KR20160011281A - The method for manufacturing rice bran or barley bran fermented material containing GABA and rice bran or barley bran fermented material containing GABA thereof - Google Patents

Abstract

The present invention relates to a manufacturing method of a rice bran or barley bran fermented material containing gamma-aminobutyric acid (GABA), and a rice bran or barley bran fermented material containing GABA produced by the manufacturing method. The manufacturing method of a rice bran or barley bran fermented material containing GABA comprises the steps of: mixing MSG, rice bran or barley bran, and water; sterilizing the mixture; cooling the mixture; inoculating a functional lactic acid bacteria starter in the mixture; secondarily fermenting the mixture; centrifuging the fermented product; and pulverizing the fermented liquid through lyophilization.

Description

Description: TECHNICAL FIELD The present invention relates to a process for producing fermented rice bran or fermented raw material containing GABA and a process for producing fermented rice bran or barley bran fermented material containing GABA material containing GABA thereof}

The present invention relates to a method for producing a fermented raw material of rice bran or mugang containing GABA and a fermented raw material for rice bran containing the GABA produced by the method, The present invention relates to a method for producing a fermented rice raw material or a fermented raw material containing gaba which is effective for anti-obesity and anti-stress, by producing a fermentation material using the strain Enterococcus faecium MMD-11 ( Enterococcus faecium MMD-11) To a fermented raw material of rice bran or manganese contained therein.

Generally, rice bran is a fine bruise which is separated from the process of removing rice hull from rice and then turning brown rice into white rice. These rice bran contains various amino acids such as iron, calcium, magnesium and vitamin E, and it is known that it improves the brain cell function, improves the memory and improves concentration, and decreases cholesterol and triglyceride, .

The Manganese is also a bark, a skin that comes out of the barley process. These manganese are low in fiber, abundant in protein, have various effects such as constipation, anti-obesity, skin beauty, and are known to be effective in preventing adult diseases.

Rice and barley belong to the four major grains of the world and have been widely used in Korea as an important grain for a long time in Korea. However, rice and barley remained after rice and barley cultivation, respectively, And it is not used for other purposes. Therefore, it is necessary to develop a technology capable of producing high value-added functional food materials from the above-mentioned rice germ and the low-utilization rice.

On the other hand, [γ-aminobutyric acid (GABA)] is a kind of non-protein amino acid distributed in nature. It is found in animals such as brain, heart, kidney and lung and accounts for about 30% of central nervous system precursor neurotransmitters. In plants, many of them are detected in green tea, cabbage roots, etc., and they are also contained in kimchi which is a traditional fermented food in Korea. It is a functional substance with various physiological functions such as inhibition of blood pressure rise, improvement of blood flow in the brain, prevention of obesity, improvement of visual acuity, anti-anxiety, anti-seizure and brain function, prevention of dementia, do. In particular, GABA is a functional substance that is attracting attention as a functional substance for examinees, as well as exhibiting a neurotic effect against insomnia, depression, and anxiety generated during menopause and menopausal period.

Korean Patent No. 10-1373252 discloses a method for producing a fermented brown rice guava leaf and a method for producing a fermented brown rice guava leaf using the fermented brown rice guava leaf material as a prior art document related to the above, Korean Patent Laid-Open Publication No. 10-2014-0046865 discloses a fermented composition containing legumes and rice bran and a method for producing the fermented composition. However, since a strain having the ability to produce GABA is not used, There is a problem that the content is remarkably lowered.

Accordingly, the present invention relates to a method for producing a fermented material by fermenting rice bran or a manganese fermented material and adding a strain capable of producing the same, To provide a raw or fermented raw material for rice bran containing ghava.

Korean Patent Publication No. 10-2014-0046865 (Apr. 21, 2014) Korean Patent No. 10-1373252 (Apr.

The present invention has been made in order to solve the above-mentioned problems of the prior art, and it is an object of the present invention to provide an enterococcus passive MMD-11 The present invention relates to a method for producing a fermented rice raw material or a fermented raw material containing gaba which is effective for anti-obesity and anti-stress by producing a fermented material using a strain Faecium MMD-11, It is an object of the present invention to provide a fermented manganese material.

