 47.32 60.41 HIS 23.91 20.16 ARG 120.85 122.56 THR 76.01 65.03 ALA 162.40 113.36 GABA 220.49 9276.38 PRO 54.98 57.00 TYR 90.36 54.20 VAL 120.84 103.98 MET 34.43 25.60 Cys2 0.00 0.00 ILE 124.03 89.63 LEU 187.52 134.91 PHE 124.78 69.33 TOTAL 16101.39 15052.27

< Example  6> Prototype of Omija lactate fermentation broth

After fermentation of lactic acid, the color of GABA - enriched Omija fermentation broth tended to be discolored due to the increase of pH of fermentation broth. Therefore, a prototype of the final Omija fermentation broth was completed after mixing the Omija broth with a certain proportion of the Omija broth. When Omiza extract was added at the level of 25 or 50%, a unique red Omiza color could be obtained. Fig. 14 shows the characteristics of the final product after stepwise mixing the Omija extract with the fermentation broth of Omija. Thus, by optimizing the fermentation conditions of useful lactic acid bacteria using Omiza berries, we completed the Omija fermentation product containing lactic acid bacterium probiotic of about 7 × 10 ^{8 in} a short period of time by lactic acid fermentation of the fruit of Omiza fruit containing a large amount of functional substance GABA.

  This is because, in general, the fermented milk of Omija obtained through a long-term fermentation aging process for 6 months or more by general sugar picking tends to avoid consumption by consumers at a high concentration of sugar, and in view of the non- Fermented LAB fermented by lactic acid fermentation is considered to be an excellent fermentation product in terms of economic, nutritional and functional properties. This fermentation technology is considered as a fermentation method that can improve the manufacturing method of enzyme food by conventional sugar - picking.

Korea Microorganism Conservation Center (overseas) KCCM11545P 20140609

Claims (5)

(1) preparing omiza diluted solution by juicing and diluting omija;
(2) adding 1 to 5 parts by weight of monosodium glutamate (MSG) and 0.1 to 1 part by weight of yeast extract to 100 parts by weight of the total of the omiza diluted solution;
(3) Inoculating the above mixture with a starter of Lactobacillus plantarum EJ2014 (KCCM11545P) having the ability to produce gamma-aminobutyric acid (GABA), culturing and fermenting Wherein the gamma-aminobutyric acid (GABA) is promoted in the presence of a reducing agent. The method according to claim 1, wherein the diluted Omija liquid of step (1) has a dilution factor of 1 to 5 times by weight. delete The method according to claim 1, wherein the fermentation in step (3) is performed at 25 to 30 DEG C for 1 to 120 hours. delete

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