KR20200142691A - Novel Bacillus velezensis L2 separated from fermented broth of anchow and Fish sauce prepared by using the same - Google Patents

Abstract

The present invention relates to Bacillus velezensis L2 (KCCM12498P), which shows a positive reaction to lactose, amygdalin, melibiose and gentiobiose and has excellent proteolytic activity, and a fish sauce prepared by using the same. The Bacillus velezensis L2 (KCCM 12498P) selected in the present invention is used as a seed for fermentation of fish sauce, can minimize the production of histamine, a component involved in allergic reactions or inflammation, and increase the amount of total nitrogen and the production of amino shape nitrogen, so as to improve the flavor.

Description

Novel Bacillus velezensis L2 separated from fermented broth of anchow and Fish sauce prepared by using the same}

The present invention relates to a novel Bacillus velezensis L2 (KCCM12498P) isolated from anchovy fermentation broth, and in more detail, lactose, amygdalin, melibiose, and gentiobiose. ) Shows a positive reaction, and has excellent proteolytic activity, Bacillus velezensis , L2 (KCCM12498P).

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

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

In general, the fermentation period of fish sauce generally takes 3 months to 24 months, so production efficiency is low. In order to increase the production efficiency of fish sauce, proteolytic enzymes with high activity proteolysis are produced. It is necessary to develop a strain that becomes.

In addition, conventional fish sauce contains more than a predetermined content of histamine, one of the biogenic amine components, and this histamine is known to be a component involved in allergic reactions or inflammation, and fish and shellfish foods containing histamine produced in large amounts can be consumed. In this case, since it has a problem that causes acute poisoning with headache, hives, and seizures, it is necessary to develop a new strain capable of minimizing the amount of histamine produced.

The present invention has been conceived by such a need, and an object of the present invention is to provide a strain that has excellent protein resolution, can increase the content of free amino acids, and minimize the amount of histamine produced by being used as a seed for fish sauce fermentation. Is to do.

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

Objects of the present invention are not limited to the above-mentioned objects, 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 as 16S rRNA, which includes the nucleotide sequence shown in SEQ ID NO: 1, which is isolated from anchovy fermentation broth and has proteolytic activity. Bacillus velezensis L2 (KCCM1 2498P) is provided.

In a preferred embodiment, the Bacillus velezensis L2 (KCCM1 2498P) exhibits a positive reaction to lactose, amygdalin, melibiose and genthiobiose.

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

In a preferred embodiment, the Bacillus velezensis L2 (KCCM1 2498P) is used as a seed for preparing fish sauce using raw materials for fish and shellfish to increase the content of total nitrogen and amino nitrogen.

In a preferred embodiment, the Bacillus velezensis L2 (KCCM1 2498P) is used as a seed for preparing fish sauce using raw materials for fish and shellfish, and increases the content of sweet and umami amino acids.

In a preferred embodiment, the Bacillus velezensis L2 (KCCM1 2498P) is used as a seed for preparing fish sauce using raw materials for fish and shellfish, and serine, glutamic acid, and valine And increasing the content of amino acids including 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. It provides a seed composition for fermentation of fish sauce containing one or more selected from the group as an active ingredient, and further provides a fish sauce prepared by adding the seed composition for fermentation of fish sauce.

The present invention has the following excellent effects.

According to the 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, so that fish sauce production When used as a seed for the purpose, it has the effect of shortening the fermentation period.

In addition, according to Bacillus velezensis L2 (KCCM1 2498P) of the present invention, the content of histamine, a component involved in allergic reactions or inflammation, and the content of volatile basic nitrogen, a component serving as an indicator of freshness, are drastically reduced. It has the advantage of providing high quality fish sauce.

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

1 is a photograph showing the culture of Bacillus velezensis L2 (KCCM1 2498P) according to the present invention in a medium.
FIG. 2 is an experimental result of generating a transparent ring showing the protein resolution of Bacillus velezensis L2 (KCCM1 2498P) according to the present invention.
3 is a photograph of a Gram staining result of Bacillus velezensis L2 (KCCM1 2498P) according to the present invention observed with an optical microscope.
Figure 4 is a diagram showing the genetic tree (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 by salt concentration.
Figure 6 is a graph confirming the number of viable cells by 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 terms used in the present invention, general terms that are currently widely used are selected, but in certain cases, some terms are arbitrarily selected by the applicant. In this case, the meanings described or used in the detailed description of the invention are considered rather than the names of simple terms. Therefore, its meaning should be grasped.