In order to accomplish the above object, the present invention provides a method of mixing MSG, rice bran, Sterilizing the mixture obtained through the mixing step at 100 to 130 DEG C and 1.2 to 1.8 atm for 10 to 20 minutes; A cooling step of cooling the mixture after the sterilization step to 40 to 45 캜; A step of inoculating 0.05 to 0.1% (w / v) of a functional lactic acid bacteria starter into the mixture after the cooling step; A fermentation step of fermenting the mixture through the inoculation step at 25 to 35 DEG C for 70 to 74 hours; Centrifuging the fermentation product obtained through the fermentation step at 11,000 to 13,000 rpm for 10 to 30 minutes; And a pulverizing step of pulverizing the fermentation liquid obtained through the centrifugal separation step by lyophilization, wherein the inoculating step is an enterococcus passive MMD-11 faecium MMD-11) is cultured in MRS medium at 28 to 32 ° C for 45 to 50 hours, and the obtained functional lactic acid bacteria starter is inoculated.

As described above, the GABA-containing rice bran or fermented raw material according to the present invention can be used as an enterococcus passive MMD-11 ( Enterococcus faecium MMD-11) produced by the present invention can be significantly improved.

As a result, it is possible to produce fermented rice bran or fermented raw material containing GABA by fermentation through a natural material, and produce and produce a fermented material having a high content of GABA.

Also, because of high content of GABA, anti-obesity and anti-stress effect due to GABA is also remarkably high.

Thereby, there is an effect that it contributes to the improvement of the health of the user by providing the rice bran or the fermented raw material containing the GABA which is excellent in nutrition and efficacy.

In addition, the use of rice and manggu which are not used for many purposes can contribute to the rice and the steel industry.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a flow chart showing a method of manufacturing a raw or fermented raw material of rice bran containing a GABA according to an embodiment of the present invention; FIG.
FIG. 2 is a graph showing the pH change of a rice bran-containing fermented rice raw material according to Example 1 of the present invention. FIG.
3 is a graph showing the pH change of a fermented mugwort containing Gabas according to Example 2 of the present invention.
4 is a graph showing changes in acidity of a rice bran-containing fermented rice raw material according to Example 1 of the present invention.
5 is a graph showing changes in acidity of a fermented mugwort containing Gabas according to Example 2 of the present invention.
FIG. 6 is a graph showing the content of goverbose of a rice bran-containing rice bran fermentation material according to Example 1 of the present invention. FIG.
FIG. 7 is a graph showing the content of Gabas in a fermented mugwort containing Gabas according to Example 2 of the present invention. FIG.
FIG. 8 is a graph showing the anti-obesity effect of the rice bran-containing rice bran fermentation material according to Example 1 of the present invention.
FIG. 9 is a graph showing the anti-obesity effect of a fermented mugwort containing Gabas according to Example 2 of the present invention. FIG.
Fig. 10 is a graph showing the anti-stress efficacy of a rice bran-containing fermented rice raw material according to Example 1 of the present invention. Fig.
FIG. 11 is a graph showing antistress efficacy of a crude fermented material containing Gaba according to Example 2 of the present invention. FIG.

Hereinafter, preferred embodiments of the present invention will be described in detail. These examples are for further illustrating the present invention, and the scope of rights of the present invention is not limited to these examples.

1 to 11 of the accompanying drawings, there is shown a method for producing a raw or raw fermented raw material containing Gaba according to the present invention and a method for producing fermented raw materials for raw or fermented raw materials containing GABA Look at the following.

In the method for manufacturing a raw or fermented raw material containing GABA according to the present invention, a mixing step (S10) of mixing MSG (monosodium glutamate), rice bran or steam, and water is performed first.

Wherein the mixing step comprises mixing 0.5 to 1.5 parts by weight of MSG and 15 to 25 parts by weight of rice bran or manganese per 100 parts by weight of water.

When MSG is less than 0.5 part by weight, the MSG content may be lowered by the glutamate decarboxylase, and when the MSG is less than 0.5 part by weight, There is a problem that the MSG is decomposed by a predetermined amount and is no longer decomposed. The rice bran or the manganese is the main constituent material to be fermented when the rice bran or the fermented rice material containing gaba is prepared. When the rice bran or the rice bran is less than 15 parts by weight, the fermentation is not smoothly performed. There is a problem in that the fermentation is no longer possible.

