KR20210076550A - Methods for preparing oat doenjang using a novel Bacillus subtilis TSD - Google Patents

Abstract

The present invention relates to a novel bacillus subtilis TSD strain and a method for preparing soybean paste using the same. The present invention provides a bacillus subtilis TSD strain and an application thereof. In addition, when fermented with steamed soybeans, the bacillus subtilis TSD strain of the present invention can prepare soybean paste with an excellent flavor by increasing the content of amino nitrogen. The bacillus subtilis TSD strain of the present invention can be variously used in preparing intestinal preparations, probiotics, food additives, and fermented products. The strain can be usefully used as a seed starter for soybean paste fermentation in particular. In addition, in oat soybean paste prepared using steamed oat powder and the bacillus subtilis TSD strain by the method of the present invention, the functionality of the oat is maximized and the palatability of the soybean paste is further enhanced.

Description

Method for preparing oat soybean paste using a novel Bacillus subtilis TSD strain {Methods for preparing oat doenjang using a novel Bacillus subtilis TSD}

The present invention relates to a method for producing oat soybean using a novel Bacillus subtilis TSD strain.

In Korea, along with the increase in national income and changes in social and cultural lifestyles, the consumption pattern of soybean paste is gradually changing from the form of making and consuming doenjang at home to the consumption pattern of purchasing and consuming commercialized soybean paste. . Due to this, soybean paste has secured a strong domestic market, and as its unique flavor and excellent anticancer effect are known to the international community, it is gradually taking its place in the international market.

Doenjang is nutritionally excellent as it contains essential amino acids, fatty acids, organic acids, minerals, and vitamins that are easily lacking in grain proteins. Useful substances related to physiological activity contained in soybean include saponin, phytic acid, lectin, oligosaccharide, and isoflavone derived from soybean. In particular, it is known that isoflavones, one of the useful components of soybeans, are converted to aglycone/glycoside by microorganisms during the fermentation process of doenjang, thereby increasing the absorption rate in the body. In addition, anti-obesity, prevention of hypertension, anticancer, antioxidant activity, antithrombosis, liver function enhancement, dementia prevention effect, and physiological activities such as osteoporosis are known. Recently, studies to improve the functionality of doenjang such as mixing functional ingredients with soybean paste is being performed in various ways.

On the other hand, according to the results of a recently reported study, some of the many European countries had a significantly lower cancer incidence. What these countries have in common is that they generally consume a lot of fermented foods, especially Finland. , In Bulgaria, there was a report that the incidence of cancer was significantly lowered due to the high intake of published foods such as yogurt. Also, in the case of Korea, it has been reported that doenjang, a representative fermented food, is consumed by most of the people, so the incidence of cancer is lower than that of other countries.

On the other hand, the present inventors tried to develop a novel strain that can improve the quality and functionality of soybean paste. As a result, Bacillus subtilis TSD strain was newly isolated and identified from soybean paste, and the present invention was completed by experimentally confirming the applicability of the strain to fermented products. In particular, in the case of producing soybean paste using the strain, it was specifically confirmed that the flavor could be improved by increasing the amino nitrogen content, so that the soybean paste with high preference could be produced.

In addition, the present invention was completed by confirming that soybean paste having a high beta-glucan content and preference while enhancing useful physiologically active ingredients of oats could be prepared when preparing soybean paste using the Bacillus subtilis TSD strain and oats.

KR 1894638 B KR 1547896 B

An object of the present invention is to provide a method for manufacturing oat soybean paste using steamed oat powder, and Bacillus subtilis TSD strain [Accession No.: KCCM12595P] or a culture solution thereof.

In order to achieve the above object, the present invention is a Bacillus subtilis ( Bacillus subtilis ) TSD strain [Accession No.: KCCM12595P] or a culture solution thereof inoculated into a mixture of steamed oat powder and steamed soybeans, followed by fermentation; It provides a method for manufacturing oat soybean paste comprising.

In addition, the present invention comprises the steps of (a) immersing meju in brine and then separating the salted meju from the brine; (b) preparing steamed oat powder by steaming the oat powder; (c) inoculating a Bacillus subtilis TSD strain [Accession No.: KCCM12595P] or a culture solution thereof in a mixture of steamed oat powder and steamed soybeans, followed by fermentation to prepare a seed fermented product; (d) preparing soybean paste by mixing the seed fermented product and the salted soybean paste; and (e) fermenting the soybean paste prepared in step (d).

The present invention provides Bacillus subtilis TSD strains and uses thereof. In addition, when the Bacillus subtilis TSD strain of the present invention is fermented with steamed soybeans, the content of amino nitrogen can be increased to produce soybean paste with better flavor.

The Bacillus subtilis TSD strain according to the present invention can be used in various ways in the preparation of intestinal tract agents, probiotics, food additives and fermented products, and in particular, it can be usefully used as a seed starter for soybean paste fermentation.

In addition, oat doenjang prepared using steamed oat powder and Bacillus subtilis TSD strain according to the method of the present invention maximizes the functionality of oats while further increasing the palatability of the doenjang.

1 is a photograph showing the cellular morphological characteristics of Bacillus subtilis (Bacillus subtilis) TSD strain (KCCM12595P).
Figure 2 shows the sequence list of Bacillus subtilis (Bacillus subtilis) TSD strain.
3 is a graph showing the number of viable cells according to the growth time of Bacillus subtilis ( Bacillus subtilis ) TSD strain.
Figure 4 is a graph showing the protease activity according to the growth time of Bacillus subtilis (Bacillus subtilis) TSD strain.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention relates to Bacillus subtilis ( Bacillus subtilis ) TSD strain [Accession No.: KCCM12595P].

Bacillus subtilis ( Bacillus subtilis ) TSD strain [Accession No.: KCCM12595P] according to the present invention is a new strain of Bacillus subtilis derived from traditional fermented foods. Although the Bacillus subtilis TSD strain in the present invention was isolated and identified from soybean paste, the acquisition route is not limited thereto.

As a result of 16S rRNA sequencing analysis for identification and classification of microorganisms, the TSD strain isolated from soybean paste was found to have the nucleic acid sequence of SEQ ID NO: 1. The nucleic acid sequence is a result of confirming the homology through the blast search program provided by NCBI GenBank (http://www.ncbi.nin.gov), Bacillus subtilis NCIB 3610 strain and It was confirmed that they were 99% identical.

Therefore, the microorganism of the present invention having the 16S rRNA base sequence of SEQ ID NO: 1 was named Bacillus subtilis TSD strain, and was deposited with the Korean Seed Association (KCCM) on October 2, 2019 (trusted). No. KCCM12595P).

