WO2022030662A1 - Novel bacillus velezensis l2 strain isolated from anchovy fermentation solution, and aekjeot prepared using same - Google Patents

Abstract

The present invention relates to: Bacillus velezensis L2 (KCCM12498P) which exhibits a positive reaction to lactose, amygdalin, melibiose and gentiobiose, and which has excellent proteolytic activity; and aekjeot prepared using same. Bacillus velezensis L2 (KCCM 12498P) screened in the present invention is used as a starter for aekjeot fermentation to minimize the generation of histamines, which are substances associated with allergic reactions or inflammation, and increase the generation amount of total nitrogen and amino nitrogen, so that flavor can be improved.

Description

A novel Bacillus belegensis L2 strain isolated from anchovy fermented broth and a fish sauce manufactured using the same

The present invention relates to a novel Bacillus velezensis L2 (KCCM12498P) isolated from anchovy fermentation broth, and more particularly, lactose, amygdalin, melibiose and gentiobiose. ), and it relates to Bacillus velezensis L2 (KCCM12498P) having excellent proteolytic activity.

Fish sauce, along with kimchi, is one of Korea's representative fermented foods and has been used since ancient times to provide high protein and salt regardless of the season.

Specifically, fish sauce is a salting product in a broad sense, obtained by fermenting fish and shellfish by salting and then separating and filtering the supernatant. Salt is added to the muscles, intestines, and eggs of fish and shellfish, and then a specific seed is added to ferment it. By doing so, various amino acids are produced to give special flavor and umami, and it is nutritionally excellent because it is rich in amino acids and inorganic components, and it is also known to have good digestion and absorption rate because it is composed of small molecules.

In general, the fermentation period of the fish sauce is 3 to 24 months, so the production efficiency is lowered. It is necessary to develop strains that become

In addition, the conventional fish sauce contains histamine, one of the biogenic amine components, in a predetermined content or more. This histamine is known to be a component involved in allergic reactions or inflammation. In this case, since it has a problem of causing acute poisoning accompanied by headaches, urticaria, and seizures, it is necessary to develop a new strain capable of minimizing the amount of histamine produced.

The present invention was devised in response to this need, and an object of the present invention is to provide a strain that has excellent protein decomposition ability, can be used as a seed strain for fish sauce fermentation, can increase the content of free amino acids, and can minimize the amount of histamine will do

Another object of the present invention is to provide a fish sauce prepared using the strain.

Objects of the present invention are not limited to the objects mentioned above, and other objects not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the above object, the present invention is Bacillus velezensis comprising the nucleotide sequence set forth in SEQ ID NO: 1 as 16S rRNA, which is isolated from anchovy fermented broth and has proteolytic activity. Gensis ( Bacillus velezensis ) L2 (KCCM1 2498P) is provided.

In a preferred embodiment, the Bacillus velezensis L2 (KCCM1 2498P) shows a positive reaction to lactose (Lactose), amygdalin (Amygdalin), melibiose (Melibiose) and gentiobiose (gentiobiose).

In a preferred embodiment, the Bacillus velezensis L2 (KCCM1 2498P) is used as a seed for the production of a fish sauce using raw materials for fish and shellfish, thereby suppressing the production of volatile basic nitrogen and histamine.

In a preferred embodiment, the Bacillus velezensis L2 (KCCM1 2498P) is used as a seed strain for manufacturing a fish sauce using a raw material for fish and shellfish, thereby increasing the total nitrogen and amino nitrogen content.

In a preferred embodiment, the Bacillus velezensis L2 (KCCM1 2498P) is used as a seed strain for the production of a fish sauce using a raw material for fish and shellfish, thereby increasing the content of sweet amino acids and umami-based amino acids.

In a preferred embodiment, the Bacillus velezensis L2 (KCCM1 2498P) is used as a seed strain for the production of a fish sauce using raw materials for fish and shellfish, and contains serine, glutamic acid, and valine. And increase the content of amino acids including proline (Proline).

