KR101917534B1 - Vegetable Brown Sauce with enhanced taste and anti-oxydant property and preparation method thereof - Google Patents

Abstract

Disclosed are: vegetable brown sauce which contains sauce stock of fermented soybeans fermented by using a specific medical herb complex extract and Bacillus amyloliquefaciens MGD02 strains, has excellent palatability by shielding the bad smell from the fermented soybeans, and contains enhanced physiologically active substances so as to improve antioxidative activity; and a preparation method thereof. The vegetable brown sauce of the present invention is based on 100% pure vegetable, is excellent in palatability, and has excellent nutrition and antioxidant activity, and thus can substitute meat sauce meeting the modern health-oriented trend, and can be good for soy meat for vegetarians.

Description

[0001] The present invention relates to a vegetable brown sauce having enhanced palatability and antioxidative activity, and a method for producing the vegetable brown sauce,

The present invention relates to a vegetable brown sauce having enhanced palatability and antioxidative activity and a method for producing the same. More particularly, the present invention relates to a vegetable brown sauce having enhanced palatability and antioxidant activity, The present invention relates to a vegetable brown sauce having enhanced antioxidative activity by enhancing physiologically active substances and a method for producing the same.

Sauce is derived from Latin salsa, meaning salt-based seasoning solution. In France, England, Korea and Japan, sauce, salsa in Italy and Spain, sosse in Germany, 'In China,' zhi 'in India,' chartni 'in India, and various sources using different materials according to social and geographical conditions. Because of the variety of sauces, the classification standard varies according to color, usage, main ingredients, etc., and reaches to about 500 kinds. Generally, there are five kinds of white sauce, brown sauce, yellow sauce, red sauce, , And brown sauce is most widely used for cooking.

Brown sauce is the most widely used source of Western meat (sauce stock) as a typical meat source in Western countries after boiling with an appropriate amount of spice and wine and then concentrating it with a thickening agent . Brown seaweed is produced by boiling and concentrating meat, bone, and vegetables for a certain period of time (Korean Patent No. 10-1254635, Korean Patent No. 10-1254643). However, brown sauce has been required to develop a vegetable brown sauce that can not replace the trend of using meat stocks to consume meats, and can replace them.

There has been an attempt to produce sauce using the chongkukjang. However, there is a problem that the odor characteristic of the chonggukjang is not completely removed and the palatability is poor. Also, as the chongkukjang is diluted with the source, the antioxidant activity and the physiologically active substance This final source product is insignificant and needs to be improved.

Accordingly, the present inventors have found that when mixed with a specific herbal composition extract and a source stock of chungkukjang fermented with Bacillus amyloliquefaciens MGD02, the odor derived from chungkukjang is shielded and excellent palatability is obtained It is possible to produce a vegetable brown sauce in which a physiologically active substance is enhanced and an antioxidative activity is enhanced, and the present invention has been accomplished.

Accordingly, an object of the present invention is to provide a herbal composition comprising a herbal composition extract of Lilium japonicum, Angelica japonica, Rhododendron japonica and Bacillus amyloliquefaciens MGD02, and a source stock of Chungkukjang fermented with a strain of Bacillus amyloliquefaciens MGD02, To provide a vegetable brown sauce having enhanced active substances and antioxidative activity, and a method for producing the same.

In order to attain the above object, the present invention provides a herbal composition extract of Jangsoobil, Angelica japonica, Doraji and Baeguso and a source stock of Chungkukjang fermented with Bacillus amyloliquefaciens MGD02, And a vegetable brown sauce having enhanced physiologically active substances and enhanced antioxidant activity.

The term " stock " in the present invention refers to the undiluted solution that underpins the manufacture of the source.

A. herbal compound extract

In the present invention, the herbal composition extract is prepared by extracting a mixed powder of Liliaceae, Angelica keiskei, Rhodiola and Rhodiola with ethanol, and adjusting the soluble solids to 3 to 5 brix.

Perilla frutescens is a herb known to have anti-inflammatory, anti-allergic, antimicrobial and antitumor effects. Pharmacologically active ingredients include phenolic compounds (phenolics), rosmarinic acid, anthocyanin, caffeic acid, .

Angelica gigas Nakai) is known to have a blood-replicating effect that replenishes blood, and its pharmacologically active components include coumarin derivatives deckacin, deacercin, umbeliferone, and β-sitosterol.

Platycodon grandiflorum ) are used for food and medicinal purposes and are root plants that are effective for bronchial diseases such as drainage, genomes, bronchitis and asthma. Platycodin A, C and D, which are triterpenoid saponins, In addition, other pharmacologically active ingredients include inulin, betulin, and stigmasterol.

Cynanchum wilfordii Hemsley) is used as a herbal medicine in herbal medicine. It has been known to be effective for the prevention of osteoporosis including osteopenia, skeletal growth and induction of insulin-like growth factor, and improvement of lipid metabolism of hyperlipidemia. A, 2,5-dihydroxyacetophenone, synanone A, and the like.

The mixed powder of herbal medicine combination extract contains 50 ~ 70% (w / w), 15 ~ 25% (w / w), 10 ~ 25% (w / w) (Example 4).

For mixed powder extraction, the composite powder is extracted with 50 to 70% ethanol and filtered to adjust the soluble solid content so that the soluble solid content is 3 to 5 brix. If the soluble solids content is outside the above range, the efficacy is weak or the herbal odor is strong, and there is a problem that the cost for concentrating may be excessively increased.

In the present invention, the herbal composition extract greatly enhances the antioxidative activity of the final sauce and the content of the physiologically active substance (Example 4).

It is preferable that the herbal composition extract is contained in the brown sauce of the present invention in an amount of 10 to 20% by weight. If the content is less than 10% by weight, the antioxidant ability is weak. If the content is more than 20% by weight, the odor of the herb is strong.

B. Cheonggukjang sauce stock

In the present invention, a source stock of chungkukjang fermented with Bacillus amyloliquefaciens MGD02 strain is used.

The present inventors have succeeded in isolating various Bacillus species from soy sauce (soy sauce) Bacillus amyloliquefaciens MGD02, which is the most suitable strain for the production of chungkukjang stock for brown sauce of the present invention, was isolated and identified through nutrition and antioxidation of fermented chongkukjang, (KACC) on Dec. 12, 2016, and received the accession number KACC92158P (Example 2).