The Bacillus subtilis TSD strain of the present invention is a Gram-positive bacterium, a facultive anaerobe capable of growth under both aerobic and anaerobic conditions, and takes the form of a bacillus (see FIG. 1).

When the Bacillus subtilis TSD strain of the present invention is fermented together with steamed soybeans, the content of amino nitrogen is increased to make soybean with better flavor, so it is very useful for the production of soybean paste.

In one embodiment, the present invention provides a starter composition for fermentation comprising the Bacillus subtilis TSD strain (Bacillus subtilis TSD) strain.

In the present invention, the term "starter (starter)" refers to a preparation containing microorganisms involved in the fermentation of food, and refers to a microorganism that predominantly grows in aged food by adding it during the manufacture of fermented food. In the present invention, the starter refers to a preparation essentially containing a microorganism, that is, a Bacillus subtilis TSD strain, which constantly controls the taste of fermented food and provides excellent sensory taste.

In one embodiment, the starter composition may be used for the production of fermented food. The fermented food may include, but is not limited to, fermented foods such as soybean paste, kimchi, and cheese, and may be prepared by various methods known in the art.

In one embodiment of the present invention, the Bacillus subtilis TSD strain may be included in a starter composition for fermentation during the manufacture of soybean paste, and when used, it helps to uniformly ripen the soybean paste while maintaining the taste of the well-aged soybean paste for a long period of time. help you keep

In the present invention, the starter composition for fermentation may be formulated as a liquid formulation, a powder formulation, etc., for example, may be used as a powder formulation. The powder formulation can be obtained by collecting the cultured strain by centrifugation or the like and freeze-drying it. When a starter strain is added to a fermented food, it is common to add a wet type of bacteria separated from a culture medium, but liquid formulations are difficult to store and difficult to maintain quality during distribution, so it is difficult to distribute them as products. On the other hand, powder formulations have the advantage of being easy to supply, maintain quality and use. The starter composition may further include not only the Bacillus subtilis TSD strain, but also ingredients typically added to the starter, such as soybean flour, skim milk, amino acids, sugars, sugar alcohols, and starch.

Another aspect of the present invention relates to a method for producing soybean paste using a Bacillus subtilis TSD strain [Accession No.: KCCM12595P] or a culture solution thereof.

In one embodiment, the present invention relates to a method for producing soybean paste comprising; inoculating the steamed soybean with a Bacillus subtilis TSD strain [Accession No.: KCCM12595P] or a culture solution thereof and then fermenting it.

In the above, the present invention may further comprise the step of mixing the steamed soybean fermented product to the meju mixture mixed with meju and brine. The meju mixture may be a mixture of meju and 13 to 16% brine. The meju is not particularly limited as long as it is prepared by a conventional method.

In another embodiment, the present invention comprises the steps of (a) immersing meju in brine and then separating the salted meju from the brine; (B) Bacillus subtilis ( Bacillus subtilis ) TSD strain [Accession No.: KCCM12595P] or a culture medium thereof inoculated and then fermented to prepare a seed fermented product; (c) preparing soybean paste by mixing the seed fermented product and the salted soybean paste; (d) fermenting the soybean paste prepared in step (c); may be a method for producing soybean paste comprising.

In the doenjang manufacturing method, the Bacillus subtilis TSD strain may be added as a strain itself or added as a culture solution. The culture solution may include the culture solution itself in which the strain is cultured in a liquid medium, a concentrate obtained by concentrating the culture solution, and the like.

^{The Bacillus subtilis TSD strain is inoculated at a concentration of 4X10 9} to 5X10 ¹⁰ CFU based on 1 kg of soybean paste. If the concentration of the strain is less than the lower limit, doenjang with improved flavor can be obtained even if a strain isolated from soybean paste is used. It is not possible, and when it exceeds the upper limit, antioxidant activity and functionality may be reduced.

When the Bacillus subtilis TSD strain is used for the production of soybean paste, compared to soybean to which the strain is not added, not only does it contain a higher amount of amino nitrogen, but also the taste and flavor such as umami, bitterness, and sweetness are further improved. can be manufactured.

In the present invention, the Bacillus subtilis TSD strain may be added as a powder formulation or a liquid formulation during the preparation of soybean paste in order to exhibit excellent sensuality and an effect of increasing the amino nitrogen content.

Another aspect of the present invention is a mixture of steamed oat powder and steamed soybeans inoculated with Bacillus subtilis TSD strain [Accession No.: KCCM12595P] or a culture solution thereof and then fermented; oats comprising; It relates to a method for manufacturing soybean paste. The steamed oat powder may be prepared by steaming oats, drying them, and then pulverizing them.

In another embodiment, the present invention comprises the steps of (a) immersing meju in brine and then separating the salted meju from the brine; (b) preparing steamed oat powder by steaming the oat powder; (c) inoculating a Bacillus subtilis TSD strain [Accession No.: KCCM12595P] or a culture solution thereof in a mixture of steamed oat powder and steamed soybeans, followed by fermentation to prepare a seed fermented product; (d) preparing soybean paste by mixing the seed fermented product and the salted soybean paste; and (e) fermenting the soybean paste prepared in step (d).

In the oat soybean paste manufacturing method of the present invention, in step (a), the meju is immersed in brine and then the salted meju is separated from the brine. At this time, the salted meju may have a salinity of 11 to 14%, preferably 11.5 to 14.5%, more preferably 12 to 13%.

In addition, in step (b), the steamed oat powder is prepared by steaming the oat powder.

The steamed oat powder is prepared by heating the oat powder at 90 to 110 ℃, preferably 90 to 100 ℃, more preferably 95 to 100 ℃ for 20 to 50 minutes, preferably for 25 to 40 minutes, more preferably 25 It can be prepared by heat treatment for 35 minutes. When the steamed oat powder prepared by the above method is used as a raw material for soybean paste, it is possible to further enhance the umami and flavor of the doenjang.

And, in step (c), a mixture of steamed oat powder and steamed soybeans is inoculated with a Bacillus subtilis TSD strain or a culture solution thereof and then fermented.

Oats are a grain belonging to the Grape family, and contain essential amino acids in a balanced way and contain 2 to 6% by weight of beta-glucan, so they have high nutritional value. The main ingredients contained in the oats include beta-carotene, beta-glucan, vitamin C, niacin, riboflavin, thiamine, retinol, dietary fiber, avenanthramide, etc., among which avenanthramide is As an antioxidant, it is known to inhibit free radicals.