In addition, the present invention consists of a culture of the Bacillus velezensis L2 (KCCM 12498P), the Bacillus velezensis L2 (KCCM 12498P), a concentrate of the culture, and a dried product of the culture Provided is a seed composition for fermentation of a fish sauce comprising at least one selected from the group consisting of an active ingredient, and a fish sauce prepared by adding the seed composition for fermentation of a fish sauce.

The present invention has the following excellent effects.

According to Bacillus velezensis L2 (KCCM1 2498P) of the present invention, compared to the standard strain Bacillus velezensis LMG 22478, it has about 1.5 times higher proteolytic activity, making it easier to manufacture fish sauce. When used as a seed bacterium for

In addition, according to Bacillus velezensis L2 (KCCM1 2498P) of the present invention, the content of histamine, which is a component involved in allergic reactions or inflammation, and the content of volatile base nitrogen, a component that is a freshness index, are dramatically reduced. It has the advantage of being able to provide high-quality fish sauce.

In addition, according to Bacillus velezensis L2 (KCCM1 2498P) of the present invention, a fish sauce with an increased content of sweet amino acids and umami-based amino acids can be prepared, which has the advantage of improving taste preference. have

1 is a photograph showing that Bacillus velezensis L2 (KCCM1 2498P) was cultured in a medium according to the present invention.

Figure 2 is a Bacillus velezensis ( Bacillus velezensis ) according to the present invention is an experiment result showing whether a transparent ring is generated showing the protein decomposition ability of L2 (KCCM1 2498P).

3 is a photograph of observing the results of Gram staining of Bacillus velezensis L2 (KCCM1 2498P) according to the present invention under an optical microscope.

4 is a diagram showing a genetic phylogenetic tree of Bacillus velezensis L2 (KCCM1 2498P) according to the present invention.

5 is a view confirming the 16S rRNA nucleotide sequence of Bacillus velezensis L2 (KCCM1 2498P) according to the present invention cultured for each salt concentration.

6 is a graph confirming the number of viable cells for each salt concentration of Bacillus velezensis L2 (KCCM1 2498P) according to the present invention.

7 is a graph confirming the proteolytic activity of Bacillus velezensis L2 (KCCM1 2498P) according to the present invention cultured by salt concentration.

As for the terms used in the present invention, general terms that are currently widely used are selected as possible, but in certain cases, there are also terms arbitrarily selected by the applicant. So the meaning should be understood.

Hereinafter, the technical configuration of the present invention will be described in detail with reference to preferred embodiments shown in the accompanying drawings.

However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Like reference numerals refer to like elements throughout.

The present invention is Bacillus velezensis comprising the nucleotide sequence set forth in SEQ ID NO: 1 as 16S rRNA, the standard strain B. velezensis Compared to LMG 22478 and isolates of the same genus, B. velezensis J3 and B. subtilis KCCM 11316, a novel strain characterized by significantly superior proteolytic activity Bacillus velezensis L2 (KCCM1 2498P) ) is provided.

In the present invention, excellent strains were screened from the anchovy fermented broth prepared by a traditional method, and it was determined that the production of transparent rings for some colonies selected among them had proteolytic ability, and the results of physiological and biochemical experiments were the most By selecting excellent strains, the present invention was completed.

The Bacillus velezensis of the present invention L2 (KCCM1 2498P), the standard strain B. velezensis Unlike LMG 22478, it was confirmed to have a positive reaction to lactose, amygdalin, melibiose and gentiobiose, that is, to have sugar utilization.

In addition, the Bacillus velezensis L2 (KCCM1 2498P) of the present invention was used as a seed strain for the production of a fish sauce using a raw material of fish and shellfish through the experimental examples to be described later, thereby suppressing the production of volatile basic nitrogen and histamine. It was confirmed that the content of total nitrogen and amino nitrogen was increased.

