KR102043917B1 - Fermented food using strain isolated from traditionally fermented soybean and preparation method thereof - Google Patents

Abstract

The present invention relates to fermented food prepared by using a strain isolated from traditional paste products, and to a method for preparing the same. By comprising a step of inoculating a strain isolated from traditional paste products in a mixture of meju (fermented soybean block) and 13 to 16% of saline at a concentration of 1 × 10^6 to 9 × 10^7 CFU/ml, followed by fermentation, it is possible to improve the flavor of commercially available traditional soybean paste.

Description

Fermented food prepared using Bacillus amyroriquipysense strain and its preparation method {Fermented food using strain isolated from traditionally fermented soybean and preparation method

The present invention relates to a fermented food prepared using Bacillus amiriquiquisense strain to improve the flavor and a method for producing the same.

 Traditional Korean food is a heritage that represents the fermented food culture of Korean people. It has unique taste, excellent nutrition and functionality, and is a food with high potential for competition with world products (Hong et al 2015).

Traditional soybeans and traditional soy sauce are fermented foods traditionally produced using salt and purified water as soybeans as main ingredients. Starters are not used, and fermentation and fermentation are carried out depending on spontaneous fermentation process by strains derived from nature through meju. It is reported that various strains such as Aspergillus, Mucor, Rizopus, Penicillium, Bacillus, Saccharomyces are mainly involved (Lee, 1995; Lee et al., 2010; Kim et al., 2011c; Hong et al., 2011, 2012). ).

The traditional doenjang and traditional soy sauce provides a variety of amino acids, free sugars, organic acids and nucleic acid components of taste, sweet and sour and umami (Kim et al; 2003; Choi et al 2000), anti-obesity, anti-cancer, anti-diabetic, Various functionalities such as immune enhancement and liver function protection have also been reported (Kwon et al., 2006; Bae et al., 2013; Jung et al, 2006; Masilamanio et al., 2012; Soomro et al; 2008).

Ingredients related to flavor and functionality of traditional soybean paste and traditional soy sauce are known to be affected by fermentation strains and their fermentation conditions. Unfavorable flavors such as bitter taste are needed to improve its research (Hong et al). As these foods can be used as raw materials for the development of various source products as ethnic foods, according to the recent trend of demand for the world's new flavors, improvement of flavor is also very important for diversification of use.

A number of studies have been reported on the analysis of volatile or nonvolatile components in relation to the flavor of enteric (Kim et al; 1992, Kim et al., 1994, Kim et al; 1998, Shin et al 1999, Park et al 2002, Kim et al 2003 Lee et al 2009, Park et al 2011, Lee et al 2016). Research on the interrelationship between fermented strains and products through information analysis has been conducted, and Web-DB has been established (Hong et al., 2017).

Ingredients related to flavor and functionality of the enteric acid are estimated to be produced through enzymatic or non-enzymatic reactions and complex steps depending on the type of microorganism and fermentation conditions (Lee et al 2011), and can produce ingredients such as GABA. Excavation of the strains was carried out mainly limited to the functionality, and research on the discovery of strains that can enhance the flavor of the intestines has not been reported yet.

On the other hand, long-term maturation samples, unlike the short-term maturation samples, the overall number of bacteria and types are reduced, but also changes in the composition, the flavor components such as ester, lactic acid, etc. are increased, especially GABA component is known to increase more than 10 times. In addition, Doenjang has been reported to have the best taste in aged when aged for 3 years.

Therefore, it is expected that the strains capable of producing useful flavor components from long-term matured berries rather than short-term matured berries are expected to be greater.

Accordingly, various strains were collected from 1-30 years of traditional soy sauce and doenjang, which have been traditionally manufactured in Korea, to isolate various strains and to select strain cultures with excellent flavor preference. .

Republic of Korea Patent No.1786938 Republic of Korea Patent No. 1814420

An object of the present invention is to provide a fermented food prepared using a new strain isolated from the traditional soybeans to improve the flavor.

In addition, another object of the present invention to provide a method for producing the fermented food.

In addition, another object of the present invention to provide a starter composition for fermentation using a novel strain isolated from the traditional Jangjang.