The sterilization step (S20) of sterilizing the mixture obtained at the mixing step at 100 to 130 DEG C and 1.2 to 1.8 atm for 10 to 20 minutes is performed.

A cooling step (S30) for cooling the mixture passed through the sterilization step to 40 to 45 DEG C is carried out.

The inoculation step (S40) of inoculating 0.05 to 0.1% (w / v) of the functional lactic acid bacteria starter into the mixture after the cooling step is performed.

The inoculation step is carried out by inoculating the obtained lactobacillus starter obtained by culturing the Enterococcus faecium MMD-11 having an ability to produce GABA in an MRS medium at 28 to 32 ° C for 45 to 50 hours.

At this time, the Enterococcus passium MMD-11 can be isolated from traditional doenjang.

Also, in the inoculation step (S40), if the functional lactic acid bacteria starter is inoculated at less than 0.05% (w / v), the GABA production capacity may be lowered. If it exceeds 0.1% (w / v) Or more.

The fermentation step (S50) of fermenting the mixture through the inoculation step at 25 to 35 DEG C for 70 to 74 hours is performed.

A centrifugation step (S60) for centrifuging the fermentation product obtained through the fermentation step at 11,000 to 13,000 rpm for 10 to 30 minutes is performed.

A pulverization step (S70) of pulverizing the fermentation broth obtained through the centrifugal separation step by lyophilization is carried out, and the production of the fermented rice bran or fermented maize containing Gaba is completed.

The fermented raw material of rice bran or the fermented raw material containing the above-mentioned processed Gabas is stored at a temperature of -1 to 5 ° C.

Hereinafter, to isolate and identify the enterococcus pathum MMD-11 strain to produce a rice bran-containing fermented rice raw material or a fermented raw material for manganese fermentation, Examples and test examples will be described in detail.

< Experimental Example >

The present invention describes the isolation and identification of Enterococcus faecium MMD-11 strains having GABA production capability in more detail below.

[Materials and Methods]

(1) From traditional miso Gabba  Productive Enterococcus ( Enterococcus ) genus Isolation of candidate strains

In order to improve the functionality of rice germ or fermenting raw material containing GABA, a lactic acid bacterium capable of producing GABA was isolated from traditional doenjang as part of a study on the ability of Enterococcus sp. The mixture was diluted to 10 ^-1 to 10 ^-8 by the number of peptones using conventional doenjang, and 0.1 mL of the diluted solution was dispensed into DE MAN, ROGOSA agar medium (Difco, Detroit, MI, USA). The medium was supplemented with 0.02% Bromocresol purple (BCP) (Sigma Chemical Co., St. Louis, USA) and cultured at 30 ° C for 48 hours to obtain colonies. Among the colonies, those with a deep yellow color without forming a circle were separated into Enterococcus candidates. The possibility of GABA production was investigated using TLC. TLC was performed using silica gel F ₂₅₄ and standard mobile phase (Sigma, USA) and mobile phase (Butanol: Acetic acid: water = 4: 1: 1).

(2) Gabba  Productive Enterococcus ( Enterococcus ) Genus Identification of the strain

Genomic DNA was isolated from candidate strains by DNeasy Blood & Tissue kit (QIAGEN, Hilden, Germany). Purification of the DNA followed the manufacturer's instructions. For cloning of 16S rDNA, Primer was subjected to PCR amplification using 5'-AGAGTTTGATCMTGGCTCAG-3 '(Forward) and 5'-ACGGGCGGGTGTGTRC-3' (Reverse). PCR amplification was performed at 95 ° C for 30 seconds with denaturation at 55 ° C for 30 seconds using annealing (annealing) at 55 ° C using 5 μl / well of 100 ng template DNA, 0.5 μM primer DNA, 2 mM dNTPs, 10 × reaction buffer and Taq polymerase ) And 35 cycles of 1 minute extension at 72 ° C were amplified with a Biometra thermocycler (Tampa, Florida, USA). PCR products were confirmed by 1% agarose gel electrophoresis. Sequence information of MMD-11 was obtained by 16S rDNA sequencing of the confirmed PCR product. Using the obtained sequence information of 16S rDNA, NCBI (www.ncbi.nlm.nih.gov) was used for the homology of DNA and the homology with the nucleotide sequence was compared and analyzed.