In addition, it was confirmed that the Bacillus velezensis L2 (KCCM1 2498P) increases the content of sweet-based amino acids and umami-based amino acids, and more specifically, serine and valine (Serine) and valine ( It was confirmed to increase the content of amino acids including valine), glutamic acid, an amino acid that gives a umami taste, and proline, an amino acid that gives a sweet and sour taste.

On the other hand, the present invention consists of a culture of the Bacillus velezensis L2 (KCCM 12498P), the Bacillus velezensis L2 (KCCM 12498P), a concentrate of the culture, and a dried product of the culture It may be provided in the form of a seed composition for fermentation of fish sauce containing one or more selected from the group as an active ingredient.

Here, the seed composition refers to a composition containing microorganisms involved in fermentation, the culture refers to culturing a specific microorganism in a culture medium or a culture medium, and the culture and the concentrate of the culture are not limited in the formulation, For example, it may be a liquid or a solid.

In addition, the present invention further provides a fish sauce prepared by adding the seed composition for fermentation of a fish sauce.

Here, the fish sauce may be, for example, anchovy fish sauce, canary fish sauce, cutlass fish sauce, shrimp fish sauce, saury fish sauce, clam fish sauce, or smelt fish sauce, and if it is a fish sauce made from seafood, the type is not limited. .

(Example 1) Bacillus velezensis L2 selection and isolation and identification with excellent proteolytic activity

1-1. Selection of strains with excellent proteolytic activity

100 μl of the fermented anchovy produced in Jeollanam-do, Korea was smeared on a TSA (Trypticase Soy Agar) plate medium, and the plated plate medium was cultured at 37° C. for 72 hours.

As shown in Figure 1, after culturing the medium for 72 hours, the dominant strain (W) was selected and named L2.

Then, the isolated L2 strain was progenitor inoculated (streaking) on a TSA (Trypticase Soy Agar) plate medium containing 1% skim milk, and then cultured for 48 hours to check whether a clear ring was generated. .

As shown in Figure 2, the isolated L2 strain was confirmed to form a clear ring around the colony by the proteolytic enzyme generated in the culture process.

1-2. Identification of selected strains

For identification of the selected isolates, morphological and biochemical investigations were primarily performed, and for final identification, 16S rRNA sequencing was performed.

The morphological investigation was performed by observing the presence of Gram staining and the morphology of bacteria and colonies using a microscope. was carried out by the method of investigating.

The results of the morphological investigation are shown in FIG. 3 , and referring to FIG. 3 , it was confirmed that the L2 strain was a Gram-positive bacilli.

In addition, the biochemical investigation results are shown in Table 1 below, after molecular biological identification of the L2 strain, B. velezensis , a standard strain of the same species The results for LMG 22478 were compared together.

Sugar	Metabolism		Sugar	Metabolism
	standard strain	L2 strain		standard strain	L2 strain
Control	-	-	Esculin	+	+
Glycerol	-	-	Salicin	-	-
Erythritol	-	-	Cellobiose	+	+
D-Arabinose	-	-	Maltose	+	+
L-Arabinose	-	-	Lactose	-	+
Ribose	+	-	Melibiose	-	+
D-Xylose	-	-	Sucrose	+	+
L-Xylose	-	-	Trehalose	+	+
Adonitol	-	-	Inulin	-	-
Methyl β-D-xylopyranoside	-	-	Melezitose	-	-
Galactose	-	-	Raffinose	+	+
Glucose	+	+	Starch	+	+
Fructose	+	+	Glycogen	+	+
Mannose	+	+	Xylitol	-	-
Sorbose	-	-	Gentiobiose	-	+
Rhamnose	-	-	D-Turanose	-	-
Dulcitol	-	-	D-Lyxose	-	-
Inositol	+	-	D-Tagatose	-	-
Mannitol	+	+	D-Fucose	-	-
Sorbitol	+	+	L-Fucose	-	-
Methyl α-D-mannopyranoside	-	-	D-Arabitol	-	-
Methyl α-D-glucoside	+	+	L-Arabitol	-	-
N-Acethyl-glucosamine	+	-	Gluconate	-	-
Amygdalin	-	+	2-Keto-gluconate	-	-
Arbutin	+	+	5-Keto-gluconate	-	-