The identification of Bacillus amyloliquefaciens MGD02 strain was carried out by Gram stain and transmission electron microscopic observation by morphological observation, and API50CHB kit for assaying physiological and biochemical characteristics was used for analysis of cell wall fatty acid (MIDI Analysis) and 16S rDNA sequencing. It was found that the Bacillus amyloliquefaciens MGD02 strain of the present invention based on the 16S rRNA base sequence belongs to the genus Bacillus (Fig. 3).

Chungkookjang fermented with the Bacillus amyloliquefaciens MGD02 strain of the present invention has excellent palatability due to its reduced characteristic odor and also has excellent nutritional and enhanced antioxidative activity and physiologically active substance and can be used as a material of Chungkookjang stock have.

In the present invention, the fermented chungkukjang is dried to prepare a dried chungkukjang, and then water is added to the dried chungkukjang, followed by boiling and concentration to prepare a chunggukjang sauce stock.

The addition amount of the chungkukjang sauce stock is preferably 50 to 70% by weight based on 100% by weight of the total weight of the brown sauce. When the content is less than 50% by weight, the flavor of the sauce is insufficient due to the fermented product.

According to still another aspect of the present invention, there is provided a method for producing a vegetable brown sauce comprising the steps of: (a) providing a vegetable brown sauce having an odor from chungkukjang, which is excellent in palatability, :

I) fermenting soybean with Bacillus amyloliquefaciens MGD02 to prepare and drying a cheonggukjang;

Ii) adding water to the dried chungkukjang, boiling and concentrating to produce a stock of chungkukjang;

Iii) adding a vegetable ingredient, roasted chickpea stock and seasoning prepared in step ii) to roasted flour, boiling, concentrating and filtering; And

Iv) preparing a herbal composition combination extract of Liliaceae, Angelica keiskei, Doraji and Bacillus subtilis and adding to the filtrate of step iii).

Step i) Chungkukjang manufacturing

The beans are fermented with Bacillus amyloliquefaciens MGD02 strain to prepare and dry the cheonggukjang.

In the present invention, soybean can be used in white beans (yellow soybeans), black beans (black beans), and snow beans (beans). The beans are preferably poured into water at a rate of 2 to 3 times, followed by being blanched for 6 to 24 hours to remove water, and then boiled at 100 to 120 ° C for 15 to 60 minutes and cooled to 35 to 45 ° C. But is not limited to.

The novel Bacillus amyloliquefaciens MGD02 according to the present invention is used as a seed culture according to a conventional method. The culture can be carried out as a culture medium in a TSB (Trytic Soy Broth) liquid medium for about 12 hours, but is not limited thereto. It is preferable that the inoculation amount is 2.5 to 5.0% (v / w). If it is more than 5.0% (v / w) Which may affect the fermentation and quality of Chungkookjang.

The fermentation is preferably performed at 35 to 45 DEG C for 2 to 5 days, but is not limited thereto.

The drying is preferably performed at 50 to 60 DEG C for 2 to 4 days, but is not limited thereto. By using the dried fermented soy sauce, it is possible to prevent the quality change by continuous fermentation of the fermented soy sauce, and to prevent the occurrence of bad smell due to over fermentation and to maintain a constant quality.

Step ii) stock  Produce

Water is added to the dried Chungkookjang, boiled and concentrated to produce Chungkukjang stock.

The amount of water to be added is not limited, but may be 5 to 10 times the volume of the dried chungkukjang.

Concentration can be carried out by a conventional method. For example, when the mixture is boiled, the mixture is boiled for a certain time and boiled for a certain time. The extracted chungkukjang extract is boiled for a certain period of time and concentrated to an appropriate concentration.

The cheonggukjang sauce stock according to the present invention is excellent in palatability due to reduced odor peculiar to Cheonggukjang (Example 5).

Step iii) source preparation

To the roasted flour (roux), the vegetable ingredient, the chungkukjang stock prepared in step ii) and the seasoning are added, boiled, concentrated and filtered.

In the above, roux (roasted flour) may be prepared by pouring olive oil into a pan at a temperature of about 130 ° C, adding flour, lowering the flame, stirring to avoid sticking, and frying until brown

The vegetable ingredient may include, but is not limited to, tomato, onion, carrot, celery, garlic, radish, kiwi, cabbage and the like.

Seasonings may include, but are not limited to, sugar, salt, plum, vinegar, wine, laurel leaf, pepper, and the like.

The amount of the chungkukjang stock added is preferably 50 to 70% by weight based on 100% by weight of the total weight of the brown sauce.

Concentration and filtration (of solids) can be carried out in a conventional manner. As an example, it is heated at a high temperature for the first time, boiled, then reduced to medium-heavy, boiled for a certain period of time and concentrated to an appropriate concentration, and filtration can be carried out using a plate.

Step iv): Addition of herb complex extract

After preparing the herbal medicine complex extract of Lilium japonicum, Angelicae japonica, Rhododendron japonica, and Bacillus subtilis, it is added to the filtrate of step iii) to complete the brown sauce.

The herbal composition extract and its preparation are as defined above.

A mixed powder of 50 to 70% (w / w) of lobulus, 15 to 25% (w / w) of Angelica japonica, 10 to 25% (w / w) of bellflower and 5 to 10% Extracted with ~ 70% ethanol and filtered to prepare a herbal composition extract by adjusting soluble solids to a solids content of 3 ~ 5 brix.

The amount of the herbal composition extract added is preferably 10 to 20% by weight based on 100% by weight of the total weight of the brown sauce.

The brown sauce according to the present invention is a vegetable source which can replace meat brown sauce which is excellent in palatability and nutritive and antioxidative activity by reducing odor of chongkukjang even though it is 100% vegetable.

The Bacillus amyloliquefaciens MGD02 strain according to the present invention enhances nutritional and antioxidative activity while reducing odor of chongkukjang, and is useful as a fermentation seed of Chungkukjang. The chungkukjang sauce stock of the present invention produces a vegetable brown sauce which is shielded from odor originating from chungkukjang, and in which palatability is remarkably improved.

The vegetable brown sauce according to the present invention contains chungkukjang sauce stock and has excellent odor from the offensive odor derived from chungkukjang, and has the antioxidant activity enhanced and enhanced by the physiologically active substance derived from the herbal composition extract.

The vegetable brown sauce according to the present invention is 100% vegetable, excellent in palatability, has superior nutritional and antioxidant activity, and can meet the modern health-oriented trend and can substitute meat sauce, and is suitable for veggies to be.