Method for producing a fermented food of the present invention for achieving the above object is inoculated with a strain of 1X10 ⁶ to 9X10 ⁷ CFU / ml of strains isolated from traditional jang in a meju mixture mixed with meju and 13 to 16% saline And fermentation.

The fermented food may be miso, soy sauce or red pepper paste.

The strain isolated from the traditionaljang is Bacillus amyloliquefaciens J108 [Accession No .: KCCM12389P], Enterococcus faecium J161 [Accession No .: KCCM12387P] may be further included. Preferably Enterococcus faecium J161 [Accession Number: KCCM12387P]: Bacillus amyloliquefaciens J108 [Accession Number: KCCM12389P] may be mixed in a weight ratio of 1: 2-5. .

In addition, the fermented food of the present invention for achieving the above another object can be prepared by inoculating a strain isolated from traditional jang in Meju at a concentration of 1X10 ⁶ to 9X10 ⁷ CFU / ml.

In addition, Enterococcus faecium J161 [Accession No .: KCCM12387P] or Bacillus amyloliquefaciens J108 [Accession No .: KCCM12389P] of the present invention for achieving the above-mentioned other objects is obtained from traditional Can be separated.

In addition, the starter composition for fermentation of the present invention for achieving the above another object may include the Bacillus amyloliquefaciens J108 (Accession Number: KCCM12389P).

The starter composition for fermentation may further include Enterococcus faecium J161 (Accession No .: KCCM12387P).

Fermented food of the present invention is prepared by using a novel strain isolated from traditional berries, it is possible to improve the flavor of the fermented food.

Figure 1a is a view showing the content of GABA measured by LC / MS of the doenjang prepared according to Examples 1 and 2 of the present invention, Figure 1b is a LC / the miso prepared according to Examples 1 and 2 of the present invention It is a view showing the content of alanine (alanine) measured by MS, Figure 1c is a view showing the content of glucose (glucose) measured by LC / MS of the doenjang prepared according to Examples 1 and 2 of the present invention.
Figure 2 is a photograph of the miso prepared in accordance with Examples 1 and 2 of the present invention.
Figure 3a is a view showing the content of GABA measured by LC / MS of the soy sauce prepared according to Examples 3 and 4 of the present invention, Figure 3b is LC / LC of the soy sauce prepared according to Examples 3 and 4 of the present invention 3 is a view showing the content of alanine (alanine) measured by MS, Figure 3c is a view showing the content of glutamic acid (glutamic acid) measured by LC / MS of the soy sauce prepared according to Examples 3 and 4 of the present invention, Figure 3d is a diagram showing the content of proline (proline) measured by LC / MS of the soy sauce prepared according to Examples 3 and 4 of the present invention.
Figure 4 is a photograph of the soy sauce prepared according to Examples 3 and 4 of the present invention.

The present invention relates to a fermented food prepared using a strain isolated from traditional soybeans and a method for producing the same to improve the flavor.

The present invention was isolated and identified from the traditional soybean paste (soybean paste or soy sauce) to fermented strains that can produce fermented foods with improved properties while improving the flavor of conventional doenjang, soy sauce or red pepper paste that does not satisfy the taste of consumers, The fermented food of the present invention inoculated with the strain is fermented, unlike the conventional traditional fermented food or improved fermented food may have a flavor that satisfies the preference of the consumer.

As a result of identifying the strains isolated from the traditional Jangjang , Bacillus amyloliquefaciens and Enterococcus faecium belong to Bacillus amyloliquefaciens J108 and Enterococcus, respectively. It was named Passococcus faecium J161 and was deposited on November 13, 2018 by the Korea Microorganism Conservation Center and assigned the accession numbers KCCM12389P and KCCM12387P, respectively.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

Fermented food of the present invention comprises the step of fermenting by inoculating a strain isolated from traditional jang in a meju mixture mixed with meju and 13 to 16% saline.

The meju is not particularly limited as long as it is produced by a conventional method.

The strain isolated from the traditional Jang as Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens ) J108 [Accession No .: KCCM12389P] and Enterococcus faecium J161 [Accession No .: KCCM12387P] In order to obtain a fermented food having excellent flavor and excellent antioxidant activity, enterococcus faecium J161 [Accession No .: KCCM12387P]: Bacillus amyloliquefaciens J108 [Accession No .: KCCM12389P] This is mixed in a weight ratio of 1: 2-5, preferably in a weight ratio of 1: 2.5-3.5. Based on Enterococcus faecium J161 [Accession No .: KCCM12387P], when the content of Bacillus amyloliquefaciens J108 [Accession No .: KCCM12389P] is less than the lower limit, the flavor and antioxidant activity are not excellent. If the upper limit is exceeded, the flavor may not be good and the functionality may be degraded.