On the other hand, beta-glucan contained in oats is a type of polysaccharide, known to have an immune-enhancing action, reduces blood sugar and blood cholesterol, improves lipid metabolism, and suppresses body fat formation and accumulation, thereby preventing obesity. reported to be effective. In addition, beta-glucan does not directly attack cancer cells, but activates the immune function of normal human cells through a non-specific immune response to suppress the proliferation and recurrence of cancer cells, and activate macrophages to enter the body of cancer cells and activate various cytokines. It activates the immune function of T cells and B cells, which are immune cells, by promoting the secretion of (Cytokine). On the other hand, the taste is sweet, and the color is dark blackish brown.

An object of the present invention is to provide a method for preparing oat soybean that can enrich the taste and flavor of soybean while easily ingesting various physiologically active ingredients of the oats by adding steamed oat powder in the process of manufacturing soybean paste.

To this end, the steamed oat powder is 3.0 to 5.0% by weight, preferably 3.25 to 4.75% by weight, more preferably 3.5 to 4.5% by weight, more preferably 3.6 to 4.4% by weight, more preferably based on the total weight of soybean paste Preferably 3.75 to 4.25% by weight, more preferably 3.8 to 4.2% by weight, most preferably 4% by weight.

In the case of adding steamed oat powder in excess of the upper limit when manufacturing soybean paste, the characteristic taste of oats is strengthened, and the bitter taste and sour taste are strengthened, thereby deteriorating the umami taste of soybean paste. In addition, the increase in the volume occupied by steamed oats resulted in poor growth of the seed bacteria.

In addition, when steamed oat powder is added below the lower limit, the effect of adding oats becomes insignificant.

The steamed oat powder and steamed beans of step (c) may be mixed in a weight ratio of 3 to 5: 10, preferably 3.5 to 4.5: 10 by weight.

In addition, the fermentation of step (c) may be a solid fermentation of the steamed oat powder and the steamed soybean mixture at 28 to 35° C. for 35 to 55 hours.

And, in step (d), doenjang is prepared by mixing the seed fermented product obtained in step (c) and the salted meju obtained in step (a).

The seed fermented product for producing oat soybean paste is 10 to 25% by weight, preferably 10 to 20% by weight, more preferably 11 to 17% by weight, more preferably 12 to 16% by weight of the total weight of soybean paste, More preferably, it may be mixed in an amount of 13 to 15% by weight to prepare soybean paste.

Specifically, the seed fermented product is the salted soybean paste in a weight ratio of 0.5 to 3: 7 to 9.5, preferably 0.5 to 2: a weight ratio of 8 to 9.5, more preferably 1 to 2: a weight ratio of 8 to 9, more Preferably 1.2 to 1.8: 8.2 to 8.8, more preferably 1.4 to 1.6: may be mixed in a weight ratio of 8.4 to 8.6.

The salted meju may be mixed at 70 to 80% by weight based on the total weight of soybean paste, and the salinity may be adjusted by appropriately adding steamed soybeans.

The doenjang prepared in step (d) is adjusted to 8 to 12%, preferably 9 to 11%, more preferably 9 to 10% of the salinity of the doenjang using steamed soybeans or salted soybean.

In the method for producing oat soybean paste according to the present invention, when the steamed oat powder is added in the form of a seed fermented product previously fermented with a Bacillus subtilis TSD strain or a culture solution thereof when oats are added, the steamed oat powder and Bacillus subtilis Compared to the soybean paste prepared by adding each TSD strain, the flavor and umami taste are further increased, and the soybean paste with higher preference can be manufactured because there is no bitter or sour taste of oats at all. In addition, when oat soybean paste is prepared by the production method according to the present invention, soybean paste having higher amino nitrogen content and beta-glucan content can be produced.

Specifically, in the following example, as a result of comparing soybean paste inoculated with the Bacillus subtilis TSD strain and not soybean paste during production of soybean paste, the strain is a dominant species in doenjang inoculated with the strain, leading to fermentation while leading the amino It was confirmed that the nitrogen content and beta-glucan content were increased. In addition, although not described in the following examples, when the above strain is added during the production of soybean paste, a specific experiment was conducted to demonstrate that the antioxidative effect, antibacterial effect and anticancer effect in doenjang can be increased by the presence of the strain as the main fermentation bacterium from the beginning of the fermentation of doenjang confirmed through.

In addition, the present invention provides, characterized in that it comprises the Bacillus subtilis ( Bacillus subtilis ) TSD strain [Accession No.: KCCM12595P] or a culture solution thereof, prepared according to the method described above, it provides oat soybean paste.

The oat soybean paste may be included in 10 to 25 wt% of a fermented seed fermented by Bacillus subtilis TSD strain of steamed oat powder and steamed soybeans, 70 to 80 wt% of salted soybean, and 2.5 to 15 wt% of steamed soybeans have. In this case, the steamed oat powder may be included in an amount of 3.0 to 5.0% by weight based on the total weight of the oat soybean paste.

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

<Example>

Example 1: Selection of fermented soybean paste

1-1. Preparation of strains

Various kinds of soybean paste prepared by a conventional doenjang manufacturing method were collected and used as a strain-isolated soybean paste sample. The soybean paste samples were collected, diluted 10-fold with 0.85% saline, and inoculated into LB agar medium by 0.1 ml each, and then smeared with a glass rod. Thereafter, the plate was incubated for 1 day in an incubator at 37°C. Each of the produced colonies was inoculated with LB agar plates, and cultured at 37° C. for 1 day to isolate strain colonies. The following various experiments were performed using the strain colonies.

1-2. Assessment of protease activity

In order to select a fermenting seed suitable for soybean paste fermentation, a strain capable of improving soybean protein degradation, that is, a strain with excellent proteolytic enzyme activity, was selected.

Accordingly, 25 strains were selected from among the various strain colonies and the protease activity was compared.

The protease activity of the strains was inoculated with each strain in tryptic soy broth (TSB) medium, cultured at 37° C. for 24 hours, and then the culture supernatant was recovered, and then measured using an Azocasein assay. The protease activity of each strain measured in this way is shown in Table 1 below.

strain name Protease activity (unit/mL) strain name Protease activity (unit/mL) SYD 523 BYD 633 HJD1 1,647 WSJD 711 SND 126 HJD2 796 YKD 588 YSD 1,176 SKD 1,664 GP6 238 KMD 1,622 GP8 462 GDD 571 J2 318 SJD 101 J3 432 EJD 715 J4 1,240 HMD 437 GG1 39 MHD 385 GG2 1,546 IGD 508 GG3 689 TSD 1,471

In Table 1, a total of 7 strains, HJD1, SKD, KMD, TSD, YSD, J4 and GG2 strains having protease activity of 1,000 unit/mL or more, were selected.