1 is an example of a process for producing vegetable brown sauce according to the present invention.
2 shows the nucleotide sequence of 16S rRNA gene of Bacillus amyloliquefaciens MGD02 strain.
3 is a phylogenetic relationship diagram of Bacillus amyloliquefaciens MGD02 strain and other Bacillus strains.
4 is a graph comparing antioxidative activities of soaked soybeans, augmented soybeans, Bacillus amyloliquefaciens Ja-8 fermented Chungkookjang, and Bacillus amyloliquefaciens MGD02 fermented Chungkukjang. a is a DPPH radical scavenging activity, b is an ABTS radical scavenging activity, and c is a hydroxyl radical scavenging activity.
5 is a graph showing the palatability evaluation of the vegetable brown sauce of the present invention. 5a is brown sauce containing 0% of herbal compound extracts (control), 5b is brown sauce containing 10% of herbal extracts, and 5c is a result of palatability evaluation of brown sauce containing 20% of herbal extracts.

The present invention will be described in more detail by the following examples. These examples are for illustrating the present invention, and the scope of the present invention should not be limited by them.

Example

Example  1. Strain isolation and selection

<Strain isolation>

1 g of doenjang (trade name: Mongolia) was suspended in 9 ml of sterilized physiological saline, vigorously shaken, left for 5 minutes, and the resulting supernatant was used as a sample for strain isolation.

The samples were diluted with sterilized physiological saline to show 30-300 colonies per plate and plated on a plate medium of TSA (Tryticase Soy Agar, DIFCO, USA), and incubated in a thermostat at 35 ° C for 48 After culturing for 72 hours, specific colonies of various Bacillus species were taken and purely separated using a plate medium of the same composition. The isolated strains were suspended and cryopreserved in TSB (Trytic Soy Broth, DIFCO) containing 20% (v / v) glycerol, respectively, and used as needed in TSA plate media.

<Odor Reduction Chungkookjang fermentation strain selection>

The pure isolates were inoculated into a 250 ml Erlenmeyer flask containing 50 g of boiled soybeans and fermented with Chongkukjang at 39 ° C in a thermostat, and the content of isoflavone-aglycone and odor generation were measured by ammonia content and sensory evaluation , And 10 strains were selected first (Table 1).

The non - glycosylated isoflavones were prepared by lyophilizing fermented chungkukjang, respectively. Powder was prepared by adding 20 ml of 50% methanol to 1 g of powder and then extracted at room temperature for 24 hours. The filtrate was filtered through 0.45 ㎛ filter and used as an analytical sample Respectively. The prepared samples were analyzed by HPLC. The mobile phase solvent was 0.2% glacial acetic acid (in water) (solution A) and 0.2% acetonitrile (in glacial acetic acid) (solution B) using a Lichrophore 100 RP C18 column (4.6 mm, Respectively. The mobile phase conditions were maintained at 0 min -100%, 15 min-90%, 25 min-80%, 35 min-75%, 45 min-65% and 50 min-65% on the A solvent basis. 20 μl of sample was injected and the mobile phase velocity was maintained at 1 ml / min at 30 ° C. The detector was detected at 254 nm using a diode array detector (DAD) and the results are shown in Table 1.

Ammonia content was analyzed using an automatic amino acid analyzer (Hitachi, L-8900, Japan). 1 ml of the sample was weighed into a test tube, and 4 ml of distilled water was added, followed by hydrolysis at 60 ° C for 1 hour. Then, 1 ml of 10% 5-sulfosalicylic acid was added, and the mixture was allowed to stand at 4 ° C for 2 hours to precipitate proteins. The filtrate was filtered through a glass filter, and the filtrate was concentrated under reduced pressure at 60 ° C to completely evaporate the water. The concentrated sample is lithium citrate buffer (pH 2.2) was analyzed liquid filtrate was filtered through a membrane filter 0.45㎛ (Dismic-25CS) was dissolved in 2ml in automatic amino acid analyzer, the sensory evaluation of the odor level is Kyungnam University of Science and Technology students and general population 30 The results are shown in Table 1. The results are shown in Table 1. &lt; tb &gt;&lt; TABLE &gt;

Isolated strain Non-glycoside isoflavone
(/ / G) Odor level Ammonia (mg / g) Sensory score Miso MGD01 223.45 208.43 4.88 MGD02 292.77 77.52 3.63 MGD03 225.83 206.86 4.62 MGD04 283.3 136.91 4.35 MGD05 312.62 114.10 4.03 MGD06 239.12 110.10 3.99 MGD07 279.83 101.67 4.05 MGD08 260.46 140.08 4.42 MGD09 239.77 106.72 4.12 MGD10 238.28 112.77 4.19 Note 1) All experiments were repeated three times and expressed as average values.
Note 2) Sensory evaluation score means that the higher score score is, the more bad odor is.

As shown in Table 1, the MGD02 strain having the least amount of ammonia and having the best sensory evaluation was finally selected.

Example  2. Bacillus Amyloliquefaciens MGD02 Identification of strain

The morphological, biochemical, fungal fatty acid composition, and molecular genetic characteristics of MGD02 strains selected in the above examples were significantly reduced.

<Morphological / biochemical characteristics>

The morphological observations were as follows: Gram-positive, rod-shaped Bacillus strains were observed with a microscope after Gram stain. The viable temperature was 20 ~ 45 ℃, the growth pH was 3.0 ~ 9.0, To 8%. Biochemical characterization of the strain using the API50CHB kit (BioMerieux, France), which is a method for testing the sugar availability of bacteria, resulted in the magnetization of 17 carbon sources including glycerol and was assumed to be a tuberous anaerobic Bacillus sp. Strain Table 2).

