KR101957828B1 - Tetragenococcus sp. having inhibition activity of biogenic amine production and anti-microbial activity in fermented soybean foods - Google Patents

Abstract

The term " Tetragenococcus " ( Tetragenococcus ), which has the ability to generate biogenic amines, The present invention also relates to a fermented food comprising the same, wherein the biogenic amine is selected from the group consisting of histamine, tyramine, putrescine and cadaverine, cadaverine, and the strain, the crushed product, the culture product, the microorganism preparation, and the fermented food have an effect of inhibiting the formation of the biogenic amine or inhibiting the growth of the harmful microorganism.

Description

Tetragenococcus sp., Which is characterized by the inhibition of the production of biogenic amines in soybean and the antimicrobial activity. having inhibition activity of biogenic amine production and anti-microbial activity in fermented soybean foods}

The present invention relates to a microorganism preparation containing any one or more of the strain, the microorganism, the crushed product thereof and the culture thereof, the strain, the crushed product thereof and the culture thereof And a fermented food comprising the fermented food and the strain.

Fermented food is an indispensable food that is responsible for the table for Koreans, and securing safety is the most important factor. However, biogenic amines have been a problem in the stability due to harmful substances generated in recent fermentation processes. Biogenic amines are basic nitrogen compounds that are formed from amino acids by the action of amino acid decarboxylase in microorganisms in foods. This material is mainly found primarily in fermented foods such as soy sauce, bacteria in the arc Pseudomonas (Acromonas) genus Bacillus (Bacillus), A sheet as bakteo (Citrobacter) genus, Clostridium (Clostridium), An Escherichia (Escherichia ), a keulrep when it is reported to have the Ella (Klebsiela) genus Lactobacillus bacteria (Lactobacillus), a Phedi O Rhodococcus (Pediococcus) genus Proteus (Proteus) genus Pseudomonas (Pseudomonas) in such an amino acid decarboxylase enzyme.

These biogenic amines are a trace amount of essential physiological substances that are introduced into foods or biosynthesized in the human body and are balanced through metabolic processes of synthetic decomposition. However, when the balance is overcome by overdose, it is transformed into a toxic substance in the human body. Especially, in fermented foods containing soybean or milk with high amino acid and protein content, biogenic amines exceeding the decomposition limit of human body during fermentation May be generated. Among these, histamine causes blood pressure drop and allergy, and tyramine causes blood pressure rise and headache. Because of these risks, several countries are conducting projects to reduce biogenic amines in fermented foods. In Korea, the recommended concentration is less than 500 ppm histamine and 200 ppm in the European Union.

Formation of biogenic amines is produced by amino acid decarboxylase enzymes, and in the case of traditional varieties, by complex metabolism of bacillus, aspergillus, lactic acid bacteria and yeast. Although the enzyme activity due to the dominance of the strains is remarkable due to the characteristics of the strains, the problems related to the production of biogenic amines are also emerging in that it is difficult to control the growth of microorganisms by other microorganisms. In fact, the distribution of biogenic amines in domestic fermented foods in 2006 showed that histamine in doenjang was 260 ~ 952 ppm and that of tyramine was 285 ~ 1431 ppm, respectively. Next, the contents of biogenic amines were higher in the order of salted and liver. As a result, it can be known that the soybean is a main source of biogenic amines, which is a health risk, and efforts to reduce biogenic amines are urgently needed.

Various studies have been carried out to reduce the biogenic amines in this area and it has been known that the amino acid decarboxylase enzyme is greatly influenced by the amino acid concentration, sugar concentration, temperature, pH, oxygen concentration, and salt concentration during the fermentation process . In this respect, the method of separating the lactic acid bacteria series and lowering the pH of the food and also producing antimicrobial substances reduces the microorganisms other than the major microorganisms, or enhances the flavor through the organic acids produced through metabolic processes, And Lucono stomata species may be used. Of course, bacteria that degrade biogenic amines include B. licheniformis , B. subtilis , B. amyloliquefaciens , histamine, tyramine, putrescine , And cadaverine. However, since it is difficult to utilize it as a starter strain according to the characteristics of Bacillus species which is maintained in spores under high salt conditions such as soy sauce, selection of fermentation strains capable of growing even in high salt such as soybean It is essential.