1-3. Fermentation flavor evaluation

In order to confirm the fermented flavor when the soybean paste was prepared using the seven selected strains, the flavor of the fermented soybean after fermenting the soybean with each strain was evaluated.

For this purpose, the soybeans were sufficiently soaked and steamed, and then ground. 1% (0.2 mL) of each strain (HJD1, SKD, KMD, TSD, YSD, J4 and GG2) culture solution was inoculated into 20 g of the ground steamed soybean and fermented at 30° C. for 3 days. Thereafter, the number of viable cells and proteolytic enzyme activity of the fermented steamed soybean were measured, and the fermented flavor was evaluated, and the results are shown in Table 2 below.

strain name proteolytic enzyme activity
(unit/g) Number of viable cells (cfu/g) flavor HJD1 2,493 2.8×10 ⁹ The smell of decaying soybean paste and ammonia SKD 3,240 2.4×10 ⁹ There is a putrefactive smell such as ammonia KMD 3,756 4.5×10 ⁹ Strong ammonia and cumulus aroma TSD 3,612 3.6×10 ⁹ Fermented soybean flavor YSD 3,635 1.2×10 ⁹ Chunky soybean flavor J4 1,236 1.3×10 ⁷ Soybean fishy smell (rarely fermented flavor) GG2 4,416 4.8×10 ⁸ Slight ammonia and cumulus odor

Referring to Table 2, the TSD strain had the best flavor and showed stable results in proteolytic activity and live cell count, so the TSD strain was selected as the fermented soybean paste.

1-4. Morphological Characteristics

For the analysis of the morphological characteristics of the TSD strain, the shape and color of the colonies of the strain were observed, and the shape was observed with an optical microscope after the strain was Gram stained (Gram stain kit, BD Co., USA), as a result. is shown in FIG. 1 .

As shown in FIG. 1 , it was confirmed that the TSD strain, which is an isolated strain, was a Gram-positive strain, and as a result of optical microscopic observation, it was in the form of a bacillus, and the color of the colony was light ivory.

1-5. Molecular biological identification of isolated strains

To investigate the molecular biological characteristics of the TSD strain selected in Examples 1-3, after chromosomal DNA was isolated from the strain culture, the chromosomal DNA was subjected to full sequencing of the 16s rRNA gene to identify microorganisms. The identified strain was investigated to be a strain belonging to Bacillus subtilis , and the nucleotide sequence (16s rRNA sequence) of the identified strain was shown in SEQ ID NO: 1 (see FIG. 2).

As a result of comparing the nucleotide sequence of the strain with other strains registered in GenBank, it showed 99% homology with the standard strain Bacillus subtilis NCIB 3610 strain, and a strain having 100% homology with SEQ ID NO: 1 exists in the past. confirmed not to do so. Therefore, the present inventors deposited these strains with the Korean Seed Association (KCCM) on October 2, 2019 and were given the following accession numbers.

Bacillus subtilis ( Bacillus subtilis ) - Accession number KCCM12595P (aka "TSD")

1-6. Optimization of culture conditions of isolated strains

It was intended to select the optimal culture conditions of the Bacillus subtilis TSD strain of the present invention. As a medium for the TSD strain, a tryptic soy broth (TSB) medium was used.

Incubation temperature of TSD strain

For the selection of the culture temperature conditions of the Bacillus subtilis TSD strain, the TSD strain was cultured at 30 ℃, 33 ℃, 35 ℃, 37 ℃, 39 ℃, 41 ℃ and 50 ℃, respectively, the number of viable cells and proteolysis of the TSD strain Enzyme activity was measured and shown in Table 3 below. At this time, the protease activity was measured by the Azocasein assay method.

Growth temperature (℃) Number of viable cells (Log cfu/mL) Protease activity (Unit/mL) 30 8.22 1,394 33 8.49 1,425 35 8.82 1,559 37 9.76 1,615 39 8.59 1,497 41 7.03 567 50 6.82 293

Looking at Table 3, the Bacillus subtilis TSD strain was relatively stable in the number of viable cells and protease activity at 30 to 39 °C, was stable at 35 to 39 °C, and exhibited the best results at 37 °C.

Incubation time of TSD strains

In order to select the culture time conditions of the Bacillus subtilis TSD strain, while the TSD strain was cultured at 37 ° C. for 120 hours, the number of viable cells and the proteolytic enzyme activity of the TSD strain according to the lapse of culture time were measured for each culture time. 4 is shown.

3 is a graph showing the number of viable cells according to the growth time of the Bacillus subtilis TSD strain, and FIG. 4 is a graph showing the protease activity according to the growth time of the Bacillus subtilis TSD strain. to be.

3 and 4, the Bacillus subtilis TSD strain was excellent in the number of viable cells and protease activity when cultured at 37 ° C. for 4 to 24 hours, and was even better when cultured for 8 to 20 hours, 16 In the case of incubation for a period of time, the best results were obtained.

From these results, it can be confirmed that the Bacillus subtilis TSD strain of the present invention is the optimal culture condition for the strain to be cultured at 37 ℃ for 16 hours.

Example 2: Preparation of soybean paste using TSD strain

2-1. preparation of meju

After immersing common meju in brine for about 40 days, meju was separated from the brine to prepare meju having a salinity of about 12%.

2-2. Preparation of strains

The Bacillus subtilis TSD strain isolated and identified in Example 1 was spread on a plate medium and cultured in an incubator at 37° C. for 48 hours to isolate a single colony. The isolated colonies were collected, added to the liquid MRS medium, and cultured for 16 hours at 37 °C and 200 rpm to activate the TSD strain.

2-3. ready roasted beans

Soybeans (soybeans) were washed twice, and water 1.4 times the volume of beans was added and immersed in water for 20 to 24 hours. Thereafter, the soaked soybeans were removed and steamed for 2-3 hours in a steaming pot at 100~150 ℃, then the water was removed, cooled to 35 ℃, and then ground.

2-4. Production of fermented seeds

^{After inoculating 1 mL of the TSD strain culture solution (2×10 9} cfu/ml) prepared in 2-2 to 100 g of the steamed soybean prepared in 2-3, it was fermented at 30° C. for 48 hours to prepare a fermented seed.

2-5. production of miso

Doenjang was prepared by mixing the seed fermented product prepared in 2-4 in a weight ratio of 10% of the total weight of doenjang. At this time, the salinity of the doenjang was adjusted to 9% using the steamed soybean prepared in 2-3 and the meju prepared in 2-1.