The strain isolated from the traditional Jangjang is inoculated at a concentration of 1X10 ⁶ to 9X10 ⁷ CFU / ml, preferably 5X10 ⁶ to 5X10 ⁷ CFU / ml, and when the concentration of the strain is less than the lower limit, Even when used, fermented foods having an improved flavor cannot be obtained, and when the upper limit is exceeded, antioxidant activity and functionality may be lowered.

Doenjang, soy sauce or red pepper paste using the strain isolated from the traditional Jangjang of the present invention is not particularly limited as long as it is a method for producing doenjang, soy sauce or red pepper paste using the strain.

Hereinafter, preferred examples are provided to aid the understanding of the present invention, but the following examples are merely for exemplifying the present invention, and it will be apparent to those skilled in the art that various changes and modifications can be made within the scope and spirit of the present invention. It is natural that such variations and modifications fall within the scope of the appended claims.

[Isolation and Selection of Microorganisms from Traditional Ganjang]

A total of 94 products of doenjang and soy sauce aged 1-30 years were purchased to isolate strains.

<Test Example_1>

Test Example 1 Identification of Strains

16S rRNA genes were isolated using 27F (5'-AGAGTTTGATCCTGGCTCAG-3 ') and 1,492R (5'-GGTTACCTTGTTACGACTT-3') as universal primers for phylogenetic identification of 16S rRNA genes in isolated strains. 1,443 bp containing the variable base region important for identification after PCR amplification was analyzed using BigDye Terminator v3.1 Cycle Sequencing kit (Applied Biosystems Inc., USA) (Chakravorty et al., 2007). These base sequences were obtained from the NCBI data base using the BLASTN program (Zhang et al., 2000) and the Ribosomal Database Project (RDP) Seqmatch program (version 3) to obtain 16S rRNA gene sequences of the standard strains, and then the interaction between the base sequences. The CLUSTAL W program (Thompson et al., 1994) was used for comparison.

Based on the nucleotide sequence of the 16S rRNA gene of the isolated strain, 120 strains of Bacillus subtilis , Bacillus sp. 35 kinds, Bacillus amyloliquefaciens and Bacillus licheniformis 28 jong 25 species, such as Bacillus with this series were the majority of the isolated strains were isolated 38 lactic acid bacteria, such as paper-based rosseoneun Sporolactobacillus nakayamae and Sporolactobacillus terrae type. On the other hand, there were only four varieties of Candida haemulonis .

The identified strains are shown in Table 1 below.

Test Example 2 Analysis of Volatile Flavor Components

Among the above strains, in order to select strains that produce organoleptically superior fragrance components, after incubating the strains of about 100 species in TSB (Tryptic soy broth), they were sensibly fragrant except for a sulky or odorous culture. Fourteen strain fermentation strains expressing excellent flavor were selected (Table 1).

The control group did not use the strain.

Extraction of Volatile Compounds of Doenjang Using HS-SPME Method

Extracting the volatile components of the selected strain fermentation was used to complement the method of Lee and Ahn (20). For extraction, 1 g of sample was placed in 30 mL of glass vial (Supelco, Bellefonte, PA, USA), 3.5 mL of distilled water and 1 g of NaCl were added and sealed with septum (silicon / Teflon, Supelco). The sealed vials were each equilibrated with stirring at 60 ° C. for 30 minutes and then collected in 50/30 μm DVB / carboxen ™ / PDMS StableFlex ™ iber (Supelco) for 30 minutes and injected into GC for 1 minute.