Here, "+" means positive and "-" means negative. Referring to Table 1, the L2 strain is glucose, fructose, mannose, mannitol, sorbitol, Methyl α-D-glucoside, Amygdalin, Arbutin, Esculin, Cellobiose, Maltose, Lactose, Melibiose, Sucrose, Trehalose, Raffinose, Starch, Glycogen, and Gentiobiose were confirmed to have sugar availability.

In addition, the standard strain B. velezensis Compared to LMG 22478, the L2 strain is different in that it has sugar availability for lactose, amygdalin, melibiose and gentiobiose.

In addition, for accurate identification of the L2 strain, genetic identification was performed through 16S rRNA gene analysis.

For 16S rRNA amplification of colony samples of L2 strain, forward primer 27F (5'-AGAGTTTGATCATGGCTCAG-3') and reverse primer 1492R (5'-GGATACCTTGTTACGACTT-3') were used at 95°C for 30 seconds, 52 After confirming the gene size of the PCR product amplified by electrophoresis after performing PCR (Polymerase chain reaction) under the conditions of 30 seconds at °C and 90 seconds at 72 °C for a total of 35 cycles, based on the results of sequencing analysis through NCBI genebank Genetic identification was performed.

As a general taxonomically , strains showing 97% or more nucleotide sequence homology can be defined as one species. named as

The identified Bacillus velezensis L2 was deposited with the Korea Microorganism Conservation Center on April 18, 2019, and was given an accession number KCCM12498P.

In addition, the identified Bacillus velezensis ( Bacillus velezensis ) The genetic phylogenetic tree (phylogenetic tree) of L2 can be shown as in FIG. 4, and the 16S rRNA base sequence is shown in SEQ ID NOs: 1 and 5.

(Experimental Example 1) Confirmation of proteolytic activity of the strain

The hydrolytic activity of Bacillus velezensis L2 isolated and identified in Example 1 was confirmed.

More specifically, the protein enzyme activity experiment was performed by Azocasein assay, Bacillus velezensis L2 strain, the standard strain Bacillus velezensis ( Bacillus velezensis ) LMG 22478 strain, isolates of the same genus / species Bacillus velezensis J3 strain and Bacillus subtilis KCCM 11316 strain were used.

Each strain was inoculated in 5ml TSB medium, cultured at 37°C for 24 hours, and then 1ml of the culture solution was centrifuged (12,000rpm, 10 minutes, 4°C), and then the supernatant was recovered.

To 50 μl of the supernatant of each strain, 150 μl of 0.5% azocasein dissolved in 100 mM sodium phosphate buffer (pH 7.0) was added and mixed, followed by reaction in a water bath at 30° C. for 1 hour.

Thereafter, after standing for 2 minutes to stop the reaction, centrifugation (12,000 rpm, 10 minutes, 4°C) was performed, and 500 μl of the supernatant was taken, and 700 μl of 525 mM NaOH was added and mixed.

Next, 1 ml of the mixture was put in a 48 well plate, absorbance was measured at 432 nm, and the proteolytic enzyme activity produced by each strain was confirmed, and the results are shown in Table 2 below.

	Bacillus velezensis ( Bacillus velezensis ) LMG 22478 strain	Bacillus velezensis L2 strain	Bacillus velezensis ( Bacillus velezensis ) J3 strain	Bacillus subtilis ( Bxacillus subtilis) KCCM 11316 strain
unit/ml	526	768	484	41

Here, the enzyme activity was defined as 1 unit/ml when the absorbance at 432 nm was increased by 0.01 at 30° C. for 1 hour. As shown in Table 2, the Bacillus velezensis L2 strain of the present invention was The activity of the proteolytic enzyme was measured to be 768 unit/ml, and it was confirmed that it has the best proteolytic ability, and the standard strain Bacillus velezensis ) Compared with the LMG 22478 strain, it was confirmed that it has about 1.5 times higher proteolytic activity.