2-6. Fermentation of miso

The prepared soybean paste was fermented for 4 weeks at 30° C. and 75% humidity.

Control: Preparation of TSD strain-free soybean paste

Doenjang was prepared in the same manner as in Example 2, but without adding a TSD strain during the manufacturing process of soybean paste.

<Test Example>

How to analyze the quality characteristics of soybean paste

1) Salinity

- After adding 15 g of distilled water to 5 g of soybean paste and shaking for 10 minutes, 3 g of water was added to 1 g of the supernatant obtained by centrifugation, and the salinity was measured with a salinity meter (ATAGO).

2) sodium content

- The sodium content of the same sample as the salinity measurement was measured using a Methrom potentiometric titrator (859 Titrotherm).

3) Amino nitrogen (AN) content

- For the amino nitrogen content, 50 mL of distilled water was added to 3 g of soybean paste by the formal titration method, filtered, and 2-3 drops of 1% phenolphthalein indicator were added and titrated with 0.1 N NaOH until pale red.

- 30 mL of formalin solution was added to another container, 2~3 drops of 1% phenolphthalein indicator were added, and titrated with 0.1 N NaOH until pale red.

- The two solutions were mixed and titrated with 0.1 N NaOH until pale red, and the amino nitrogen content was calculated using the following formula.

AN (mg%) = (0.1 N NaOH consumed × 0.0014 × 100 × 1000 × dilution factor) / sample amount (g)

4) pH

- The sample prepared in the same way as the salinity measurement was measured using a pH meter (SevenMulti).

5) Chromaticity

- After correcting with a standard color plate (L:96.85, a:-0.28, b: -0.30) using a colorimeter (Chroma meter, Cm-3500d, Monolta, Japan), L value (brightness), a value (red Figure) and b value (yellowness) were measured 5 or more times, respectively, and expressed as the average and deviation.

6) Number of microorganisms (general bacteria, yeast)

-The number of viable cells of general bacteria and yeast was analyzed, and film medium (3M) was used.

- For general bacteria, AC (for general bacteria) film medium was used, and for yeast, YM (for fungi) film medium was used.

- For the analysis of the number of microorganisms, 1 g of soybean paste was taken and a 10-fold diluted solution was prepared using distilled water. After inoculating 1 mL of the diluent on the film medium and pressing with a press plate to harden, the number of colonies was counted by culturing for 24 hours at 37 °C for general bacteria and 3 days for yeast at 30 °C.

7) Beta glucan content

- The beta-glucan content of oats and doenjang was analyzed using the McCleary method applied beta-glucan analysis kit (Megazyme Mixed-linkage Beta-glucan (K-BGLU)).

Test Example 1: Evaluation of soybean paste prepared using TSD strain

1-1. Analysis of quality characteristics of doenjang prepared using TSD strain

In order to compare the quality characteristics of the control, soybean paste prepared in Example 2, the pH, amino nitrogen content (mg%), total number of bacteria, and chromaticity of each soybean sample were measured and shown in Table 4 below.

division pH amino nitrogen
(mg%) total bacteria
(cfu/mL) chromaticity Brightness (L value) Redness (a value) Yellowness (b value) control 4.40 217 1×10 ¹¹ 52.83 10.02 21.75 Example 2 4.43 278 9×10 ¹⁰ 50.60 10.70 22.80

Looking at Table 4, it can be seen that in the case of the doenjang according to Example 2 of the present invention, the amino nitrogen content is significantly higher than that of the control (general doenjang).

1-2. Sensory evaluation of doenjang prepared using TSD strain

For the control group and the soybean paste prepared in Example 2, sensory evaluation was performed, and it is shown in Table 5 below.

For the sensory evaluation, 10 experts in the taste evaluation of soybean paste were composed of a panel, and the evaluation was performed according to the 9-point scale method (strength: 1. very bad; 5. average; 9. very good). At this time, statistical processing was performed using the SPSS 12.0 statistical program, and significance was verified at the significance level p<0.05 by the DMR-test (Duncan multiple range test).

division color flavor incense salty umami bitter sweetness Comprehensive symbol map control 6.6 5.6 5.8 6.4 4.8 4.8 5.6 5.0 Example 2 6.8 6.4 6.6 6.8 6.2 6.0 6.2 6.7

Looking at Table 5 above, in the case of the doenjang according to Example 2 of the present invention, the overall preference was increased compared to the control (general soybean paste), in particular, the umami taste was significantly improved, the flavor was further improved, and the taste such as bitter taste and sweet taste was also You can see the improvement.

Test Example 2: Quality characteristics of soybean paste prepared by adding oats

1-1. Quality characteristics of soybean paste according to oat pretreatment method

1) Raw material analysis and pretreatment method of oats

For oats, oats from Gangjin were purchased in the form of grains, ground with a mixer, and then powder of 30 mesh or less was used as a raw material. The general components and beta-glucan content contained in the oat powder were analyzed and shown in Table 6 below.

sample moisture(%) Views (%) Crude protein (%) Crude fat (%) carbohydrate(%) Beta glucan (%) strong oats 6.26 1.33 12.90 10.22 69.29 2.06

In addition, various pretreatment methods of oats for addition to soybean paste are summarized in Table 7 below.

sample Pretreatment method oat powder The oats were ground with a mixer to prepare a powder. increased oats Oat powder was prepared by steaming at 100° C. for 30 minutes. roasted oats Oats were roasted in a pan at 180° C. for 3 minutes and then pulverized to prepare roasted oat powder. singer luxury oats It was prepared by adding three times the weight of water to the oat powder, and heat-treating it for 20 minutes at low heat to make it gelatinous.

2) Manufacture of soybean paste with oats

The purpose of this study was to investigate the optimal oat pretreatment method that can harmonize the taste and aroma of oats and soybean paste in the case of manufacturing doenjang using oats. For this purpose, in the doenjang prepared using oats, the composition of the doenjang according to the oat pretreatment method is summarized in Table 8 below.

division Doenjang Raw Material (Meju) (%) Steamed Beans (%) oat(%) Sum(%) control 75 25 0 100 oat powder 75 21 4.0 100 increased oats 75 21 4.0 100 roasted oats 75 21 4.0 100 singer luxury oats 75 17.2 7.8
(4% based on oat powder) 100

3) Analysis of the quality characteristics of soybean paste according to the oat pretreatment method

After preparing oat soybean paste with the above composition, it was fermented for 3 weeks at 30° C. and 75% humidity, followed by analysis.