Qualitative and quantitative analysis of fragrance components using GC / MS

Volatile aromatic components of the strain fermentation extracted by the HS-SPME were analyzed by GC / MS (Agilent 6890 gas chromatography coupled with 5973 mass selective detector, Agilent Co., Palo Alto, Ca, USA). Stabilwax DA (30 m length × 0.25 mm i.d.X0.25 μm film thickness: Restek, Bellefonte, PA, USA) was used for the analysis column. The Oven temperature was maintained at 40 ° C. for 3 minutes and then heated up to 210 ° C. at a rate of 4 ° C./min. Injector temperature was analyzed in splitless mode at 250 ° C. Carrier gas was maintained at 0.8 mL / min using helium. MSD (Agilent 5975C) conditions were capillary direct interface temperature 250 ℃, ion source temperature 230 ℃, EI ionization voltage 70 eV, mass range 40 ~ 550 a.m.u and scan rate 2.2 scans / s. Volatile compounds were identified by comparing retention indices (RI), mass spectrum library (NIST05A) and aromatic properties (Lee et al 2008).

 Strain Ethyl alcohol Acetone 2-methyl propanol 2,3-butanedione 3-methyl butanal Acetic acid 1-butanol 3-hydroxy-2-butanone 1-pentanol Control 4285027 544832339 160239025 235337097 834664748 356411264 29616920 51219727 13386981 B. sonorensis 72414643 176462469 34739970 197815192 338446453 207266394 353241346 341179458 36829085 B. subtilis -H1 - 192452105 24318547 181841901 305739772 127905158 41759920 440730751 18277901 B. subtilis - H2 - 122545247 9662221 238479419 190881587 150954541 241301751 1.19E + 09 27168972 B. subtilis - H3 - 128672854 12303367 235100624 160282342 135946794 300629055 1.08E + 09 27215916 B. subtilis - H4 - 125458093 11155125 197385145 196779397 135716809 257748372 788603454 15111305 B. subtilis - H5 - 125262964 13290743 184336567 191940364 129766303 273332173 837949141 30343291 B. amyloliquefacience - J1 - 174002502 21417266 214276694 294063087 128620722 72803264 641626104 31872242 B. amyloliquefacience - J108 - 133698250 11509403 193854777 195169898 106243851 160330574 861328590 18800794 B. sp - K1 60386661 120819541 21976953 188339173 292220044 156787219 308094823 623242777 18381901 B. sp - K2 42419235 126770408 21185279 207619813 285455729 154877453 315230371 797039152 17680433 B. sp - K3 13865062 180920826 36418876 149377668 442013588 193334231 324049202 75760709 21683094 B. sp - K4 53859962 121398989 20803246 191416223 274038834 150980280 291945818 765571372 17712857 Brevibac . frigoritoleranc e 373375 209986966 63313772 152340175 677356446 224802228 30175709 48098885 17943173

 Strain 2-methyl-propanoic acid methyl pyrazine butanoic acid 2,5-dimethyl pyrazine benzaldehyde trimethyl pyrazine Phenylethyl slcohol Control 12937027 82157110 14077387 258110115 488707836 75901208 552411 B.sonorensis 14887892 79637693 20110081 326304984 41753379 43635894 2858400 B. subtilis -H1 11146079 78528841 9587173 348402876 39502460 46018700 8053258 B. subtilis - H2 34033185 81728409 14841209 324097567 49820494 58446871 9939930 B. subtilis - H3 27866416 83772215 18177185 332069939 34797141 46694118 9833732 B. subtilis - H4 28294434 85795609 17500539 381252640 39810424 51347295 9117986 B. subtilis - H5 16896581 86004426 17656327 377133262 42495446 51322997 13113223 B. amyloliquefacience - J1 16331307 82999789 12687120 352544496 36924612 47447415 8001346 B. amyloliquefacience - J108 20088255 88312573 11965372 383861817 42738815 54208164 10897488 B. sp - K1 26592887 88611076 16621879 407112784 50808954 54959770 11741418 B. sp - K2 29367285 85593964 16267739 48897143 48584455 51060156 14250340 B. sp - K3 20714654 81886600 13884973 388366963 84660699 49372247 9817513 B. sp - K4 27401989 85510354 17133695 383226196 47124894 52458934 9828641 Brevibac . frigoritolerance 14903999 85002484 3768137 406671315 125833202 50874968 5481754

When volatile components of the fermented product were analyzed by GC / MS, ethyl alcohol, acetone, 2-methyl propanol, 2,3-butanedione, 3-methyl butanal, acetic acid, 1-butanol, 3-hydroxy-2-butanone Sixteen species were analyzed, 1-pentanol, 2-methyl-propanoic acid, methyl pyrazine, butanoic acid, 2,5-dimethyl pyrazine, benzaldehyde, trimethyl pyrazine and phenylethyl alcohol.