(3) GABA-containing Rice bran  or Mangjiang  Starters and traditional miso for manufacturing fermented materials As a result, Detached Enterococcus ( Enterococcus ) genus Characteristics of strains

The API 50 CHL kit (BioMerieux sa, Marcy-L'Etoile, France) was used according to the manufacturer's instructions to investigate the sugar fermentation pattern and biochemical characteristics of the isolated strains. (Lee, JS, KC Lee, JS Ahn, TI Mheen, YR Byun and YH Park. 2002. Pediococcus. Koreensis sp. nov., isolated from kimchi. Int'l J. Syst . Evol. Microbial . 52, 1257-1261.). The strains were inoculated with MRS medium (Difco, Detroit, MI, USA, pH 6.5) supplemented with 1% MSG and cultured at 30 ° C for 30-48 hours. After centrifugation for minutes, the cells and the culture broth were separated and analyzed for Gabba respectively.

(4) Gabba  Containing Rice bran  or Mangjiang  Fermentation material manufacturing

The preparation of raw or fermented raw materials containing GABA was carried out by high pressure sterilization of 1% MSG + 20% raw beef or manganese, and the enterococcus passive MMD-11 ( Enterococcus 0.1% (w / v) of faecium MMD-11) and fermented at 30 ° C for 72 hours. After the fermentation, the fermentation broth was extracted by centrifugation at 12,000 rpm for 20 minutes. After extraction, freeze-drying was performed to produce a fermented rice bran or a fermented maize containing gaba (see FIG. 1).

(5) Gabba  analysis

The GABA assay was performed by mixing 800 μl of a solvent (methanol: chloroform: water = 12: 5: 3 (v / v)) with 200 μl of a fermentation sample. The supernatant, which is an aqueous solution layer, was firstly recovered by centrifugation (12,000 rpm, 4 ° C, 15 minutes). To the organic solvent layer, 600 μl of a mixture of chloroform and water (1: 2 (v / v) And the supernatant was recovered secondarily. The supernatant was recovered by lyophilization. The supernatant was dissolved in ultrapure water and passed through a 0.45 μm PVDF membrane to be used as a sample for HPLC analysis. The samples were derivatized with 6-aminoquilio-N-hydroxysuccinimidyl carbonate (AQC) for analytical HPLC (Waters, Milford, Mass., USA) and analyzed with a 3.9x150 mm AccQ Tag ^TM (Nova-Pak ^TM C18, Waters) was isolated (Seok, JH, KB Park KB , YH Kim, MO Bae, MK Lee and SH Oh. 2008 Production and characterization of Kimchi with enhanced levels of -aminobutyric acid. Food Sci . Biotechnol . 17, 940-946.). The content of GABA was calculated using a standard curve based on the results of HPLC analysis of standard GABA.

[result]

(One) Gabba  Containing Rice bran  or Mangjiang  From traditional doenjang using starter for fermentation material production Gabba  produce Enterococcus ( Enterococcus ) Isolation of Candidate Candidate Strains

In order to isolate lactic acid bacteria producing GABA from traditional doenjang containing starch for the production of rice bran or fermented maize containing GABA, colonies grown on MRS medium supplemented with bromophenol blue were selected as candidates that did not form a ring but were dark yellow . The possibility of GABA production of candidate strains was investigated using TLC. TLC was used to select the strains with excellent GABA production capacity. The morphological and biochemical characteristics of MMD-11 from traditional doenjang were investigated. The isolated strains were Gram-positive strains. The isolated lactic acid strains were L-arabinose, D-xylose, D-galactose Decomposed to produce an acid, and inositol, glucogen, L-xylose and the like were not degraded (see Table 1).

(Table 1) Enterococcus passum MMD-11 ( Enterococcus Morphological and biochemical characteristics of faecium MMD-11)

In Table 1, + means positive and - means negative.