(Experimental Example 2) Measurement of growth and proteolytic enzyme activity by salt concentration

Using the Bacillus velezensis L2 strain isolated and identified in Example 1, growth rate by salt concentration and proteolytic activity by salt concentration were confirmed.

In more detail, 5 ml of TSB medium in which NaCl is differently added at 0, 5, 10, 15% was prepared and prepared, and then the Bacillus velezensis L2 strain was inoculated into each medium, 37 ° C, After culturing for 24 hours, the number of viable cells and proteolytic enzyme activity were evaluated.

In the experiment to confirm the number of viable cells, the cultured medium was diluted 10-fold, spread on 100 μl of TSA medium, and then cultured at 37° C. for 24 hours, and the number of colonies was measured, and the results are shown in FIG. 6 .

As shown in Figure 6, the growth rate of each salt concentration of the Bacillus velezensis L2 strain of the present invention was confirmed to be most suitable at a salt concentration of 5%.

In addition, the protein-protein enzyme activity test was performed in the same manner as the Azocasein assay tested in Experimental Example 1 using the cultured medium, and the results are shown in FIG. 7 .

7, the Bacillus velezensis L2 strain of the present invention was confirmed to have the best proteolytic activity when cultured in 5% NaCl culture conditions.

(Experimental Example 3) Check the quality of anchovy fish sauce

Anchovy fish sauce was prepared using the Bacillus velezensis L2 strain isolated and identified in Example 1, and the quality of the prepared anchovy fish sauce was checked.

For quality comparison with the anchovy fish sauce (test group) prepared using the Bacillus velezensis L2 strain, an anchovy fish sauce was prepared without using a separate strain as a control group, and as a comparison group, an anchovy fish sauce was prepared. sis ( Bacillus velezensis ) Anchovy fish sauce was prepared using LMG 22478 strain.

More specifically, fresh anchovies with a length of about 17 cm caught in the southern coast as a raw material for aquatic products were washed twice with brine, and then semi-dried to a moisture content of 30% to 55% to prepare a raw material. Thereafter, 5 parts by weight of salt was added to the semi-dried anchovy based on 100 parts by weight of the raw material and mixed. Next, the Bacillus velezensis L2 strain was cultured and then added so that the salt was mixed with the raw material to 1 part by weight based on 100 parts by weight of the raw material, and then fermented at 50 ° C., anaerobic conditions for 8 weeks. A fermented product was produced, and then the resulting fermented product was filtered to remove residues to prepare a fish sauce.

On the other hand, a separate strain was not used as a control group, and as a comparison group, Bacillus velezensis ( Bacillus velezensis ) A fish sauce was prepared using the LMG 22478 strain. Except for this, the manufacturing process of the fish sauce was the same as described above to prepare the control fish sauce and the control group fish sauce, respectively.

3-1. Check total nitrogen, amino nitrogen, volatile nitrogen, histamine content

For a total of 8 weeks during the fermentation period, 5 ml of fish sauce samples for the test group, control group, and comparison group were each taken at 2-week intervals. Basic Nitrogen) content and histamine content were checked, and the results are shown in Tables 3 to 5 below.

	Control (with strain X)
Fermentation period	0 parking	2nd week	4 weeks		Week 6	Week 8
Total nitrogen (%)	0.83	1.28	1.32	1.35	1.35
Amino nitrogen (AN, mg/100g)	990	1392	1414	1436	1437
Volatile basic nitrogen (mg%)	20.8	21.6	21.4	21.2	21.2
Histamine (ppm)	123.4	128.8	136	149.1	155.5