As shown in Tables 1 and 2 above, Bacillus amyloliquefaciens J108 was confirmed to have a higher content of 3-hydroxy-2-butanone known as a fragrance component of cheese than other strain cultures. In addition, except for some strain fermented products, 2, 3-butanedione, which is a fragrance such as 3-methyl butanal, margarine, etc. used in coffee flavor, and high acetic acid, .

In particular, Bacillus amyloliquefaciens J108 is very low in the content of butanoic acid associated with odor, so these strains are expected to be of great value in the production of superior flavored intestines.

Test Example 3 Enzyme Activity Analysis

Protese extracellular secretion was measured by modifying the Kuniz method. 0.25 mL of the 0.6% casein solution (pH 7.0) dissolved in 0.5 M phosphate buffer was mixed with 0.25 mL of the sample, and then reacted in a water bath at 37 ° C. for 30 minutes. Then, 0.5 mL of 0.44 M Trichloroacetic acid (TCA) was added thereto and left at room temperature for 30 minutes. The reaction was stopped. Subsequently, the reaction solution was centrifuged at 3,000 xg, 10 min, 50 μl of supernatant, 125 μl of 0.55 M sodium carbonate anhydrous, and 25 μl of diluted folin ciocalteu solution were mixed and developed at 37 ° C. for 30 minutes, followed by 660 nm. Absorbance was measured. The standard curve was prepared by quantifying tyrosine, and the active unit was defined as the amount of enzyme that produces 1 μg of tyrosine per mL.

In addition, amylase was measured by the DUN (Dextrinogenic Unit of Nagase) method. 1 ml of coenzyme solution was added to 3 ml of 1% starch liquor (pH 7.0), and the reaction was stopped by adding 10 ml of 0.1 M HCL in a water bath at 40 ° C. for 10 minutes. 10 ml of 0.005% I ₂ and 0.05% KI solution was added to the filtrate of the reaction stop solution, and the absorbance was measured at 660 nm. Enzyme activity was calculated according to the following equation.

[Equation 1]

D.U.N = D-D '/ D x 100/10 x n

(D: control absorbance, D ': absorbance of reaction solution, n: enzyme dilution factor)

In addition, lipase was added to 1.0 ml of 0.5 mM KCl, 1.0 ml of 5 mM CaCl ₂ , and 7.3 ml of distilled water to 1.0 ml of coenzyme solution. The mixture was heated at 35 ° C. for 15 minutes, and 0.2 ml of tributyrin was added as a substrate. After boiling for 2 minutes at 150 ℃ to stop the enzyme action was titrated fatty acid with 0.01 N NaOH. Activity was defined as 1 unit of enzyme that liberates 1 μg equivalent of acid per minute.

[Equation 2]

Lipase activity (unit / g) = (sample titration (ml)-control amount corrected (ml)) / sample amount in sample solution (ml) (g)

Strain Enzyme (units / ml) lipase amylase protease B.sonorensis 986.8 ± 53.4 0 ± 47.2 216.8 ± 28.7 B.subtilis-H1 1105.3 ± 48.8 164 ± 51.6 275.5 ± 38.3 B.subtilis-H2 1042.4 ± 71.3 0 ± 77.2 285.6 ± 17.4 B.subtilis-H3 989.8 ± 77.3 0 ± 45.2 277.3 ± 46.8 B.subtilis-H4 1035.1 ± 54.8 0 ± 53.3 230.1 ± 26.6 B.subtilis-H5 1038.0 ± 41.5 0 ± 79.8 261.6 ± 47.0 B.amyloliquefacience-J1 1244.2 ± 20.3 0 ± 51.4 282.5 ± 71.3 B. amyloliquefacience - J108 1328.9 ± 35.1 243.6 ± 51.6 328.3 ± 41.1 B. sp - K1 2460.5 ± 82.2 0 ± 63.4 197.0 ± 34.7 B. sp - K2 1641.8 ± 58.4 0 ± 65.3 306.1 ± 7.9 B. sp - K3 1567.3 ± 75.6 836.8 ± 117.3 307.7 ± 19.1 B. sp - K4 1578.9 ± 107.7 272.9 ± 69.2 348.4 ± 52.4 Brevibac . frigoritolerance 913.7 ± 39.3 0 ± 55.9 226.1 ± 30.6

The production of good fragrance components of the strain culture was also predicted to be related to the enzyme activity to evaluate the activity levels of lipase, amylase and protease.