(2) 16S rDNA  Identification of the strain by sequence analysis

The 16S rDNA sequence was analyzed to identify the selected strains. Selectococcus sp. The 16S rDNA sequence of MMD-11 was 98% homologous to the 16S rDNA sequence of Enterococcus faecium strain F6S1 (Access No. KF245564.1) (see Table 2). (Sharpe, ME, TF Fryer, and DG Smith, 1996. Identification of lactic acid bacteria, pp. 65-79, in BM Gibbs and FA Skinner, ed.) Identification Methods for Microbiologists:. part A. Academic Press, Inc., New York, USA) a reference identifying the strains with a Enterococcus faecium strain, and Enterococcus faecium MMD-11. &Lt; / RTI &gt;

(Table 2) Enterococcus faecium MMD-11 base sequence

< Example  1> Enterococcus Passion MMD -11 ( Enterococcus faecium MMD -11) strain Gabba  Containing Rice bran  Manufacture of fermentation materials

1 part by weight of MSG and 20 parts by weight of rice bran are mixed with 100 parts by weight of water to obtain a mixture.

The mixture is sterilized at 121 占 폚 and 1.5 atm for 15 minutes.

The sterilized mixture is cooled to 45 &lt; 0 &gt; C.

Enterococcus to the cooled mixture Lactococcus passive help MMD-11 (Enterococcus faecium MMD-11) was cultured in MRS medium at 30 ° C for 48 hours, and the obtained lactic acid bacteria starter was inoculated at 0.1% (w / v).

The mixture inoculated with the functional lactic acid bacteria starter was fermented at 30 DEG C for 72 hours to obtain a fermented product.

The fermented product is centrifuged at 12,000 rpm for 20 minutes to obtain a fermentation broth.

The above fermentation broth is lyophilized and pulverized to complete the production of a fermented raw material containing GABA.

< Example  2> Enterococcus Passion MMD -11 ( Enterococcus faecium MMD -11) strain Gabba  Containing Mangjiang  Manufacture of fermentation materials

To 100 parts by weight of water, 1 part by weight of MSG and 20 parts by weight of manganese are mixed to obtain a mixture.

The mixture is sterilized at 121 占 폚 and 1.5 atm for 15 minutes.

The sterilized mixture is cooled to 45 &lt; 0 &gt; C.

Enterococcus to the cooled mixture Lactococcus passive help MMD-11 (Enterococcus faecium MMD-11) was cultured in MRS medium at 30 ° C for 48 hours, and the obtained lactic acid bacteria starter was inoculated at 0.1% (w / v).

The mixture inoculated with the functional lactic acid bacteria starter was fermented at 30 DEG C for 72 hours to obtain a fermented product.

The fermented product is centrifuged at 12,000 rpm for 20 minutes to obtain a fermentation broth.

The above fermentation broth is lyophilized and pulverized to complete the production of a fermented fermented muggar containing GABA.

< Test Example  1> pH  change

The pH change of each of the above-prepared Example 1 and Example 2 was measured for each day when fermentation was performed for 3 days after inoculation of the strain.

The pH change in Example 1 was 6.5 at day 0, decreased from day 1 to 5.88, and decreased to 5.06 at day 3 (see FIG. 2). The pH change of Example 2 was 6.04 at day 0, and there was no significant change by day 1 and decreased to 4.82 at day 3 (see FIG. 3).

< Test Example  2> Change of pH

The fermentation was carried out for 3 days after inoculation of the strains prepared in Example 1 and Example 2, and acidity changes were measured for each day.

The acidity change of Example 1 was 0.32 on day 1 and increased to 0.75 on day 3 (see FIG. 4). The acidity change of Example 2 was 0.31 on day 1 and increased to 0.49 on day 3 (see FIG. 5).

< Test Example  3> Gabba  content

For the above prepared Example 1 and Example 2, when the fermentation was carried out for 3 days after inoculation of the strain, the amount of goverberry was measured.

It can be seen that in Examples 1 and 2, the amount of the gas is increased over time (see FIGS. 6 and 7). This shows that it is possible to produce fermented rice bran or fermented raw material containing GABA by fermentation through a natural material, and can produce a high-content FG fermented material.