Japanese Patent No. 4135846 Korean Patent No. 10-1300191 Korean Patent No. 10-1379230 Korean Patent No. 10-1516588 Korean Patent Publication No. 10-2015-0091789 Korean Patent Publication No. 10-2016-0036784

In view of the above problems, the inventors of the present invention have found that a salt tolerant lactic acid bacterium that can grow even under 16% salt conditions of a soybean and has excellent fermentation ability inhibits the production of biogenic amines and inhibits the growth of harmful microorganisms The aim of this study was to select the best strains that can be used.

One aspect of the present invention is to provide a strain of Tetragenococcus halophilus having an ability to produce a biogenic amine.

One aspect of the present invention is to provide a microorganism preparation having the ability to produce a biogenic amine.

One aspect of the present invention is to provide a fermented food containing a small amount of a biogenic amine.

One aspect of the present invention is to provide a fermented food in which the growth of harmful microorganisms is inhibited.

One aspect of the present invention relates to a method for the production of biogenic amines from Tetragenococcus halophilus ).

In one aspect of the present invention, the biogenetic amine provides a strain of Tetranynococus halophilus, which is at least one of histamine, tyramine, putrescine, and cadaverine .

In one aspect of the present invention, the strain has an ability to inhibit an amino acid decarboxylase of one or more of histidine decarboxylase and tyrosine decarboxylase, Lt; / RTI >

In one aspect of the present invention, the strain provides a strain of Tetranychococcus halophilus having the ability to inhibit the growth of harmful microorganisms.

In one aspect of the present invention, the strain provides Tetranyococcus halofillus strain capable of growing under a 16% salt concentration.

In one aspect of the present invention, the strain provides Tetragenococcus halophilus strain Tetragenococcus halophilus SMB688.

In one aspect of the present invention, the strain provides a strain of Tetranynococus halofilus, wherein the accession number is KCTC 13240BP.

Another aspect of the present invention provides a microorganism preparation comprising any one or more of any of the strains, the lysate thereof, and the culture thereof as an active ingredient.

Yet another aspect of the present invention provides a fermented food comprising any one of the strains, the lysate thereof, and the culture thereof.

According to another aspect of the present invention, there is provided a fermented food, wherein the fermented food is a soup comprising at least one of soy sauce, miso, kochujang, chunchon, chungkukjang and a mixture thereof.

In another aspect of the present invention, the fermented food comprises not more than 300 ppm histamine based on the total weight of the fermented food; Or less than 200 ppm of tyramine based on the total weight of the fermented food.

In yet another aspect of the present invention, the fermented food comprises less than 100 ppm histamine based on the total weight of the fermented food product; Or less than 50 ppm of tyramine based on the total weight of the fermented food.

The strain according to one aspect of the present invention has an effect of inhibiting the production of a biogenic amine.

The strain according to one aspect of the present invention has antagonistic action against harmful microorganisms.

The microbial agent according to another aspect of the present invention has an effect of inhibiting the production of a biogenic amine.

The microbial agent according to another aspect of the present invention has an antagonistic action against harmful microorganisms.

The microbial agent according to another aspect of the present invention can be very usefully used as a food, a food additive, a feed additive, or a medicine.

The fermented food according to another aspect of the present invention has a very low content of biogenic amine.

The fermented food according to another aspect of the present invention has a very low content of harmful microorganisms.

The fermented food according to another aspect of the present invention is a safe fermented food without toxicity.

The fermented food according to another aspect of the present invention has an effect of inhibiting the production of biogenic amines during fermentation.