As shown in Table 3 above, Bacillus amyloliquefaciens J108 was confirmed to have higher activity levels of both lipase, amylase and protease compared to other strain cultures. In particular, it was confirmed that the activity level of amylase was remarkably high.

Bacillus (Bacillus) is amyl raised and generates various enzymes of the protease, such as the process proceeds the fermentation grant traditional soy sauce unique taste and flavor, and the proteins in the soybean is hydrolyzed into peptide by the protease step by step through the aa It splits into four, first forming amine and second forming aldehyde and then acid and alcohol. Thirdly, the pyrazine forms pyrroles through the maillard reaction with the reductones. Finally, the aromatic aa forms phenol, phenethyl alcohol and benzaldehyde.

Therefore, strain cultures having high activity of lipase, amylase and protease as described above are considered to be highly associated with the production of various flavor components.

[Manufacturing of miso]

Control group 1.

The control group 1 used doenjang prepared conventionally by adding traditional meju and brine.

Example 1. Bacillus amiriquifaciens ( Bacillus amyloliquefaciens ) J108 [Accession Number: KCCM12389P]

After mixing 20 kg of conventional meju and 60 kg of 15% saline solution, Bacillus amyloliquefaciens J108 [Accession No .: KCCM12389P] was inoculated at a concentration of ⁷ × 10 ⁶ and then 40 to 100 days at 12 to 18 ° C. Fermented for 2 to 10 months at 20 to 25 ℃ and then soybeans were separated from soy sauce to obtain miso.

Example 2. Bacillus amiriquifaciens ( Bacillus amyloliquefaciens ) J108 [Accession No .: KCCM12389P] + Enterococcus fascium ( Enterococcus faecium ) J161 [Accession No .: KCCM12387P]

In the same manner as in Example 1, Bacillus amyloliquefaciens J108 instead of Bacillus amyloliquefaciens (J108) J108 and Enterococcus faecium ( Enterococcus faecium ) J161 in a weight ratio of 1: 3. Doenjang was prepared by mixing.

Comparative Example 1. Bacillus subtilis ( Bacillus subtilis ) H5

In the same manner as in Example 1, Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens ) J108 instead of Bacillus subtilis ( Bacillus subtilis ) H5 was prepared using.

<Test Example_2>

Test Example 4 General Component Analysis

4-1. pH and titratable acidity: 5 g of the sample was taken and measured with an automatic titrator T50 (Mettler Toledo GmbH, Switzerland).

4-2. Amino nitrogen: The amino nitrogen content was analyzed using T50 (Mettler Toledo GmbH, Switzerland) according to Formol titration. 2 g of the sample was taken, 100 ml of distilled water was added thereto, stirred for 1 hour, and titrated to pH 8.4 with 0.1N NaOH solution. 20 ml of neutral formalin solution was added thereto, and titrated again to pH 8.4 with 0.1N NaOH solution, and a background test was performed on distilled water to determine amino nitrogen content.

division pH Titratable acidity (0.1N NaOH ml / g) Amino nitrogen (mg%) Control group 1 8.15 0.05 420.4 Example 1 8.31 0.02 591.0 Example 2 8.32 0.03 701.5 Comparative Example 1 8.09 0.06 411.3

As shown in Table 4 above, the pH and titratable acidity of the doenjang prepared in Examples 1 and 2 of the present invention, the doenjang of the control group 1 and the doenjang of Comparative Example 1 were similar.

In all treatments, the content of amino nitrogen was more than 400 mg%, but the content of amino nitrogen of Examples 1 and 2 was much higher than that of Control 1 and Comparative Example 1.