< Test Example  4> Anti-obesity  efficacy

In Example 1 and Example 2, 3T3-L-1 cell anti-obesity effect was measured after fermentation for 3 days after strain inoculation.

The 3T3-L-1 cells were adipocytes, and control (control) in which 3T3-L-1 cells were not treated with the 3T3-L-1 cells, When the fermented raw material of rice bran showing the result of treating the rice bran and the 3T3-L-1 cells with the rice bran fermented material of Example 1 was examined, the rice bran in the measured 3T3-L- While the rice bran fermented material shows a significantly lower value than the Control.

In addition, control (control) in which 3T3-L-1 cells were not treated with the above 3T3-L-1 cells, As for the 3T3-L-1 cell counts measured in the fermented Manganese fermented material showing the result of treating the Fermented Manganese fermented material of Example 2, the fermented manganese showed no significant difference from the Control in the same manner as in Example 1, The fermented material shows a significantly lower value than the above Control.

As a result, it can be seen that the rice bran fermentation material (Example 1) and the fermented rice material (Example 2) inhibit adipocyte differentiation and fat accumulation and have anti-obesity effect.

< Test Example  5> Antistress  efficacy

The cells were fermented for 3 days after the inoculation of the strains in Example 1 and Example 2, and the cell antistressing activity was measured.

10, control (control) in which the cells were subjected to no treatment, rice bran showing the result of treatment with only rice bran in the cells, and fermentation inoculated with the strain having the ability to produce GABA in the rice bran of Example 1 In the case of the rice gangue + garbage showing the result of treatment of the material, the rice gangue did not show much difference from the control in the measured cell viability, but the rice gangue + garbage showed a significantly higher value than the control.

11, control (control) in which the cells were subjected to no treatment, manganese showing a result of treating only the manganese in the cells, and a strain in which the manganese of Example 2 was capable of producing GABA was inoculated As for the measured cell viability of the Manganese + GABA showing the result of treating the fermented material, the Makkang had no significant difference from the Control in the same manner as in Example 1, while the Makkang + GABA had a significantly higher value than the Control Respectively.

As a result, the cell viability was enhanced when the rice gangue + gaba (Example 1) and the rice gangue + gaba (Example 2) were treated, indicating that there is an anti-stress effect against the stress.

Claims (5)

A mixing step (S10) of mixing rice bran or steam, MSG and water;

(S20) sterilizing the mixture obtained through the mixing step at 100 to 130 DEG C and 1.2 to 1.8 atm for 10 to 20 minutes;

A cooling step (S30) of cooling the mixture after the sterilization step to 40 to 45 DEG C;

An inoculation step (S40) of inoculating 0.05 to 0.1% (w / v) of the functional lactic acid bacteria starter into the mixture after the cooling step;

A fermentation step (S50) of fermenting the mixture after the inoculation step at 25 to 35 DEG C for 70 to 74 hours;

A centrifuging step (S60) of centrifuging the fermentation product obtained through the fermentation step at 11,000 to 13,000 rpm for 10 to 30 minutes;

And a pulverization step (S70) of pulverizing the fermentation liquid obtained through the centrifugal separation step by lyophilization.
The method according to claim 1,
The mixing step
A process for producing a fermented raw material for maize or manganese, which comprises mixing 0.5 to 1.5 parts by weight of MSG with 15 to 25 parts by weight of rice bran or manganese per 100 parts by weight of water.
The method according to claim 1,
The step of inoculation
Wherein the functional lactic acid bacteria starter is composed of Enterococcus faecium MMD-11 having an ability to produce GABA.
The method of claim 3,
The step of inoculation
The enterococcus passive MMD-11 ( Enterococcus The present invention also relates to a method for producing a fermented rice raw material or a fermented manganese fermented product, comprising the step of inoculating the obtained lactic acid bacteria starter with the fermented lactic acid bacteria starter, which is obtained by cultivating faecium MMD-11 in an MRS medium at 28 to 32 ° C for 45 to 50 hours.
6. A fermented raw material for rice bran or manganese containing the gabas prepared by the method of any one of claims 1 to 4.

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