The fermented food according to another aspect of the present invention has an effect of inhibiting the growth of harmful microorganisms during the fermentation process.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a sequence listing of the nucleotide sequence of 16S rRNA of the SMB688 strain of the present invention. FIG.
FIG. 2 is a genealogical diagram (genealogical relationship diagram) created during the genetic identification process of the SMB688 strain of Example 2 of the present invention.
Fig. 3 is a digital microscope photograph (750x) in which the morphological characteristics of the SMB688 strain of Example 3 were observed under an optical microscope.
Fig. 4 is a photograph of the results of confirmation of the culture medium for biosgenic amine production in (1) of Example 3. Fig.
Fig. 5 shows the results of experiments in which the expression of the biogenic amine-related enzyme of (3) of Example 3 was confirmed.
Fig. 6 shows experimental results of the biogenic amine production of Example 4 (4) and the ability of the Example 4 to inhibit microbial growth.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention relates to a method for the production of biogenic amines from Tetragenococcus halophilus ).

As used herein, the term " biorenic amines " refers to basic nitrogen compounds that are formed from amino acids by the action of amino acid decarboxylase of microorganisms in foods and are toxic substances.

In one aspect of the present invention, the biogenetic amine provides a strain of Tetranynococus halophilus, which is at least one of histamine, tyramine, putrescine, and cadaverine . Among these, histamine causes blood pressure drop and allergy, and tyramine causes blood pressure rise and headache.

In one aspect of the present invention, the strain has an ability to inhibit an amino acid decarboxylase of one or more of histidine decarboxylase and tyrosine decarboxylase, Lt; / RTI > Formation of biogenic amines is produced by amino acid decarboxylase enzymes, and in the case of traditional varieties, by complex metabolism of bacillus, aspergillus, lactic acid bacteria and yeast. The biogenic amine of the present invention is formed by the action of the amino acid decarboxylase of a microorganism. When the biogenic amine is consumed in the food beyond the degradation threshold of the human body, symptoms such as rash, local skin inflammation, allergy, vomiting and diarrhea ≪ / RTI >

In one aspect of the present invention, the strain provides a strain of Tetranychococcus halophilus having the ability to inhibit the growth of harmful microorganisms. The harmful microorganisms may be selected from the group consisting of Staphylococcus sp ., Streptococcus sp ., Weissella sp ., Bacillus sp ., Macrococcus sp . sp .) and Lactococcus sp . & Lt ; / RTI > However, in the specification of the present invention, harmful microorganisms are not limited to the above strains.

In one aspect of the present invention, the strain provides Tetranyococcus halofillus strain capable of growing under a 16% salt concentration.

In one aspect of the present invention, the strain provides Tetragenococcus halophilus strain Tetragenococcus halophilus SMB688. The strain is a lactic acid bacterium isolated from soy sauce and has the ability to inhibit the production of bio-genic amines, and has antagonistic action against harmful microorganisms, Tetrogenococcus halophilus strain SMB688. The SMB688 strain was identified as a tetrazenocosugous halophilus known as GRAS (Generally Recognized As Safe) as a result of 16S rRNA gene sequence and API analysis, and this strain is known as a starter lactic acid bacteria used for the purpose of enhancing the flavor of soy sauce.

In one aspect of the present invention, the strain provides a strain of Tetranynococus halofilus, wherein the accession number is KCTC 13240BP.

Another aspect of the present invention provides a microorganism preparation comprising any one or more of any of the strains, the lysate thereof, and the culture thereof as an active ingredient. The microbial agent can be very useful as a food, a food additive, a feed additive or a medicine. The microbial formulation according to the present invention can be prepared as a solution, a powder, a suspension, a dispersion, an emulsion, an oil dispersion, a paste or a granule, but is not limited thereto.

Yet another aspect of the present invention provides a fermented food comprising any one of the strains, the lysate thereof, and the culture thereof.

According to another aspect of the present invention, there is provided a fermented food, wherein the fermented food is a soup comprising at least one of soy sauce, miso, kochujang, chunchon, chungkukjang and a mixture thereof.

In another aspect of the present invention, the fermented food comprises not more than 300 ppm histamine based on the total weight of the fermented food; Or less than 200 ppm of tyramine based on the total weight of the fermented food.

In yet another aspect of the present invention, the fermented food comprises less than 100 ppm histamine based on the total weight of the fermented food product; Or less than 50 ppm of tyramine based on the total weight of the fermented food.