Characteristic reaction Characteristic reaction shape   D-Tagatose -   shape Rod   D-Fucose -   Gram stain +   Ribose +   Cell dimension 0.5-10 탆   D-Xylose - Physiological properties   L-Xylose -   Aerobic growth +   Adonitol -   Growth at   β-Methyl-xyloside -    10 ℃ -   Galactose -    20 ℃ w   D-Glucose +    25 ℃ +   D-Fructose +    30 ℃ +   D-Mannose +    35 ℃ +   L-sorbose -    40 ℃ +   Rhamnose -    45 ° C +   Dulcitol -    50 ℃ -   Inositol -    55 ° C -   Mannitol -   Growth in NaCl   Sorbitol -    0% +   alpha methyl-D-mannoside -    2% +   alpha methyl-D-glucoside -    4% w N Acetyl glucosamine +    6% w   Amygdaline +    8% w   Arbutine +    10% -   Esculine +    12% -   Salicine +    14% -   Cellobiose +   pH   Maltose +    3 w   Lactose +    5 +   Melibiose -    7 +   Saccharose +    9 w   Trehalose +    11 -   Inuline - carbohydrate   Melezitose -   Control -   D-Raffionse +   Glycerol +   Amidon -   Ertythritol -   Glycogene -   D-Arabinose -   L-Fucose -   L-Arabinose -   D-arabitol -   Xylitol -   L-Arabitol -   β Gentiobiose +   Gluconate -   D-Turanose -   2 ceto-gluconate -   D-Lyxose -   5 ceto-gluconate - +, Positive; -, negative reaction; w, weak reaction.

&Lt; Analysis of fatty acid composition of fungi &

For the analysis of the fatty acid composition, 1 ml of MGD02 culture medium was taken in a test tube, 4 ml of 0.5 N methanolic NaOH was added, and the mixture was heated for about 10 minutes using a heating block at 100 ° C to hydrolyze the fatty acid and glycerol. Then, 2 ml of boron trifluoride (BF ₃ ) was added and stirred, and the mixture was heated again for 30 minutes to proceed the methyl esterification of the fatty acid. After completion of the reaction, 1 ml of isooctane was added, and the mixture was vigorously shaken. Then, centrifugation and only the isooctane layer were collected, dehydrated with anhydrous sodium sulfate, and then filtered through a 0.45 μm membrane filter (Dismic-25CS) to obtain GC and phenylmethyl silicone fused silica capillary Fatty acids were detected using a column (2.5 mm, Hewlett Packard) and finally analyzed using a MIDI program.

As a result of the analysis, the C15: 0 anteiso contained in the Bacillus sp. Strain was 42.71%, indicating that the MGD02 strain of the present invention was also found to be Bacillus spp. (Table 3).

fatty acid content (%) Linearly saturated   C14: 0 iso 1.33   C14: 0 1.38   C15: 0 iso 15.97   C15: 0 anteiso 42.71   C16: 0 iso 3.16   C16: 0 12.06   C15: 0 iso 3OH 0.45   C15: 0 2OH 0.42   C17: 0 iso 6.91   C17: 0 anteiso 11.59   C18: 0 1.44 Single-unsaturated C16: 1 w 11 c 1.16 C17: 1 iso w 10 c 0.38 C18: 3 w 6 c (6, 9, 12) 0.57 Summed feature 2 * 0.46 * Summed feature 2: C16: 1 iso I or C14: 0 3OH

<Molecular Genetic Characterization: 16S rRNA Sequence Analysis>

Molecular genetic identification of MGD02 strain was carried out by shaking culture of MGD02 in TS liquid medium for 2 days and isolating genomic DNA using Intron Genomic DNA purification kit (Intron Botechnology Co., Suwon, Korea) rDNA sequence was determined.

The PCR reaction was denatured at 95 ° C for 5 minutes, DNA annealed at 49 ° C for 30 seconds, and elongated at 72 ° C for 1 minute. This reaction was repeated 30 times. The primer production was made to be about 1,500 bp in 16S rDNA preservation area of bacteria. The forward primer used was 5'-CGGAGAGTTTGATCCTGG-3 ', and the reverse primer used was 5'-TACGGCTACCTTACGAC-3'. After completion of the PCR reaction, the 16S rRNA fragment was confirmed by electrophoresis and DNA was separated from the agarose gel according to the method described in the Total Fragment DNA Purification Kit (Intron Biotechnology). The nucleotide sequence of the purified 16S rRNA fragment was determined as a template. The nucleotide sequence was determined by the dideoxy chain termination method using a PRISM Ready Reaction Dye terminator / primer cycle sequencing kit (Perkin-Elmer Corp., Norwalk, Conn., USA). As a result, a nucleotide sequence of 1,517 bp was determined ).

The determined nucleotide sequence was compared with 16s rDNA of another bacterium. The similarity value was calculated from the alignment and evolution distance using a DNAMAN analysis system (Lynnon Biosoft), and a phylogenetic tree was created. 3.

As shown in Fig. 3, it showed 99% similarity with Bacillus amyloliquefaciens strains.

The MGD02 strain was named as the final Bacillus amyloliquefaciens WCP02 in consideration of morphological, biochemical, fats composition and molecular genetic characteristics, and was deposited on December 12, 2016 at the National Institute of Agricultural Science and Technology (KACC) And received grant number KACC 92158P.

Example  3. Bacillus Amyloliquefaciens MGD02  Production of fermented Chungkukjang and dried chungkukjang powder

The fermented chungkukjang and dried chungkukjang powder fermented with the Bacillus amyloliquefaciens MGD02 strain of the present invention were prepared and the fermented soybeans before and after fermentation and the Bacillus amyloliquefaciens Ja-8 strain for the preparation of the conventional source chungkukjang (accession number: KACC92015P ) Were compared with those of fermented Chungkookjang.

<Cheongkukjang and dried chungkukjang>

1 kg of soybeans was immersed in about 2 times of water for about 12 hours and then drained for about 2 hours. The mixture was heated at 120 ° C for 30 minutes and then cooled to about 40 ° C. The soybeans were put into a fermentation mold, 2.5% (55 ml) of Bacillus amyloliquefaciens MGD02 cultured in the medium for 12 hours was inoculated and fermented at 37 ° C for 72 hours to prepare chungkukjang. The chungkukjang was dried at 50 ~ 60 ℃ for 2 days and dried Cheonggukjang was prepared. On the other hand, as a comparative example, Chungkukjang and dried chungkukjang were also prepared by using Bacillus amyloliquefaciens Ja-8 strain.

<Sample Preparation>

Twenty times (200 ml) of methanol was added to 10 g of each dried chungkukjang powder and extracted at room temperature for 12 hours. The extract was centrifuged and the supernatant was filtered with a 0.45 μm membrane filter (Dismic-25CS), and a portion of the supernatant was used to measure total phenolics and isoflavone content. The remaining extracts were dissolved in extraction solvent to give 0.25, 0.5 and 1.0 mg / ml And used for the evaluation of radical scavenging activity. As a comparative example, a sample was also prepared in the same manner as the soaked soybean and the soybean soybeans before fermentation.