Hereinafter, the technical configuration of the present invention will be described in detail with reference to exemplary 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. The same reference numerals denote the same elements throughout the specification.

The present invention is a 16S rRNA as Bacillus velezensis containing the nucleotide sequence shown in SEQ ID NO: 1, which is a standard strain, B. velezensis Compared with LMG 22478 and isolates of the same genus, B. velezensis J3 and B. subtilis KCCM 11316, a novel strain characterized by having remarkably excellent proteolytic activity, Bacillus velezensis L2 (KCCM1 2498P) ).

In the present invention, excellent strains were screened from anchovy fermented broth prepared by the traditional method, and it was determined that there is protein decomposition ability to generate transparent rings for some selected colonies, and the results of physiological and biochemical experiments are the most. By selecting excellent strains, the present invention was completed.

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

In addition, the Bacillus velezensis L2 (KCCM1 2498P) of the present invention is used as a seed for preparing fish sauce using fish and shellfish raw materials through experimental examples to be described later, and suppresses the production of volatile base nitrogen and histamine. And it was confirmed that the content of total nitrogen and amino nitrogen was increased.

In addition, the Bacillus velezensis L2 (KCCM1 2498P) was confirmed to increase the content of sweet-based amino acids and umami-based amino acids, more specifically, serine and valine ( Valine), it was confirmed to increase the content of amino acids including glutamic acid, an amino acid that gives off a umami taste, and Proline, an amino acid that gives off 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 at least one selected from the group as an active ingredient.

Here, the seed composition means a composition containing microorganisms involved in fermentation, and the culture means that a specific microorganism is cultivated in a culture medium or a culture medium, and the culture and the concentrate of the culture are not limited in their formulation, For example, it may be liquid or solid.

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

Here, the fish sauce may be, for example, anchovy fish sauce, canary fish sauce, blackfish fish sauce, shrimp fish sauce, saury fish sauce, clam fish sauce, or smelt fish sauce, and if it is a fish sauce made of fish and shellfish as a raw material, the type is not limited thereto. .

(Example 1) Selection and isolation of Bacillus velezensis L2 excellent in proteolytic activity

1-1. Selection of strains with excellent proteolytic activity

100 µl of anchovy fermentation broth produced in the Jeonnam region of Korea was plated on TSA (Trypticase Soy Agar) plate medium, and the plated plate medium was cultured at 37°C for 72 hours.

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

In addition, the isolated L2 strain was inoculated (streaking) in a TSA (Trypticase Soy Agar) plate medium containing 1% skim milk, and then cultured for 48 hours to confirm whether or not a transparent ring was produced. .

As shown in FIG. 2, it was confirmed that the isolated L2 strain formed a transparent ring around the colony by the proteolytic enzyme produced in the culture process.

1-2. Identification of selected strains

For the 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 morphology of bacteria and colonies using a microscope and whether or not Gram staining was performed, and the biochemical investigation was performed using API 50 CHB Kit (BioMerieux SA) to determine whether or not substrate fermentation was performed by sugar type. It was carried out in a way to investigate.

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 bacillus.

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. In addition, it was confirmed that it has sugar utilization for Gentiobiose.

Also, the standard strain, B. velezensis Compared with LMG 22478, the L2 strain is different in that it has sugar availability for lactose, amygdalin, melibiose, and genthiobiose.

In addition, genetic identification was performed through 16S rRNA gene analysis in order to accurately identify the L2 strain.

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 to amplify 16S rRNA at 95°C for 30 seconds, 52 After performing PCR (polymerase chain reaction) under the conditions of a total of 35 cycles for 30 seconds at ℃ and 90 seconds at 72 ℃, check the gene size of the PCR product amplified by electrophoresis, and then based on the sequencing analysis result through the NCBI genebank. Genetic identification was carried out.