The histamine may be 300 ppm or less, 250 ppm or less, 200 ppm or less, 150 ppm or less, 100 ppm or less, 80 ppm or less, 60 ppm or less, 55 ppm or 50 ppm or less based on the total weight of the fermented food. The histamine may be 0 ppm or more, 1 ppm or more, 3 ppm or more, 7 ppm or more, 10 ppm or more, 20 ppm or more or 30 ppm or more based on the total weight of the fermented food.

The amount of tyramine may be less than 200 ppm, less than 150 ppm, less than 100 ppm, less than 80 ppm, less than 60 ppm, less than 55 ppm, or less than 50 ppm based on the total weight of the fermented food. The above-mentioned tyramine may be 0 ppm or more, 1 ppm or more, 3 ppm or more, 7 ppm or more, 10 ppm or more or 12 ppm or more based on the total weight of the fermented food.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Example  1. Sample selection and strain isolation

A total of 46 kinds of lactic acid bacteria were selected from primary fermentation fermentation tanks of Sam - Pyo Foods through sampling of fermented fermentation tanks. The selected strains were further screened again using a biogenetic amine production medium to select only 16 non-biogenic amines. One of the remaining 16 strains was named as SMB688 strain after the 3rd salt selection after confirming the salt tolerance. Specifically, it was separated in the following manner.

( 1) Badge  And culture conditions

In order to screen only resistant fluconobacteria species in soy sauce fermented agar of Sampoi Food, cycloheximide was added to inhibit the growth of fungi in the lactic acid bacteria selection medium and cultured in the medium composition shown in Table 1 below.

division Configuration Concentration Remarks Primary selection medium
Lactobacilli MRS 5% pH 5.8
2% Agar addition Sodium chloride 5% Cycloheximide 50 μg / ml

Further, in order to confirm the presence or absence of biogenamic amine production, a biogennic amine activity assay medium as shown in Table 2 below was used

division Configuration Concentration (%) Remarks Secondary selection medium
(Improved medium *) Tryptone 0.5 pH 5.3
2% Agar addition Yeast extract 0.5 Beef extract 0.5 Sodium chloride 0.25 Glucose 0.05 Tween 80 0.1 MgSO ₄ 0.02 MnSO ₄ 0.005 FeSO ₄ 0.004 Ammonium citrate 0.2 Thiamine 0.001 K ₂ HPO ₄ 0.2 CaCO ₃ 0.3 Pyridoxal-5-phosphate 0.005 Histidine 0.3 Bromcresol purple 0.006

The isolated microorganisms were identified by culturing using the lactic acid bacteria-derived soy sauce medium shown in Table 3 for the purpose of examining the utilization in soy sauce (confirming salt tolerance).

Composition of raw soy sauce medium to verify salt tolerance of isolated lactic acid bacteria division Configuration Concentration (%) Remarks Raw soy sauce medium Raw soy sauce 25 pH 7.0
1.5% agar added
(Final salt concentration 16%) Sodium Chloride 12 Glucose 2

( 2)  detach

In order to isolate the lactic acid bacteria in the soy sauce using the medium of the above (1), the fermentation maturity was sampled according to the kind of the soy sauce and the fermentation period, the appropriate amount was diluted with 0.85% sterilized saline, And 46 strains were isolated (primary screening).

In order to isolate strains lacking the ability to produce biorenic amines from the strains isolated from the primary selection medium, each strain was subcultured in a secondary selection medium and cultured for 30 and 48 hours to transform the strain into a purple color 16 species were isolated (secondary screening).

The isolate was screened by the second screening process and tested for resistance to salt in the broth of lactic acid bacteria and labeled SMB688 (tertiary screening).

Example  2. Identification of selected strains

(One) Of the SMB688 strain  Genetic identification

In order to identify the SMB688 strain, 16S rRNA gene sequences were identified. For this purpose, 27F and 1492R were used as universal primers, and the corresponding fragments were amplified by PCR reaction and decoded. The decrypted sequence was confirmed using the BLAST program of NCBI using the win the 16S rRNA gene sequence of strains, CLUSTALW for mutual comparison of the nucleotide sequence SMB688. In the phylogenetic analysis, the 16S rRNA gene sequences of the representative lactic acid parent strains were sorted, and statistical reliability for each branch was analyzed for each phylogenetic tree.