In order to compare the physicochemical quality characteristics of doenjang prepared by different oat pretreatment methods after the fermentation, pH, amino nitrogen content (mg%), total number of bacteria, color and beta-glucan content of each doenjang sample were measured. Then, it is shown in Table 9 below. At this time, the doenjang was measured to have a salinity of about 9% and a sodium content of 3.8% in common.

division pH amino nitrogen
(mg%) total bacteria
(cfu/mL) chromaticity beta glucan
(mg%) brightness redness yellowness control 4.38 302 1.0×10 ¹¹ 57.08 10.35 23.08 - oat powder 4.24 280 8.7×10 ⁹ 58.39 9.72 23.79 93.01 increased oats 4.33 292 4.8×10 ¹⁰ 58.07 9.54 23.09 87.70 roasted oats 4.30 257 7.3×10 ¹⁰ 55.49 10.24 23.51 86.90 singer luxury oats 4.18 307 5.7×10 ⁹ 56.46 10.15 23.18 88.82

Looking at Table 9, the pH was a little lower in the hydrogelled oats added group, and the amino nitrogen content was the lowest in the roasted oat added group. The beta-glucan content was slightly higher in the oat powder-added group, but there was no significant difference between all samples.

4) Sensory evaluation of soybean paste according to the oat pretreatment method

For each doenjang prepared by changing the pretreatment method of oats, sensory evaluation was performed and shown in Table 10 below.

For the sensory evaluation, 10 experts in the taste evaluation of soybean paste were composed of a panel, and the evaluation was performed according to the 9-point scale method (strength: 1. very bad; 5. average; 9. very good). At this time, statistical processing was performed using the SPSS 12.0 statistical program, and significance was verified at the significance level p<0.05 by the DMR-test (Duncan multiple range test).

division color flavor incense salty umami bitter sweetness Synthesis
symbol control 6.0 5.2 5.8 5.4 5.2 5.6 5.2 5.2 oat powder 6.0 5.6 6.2 6.0 5.6 6.0 5.6 5.5 increased oats 6.0 6.8 6.8 6.4 6.8 6.6 6.4 6.6 roasted oats 7.0 6.2 6.2 6.6 6.4 5.4 6.0 6.2 singer luxury oats 6.0 5.8 5.8 6.0 5.6 5.8 6.0 5.6

As shown in Table 10, it was confirmed that the doenjang of the control group had a weak umami taste, a salty taste, and a strong taste and aroma. Doenjang prepared by adding oat powder had increased umami and flavor compared to the control group. Doenjang prepared by adding roasted oats had a savory taste, but had a burnt aftertaste, and doenjang prepared by adding hydrolyzed oats had a weak sweetness and umami taste. Overall, it was confirmed that the umami taste and savory taste were increased when oats were added compared to the control group.

On the other hand, soybean paste prepared by adding steamed oat powder had a strong umami taste unique to oats and had a savory aftertaste, so it had the best sensory preference. Since the group with the added steamed oats had the highest overall preference, the steaming method was selected as the oat pretreatment method during the production of soybean paste.

1-2. Quality characteristics of soybean paste according to the amount of steamed oat powder added

1) Preparation of soybean paste with steamed oat powder

In the case of making soybean paste using oats, in order to find out the optimal range of oat addition amount in which the taste and flavor of oats and soybean paste can be most harmonious, prepare oat soybean paste with the following composition, After fermentation for a week, the analysis was performed. For this purpose, in the doenjang prepared by adding steamed oat powder, the composition of the doenjang according to the added amount of steamed oat powder is summarized in Table 11 below. In Table 10 below, the fermented seed means 1 mL of the TSD strain culture solution in 100 g of steamed soybeans and then cultured at 30° C. for 48 hours.

division Meju (%) Steamed Beans (%) soybean raw material
Sum(%) increased oats
powder(%) Seed (TSD strain) fermented product (%) Sum(%) control 75 25 100 - - 100 Steamed oats 2.0% 75 13 88 2.0 10 100 Steamed oats 4.0% 75 11 87 4.0 10 100 Steamed oats 6.0% 75 9 85 6.0 10 100

2) Analysis of quality characteristics of soybean paste according to the amount of steamed oats added

After preparing oat soybean paste with the above composition, it was fermented for 3 weeks at 30° C. and 75% humidity, followed by analysis.

In order to compare the physicochemical quality characteristics of soybean paste according to the amount of steamed oats added, the pH, amino nitrogen content (mg%), total number of bacteria and beta-glucan content of each doenjang sample were measured, respectively, and are shown in Table 12 below. At this time, the doenjang was measured to have a salinity of about 9% and a sodium content of 3.8% in common.

division pH amino nitrogen
(mg%) total bacteria
(cfu/mL) beta glucan
(mg%) control 4.55 278 2.7×10 ¹⁰ - Steamed oats 2.0% 4.37 320 2.5×10 ¹⁰ 59.40 Steamed oats 4.0% 4.26 339 3.0×10 ¹⁰ 96.29 Steamed oats 6.0% 4.38 343 2.8×10 ¹⁰ 110.38

Looking at Table 12, it can be seen that the amino nitrogen content and beta-glucan content of the soybean paste sample increased in proportion to the amount of oats added.

3) Sensory evaluation of soybean paste according to the added amount of steamed oats

For each doenjang prepared by varying the amount of steamed oats added, a sensory evaluation was performed and shown in Table 13 below.

For the sensory evaluation, 10 experts in the taste evaluation of soybean paste were composed of a panel, and the evaluation was performed according to the 9-point scale method (strength: 1. very bad; 5. average; 9. very good). At this time, statistical processing was performed using the SPSS 12.0 statistical program, and significance was verified at the significance level p<0.05 by the DMR-test (Duncan multiple range test).

division color flavor incense salty umami bitter sweetness Synthesis
symbol control 6.4 5.2 5.4 5.2 5.2 5.2 5.2 5.2 Steamed oats 2.0% 6.6 6.6 6.4 6.4 6.4 6.4 6.4 6.8 Steamed oats 4.0% 6.6 7.2 6.6 6.8 7.0 6.6 7.0 7.2 Steamed oats 6.0% 5.8 5.6 6.0 6.0 5.8 5.6 6.0 5.8

As shown in Table 13, the doenjang of the control group lacked umami and flavor, and it was confirmed that the doenjang prepared by adding steamed oat powder had superior acceptance compared to the control group. The group added with 2.0% steamed oats felt savory and sweet compared to the control group, the group added with steamed oats with 4.0% had the best umami taste and had excellent sweetness and flavor, and the group with steamed oats added with 6.0% showed umami taste and preference due to the bitterness and sourness. was lowered

Since the doenjang with steamed oats added at 4.0% by weight had high scores in all evaluation items, the amount of oat added during the production of oat soybean was selected under the condition of 4%.