	Comparative group (using Bacillus belegensis LMG 22478 strain)
Fermentation period	0 parking	2nd week	4 weeks		Week 6	Week 8
Total nitrogen (%)	0.86	1.48	1.51	1.55	1.56
Amino nitrogen (AN, mg/100g)	995	1573	1609	1620	1620
Volatile basic nitrogen (mg%)	20.7	19.4	17.6	17	16.8
Histamine (ppm)	123.7	113.3	106.4	102.4	102.1

	Test group (using Bacillus belegensis L2 strain)
Fermentation period	0 parking	2nd week	4 weeks		Week 6	Week 8
Total nitrogen (%)	0.84	1.81	1.86	1.89	1.89
Amino nitrogen (AN, mg/100g)	989	1890	1928	1931	1932
Volatile base nitrogen (mg%)	21	18.3	15.2	14.6	14.4
Histamine (ppm)	124.1	75	55.6	49.9	48

As shown in Tables 3 to 5, when an anchovy fish sauce is prepared using the Bacillus velezensis L2 strain, the fermentation is completed in about 4 weeks, so the fermentation period can be shortened, and total nitrogen , amino nitrogen content is high, and the content of histamine, which is a component involved in allergic reactions or inflammation, and the content of volatile basic nitrogen, a component that is an indicator of freshness, has the advantage of being provided as a high-quality fish sauce.

3-2. sensory test

As an evaluation of the sensory palatability of fish sauces (control group, comparison group, test group) prepared by fermentation for 8 weeks, a sensory test was performed and shown in Table 6 below.

Here, the evaluation items were overall color, aroma, taste, umami taste, fishy taste, aftertaste, and overall preference, and a 9-point scale method was applied to 30 target consumer panels who actually used the product, and a sensory test was conducted.

	Control (with strain X)	Comparative group (using Bacillus belegensis LMG 22478 strain)	Test group (using Bacillus belegensis L2 strain)
color	6.5	7.0	8.0
incense	4.8	6.0	7.4
taste	3.3	5.6	7.5
umami	4.8	5.0	7.8
fishy taste	4.8	6.2	7.5
cove	4.1	6.1	7.7
Comprehensive symbol map	4.3	5.3	7.5

As a result, as shown in Table 6, the anchovy fish sauce prepared by fermenting with the Bacillus velezensis L2 strain of the present invention had color, flavor, It was excellent in all indicators for taste, umami, fishy taste, and aftertaste, and was investigated to be excellent in overall palatability.

3-3. Free Amino Acid Analysis

Analysis of free amino acids was performed using fish sauce (control group, comparative group, test group) prepared by fermentation for 8 weeks, and the results are shown in Table 7 below.

Here, the free amino acid analysis was confirmed using an automatic amino acid analyzer (Hitach Amino Acid Analyzer L-8900).

Amino acid (mg%)	Control (with strain X)	Comparative group (using Bacillus belegensis LMG 22478 strain)	Test group (using Bacillus belegensis L2 strain)	Remarks (compared to standard strain comparison group)
Taurine	88.0	56.5	128.0
Aspartic acid	10.0	12.8	24.3
Threonine	0.0	0.0	87.9
Serine	153.8	190.0	471.9	2.5 times increase, sweet amino acid
Asparagine	0.0	0.0	17.3
Glutamic acid	423.8	554.0	923.9	1.7 times increase, umami taste
Glutamine	0.0	0.0	0.0
Glycine	139.6	199.1	3.8
Alanine	338.5	420.6	286.6
Valine	276.4	374.8	790.6	1.9 times increase, sweet amino acid
Methionine	127.0	184.7	37.4
Isoleucine	168.2	270.9	491.1
Leucine	284.2	439.8	618.9
Tyrosine	94.9	101.7	227.4
Phenylalanine	125.3	183.8	389.2
γ-Amino-n-butyric acid	2.7	0.0	7.9
Lysine	388.2	418.4	540.0
Histidine	106.1	160.2	223.6
Arginine	5.9	124.3	478.2
Hydroxyproline	304.1	488.5	34.9
Proline	99.4	109.6	365.0	3.3 times increase, sweet and sour taste
Total	3,136.1	4,289.7	6,147.9