Primers 27F and 1492R used in the identification of strains are shown in SEQ ID NOS: 1 and 2, respectively.

SEQ ID NO: 1: forward_27F (AGAGTTGATCMTGGCTCAG)

SEQ ID NO: 2: reverse direction 1492R (TACGGYTACCTTGTTACGACTT)

The 16S rRNA base sequence of the strain of the present invention was as shown in SEQ ID NO: 3. SEQ ID NO: 3 is shown in Fig.

The results of the systematic analysis were as shown in FIG. As a result of the phylogenetic analysis, the isolated strain SMB688 showed 99% homology with Tetranyjnococus halophilus ATCC 33315 in the closest relation.

(2) Of the SMB688 strain  Biochemical identification

For biochemical identification, the API 50 CHL kit (BioMeReux, Marcy, France) test was used. The biochemical properties obtained were compared with the taxonomic biochemical characteristics of the apiweb software database.

The experimental results are shown in Table 4 below.

Biochemical identification (API analysis) results Control (Blank) Glycerol Erythritol D-Arabinose L-Arabinose 0 15 0 0 100 Ribose D- Xylose L- Xylose Adonitol beta -Methyl-D-xylose 0 0 0 0 0 Galactose Glucose Fructose Mannose Sorbose 100 100 100 100 0 Rhamnose Dulcitol Inositol Mannitol Sorbitol 0 0 0 20 0 alpha -Methyl-D-mannoside alpha -Methyl-D-glucoside N-Acetyl glucosamine Amygdalin Arbutin 0 15 100 90 100 Esculin Salicin Cellobiose Maltose Lactose 100 100 100 90 11 Melibiose Sucrose Trehalose Inulin Melezitose 10 6 100 0 5 Raffinose Starch Glycogen Xylitol Gentiobiose 10 0 0 0 100 D- Thranose D- Lyxose D- Tagatose D- Fucose L- Fucose 90 0 100 0 0 D- Arabitol L- Arabitol Gluconate 2- Keto -gluconate 5- Keto -gluconate 10 0 0 0 0

As a result of the analysis, it was found that the monosaccharide such as L-arabinose, Galactose, Glucose, Fructose and Manose could be used as a carbon source and the polysaccharide such as Cellobiose, Maltose and Trehalose could be used as a carbon source .

(3) Of the SMB688 strain  Morphological identification

For morphological identification, SMB688 strain and Tetranynococus halophilus ATCC 33315 were cultured in 50 ml of lactic acid bacteria liver medium for 30 to 72 hours. After centrifugation at 4000 rpm for 5 minutes, the supernatant was removed. To the pellet, 5 ml of 0.85% Was added to dissolve the pellet well and observed through a digital microscope (Olympus DX200).

The observation results were as shown in Fig. As a result of observing the morphological characteristics of the bacterium using a microscope, the identified bacterium has coagulability characteristics in which several bacterium are clustered with a structure of tetrad or more as a bacterium. The single bacterium is about 0.8-1 μm and the complex bacterium is 4-6 μm And it was confirmed that it showed a large size.

Example  3. Biogenic Amine Non-production  Confirm

(One) Biogenic Amine Generation  Review badge for review

In order to confirm the presence or absence of the biogenic amine production of the SMB688 strain, histidine and tyrosine, which are the precursor amino acids of the biogenic amine, were put into the MRS medium containing the pH indicator bromocresol purple (0.005%) and confirmed after inoculation of the strain . As a control, Tetragonococcus halophilus ATCC 33315 was used.

The results of the experiment were as shown in Fig. 4. As a result, it was confirmed that the medium of SMB688 did not change to purple.

(2) HLPC  analysis

3% Histidine, 3% Tyrosine and 16% NaCl were added to the MRS medium for 30, 72 hours to confirm the biogenic amine production ability of the SMB688 strain. Histidine and tyrosine were added to histamine and tyramine.