<Antioxidant Activity Test>

Antioxidant activity was determined by measuring the scavenging activity of DPPH (1,1-diphenyl-2-picrylhydrazyl), ABTS (2,2'-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid) and hydroxyl .

DPPH radical scavenging activity was carried out by adding 0.8 ml of DPPH solution (1.5 ^-4 M) to 0.2 ml of each of the samples prepared above, uniformly mixing, leaving for 30 minutes, and measuring the absorbance at 525 nm. The negative control of the DPPH radical scavenging activity proceeded in the same manner using distilled water instead of the sample, and the difference in absorbance was calculated as a percentage (%) according to the following equation, and the result is shown in FIG.

Radical scavenging activity (%) = [1- (negative control absorbance / experiment absorbance)] × 100

The ABTS radical scavenging activity was obtained by mixing 5 ml of ₇ mM ABTS reagent and 5 ml of 140 mM K ₂ S ₂ O ₈ (FW 270.3, Sigma 9392) in a dark place for 14 to 16 hours to generate cation radicals. The absorbance value of the ABTS solution was adjusted to 0.7 ± 0.02. 0.1 ml of each sample and 0.9 ml of ABTS solution were mixed and reacted for 3 minutes and absorbance was measured at 732 nm. The ABTS radical inhibitory activity was also measured by the same method using distilled water instead of the sample as the negative control, and the difference in absorbance was calculated as a percentage (%) by the above equation. The results are shown in FIG.

Hydroxyl radical scavenging activity was measured 10mM FeSO _{4. 7H 2 0-EDTA 0.2ml, 10mM} 2- deoxyribose After reacting for 4 hours at _{0.2ml, 10mM H 2 O 2 0.2ml} , 1.4ml extract was mixed after 37 ℃ made of a mixture of 1% in this mixed solution, thio 1 ml of each of barbitic acid (in DW) and 2.8% trichloroacetic acid (in DW) was added thereto, followed by color development at 100 ° C for 20 minutes. After cooling, the absorbance was measured at 520 nm. As a negative control, PBS (8.76 g of 1 L NaCl, 0.11 g of NaH ₂ PO ₄ , 0.596 g of Na ₂ HPO ₄ ) was used instead of the sample. The hydroxyl radical scavenging activity was calculated by the above equation using the above equation, and the results are shown in FIG. 4, where the difference in absorbance between the addition of the sample solution and the no addition was calculated.

As shown in FIG. 4, the DPDH radical scavenging activity, the ABTS radical scavenging activity, and the hydroxyl radical scavenging activity of the MGD02 fermentation chungkukjang according to the present invention at all concentrations, Was not only remarkably improved but also showed a much higher activity than that of the conventional Ja-8 strain fermentation chungkukjang.

&Lt; Determination of physiologically active substance content >

The contents of physiologically active substances were analyzed by measuring the contents of total phenolics, total flavonoids, phenolic acid, flavonol and isoflavone.

Total phenolic content was determined by slightly modifying the Folin-Denis method. 0.5 ml of each sample was dispensed into a test tube and diluted with 25% Na ₂ CO ₃ 0.5 ml was added and allowed to stand for 3 minutes. After that, 0.25 ml of 2 N Folin-Ciocalteu phenol solution was added and mixed, followed by color development at 30 ° C for 1 hour. The absorbance of the color-developed sample was measured at 750 nm using a spectrophotometer (Spectronic 2D), and the value was calculated from a standard calibration curve prepared using gallic acid. The results are shown in Table 4.

Total flavonoid contents were measured by Davis method. 0.5 ml of each sample was taken in two test tubes as a control and test solution, and 1.0 ml of diethylene glycol and 0.01 ml of 1 N NaOH were added, and the mixture was allowed to stand in a constant temperature water bath at 37 ° C for 1 hour. Then, the absorbance was measured at 420 nm using a spectrophotometer (Spectronic 2D), and the value was calculated from the standard calibration curve prepared using the routine. The results are shown in Table 4.

sample Analysis item Total phenolics (mg / g) Total flavonoid (mg / g) Comparative Example 1 (Soaking bean) 6.74 0.04 Comparative Example 2 (Extension bean) 9.9 0.22 Comparative Example 3 (Ja-8) 23.85 0.5 Test example (MGD02) 26.66 0.77

As shown in Table 4, the total phenolic acid and total flavonoid content of the MGD02 fermented soybean curd according to the present invention were increased about 4 times and 20 times as compared with that of soaked soybeans and soy sauce soybean (Comparative Examples 1 and 2) The activity was higher than that of the conventional fermented Cheonggukjang of Ja-8 strain.

Analysis of phenolic acid and flavonol substances was performed by high pressure liquid chromatography (HPLC, Agilent 1200 series, Agilent Co) by modifying the method of Cho et al. (2011). The mobile phase solvent was analyzed with 2.0% glacial acetic acid (solution A) and 2.0% acetonitrile (in glacial acetic acid) (solution B). The mobile phase conditions were 10, 15, 20, 25, Were maintained at 15%, 5%, 15%, 5%, 10%, 50%, 50%, 60%, 80% and 90% for 30, 35, 40, 45, 55 and 60 minutes. 20 μL of sample was injected and 20 μL of sample was injected. The mobile phase velocity was maintained at 1 ml / min at 30 ° C. Phenolic acid material was absorbed by the diode array UV detector (Agilent 1200 series, Agilent Co.) 280 nm, and the flavonols were quantified at an absorbance of 270 nm. The results are shown in Tables 5 and 6, respectively.

Content (/ / g) sample Comparative Example 1
(Soymilk) Comparative Example 2
(Increased beans) Comparative Example 3
(Ja-8) Test Example
(MGD02) Gallic acid nd nd 1.09 1.35 Protocatechuic acid 0.07 0.04 0.21 0.24 Chlorogenic acid 2.84 2.67 5.88 6.24 p- Hydrobenzoic acid 0.12 0.05 0.18 0.24 Vanilic acid nd nd 1.36 1.47 p -Coumaric acid 0.04 0.05 0.09 0.08 Ferulic acid 0.07 0.04 0.23 0.15 Ventaric acid 0.39 0.77 0.30 0.27 trans -Cinnamic acid 0.03 0.01 0.04 0.07 sum 3.56 3.63 9.38 10.11 nd: not detected

Content (/ / g) sample Comparative Example 1
(Soymilk) Comparative Example 2
(Increased beans) Comparative Example 3
(Ja-8) Test Example
(MGD02) Epigallocatechin 15.71 17.60 18.79 19.10 Vaniline 8.0 7.87 7.39 7.98 Routine 12.61 13.23 17.64 24.55 Catechin gallate 10.07 13.24 13.24 13.24 Naringin 14.08 15.96 26.77 32.05 Quercetin 13.68 13.04 16.25 16.36 Naringenin 9.46 10.52 19.94 19.96 sum 83.61 91.46 120.02 133.24

As shown in Table 5, gallic acid and vanilic acid were detected in the chungkukjang according to the present invention, and the content of total phenolic acid was remarkably increased. The main phenolic acids of chungkukjang according to the present invention were gallic acid, chlorgenic acid, and vanillic acid, and were 1.35 ㎍ / g, 6.24 ㎍ / g, and 1.47 ㎍ / g, respectively.