Test Example 5. Sensory and Chemical Fragrances

5-1. Sensory fragrance: For the doenjang prepared in Examples and Comparative Examples, the sensory characteristics were confirmed through descriptive analysis of 10 sensory test panel in the sensory test method.

division Fragrance description Control group 1 Soybean miso flavor, ammonia odor medicine, cumum flavor, salty smell Example 1 Good miso flavor, slightly sour and sweet flavor Example 2 Soft miso Comparative Example 1 Soybean miso flavor, ammonia odor river

As shown in Table 5, it was confirmed that the flavor of the doenjang is superior to the doenjang prepared in Examples 1 and 2 of the present invention and the control 1 and doenjang of Comparative Example 1.

5-2. Chemical fragrance: It was tested in the same manner as in Test Example 1.

division trimethyl pyrazine butanoic acid benzaldehyde 3-hydroxy-2-butanone Control group 1 49485301 13564821 418645267 61234580 Example 1 69152756 3796842 56042812 480126842 Example 2 85102691 2946585 42068414 611159507 Comparative Example 1 30001590 15624568 406548952 78456807

As shown in Table 6, compared to the doenjang prepared in Examples 1 and 2 of the present invention and the doenjang of the control group 1 and doenjang of Comparative Example 1 has a low butanoic acid content showing a peculiar, smoky smell peculiar to the doenjang In addition to the high content of benzaldehyde and trimethyl pyrazine, which are fragrant properties such as miso, savory, and fruity, it was confirmed that the content of 3-hydroxy-2-butanone, which is known as a fragrance of cheese, was high.

Test Example  6. Component Analysis Using LC / MS

Figure 1a is a view showing the content of GABA measured by LC / MS of the doenjang prepared according to Examples 1 and 2 of the present invention, Figure 1b is a LC / the miso prepared according to Examples 1 and 2 of the present invention It is a view showing the content of alanine (alanine) measured by MS, Figure 1c is a view showing the content of glucose (glucose) measured by LC / MS of the doenjang prepared according to Examples 1 and 2 of the present invention.

As shown in Figure 1, the doenjang prepared according to Examples 1 and 2 of the present invention was confirmed that the content of GABA, alanine (alanine) and glucose (glucose) is high.

Test Example 7 Visual Measurement

Figure 2 is a photograph of the miso prepared in accordance with Examples 1 and 2 of the present invention.

As shown in Figure 2, the control 1 and doenjang prepared according to Examples 1 and 2 of the present invention was confirmed that the color is similar.

[Manufacture of Soy Sauce]

Control 2.

In the control group 2, soy sauce prepared by adding traditional meju and brine was used.

Example 3. Bacillus amiriquifaciens ( Bacillus amyloliquefaciens ) J108 [Accession Number: KCCM12389P]

After mixing 20 kg of conventional meju and 60 kg of 15% saline solution, Bacillus Bacillus amyloliquefaciens J108 [Accession No .: KCCM12389P] was inoculated at a concentration of ⁷ × 10 ⁶ and then 40 to 100 at 12 to 18 ° C. After fermentation for one day and soaked in brine and aged at 20 to 25 ℃ for 6 to 10 months to obtain soy sauce.

Example 4. Bacillus amiriquifaciens ( Bacillus amyloliquefaciens ) J108 [Accession No .: KCCM12389P] + Enterococcus fascium ( Enterococcus faecium ) J16 [Accession No .: KCCM12387P]

But the same manner as in Example 3, the Bacillus amino Lowry quinolyl Pacific Enschede (Bacillus amyloliquefaciens) J108 instead Bacillus amino Lowry quinolyl Pacific Enschede the (Bacillus amyloliquefaciens) J108 and Enterococcus passive help (Enterococcus faecium) J16 1: in a weight ratio of 3 Soy sauce was prepared by mixing.

Comparative Example 2. Bacillus subtilis ( Bacillus subtilis ) H5

The same procedure as in Example 3, except that Bacillus amyloliquefaciens J108 instead of Bacillus subtilis ( Bacillus subtilis ) H5 was prepared using the soy.

<Test Example_3>

Test Example 8 Sensory and Chemical Fragrances

It carried out in the same manner as in Test Example 5.

Sensual fragrance

division Fragrance description Control 2 Soy soy sauce, odor medicine Example 3 Good soy flavor, slightly sour and sweet Example 4 Soft soy sauce Comparative Example 2 Soy Soy Sauce, Ammonia

As shown in Table 7, the soy sauce prepared according to Examples 3 and 4 of the present invention was confirmed that the flavor of the liver is superior to the soy of the control 2 and the soy of Comparative Example 2.