Basic setting value of LC device for analyzing biogenic amine Sample preprocessing  Way LC device name Detector Column Histamine analysis of food aquatic aquatic products
(Extraction-derivative) Shiseido UV (254 nm) C18 column Solvent Gradient presence Flow rate Injection vol. Water, ACN U* 1 ml / min 20 μl

※ * Gradient condition Time Initial 10 15 20 25 30 35 45 Solvent A ( ACN ) 55 55 65 80 80 90 90 55 Solvent B (Water) 45 45 35 20 20 10 10 45

As a result of HPLC analysis, it was confirmed that no biogenic amine was produced.

( 3) Molecules  At the biological level Biogenic Amine Non-production  Verification

Molecular biology of T. halophilus type strain ATCC 33315 (bacterium - producing microorganism) was carried out with SMB688 as a control and to verify biosgenic amine production inhibition of the strain. Specifically, Quantitative Real Time (PCR) was performed for the production of enzymes (proteins) by confirming the expression levels of histamine and tyramine production-related genes and aminolytic enzyme-related genes at the RNA level.

The experimental results are shown in Fig. In the case of SMB688 strain, it was confirmed that it did not produce a biogenic amine production-related enzyme different from ATCC 33315.

(4) Depending on the starter strain Biogenic Amine  produce Inhibition  Verification

Based on the results of (1) to (3) above, in order to evaluate the inhibitory effect on the production of biogenic amines when applied to fermented foods, brewing soy sauce having a pilot scale scale was immersed to inhibit the production of biogenic amines. The selection strain (SMB688) and the control strain (ATCC 33315; T. halophilus Type strain) was used as a starter strain in brewing soy sauce, respectively. In the analysis for monitoring, the generation of biogenic amines such as histamine and tyramine was confirmed, and the ability to inhibit the production of biogenic amines was confirmed.

The experimental results are shown in the left graph of Table 7 and FIG.

Analysis of Organic Acid and Biogenic Amine by Ripening Date division Biogenic Amine
(ppm) Aging date 0 15 30 60 90 SMB688 Histamine 2 One 5 21 36 Tyramine One 2 4 11 14 ATCC  33315 Histamine 0 0 8 330 1,592 Tyramine 0 2 8 238 443

As a result, it was confirmed that T. halophilus SMB688 was effective in reducing histamine 44 times and tyramine 31 times compared with ATCC 33315.

Example  4. Ability to inhibit the growth of harmful microorganisms monitoring

The aging was carried out in the same manner as in (4) of Example 3, but the growth inhibitory ability of the harmful microorganisms was monitored. In order to confirm the ability of microorganism to inhibit the growth of microorganisms, microbial changes were analyzed by Pyrosequencing.

The experimental results are shown in the right graph of FIG.

As a result of the analysis of OTUs (Operational Taxonomic Unit), SMB688 strain showed a remarkable inhibitory effect on the growth of harmful microorganisms from the early aging period. In the case of the control strain (ATCC 33315), since harmful microorganisms were reduced after 15 days of aging, It can be confirmed that there is difficulty in controlling the microorganism.

It was confirmed that SMB688 was superior to ATCC 33315 in inhibiting the formation of biogenic amine and inhibiting the growth of harmful microorganisms.

The strain SMB688 showing the above-mentioned efficacy was deposited on April 6, 2017 at the Korea Biotechnology Research Institute, and the deposit number is KCTC 13240 BP.