In general taxonomically, strains showing more than 97% nucleotide sequence homology can be defined as one species. As a result of the analysis, it was identified as Bacillus velezensis with more than 99% homology, and " Bacillus velezensis L2" It was named as.

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

In addition, the identified Bacillus velezensis ( Bacillus velezensis ) L2 genetic phylogenetic tree (phylogenetic tree) can be represented as shown in Figure 4, the 16S rRNA nucleotide sequence is shown in SEQ ID NO: 1 and 5.

(Experimental Example 1) Confirmation of proteolytic activity of 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, and the standard strain Bacillus velezensis LMG 22478 strain, Bacillus velezensis J3 strain and Bacillus subtilis KCCM 11316 strain, which are isolates of the same genus/species were used.

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

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

Thereafter, the reaction was stopped by standing for 2 minutes, followed by centrifugation (12,000 rpm, 10 minutes, 4° C.), 500 μl of the supernatant was taken, and 700 μl of 525 mM NaOH were added to mix.

Next, 1 ml of the mixed solution was put into a 48 well plate, and the absorbance was measured at 432 nm to confirm the proteolytic enzyme activity generated by each strain, and the results are shown in Table 2 below.

Bacillus velezensis LMG 22478 strain Bacillus velezensis L2 strain 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 increasing the absorbance of 0.01 at 432 nm at 30° C. for 1 hour.

As shown in Table 2, the Bacillus velezensis L2 strain of the present invention was found to have the best proteolytic activity as measured at 768 units/ml of protease activity, and the standard strain Bacillus velezensis Bacillus velezensis Compared to the LMG 22478 strain, it was confirmed to have about 1.5 times higher proteolytic activity.

(Experimental Example 2) Measurement of growth rate and protease activity by salt concentration

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

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

In the experiment to confirm the number of viable cells, the cultured culture medium was diluted 10 times, plated with 100 µl of TSA medium, and cultured at 37° C. for 24 hours, and the number of colony was measured, and the results are shown in FIG. 6.

As shown in FIG. 6, it was confirmed that the growth rate of the Bacillus velezensis L2 strain of the present invention is most suitable at a salt concentration of 5%.

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

As shown in FIG. 7, it was confirmed that the Bacillus velezensis L2 strain of the present invention has the best proteolytic activity when cultured in 5% NaCl culture conditions.

(Experimental Example 3) Quality check 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 confirmed.

In order to compare the quality 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. Cis ( Bacillus velezensis ) Anchovy fish sauce was prepared using LMG 22478 strain.

In more detail, a raw material was prepared by washing twice with brine using fresh anchovies with a length of about 17 cm caught in the south coast as a raw material for aquatic products, and then semi-drying at a moisture content of 30% to 55%. 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 added so that 1 part by weight based on 100 parts by weight of the raw material was added to the raw material mixed with the salt, and fermented for 8 weeks at 50°C and anaerobic conditions. A fermented product was produced, and the fermented product was then filtered to remove the residue to prepare a fish sauce.

On the other hand, a separate strain was not used as a control group, and Bacillus velezensis as a control group LMG 22478 strain was used to prepare fish sauce, except for this, the manufacturing process of fish sauce was carried out in the same manner as described above, to prepare a control fish sauce and a control fish sauce, respectively.

3-1. Total nitrogen, amino nitrogen, volatile basic nitrogen, histamine content check

5 ml of fish sauce samples for the test group, control group, and control group were taken at intervals of 2 weeks for a total of 8 weeks during the fermentation period. Using these fish sauce samples, the total nitrogen content, amino nitrogen content, and volatile base nitrogen (VBN, Volatile Basic Nitrogen) content and histamine content were confirmed, and the results are shown in Tables 3 to 5 below.

Control (strain X) Fermentation period Week 0 2nd week Week 4 Week 6 8th week 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 bellegensis LMG 22478 strain) Fermentation period Week 0 2nd week Week 4 Week 6 8th week 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 Week 0 2nd week Week 4 Week 6 8th week Total nitrogen (%) 0.84 1.81 1.86 1.89 1.89 Amino nitrogen (AN, mg/100g) 989 1890 1928 1931 1932 Volatile basic 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, in the case of manufacturing anchovy fish sauce using Bacillus velezensis L2 strain, 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, a component involved in allergic reactions or inflammation, and the content of volatile basic nitrogen, a component that serves as an indicator of freshness, are drastically reduced, so that it can be provided as a high-quality fish sauce.