As shown in Table 6, flavonol compounds were also significantly enhanced in the chitinookjang according to the present invention. The major flavonol compounds are epigallocatechin, rutin, and naringin.

The isoflavone compound content was measured in the same manner as in Example 1 for each sample, and the results are shown in Table 7.

Content (/ / g) sample Comparative Example 1
(Soymilk) Comparative Example 2
(Increased beans) Comparative Example 3
(Ja-8) Test Example
(MGD02) Glycosides  Daidzin 146.22 638.94 171.29 181.16  Glycitin 121.95 262.51 253.89 195.59  Genistin 503.16 709.11 206.72 198.77  sum 771.34 1610.56 631.89 575.52 Aglycones (non-glycoside isoflavones)  Daidzein 67.49 20.74 335.71 390.83  Glycitein 7.33 78.75 38.50 32.15  Genistein 65.69 16.99 103.06 202.64  sum 140.51 116.48 477.28 625.62

As shown in Table 7, the non-glycosyl isoflavone content of the cheonggukjang according to the present invention was remarkably improved.

Example  4. Herb complex extract

 <Preparation of herb complex extract>

example Characters used in the invention leaflets (PF, Perilla frutescens there is. acuta KVDO), Angelica gigas (AG, Angelica gigas ), Platycodon (PG, Platycodon grandiflorum ) and bacillus (CW, Cynanchum wilfordii ) was purchased from Gyeongnam Agricultural Medicinal Agricultural Cooperative Association in dry condition.

 The herbal mixture powder was prepared by pulverizing and finely grinding lobsters, Angelicae japonica, Rhododendrons and Rhododendrons, and mixing them in the same ratio as in Table 8:

Herbal mixed powder (g) Lobular leaf Angelica Bellflower Baewooo Oh A 5 2.5 2 0.5 B 6 2 1.5 0.5 C 7 1.5 One 0.5

200 ml of 70% ethanol was added to 10 g of each herbal composite powder, and the resulting mixture was diluted 20 times at a final concentration of 600 rpm at 70 ° C for 5 hours. The extract was filtered twice with filter paper (No. 2, Whatman, Tokyo Roshi Kaisha, Ltd.), and finally concentrated to obtain a brix of soluble solids to prepare a herbal composition extract.

<Sample Preparation>

The herbal composition extract prepared above was concentrated under reduced pressure at 60 ° C using a vacuum condenser (EYELA, Tokyo, Rikakikai Co.) to obtain an extract, followed by freeze-drying to prepare a dry powder. Dry powder samples were prepared at a concentration of 1.0 mg / ml as an extraction solvent to evaluate the radical scavenging activity.

For analysis of total phenolic compounds, total flavonoids, phenolic acids and flavonols, 10 ml of 70% fermented alcohol was added to 5 g of each dry powder, and the mixture was heated at 70 ° C for 5 hours at 600 rpm And then filtered through a 0.45 μm membrane filter.

<Antioxidant Activity Test>

DPPH radical scavenging activity, ABTS radical scavenging activity and hydroxyl radical scavenging activity of the samples prepared above were measured in the same manner as described in Example 3. The results are shown in Table 9.

Radical scavenging activity (%) DPPH radical ABTS radical Hydroxyl radical A 54 70.62 23.42 B 66.77 85.79 31.55 C 78.35 97.59 38.03

As shown in Table 9, all of the herbal composition extracts A, B, and C having the range of the present invention exhibited excellent radical scavenging ability and high antioxidant activity, and the herbal composition extract C was the most excellent.

<Analysis of physiologically active substance content>

The contents of total phenolic compounds, total flavonoids, phenolic acid, and flavonol compounds as physiologically active substances were measured in the same manner as described in Example 3, and the results are shown in Tables 10, 11, and 12 .

Herbal compound extract Analysis item Total phenolics (mg / g) Total flavonoid (mg / g) A 12.66 1.65 B 13.19 1.70 C 13.69 1.93

As shown in Table 10, the contents of total phenolic compounds and total flavonoids in the herb complex extracts A, B, and C of the present invention range were high, ).

Content (/ / g) Herbal compound extract A B C Gallic acid 23.44 59.78 92.84 Protocatechuic acid 54.23 80.11 128.46 Chlorgenic acid 213.47 228.20 234.59 p- Hydrobenzoic acid 212.95 216.16 231.64 Vanilic acid 187.66 211.66 240.54 trans -Cinnamic acid 27.90 20.34 24.51 sum 719.65 816.25 952.58

As shown in Table 11, the contents of total herbicide extracts A, B, and C in the herb composition extracts of the present invention were higher than those of herb extracts of the present invention. Gallic acid, protocatechuic acid, chlorgenic acid, and vanilic acid were increased

Content
(/ / G) Herbal extract compound sample A B C Epigallocatechin 130.73 169.43 197.27 Catechin 170.00 185.76 213.96 Vaniline 31.43 20.31 29.27 Routine 24.89 43.66 67.84 Catechin gallate 141.85 154.30 143.21 Naringin 84.35 100.08 133.04 Naringenin 67.36 57.74 42.65 sum 650.61 731.28 827.24

As shown in Table 12, all of the herbal composition extracts A, B, and C prepared in the scope of the present invention had a high content of flavonol compounds, and the higher the content of the natural lobules, the higher the content (C) Epigallocatechin, catechin, gallocatechin gallate, rutin, and naringin contents were increased.

Example  5. Vegetable Brown sauce  Produce

<Production of Chongkukjang sauce stock>

5 kg of drinking water was added to 1 kg of the dried chungkukjang prepared in Example 3, and the mixture was boiled for a long time. Then, the mixture was boiled for 1 hour and then extracted into a medium size chungkookjang extract. After boiling for 2 minutes, the chungkukjang extract was extracted twice to obtain 5 L of Chungkookjang extract. Then, the mixture was boiled and concentrated to prepare chungkukjang sauce stock.