As shown in Table 7, the anchovy fish sauce prepared by fermenting with the Bacillus velezensis L2 strain of the present invention was compared with the control group and the comparative group, sweet-sweet amino acids and umami-based amino acids. It is thought that it is possible to prepare a fish sauce with an increased content of , so it is possible to improve the taste preference. More specifically, anchovy produced by fermenting with the Bacillus velezensis L2 strain of the present invention. Compared to the fish sauce fermented by the standard strain Bacillus belegensis LMG 22478, the fish sauce contains about 2.5 times higher content of serine, an amino acid that gives a sweet taste, and glutamic acid, an amino acid that gives a umami taste. ), the content is about 1.7 times higher, in the case of valine, a sweet amino acid, the content is about 1.9 times higher, and in the case of proline, an amino acid that gives a sweet and sour taste, the content is about 3.3 times higher. Confirmed.

As described above, according to the Bacillus velezensis L2 (KCCM1 2498P) strain of the present invention and the fish sauce prepared by the strain, compared to the standard strain Bacillus velezensis LMG 22478, It has about 1.5 times higher proteolytic activity, so when used as a seed strain for the production of fish sauce, the fermentation period can be shortened. It is possible to provide a high-quality fish sauce with a remarkably reduced content and, at the same time, an increased content of sweet amino acids and umami-based amino acids.

As described above, the present invention has been illustrated and described with reference to preferred embodiments, but it is not limited to the above-described embodiments, and those of ordinary skill in the art to which the present invention pertains within the scope not departing from the spirit of the present invention Various changes and modifications will be possible.

[Accession number]

Name of deposit institution: Korea Microorganism Conservation Center (Overseas)

Accession number: KCCM12498P

Deposit date: 20190418

It can be used as a fermentation seed for various fermented foods, and in particular, it can be used as a fermentation seed for use in the production of fish sauce.