3-2. Sensory test

As an evaluation of the sensory palatability of fish sauce (control group, control group, test group) prepared by fermentation for an 8-week period, a sensory test was conducted and shown in Table 6 below.

Here, the evaluation items were overall color, scent, taste, umami, fishy taste, tail, and general preference, and a sensory test was conducted using a 9-point scale method for 30 target consumer panels who actually use the product.

Control
(Use strain X) Comparative group
(Use of Bacillus belegensys LMG 22478 strain) Test group
(Use of Bacillus belegensis L2 strain) color 6.5 7.0 8.0 incense 4.8 6.0 7.4 flavor 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 Chart 4.3 5.3 7.5

As a result, as shown in Table 6, the anchovy fish sauce prepared by fermentation by the Bacillus velezensis L2 strain of the present invention was compared with the fish sauce of the control group and the fish sauce of the comparative group, It was found to be excellent in all indicators for taste, umami, fishy taste, and aftertaste, and was found to be excellent in the overall acceptability.

3-3. Free amino acid analysis

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

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

Amino acid (mg%) Control
(Use strain X) Comparative group
(Use of Bacillus belegensys LMG 22478 strain) Test group
(Use of Bacillus belegensis L2 strain) Remark
(Compared to the standard strain control 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 Total 3,136.1 4,289.7 6,147.9

As shown in Table 7, the anchovy fish sauce prepared by fermentation by the Bacillus velezensis L2 strain of the present invention was compared with the fish sauce of the control group and the fish sauce of the control group, and sweet amino acids and umami amino acids It is believed that it was possible to manufacture fish sauce with an increased content of, thereby improving taste preference.

More specifically, the anchovy fish sauce prepared by fermentation by the Bacillus velezensis L2 strain of the present invention, compared with the fish sauce fermented by the standard strain Bacillus bellezensis LMG 22478, gives a sweet taste. Serine, the amino acid, is about 2.5 times higher in content, about 1.7 times higher in glutamic acid, the amino acid that gives a umami taste, and about 1.9 times in the case of Valine, a sweet amino acid. In the case of Proline, an amino acid that has a high pear content and has a sweet and sour taste, it has been confirmed that its content is about 3.3 times higher.

As described above, according to the Bacillus velezensis L2 (KCCM1 2498P) strain of the present invention and the fish sauce prepared by this strain, compared with the standard strain Bacillus velezensis LMG 22478, It has about 1.5 times higher protein decomposition activity, and when used as a seed for fish sauce production, it can shorten the fermentation period. The content of histamine, an ingredient involved in allergic reactions or inflammation, and volatile basic nitrogen, an ingredient that is an indicator of freshness. It is possible to provide a high-quality fish sauce in which the content is drastically reduced, and at the same time, the content of the sweet amino acid and the umami amino acid is increased.

As described above, the present invention has been illustrated and described with a preferred embodiment, but is not limited to the above-described embodiment, and within the scope of the spirit of the present invention, those of ordinary skill in the art Various changes and modifications will be possible.