Korea Biotechnology Research Institute KCTC13240BP 20170406

<110> SEMPIO FOODS COMPANY <120> Tetragenococcus halophilus having inhibitory activity of biogenic          amine production and growth of pathogenic microorganism <130> 17P196 / IND <160> 3 <170> KoPatentin 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer_27F (forward) <400> 1 agagtttgat cmtggctcag 20 <210> 2 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Primer_1492R (reverse) <400> 2 tacggytacc ttgttacgac tt 22 <210> 3 <211> 1512 <212> DNA <213> Tetragenococcus halophilus <220> <221> rRNA &Lt; 222 > (1) <223> 16S rRNA of Tetragenococcus halophilus SMB688 <400> 3 tcctggctca ggacgaacgc tggcggcgtg cctaatacat gcaagtcgaa cgctgcttaa 60 gaagaarctt cggttttttc ttaagyggag tggcggacgg gtgagtaaca cgtggggaac 120 ctatcyatca gcgggggata acacttggaa acaggtgcta ataccgcata yggctttttt 180 tcacctgaaa gaaagctcaa rggcgcttta cagcgtcact gatggctggt cccgcggtgc 240 attagccagt tggtgaggta acggctcacc aaagcaacga tgcatagccg acctgagagg 300 gtgatcggcc acactgggac tgagacacgg cccagactcc tacgggaggc agcagtaggg 360 aatcttcggc aatggacgya agtctgaccg agcaacgccg cgtgagtgaa gaaggttttc 420 ggatcgtaaa gctctgttgt cagcaaagaa caggagaaag aggmaatgct ttttcyatga 480 cggtagctga ccagaaagcc acggctaact acgtgccagc agccgcggta atacgtaggt 540 ggcaagcgtt gtccggattt attgggcgta aagcgagcgc aggcggtgat ttaagtctga 600 tgtgaaagcc cccagctcaa ctggggaggg tcattggaaa ctggatcact tgagtgcaga 660 agaggagagt ggaattccat gtgtagcggt gaaatgcgta gatatatgga ggaacaccag 720 tggcgaaggc ggctctctgg tctgtaactg acgctgagrc tcgaaagcgt gggtagcaaa 780 caggattaga taccctggta gtccacgccg taaacgatga gtgctaagtg ttggagggtt 840 tccgcccttc agtgctgcag ttaacgcatt aagcactccg cctggggagt acgaccgcaa 900 ggttgaaact caaaggaatt gacgggggcc cgcacaagcg gtggagcatg tggtttaatt 960 cgaagcaacg cgaagaacct taccaggtct tgacatcctt tgaccgccct agagataggg 1020 tttccccttc gggggcaaag tgacaggtgg tgcatggttg tcgtcagctc gtgtcgtgag 1080 atgttgggtt aagtcccgta acgagcgcaa cccttattgt tagttgccag cattgagttg 1140 ggcactctag caagactgcc ggtgacaaac cggaggaagg cggggatgac gtcaaatcat 1200 catgcccctt atgacctggg ctacacacgt gctacaatgg gaagtacaac gagcaagcca 1260 agccgcaagg cctagcgaat ctctgaaagc ttctctcagt tcggattgca ggctgcaact 1320 cgcctgcatg aagccggaat cgctagtaat cgcggatcag catgccgcgg tgaatccgtt 1380 cccgggcctt gtacacaccg cccgtcacac cacgagagtt tgtaacaccc aaagtcggtg 1440 cggcaaccct tmggggagcc agccgcctaa ggtgggacga atgattgggg tgaagtcgta 1500 acaaggtaac ca 1512

Claims (12)

Tetragenococcus halophilus strain SMB688 with accession number KCTC 13240BP, which has the ability to inhibit the production of biogenic amines histamine or tyramine. delete The method according to claim 1,
Wherein the strain has an inhibitory effect on any one or more of amino acid decarboxylase of histidine decarboxylase and tyrosine decarboxylase. delete The method according to claim 1,
The strain is capable of growing under a condition of 16% salt concentration, and the strain of Tetranyococcus halofilus. delete delete A microbial preparation comprising at least one of the strain of any one of claims 1, 3, and 5, a lysate thereof, and a culture thereof as an active ingredient. A fermented food comprising at least one of the strain of any one of claims 1, 3, and 5, a lysate thereof, and a culture thereof. 10. The method of claim 9,
Wherein the fermented food comprises up to 300 ppm histamine based on the total weight of the fermented food product; or
Fermented food containing less than 200 ppm of tyramine based on the total weight of the fermented food. 11. The method of claim 10,
Wherein the fermented food comprises less than 100 ppm histamine based on the total weight of the fermented food product; or
Fermented food containing less than 50 ppm of tyramine based on the total weight of the fermented food. 10. The method of claim 9,
Wherein the fermented food is a fermented food containing one or more of soy sauce, miso, kochujang, chunchon, chungkukjang, and a mixture thereof.

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