TTAGAGTTTG ATTCAGGCTC AGGACGAACG CTGGCGGCGT GCCTAATACA TGCAAGTCGA

GCGGACAGAT GGGAGCTTGC TCCCTGATGT TAGCGGCGGA CGGGTGAGTA ACACGTGGGT

AACCTGCCTG TAAGACTGGG ATAACTCCGG GAAACCGGGG CTAATACCGG ATGGTTGTTT

GAACCGCATG GTTCAGACAT AAAAGGTGGC TTCGGCTACC ACTTACAGAT GGACCCGCGG

CGCATTAGCT AGTTGGTGAG GTAACGGCTC ACCAAGGCAA CGATGCGTAG CCGACCTGAG

AGGGTGATCG GCCACACTGG GACTGAGACA CGGCCCAGAC TCCTACGGGA GGCAGCAGTA

GGGAATCTTC CGCAATGGAC GAAAGTCTGA CGGAGCAACG CCGCGTGAGT GATGAAGGTT

TTCGGATCGT AAAGCTCTGT TGTTAGGGAA GAACAAGTGC CGTTCAAATA GGGCGGCACC

TTGACGGTAC CTAACCAGAA AGCCACGGCT AACTACGTGC CAGCAGCCGC GGTAATACGT

AGGTGGCAAG CGTTGTCCGG AATTATTGGG CGTAAAGGGC TCGCAGGCGG TTTCTTAAGT

CTGATGTGAA AGCCCCCGGC TCAACCGGGG AGGGTCATTG GAAACTGGGG AACTTGAGTG

CAGAAGAGGA GAGTGGAATT CCACGTGTAG CGGTGAAATG CGTAGAGATG TGGAGGAACA

CCAGTGGCGA AGGCGACTCT CTGGTCTGTA ACTGACGCTG AGGAGCGAAA GCGTGGGGAG

CGAACAGGAT TAGATACCCT GGTAGTCCAC GCCGTAAACG ATGAGTGCTA AGTGTTAGGG

GGTTTCCGCC CCTTAGTGCT GCAGCTAACG CATTAAGCAC TCCGCCTGGG GAGTACGGTC

GCAAGACTGA AACTCAAAGG AATTGACGGG GGCCCGCACA AGCGGTGGAG CATGTGGTTT

AATTCGAAGC AACGCGAAGA ACCTTACCAG GTCTTGACAT CCTCTGACAA TCCTAGAGAT

AGGACGTCCC CTTCGGGGGC AGAGTGACAG GTGGTGCATG GTTGTCGTCA GCTCGTGTCG

TGAGATGTTG GGTTAAGTCC CGCAACGAGC GCAACCCTTG ATCTTAGTTG CCAGCATTCA

GTTGGGCACT CTAAGGTGAC TGCCGGTGAC AAACCGGAGG AAGGTGGGGA TGACGTCAAA

TCATCATGCC CCTTATGACC TGGGCTACAC ACGTGCTACA ATGGGCAGAA CAAAGGGCAG

CGAAACCGCG AGGTTAAGCC AATCCCACAA ATCTGTTCTC AGTTCGGATC GCAGTCTGCA

ACTCGACTGC GTGAAGCTGG AATCGCTAGT AATCGCGGAT CAGCATGCCG CGGTGAATAC

GTTCCCGGGC CTTGTACACA CCGCCCGTCA CACCACGAGA GTTTGTAACA CCCGAAGTCG

GTGAGGTAAC CTTTTAGGAG CCAGCCGCCG AAGGTGGGAC AGATGATTGG GGTGAAGTCG

TAACAGGGTA AACCGTA

Claims (8)

As Bacillus velezensis comprising the nucleotide sequence set forth in SEQ ID NO: 1 as 16S rRNA,

Isolated from the anchovy fermentation broth, Bacillus velezensis characterized in that it has proteolytic activity ( Bacillus velezensis ) L2 (KCCM1 2498P).

The method of claim 1,

The Bacillus velezensis ( Bacillus velezensis )

Lactose (Lactose), amygdalin (Amygdalin), melibiose (Melibiose) and gentiobiose (gentiobiose) Bacillus velezensis characterized in that it shows a positive reaction ( Bacillus velezensis ) L2 (KCCM 12498P).

The method of claim 1,

The Bacillus velezensis ( Bacillus velezensis ) is,

Bacillus velezensis L2 (KCCM 12498P), characterized in that it is used as a seed for the production of fish sauce using raw materials for fish and shellfish and suppresses the production of volatile basic nitrogen and histamine.

The method of claim 1,

The Bacillus velezensis ( Bacillus velezensis ) is,

Bacillus velezensis L2 (KCCM 12498P), characterized in that it is used as a seed strain for the production of fish sauce using raw materials for fish and shellfish to increase the total nitrogen and amino nitrogen content.

The method of claim 1,

The Bacillus velezensis ( Bacillus velezensis ) is,

Bacillus velezensis L2 (KCCM 12498P), which is used as a seed strain for the production of fish sauce using raw materials for fish and shellfish to increase the content of sweet amino acids and umami-based amino acids.

6. The method of claim 5,

The Bacillus velezensis ( Bacillus velezensis ) is,

Bacillus belegensis, characterized by increasing the content of amino acids including serine, glutamic acid, valine and proline by being used as a seed strain for the production of fish sauce using raw materials for fish and shellfish ( Bacillus velezensis ) L2 (KCCM 12498P).

Claims 1 or 2 of the Bacillus velezensis ( Bacillus velezensis ) L2 (KCCM 12498P), the Bacillus velezensis ( Bacillus velezensis ) L2 (KCCM 12498P) of a culture, a concentrate of the culture and a dried product of the culture A seed composition for fermentation of fish sauce comprising at least one selected from the group consisting of as an active ingredient.

A fish sauce prepared by adding the seed composition for fermentation of a fish sauce of claim 7.

Images