Korea Microorganism Conservation Center (overseas) KCCM12498P 20190418

<110> Jeonnam Bioindustry Foundation <120> Novel Bacillus velezensis L2 separated from fermented broth of anchow and Fish sauce prepared by using the same <130> DP19-0036 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 1517 <212> RNA <213> Unknown <220> <223> Bacillus velezensis <400> 1 ttagagtttg attcaggctc aggacgaacg ctggcggcgt gcctaataca tgcaagtcga 60 gcggacagat gggagcttgc tccctgatgt tagcggcgga cgggtgagta acacgtgggt 120 aacctgcctg taagactggg ataactccgg gaaaccgggg ctaataccgg atggttgttt 180 gaaccgcatg gttcagacat aaaaggtggc ttcggctacc acttacagat ggacccgcgg 240 cgcattagct agttggtgag gtaacggctc accaaggcaa cgatgcgtag ccgacctgag 300 agggtgatcg gccacactgg gactgagaca cggcccagac tcctacggga ggcagcagta 360 gggaatcttc cgcaatggac gaaagtctga cggagcaacg ccgcgtgagt gatgaaggtt 420 ttcggatcgt aaagctctgt tgttagggaa gaacaagtgc cgttcaaata gggcggcacc 480 ttgacggtac ctaaccagaa agccacggct aactacgtgc cagcagccgc ggtaatacgt 540 aggtggcaag cgttgtccgg aattattggg cgtaaagggc tcgcaggcgg tttcttaagt 600 ctgatgtgaa agcccccggc tcaaccgggg agggtcattg gaaactgggg aacttgagtg 660 cagaagagga gagtggaatt ccacgtgtag cggtgaaatg cgtagagatg tggaggaaca 720 ccagtggcga aggcgactct ctggtctgta actgacgctg aggagcgaaa gcgtggggag 780 cgaacaggat tagataccct ggtagtccac gccgtaaacg atgagtgcta agtgttaggg 840 ggtttccgcc ccttagtgct gcagctaacg cattaagcac tccgcctggg gagtacggtc 900 gcaagactga aactcaaagg aattgacggg ggcccgcaca agcggtggag catgtggttt 960 aattcgaagc aacgcgaaga accttaccag gtcttgacat cctctgacaa tcctagagat 1020 aggacgtccc cttcgggggc agagtgacag gtggtgcatg gttgtcgtca gctcgtgtcg 1080 tgagatgttg ggttaagtcc cgcaacgagc gcaacccttg atcttagttg ccagcattca 1140 gttgggcact ctaaggtgac tgccggtgac aaaccggagg aaggtgggga tgacgtcaaa 1200 tcatcatgcc ccttatgacc tgggctacac acgtgctaca atgggcagaa caaagggcag 1260 cgaaaccgcg aggttaagcc aatcccacaa atctgttctc agttcggatc gcagtctgca 1320 actcgactgc gtgaagctgg aatcgctagt aatcgcggat cagcatgccg cggtgaatac 1380 gttcccgggc cttgtacaca ccgcccgtca caccacgaga gtttgtaaca cccgaagtcg 1440 gtgaggtaac cttttaggag ccagccgccg aaggtgggac agatgattgg ggtgaagtcg 1500 taacagggta aaccgta 1517

Claims (8)

As 16S rRNA, Bacillus velezensis comprising the nucleotide sequence set forth in SEQ ID NO: 1,
Bacillus velezensis L2 (KCCM1 2498P), characterized in that it is isolated from anchovy fermentation broth and has proteolytic activity.
The method of claim 1,
The Bacillus velezensis is
Bacillus velezensis L2 (KCCM 12498P), characterized in that it exhibits a positive reaction to lactose, amygdalin, melibiose and genthiobiose.
The method of claim 1,
The Bacillus velezensis is,
Bacillus velezensis ( Bacillus velezensis ) L2 (KCCM 12498P), characterized in that it is used as a seed for the production of fish sauce using raw fish and shellfish, and suppresses the production of volatile basic nitrogen and histamine.
The method of claim 1,
The Bacillus velezensis is,
Bacillus velezensis ( Bacillus velezensis ) L2 (KCCM 12498P), characterized in that it is used as a seed for the production of fish sauce using raw fish and shellfish, and increases the content of total nitrogen and amino nitrogen.
The method of claim 1,
The Bacillus velezensis is,
Bacillus velezensis ( Bacillus velezensis ) L2 (KCCM 12498P), characterized in that it is used as a seed for the production of fish sauce using raw fish and shellfish, and increases the content of sweet and umami amino acids.
The method of claim 5,
The Bacillus velezensis is,
Bacillus belegensis characterized by increasing the content of amino acids including serine, glutamic acid, valine and proline, which is used as a seed for fish sauce production using raw fish and shellfish materials. ( Bacillus velezensis ) L2 (KCCM 12498P).
As a culture of claim 1 or 2 Bacillus velezensis L2 (KCCM 12498P), Bacillus velezensis L2 (KCCM 12498P), a concentrate of the culture and a dried product of the culture A seed composition for fermentation of fish sauce containing 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 the fish sauce of claim 7.

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