<Brown sauce manufacturing>

After pouring olive oil into the pan at 130 ° C and putting the flour into the pan, lower the flame and stir to prevent sticking. 24 g of rouge, carrot, celery, onion, tomato paste (45 g each), 20 ml of wine, sugar, salt and appropriate amount of salt were added to 400 ml of the prepared chungkukjang sauce stock and boiled at 100 ° C, (Control), 10 and 20% by weight, respectively, and the mixture was boiled at 100 ° C for 30 minutes, and then cooled to obtain a safflower extract. Vegetable brown sauce was prepared (Fig. 1).

<Physicochemical properties of vegetable brown sauce>

The pH, acidity, browning, reducing sugar and water soluble protein of vegetable brown sauce were measured and the results are shown in Table 13.

medicinal herbs
Complex extract
Addition amount (%) Analysis item pH Acidity
(Based on lactic acid) Browning
(420 nm) Reducing sugar
(g / L) Soluble protein
(mg / ml) 0 (control example) 5.15 0.08 1.09 26.11 33.04 10 5.12 0.07 1.18 27.19 38.56 20 5.12 0.07 1.24 26.57 36.92

As shown in Table 13, even after addition of the herbal composition extract, the excellent nutritional value derived from chungkookjang remained unchanged, and the degree of browning tended to be further enhanced.

<Sample Preparation>

50 ml of each brown sauce was mixed with 100% methanol in the same volume, and extracted at 300 rpm for 24 hours. The mixture was filtered with a 0.45 ㎛ membrane filter (Dismic-25CS), and some of them were used for measuring the contents of total phenolics and isoflavones The extracts were dissolved in extraction solvent and prepared at concentrations of 0.25, 0.5 and 1.0 mg / ml, respectively, and used for the evaluation of radical scavenging activity.

<Antioxidant activity of vegetable brown sauce>

DPPH radical scavenging activity, ABTS radical scavenging activity and hydroxyl radical scavenging activity of the samples prepared above were measured in the same manner as described in Example 3, and the results are shown in Fig.

As shown in FIG. 5A, the DPPH radical scavenging activity was enhanced with the addition of the herbal composition extract. In the low concentration (0.25 mg / ml), the radical scavenging activity of the control (0% % Of the brown sauce of the present invention exhibited a scavenging activity of 20% or more and exhibited an excellent scavenging activity of 81.72% at the concentration of 1.0 mg / ml.

As shown in FIG. 5B, the ABTS radical scavenging activity was enhanced with the addition of the herbal composition extract, and the brown sauce (10% and 20%) of the present invention was about 2 to 4 times as much as the control (0% And exhibited enhanced scavenging activity.

As shown in FIG. 5C, the hydroxyl radical scavenging activity was also increased as the addition of the herbal composition extract was increased, and the brown sauce (10% and 20%) of the present invention was about 2 to 5 And exhibited markedly enhanced scavenging activity.

&Lt; Contents of physiologically active substance in brown sauce >

Total phenolic compounds, total flavonoids, phenolic acids, flavonols and isoflavones, which are physiologically active substances of brown sauce, were measured in the same manner as in Example 3, and the results are shown in Table 14, Table 15 and Table Respectively.

Herbal compound extract
Addition amount (%) Analysis item Total phenolics (mg / ml) Total flavonoid (mg / ml) 0 (control example) 3.59 0.09 10 3.79 0.12 20 3.94 0.14

As shown in Table 14, total phenolics and total flavonoid contents were increased as the amount of the herbal composition extract was increased.

(쨉 g / ml) The amount of herbal medicine combined extract (%) 0 10 20 Gallic acid 19.05 21.78 39.40 Protocatechuic acid 11.39 30.21 52.37 Chlorogenic acid 18.61 52.45 74.07 p- Hydrobenzoic acid 4.36 17.08 34.94 Vanilic acid 6.05 10.91 22.32 p -Coumaric acid 1.98 8.28 14.24 Vertaric acid 2.41 9.78 18.64 trans -Cinnamic acid 0.88 4.94 8.30 sum 64.73 155.43 264.28

As shown in Table 15, the content of total phenolic acid compound increased with the addition of the herbal composition extract and increased by about 2.5 to 4 times as compared with the control (0%). The major compound, crogenic acid, was increased from 18.61 ㎍ / ml to 52.45 ㎍ / ml and 74.07 ㎍ / ml, respectively.

(쨉 g / ml) The amount of herbal medicine combined extract (%) 0 10 20 Epigallocatechin 9.17 37.02 58.33 Epigallocatechin gallate 3.48 9.25 10.68 Vaniline 0.74 4.49 7.60 Routine 1.48 9.99 15.98 Catechin gallate trace 2.65 5.11 Naringin 42.47 86.04 117.64 Quercetin 3.98 9.31 10.28 Naringenin 2.87 79.19 92.57 sum 64.19 237.94 318.19

As shown in Table 16, the content of flavonol compounds increased with addition of the herbal composition extract and increased about 3.5 to 5 times as compared with the control (0%). Major flavonolonarinine was detected and increased from 64.19 ㎍ / ml to 237.94 ㎍ / ml and 318.19 ㎍ / ml, respectively.

&Lt; Evaluation of palatability of brown sauce >

For the evaluation of palatability for each brown sauce (A, B, C), 20 young men and women (8 males and 12 females) The score of the lowest intensity was taken as 1 point, and when a high intensity incense or taste was recognized, it was possible to give up to 9 points, and the result is shown in FIG.

As shown in FIG. 6, as the amount of the herbal composition extract was increased, the perception of flavor was decreased. The brown sauce of the present invention containing 10 to 20% by weight of the herbal composition combination extract showed improved palatability due to harmony of flavor, color, savory taste, sweet taste and bitter taste, Was evaluated as having a strong flavor of Chongkukjang.

Based on these results, it can be seen that the vegetable brown sauce according to the present invention is a 100% vegetable source not only having excellent palatability, but also having enhanced physiologically active substances and enhanced antioxidative activity.