Chemical fragrance

division trimethyl pyrazine butanoic acid benzaldehyde 3-hydroxy-2-butanone Control 2 40135945 11594568 341568942 50678468 Example 3 4915236 3167481 40956135 425846807 Example 4 6220945 2706481 38507814 580684012 Comparative Example 2 25164856 13684245 355648942 55184561

As shown in Table 8 above, the soy sauce prepared according to Examples 3 and 4 of the present invention has a lower butanoic acid content, which shows a distinctive soy, smoky smell, compared to the soy sauces of Control 2 and Comparative Example 2. In addition to the high content of benzaldehyde and trimethyl pyrazine, which indicate aroma and fruit flavor, it was confirmed that the content of 3-hydroxy-2-butanone, which is known as a fragrance component of cheese, was high.

Test Example 9 Component Analysis Using LC / MS

Figure 3a is a diagram showing the content of GABA measured by LC / MS of the soy sauce prepared according to Examples 3 and 4 of the present invention, Figure 3b is LC / LC of the soy sauce prepared according to Examples 3 and 4 of the present invention Figure 3 shows the alanine content measured by MS, Figure 3c is a diagram showing the content of glutamic acid (glutamic acid) measured by LC / MS of the soy sauce prepared according to Examples 3 and 4 of the present invention, Figure 3d is a diagram showing the content of proline (proline) measured by LC / MS of the soy sauce prepared according to Examples 3 and 4 of the present invention.

As shown in Figure 3, the soybean paste prepared according to Examples 3 and 4 of the present invention was confirmed that the content of GABA, alanine (alanine), glutamic acid (glutamic acid) and proline (proline) is high.

Test Example 10 Visual Measurement

Figure 4 is a photograph of the soy sauce prepared according to Examples 3 and 4 of the present invention.

As shown in Figure 4, the control 2 and the soy sauce prepared according to Examples 3 and 4 of the present invention were all confirmed that the color is similar.

Korea Microorganism Conservation Center (overseas) KCCM12389P and KCCM12387P 20181113

Claims (13)

It comprises a step of inoculating the strain isolated from traditional jang in a meju mixture mixed with meju and brine at a concentration of 1X10 ⁶ to 9X10 ⁷ CFU / ml and fermentation;
The strain isolated from the traditional Jang Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens ) J108 [Accession No .: KCCM12389P] The method of producing a fermented food, characterized in that. The method of claim 1, wherein the fermented food is soybean paste, soy sauce or red pepper paste. delete The method of claim 1, wherein enterococcus faecium J161 (Accession No .: KCCM12387P) is further added as a strain isolated from the traditional Jangjang. According to claim 4, wherein the strain isolated from the traditional enterococcus faecium ( Enterococcus faecium ) J161 [Accession Number: KCCM12387P]: Bacillus amyloliquefaciens J108 [Accession Number: KCCM12389P] is 1: Method for producing a fermented food, characterized in that mixed in a weight ratio of 2-5. Inoculated strains isolated from traditional Jangju in Meju at a concentration of 1X10 ⁶ to 9X10 ⁷ CFU / ml,
The strain isolated from the traditional Jangjang Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens ) J108 [Accession number: KCCM12389P] characterized in that the fermented food. delete 7. The fermented food product of claim 6, further comprising Enterococcus faecium J161 (Accession No .: KCCM12387P) as a strain isolated from the traditional jang. According to claim 8, wherein the strain isolated from the traditional enterococcus faecium ( Enterococcus faecium ) J161 [Accession Number: KCCM12387P]: Bacillus amyloliquefaciens J108 [Accession Number: KCCM12389P] is 1: Fermented food, characterized in that mixed in a weight ratio of 2-5. The fermented food according to claim 6, wherein the fermented food is miso, soy sauce or red pepper paste. Bacillus amyloliquefaciens J108 (Accession No .: KCCM12389P), isolated from traditional berries. A starter composition for fermentation comprising Bacillus amyloliquefaciens J108 (Accession No .: KCCM12389P) according to claim 11. 13. The fermentation starter composition of claim 12, wherein the starter composition for fermentation further comprises Enterococcus faecium J161 [Accession No .: KCCM12387P].

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