Example 3: Preparation of oat soybean paste using oats and fermented seeds

It was carried out in the same manner as in Example 2, but 2-4. In the step of adding the TSD strain culture solution to the steamed soybean of the step, the steamed oat powder was added together to prepare oats and fermented seeds. At this time, 2-4. The steamed oat powder in the step was added in an amount of 40 parts by weight (4% by weight based on the total weight of soybean paste) based on 100 parts by weight of the steamed soybean.

In addition, in the mixing step of 2-5. the fermented seed and soybean paste, oat and soybean paste was prepared by mixing the oats and the fermented seed broth in an amount of 14% by weight of the total weight of the soybean paste.

Comparative Example 1: Preparation of oat miso using steamed oat powder

It was carried out in the same manner as in Example 2,

The step of preparing the seed fermentation product of 2-4. is not carried out,

2-5. In step 2-4, instead of the seed fermented product prepared in step 2-4, the steamed soybean prepared in 2-3 is mixed with the soybean prepared in 2-1 to prepare doenjang, and then steamed oat powder in an amount of 4% by weight based on the total weight of the doenjang. were mixed to prepare oat soybean paste.

Comparative Example 2: Preparation of oat soybean paste using steamed oat powder and TSD strain

It was carried out in the same manner as in Example 2,

In the step of mixing the fermented seeds and meju in 2-5., 10% by weight of the fermented seeds and 4% by weight of steamed oat powder were mixed together to prepare doenjang based on the total weight of soybean paste.

Comparative Example 3: Preparation of oat soybean paste using sterilized oats and fermented seeds

It was carried out in the same manner as in Example 3,

Oat doenjang was prepared by sterilizing the oats and seed fermented product prepared in 2-4. at 121° C. for 15 minutes.

The manufacturing method and composition of the oat soybean paste of Examples and Comparative Examples 1 to 3 are briefly summarized in Tables 14 and 15 below.

division remark control Normal miso with 9% salinity Example 2 In the production of soybean paste, the fermented seed fermented soybean with TSD strain is added. Example 3 When making soybean paste, steamed oat powder and steamed soybeans are fermented with TSD strains, and fermented oats and seed fermented products are added. Comparative Example 1 Added steamed oat powder when making soybean paste Comparative Example 2 When making soybean paste, steamed oat powder and TSD seed fermented product are added, respectively. Comparative Example 3 When making soybean paste, steamed oat powder and steamed soybeans are fermented with TSD strain and added after sterilization of oats and seed fermented products

division Meju (%) Steamed Beans (%) Total soybean raw materials (%) Steamed oat powder (%) Seed (TSD strain) fermented product (%) Oat & Seed Fermentation (%) Sum(%) control 75 25 100 - - - 100 Example 2 75 15 90 - 10 - 100 Example 3 75 11 86 - - 14 100 Comparative Example 1 75 21 96 4 - - 100 Comparative Example 2 75 11 86 4 10 - 100 Comparative Example 3 75 11 86 - - 14 100

Comparative Examples 4-9: Preparation of oat soybean paste with different strains

It was carried out in the same manner as in Example 3, but instead of the TSD strain, a Bacillus subtilis strain or an Aspergillus duckjae strain showing an excellent effect on soybean paste was sold from the Korea Microbial Conservation Center or Agricultural Biotechnology Research Institute to prepare oat soybean paste. . The strains used in Comparative Examples 4 to 9 of the present invention are summarized in Table 16 below.

division strain strain name deposit number Organization name Example 3 Bacillus subtilis TSD KCCM12595P Korea Microorganism Conservation Center Comparative Example 4 Bacillus subtilis J46 KCCM12388P Korea Microorganism Conservation Center Comparative Example 5 Bacillus subtilis CSY388 KACC91564P Agricultural Biotechnology Research Institute Comparative Example 6 Bacillus subtilis AFY-2 KACC91988P Agricultural Biotechnology Research Institute Comparative Example 7 Aspergillus duck M385 KACC93210P Agricultural Biotechnology Research Institute Comparative Example 8 Aspergillus duck CJ KY KCCM11302P Korea Microorganism Conservation Center Comparative Example 9 Aspergillus duck CJ KG KCCM11301P Korea Microorganism Conservation Center

Test Example 2: Characterization of oat soybean paste using steamed oats and TSD strains

Each of the oat soybean paste samples prepared in the control group, Examples 3 and Comparative Examples 1 to 9 were fermented at 30° C. (water 75%) for 4 weeks, and then quality characteristics were analyzed.

2-1. Analysis of quality characteristics of oat miso

pH, amino nitrogen content (mg%), total number of bacteria, seed dominance (%) and beta-glucan content (mg%) were measured using the oat soybean paste samples of the control group, Examples 3 and Comparative Examples 1 to 9, respectively. Thus, it is shown in Table 17 below.

division pH amino nitrogen
(mg%) total bacteria
(cfu/mL) Seed dominance
(%) beta glucan
(mg%) control 4.55 278 2.7×10 ¹⁰ - - Example 3 4.26 342 3.1×10 ¹⁰ 87 105.70 Comparative Example 1 4.33 292 4.8×10 ¹⁰ - 87.70 Comparative Example 2 4.34 322 8.5×10 ¹⁰ 62 93.92 Comparative Example 3 4.36 284 3.3×10 ¹⁰ - 89.57 Comparative Example 4 4.38 279 2.4×10 ⁹ - 85.34 Comparative Example 5 4.31 281 3.1×10 ⁹ - 85.67 Comparative Example 6 4.43 260 1.8×10 ¹⁰ - 83.21 Comparative Example 7 4.51 259 5.4×10 ⁸ - 86.52 Comparative Example 8 5.18 278 1.6×10 ⁹ - 88.30 Comparative Example 9 4.32 245 2.9×10 ⁹ - 84.78

Looking at Table 17, in the case of the oat soybean of Example 3, it can be seen that the content of amino nitrogen is higher than that of the oat soybean of Comparative Examples 1 and 3 prepared by different manufacturing methods. In addition, in Comparative Example 2, in which steamed oat powder and TSD strain were fermented in advance and mixed with meju instead of mixed with meju to prepare oat doenjang, the seed dominance rate and beta-glucan content were lower than those of the example.

In addition, in the case of the oat doenjang of Example 3, the amino nitrogen content and beta-glucan content were significantly higher than the oat doenjang of Comparative Examples 4 to 9 prepared using other strains instead of the TSD strain.