Agricultural Biotechnology Research Institute KACC92158P 20161212

<110> Gyeongnam National University of Science and Technology Industry-Academic Cooperation Foundation <120> Vegetable Brown Sauce with enhanced taste and anti-oxydant          property and preparation method thereof <130> p10189 <160> 3 <170> KoPatentin 3.0 <210> 1 <211> 1517 <212> RNA <213> Bacillus amyloliquefaciens MGD02 <220> <221> rRNA &Lt; 222 > (1) .. (1517) <223> 16s rRNA <400> 1 cggagagttt gatcctggct caggacgaac gctggcggcg tgcctaatac atgcaagtcg 60 agcggacaga tgggagcttg ctccctgatg ttagcggcgg acgggtgagt aacacgtggg 120 taacctgcct gtaagactgg gataactccg ggaaaccggg gctaataccg gatgcttgtt 180 tgaaccgcat ggttcagaca taaaaggtgg cttcggctac cacttacaga tggacccgcg 240 gcgcattagc tagttggtga ggtaacggct caccaaggcg acgatgcgta gccgacctga 300 gagggtgatc ggccacactg ggactgagac acggcccaga ctcctacggg aggcagcagt 360 agggaatctt ccgcaatgga cgaaagtctg acggagcaac gccgcgtgag tgatgaaggt 420 tttcggatcg taaagctctg ttgttaggga agaacaagtg ccgttcaaat agggcggcac 480 cttgacggta cccaaccaga aagccacggc taactacgtg ccagcagccg cggtaatacg 540 taggtggcaa gcgttgtccg gaattattgg gcgtaaaggg ctcgcaggcg gtttcttaag 600 tctgatgtga aagcccccgg ctcaaccggg gagggtcatt ggaaactggg gaacttgagt 660 gcgaagagg agagtggaat tccacgtgta gcgatgaaat gcgtagagat gtggaggaac 720 accagtggcg aaggcgactc tctggtctgt aactgacgct gaggagcgaa agcgtgggga 780 gcgaacagga ttagataccc tggtagtcca cgccgtaaac gatgagtgct aagtgttagg 840 gggtttccgc cccttagtgc tgcagctaac gcattaagca ctccgcctgg ggagtacggt 900 cgcaagactg aaactcaaag gaattgacgg gggcccgcac aagcggtgga gcatgtggtt 960 taattcgaag caacgcgaag aaccttacca ggtcttgaca tcctctgaca atcctagaga 1020 taggacgtcc ccttcggggg cagagtgaca ggtggtgcat ggttgtcgtc agctcgtgtc 1080 gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctt gatcttagtt gccagcattc 1140 agttgggcac tctaaggtga ctgccggtga caaaccggag gaaggtgggg atgacgtcaa 1200 atcatcatgc cccttatgac ctgggctaca cacgtgctac aatggacaga acaaagggca 1260 gcgaaaccgc gaggttaagc caatcccaca aatctgttct cagttcggat cgcagtctgc 1320 aactcgactg cgtgaagctg gaatcgctag taatcgcgga tcagcatgcc gcggtgaata 1380 cgttcccggg ccttgtacac accgcccgtc acaccacgag agtttgtaac acccgaagtc 1440 ggtgaggtaa ccttttagga gccagccgcc gaaggtggga cagatgattg gggtgaagtc 1500 gtaacaaggt agccgta 1517 <210> 2 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 2 cggagagttt gatcctgg 18 <210> 3 <211> 17 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 3 tacggctacc ttacgac 17

Claims (7)

And a cheonggukjang sauce stock fermented with a Bacillus amyloliquefaciens MGD02 strain. The present invention relates to a method for enhancing and enhancing a palatable, physiologically active substance enhanced in taste by shielding odor derived from chungkukjang As a vegetable brown sauce with antioxidant activity,
Wherein the herbal composition extract is contained in an amount of 10 to 20 wt% based on the total weight of brown sauce, and the chungkukjang sauce stock is contained in an amount of 70 to 80 wt%.
[3] The herb medicine composition according to claim 1, wherein the herbal composition extract comprises 50-70% by weight of self-lobulated leaves, 15-25% by weight of Angelica keiskei, 10-25% by weight of platycodon and 5-10% A vegetable brown sauce having enhanced physiologically active substance with enhanced palatability and enhanced antioxidant activity.
The herbal composition extract according to claim 1, wherein the mixed powder of 70% by weight of the baby lice, 15% by weight of Angelica japonica, 10% by weight of Bellflower and 5% by weight of Bacillus sp. Is extracted with 50 to 70% ethanol and filtered to obtain a soluble solid Is adjusted to be 3 to 5 Bricks, and the vegetable brown sauce having enhanced physiologically active substance with enhanced palatability and enhanced antioxidative activity, which is shielded from odor derived from chungkukjang.
4. The method according to any one of claims 1 to 3, wherein the physiologically active substance is at least one selected from the group consisting of phenolics, flavonoids, phenolics, flavonols, and isoflavones, Materials and vegetable brown sauce with enhanced antioxidant activity.
A method for producing a vegetable brown sauce having an enhanced physiologically active substance with enhanced palatability and an enhanced antioxidative activity by shielding odor derived from chungkukjang,
I) fermenting soybean with Bacillus amyloliquefaciens MGD02 to prepare and drying a cheonggukjang;
Ii) adding water to the dried chungkukjang, boiling and concentrating to produce a stock of chungkukjang;
Iii) adding a vegetable ingredient to the roasted flour, adding the chongkukjang stock and seasoning prepared in step ii), boiling, concentrating and filtering; And
Iv) preparing a herbal composition combination extract of Japanese Lilium, Angelica japonica, Rhododendron japonica and Bacillus subtilis and adding it to the filtrate of Step iii)
Wherein the herbal composition extract is added in an amount of 10 to 20 wt% based on the total weight of the brown sauce, and the chungkukjang sauce stock is added in an amount of 70 to 80 wt%.
[7] The herbal composition extract according to claim 5, wherein the mixed powder of 70% by weight of the baby lice, 15% by weight of Angelicae japonica, 10% by weight of the bellflower and 5% by weight of BSA is extracted with 50 to 70% ethanol, And the solid content is adjusted so as to be 3 to 5 Bricks. The method for producing vegetable brown sauce having enhanced physiologically active substance with enhanced palatability and enhanced antioxidative activity by shielding odor derived from chungkukjang.
A Bacillus amyloliquefaciens MGD02 strain of a vegetable brown sauce, Chungkukjang sauce stock fermented seedlings, deposited with the accession number KACC92158P and having odor originating from Chongkukjang, is shielded and promoted.

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