2-2. Sensory evaluation of oat soybean paste

The sensory evaluation was performed on each of the oat soybean paste samples prepared in the control group, Examples 3 and Comparative Examples 1 to 9, and is shown in Table 18 below.

For sensory evaluation, a panel of 10 experts in soybean paste taste evaluation was conducted, and the evaluation was performed according to the 9-point scale method (strength: 1. Very poor; 5. Moderate; 9. Very good). At this time, statistical processing was performed using the SPSS 12.0 statistical program, and significance was verified at the significance level p<0.05 by the DMR-test (Duncan multiple range test).

division color flavor incense salty umami bitter sweetness Synthesis
symbol control 6.0 5.2 5.8 5.4 5.2 5.6 5.2 5.2 Example 3 7.2 7.9 7.0 7.0 7.6 7.0 7.4 7.8 Comparative Example 1 6.0 6.8 6.8 6.4 6.8 6.6 6.4 6.6 Comparative Example 2 6.6 7.2 6.6 6.8 7.0 6.6 7.0 7.2 Comparative Example 3 7.0 6.2 6.0 6.0 6.2 6.0 6.0 6.2 Comparative Example 4 6.4 6.2 6.6 6.8 6.2 5.8 5.8 6.9 Comparative Example 5 6.0 6.8 6.2 6.6 6.0 5.8 5.4 6.7 Comparative Example 6 7.2 7.1 6.7 6.6 6.8 6.0 6.0 7.2 Comparative Example 7 6.8 7.1 7.1 6.8 7.1 5.9 6.0 7.3 Comparative Example 8 6.8 6.8 7.2 6.4 6.8 6.2 5.8 7.1 Comparative Example 9 6.6 6.5 6.9 6.5 6.6 5.7 5.3 6.8

Looking at Table 18, in the case of the oat doenjang of Example 3, compared to the oat doenjang of Comparative Examples 1 to 3 prepared by different manufacturing methods, and the oat doenjang of Comparative Examples 4 to 9 prepared using other strains instead of the TSD strain, The umami and flavor are very good, and it can be seen that the aged taste is excellent compared to the fermentation period.

Although the present invention has been described as the above-mentioned preferred embodiment, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also intended that the appended claims cover such modifications and variations as fall within the scope of the present invention.

<110> Jeonnam Bioindustry Foundation <120> Methods for preparing oat doenjang using a novel Bacillus subtilis TSD <130> HPC-9118 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1509 <212> DNA <213> Bacillus subtilis <400> 1 acgctggcgg cgtgcctaaa tacatgcaag tcgagcggac agatgggagc ttgctccctg 60 atgtttagcg gcggacgggt gagtaacacg tgggtaacct gcctgtaaga ctgggataac 120 tccgggaaac cggggctaat accggatggt tgtttgaacc gcatggttca gacataaaag 180 gtggcttcgg ctaccactta cagatggacc cgcggcgcat tagctagttg gtgaggtaac 240 ggctcaccca aggcgacgat gcgtagccga cctgagaggg tgatcggcca cactgggact 300 gagacacggc ccagactcct acgggaggca gcagtaggga atcttccgca atggacgaaa 360 gtctgacgga gcaacgccgc gtgagtgatg aaggttttcg gatcgtaaag ctctgttgtt 420 agggaagaac aagtgccgtt caaatagggc ggcaccttga cggtacctaa ccagaaagcc 480 acggctaact acgtgccagc agccgcggta aatacgtagg tggcaaagcg ttgtccggaa 540 ttattgggcg taaaagggct cggcaggcgg tttcttaagt ctgatgtgaa agccccccggc 600 tcaaccgggg agggtcattg gaaactgggg aacttgagtg cagaagagga gagtggaatt 660 ccacgtgtag cggtgaaatg cgtagagatg tggaggaaca ccagtggcga aggcgactct 720 ctggtctgta actgacgctg aggagcgaaa gcgtggggag cgaacaggat tagataccct 780 ggtagtccac gccgtaaacg atgagtgcta agtgttaggg ggtttccgcc ccttagtgct 840 gcagctaacg cattaagcac tccgcctggg gagtacggtc gcaagactga aactcaaagg 900 aattgacggg ggcccgcaca agcggtggag catgtggttt aattcgaagc aacgcgaaga 960 accttaccag gtcttgacat cctctgacaa tcctagagat aggacgtccc cttcgggggc 1020 agagtgacag gtggtgcatg gttgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc 1080 cgcaacgagc gcaacccttg atcttagttg ccagcattca gttgggcact ctaaggtgac 1140 tgccggtgac aaaccggagg aaggtgggga tgacgtcaaa tcatcatgcc ccttatgacc 1200 tgggctacac acgtgctaca atggacagaa caaagggcag cgaaaccgcg aggttaagcc 1260 aatcccacaa atctgttctc agttcggatc gcagtctgca actcgactgc gtgaagctgg 1320 aatcgctagt aatcgcggat cagcatgccg cggtgaatac gttcccgggc cttgtacaca 1380 ccgcccgtca caccacgaga gtttgtaaca cccgaagtcg gtgaggtaac cttttaggag 1440 ccagccgccg aaggtgggac agatgattgg ggtgaagtcg taacaaggta gccgtatcgg 1500 aaggtgcgg 1509

Claims (5)

Bacillus subtilis ( Bacillus subtilis ) TSD strain [Accession No.: KCCM12595P] or a culture solution thereof inoculated into a mixture of steamed oat powder and steamed soybeans, and then fermented; oat soybean paste manufacturing method comprising a. (a) separating the salted meju from the brine after immersing the meju in brine;
(b) preparing steamed oat powder by steaming the oat powder;
(c) inoculating a Bacillus subtilis TSD strain [Accession No.: KCCM12595P] or a culture solution thereof to a mixture of the steamed oat powder and steamed soybeans, followed by fermentation to prepare a fermented seed;
(d) preparing doenjang by mixing the seed fermented product and the salted soybean paste; and
(e) fermenting the soybean paste prepared in step (d); oat soybean paste manufacturing method comprising a. 3. The method of claim 2,
The steamed oat powder is a method for producing oat soybean, characterized in that it is added in an amount of 3.0 to 5.0% by weight based on the total weight of the soybean paste. 3. The method of claim 2,
The steamed oat powder and steamed soybeans in step (c) are mixed in a weight ratio of 3 to 5: 10. A method for producing oat soybean paste. 3. The method of claim 2,
The fermentation in step (c) is a method for producing oat soybean, characterized in that the solid-state fermentation is performed at 28 to 35 ℃ for 35 to 55 hours.

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