KR20220050257A - Novel starter of Lactobacillus fermentum EFEL6800 with probiotic activity - Google Patents

Abstract

The present invention relates to a novel Lactobacillus fermentum EFEL6800 fermentation starter with probiotics activity. The Lactobacillus fermentum EFEL6800 (KACC 81106BP), discovered in the present invention, showed suitability as a fermentation starter and excellent probiotics properties. The strain of the present invention can grow even at low temperature (15℃) and produce an appropriate amount of acid and abundant aroma components and metabolites, and thus is determined to be suitable as a fermentation starter. In addition, the strain has resistance against bile acid and probiotics properties with high antioxidant activity. In addition, the strain of the present invention does not contain a biogenic amine gene and is not haemolytic, thereby being safe for use in food fermentation. In conclusion, the Lactobacillus fermentum EFEL6800 is suitably used as a fermentation strain for vegetables, fruits, grains, and the like and can be used as a healthy functional probiotics starter with anti-inflammatory activity.

Description

Novel starter of Lactobacillus fermentum EFEL6800 with probiotic activity

The present invention is a novel Lactobacillus fermentum ( Lactobacillus fermentum ) It relates to EFEL6800 (KACC 81106BP), and more particularly, to a novel Lactobacillus fermentum EFEL6800 (KACC 81106BP) fermenting seed having a probiotic function.

Functional food is defined as a food or dietary ingredient that not only provides the basic function of providing nutrition, but also provides health benefits. Among the many functional foods, the field of foods containing probiotics is growing rapidly.

Probiotics are defined as live microorganisms that, when administered in appropriate amounts, provide health benefits to the host, and provide a variety of health benefits, including prevention of pathogen growth, enhancing immunity, anticancer effects, prevention of diarrhea and alleviation of irritable bowel syndrome.

Although fermented milk products play an important role as carriers of probiotics, lactose intolerance and the allergenic potential due to their high fat and cholesterol content are considered major disadvantages associated with fermented dairy products. In addition, vegetarianism is on the rise and modern consumers are increasingly interested in consuming functional foods based on vegetables, fruits and grains that are perceived as having beneficial nutritional value. Therefore, non-dairy probiotic products, including foods from vegetables, fruits and grains, are being evaluated as healthy functional food ingredients today.

Microorganisms used in fermented foods such as fermented milk and yogurt are largely divided into two types based on their roles. The first is a starter that provides the unique flavor of fermented milk through lactic acid fermentation, and the second is probiotics that settle in the intestines of the human body and provide various health functions. Most lactic acid bacteria have been developed as starters or probiotics and are used in fermented foods for different purposes, and the development of 'probiotic starters', which has both the role of a starter for flavor and a role of probiotics for health, is rare. not.

Kimchi is a traditional Korean fermented vegetable made by fermenting cabbage and radish with various ingredients such as red pepper powder, garlic, and ginger. When kimchi is manufactured by natural fermentation using non-sterilized raw materials, various microorganisms proliferate, so the quality is not uniform, and there is a problem of lack of flavor because off-flavor is already generated. On the other hand, the use of a starter for kimchi manufacturing can expect advantages such as uniform quality, improvement of sensory characteristics, extension of shelf life, and improvement of functional characteristics of kimchi products.

Recently, the use of probiotics with health and functional properties in kimchi fermentation has been reported. It produces CLA (conjugated linoleic acid) to improve the functionality of kimchi.Bifidobacterium spp. (Min SG, Kim JH, Kim SM, Shin HS, Hong GH, Oh DG, Kim KN. Manufactures of functional kimchi usingBifidobacterium strain producing conjugated linoleic acid (CLA) as starter. 2003. Korean J Food Sci Technol. 2003; 35(1): 111-114.) or γaminobutyric acid (GABA)Lactobacillus spp. (Seok JH, Park KB, Kim YH, Bae MO, Lee MK, Oh SH. Production and characterization of kimchi with enhanced levels of γ-aminobutyric acid. Food Sci Biotechnol. 2008; 17(5): 940-946.) has been developed. In addition, there is a case of developing mixed probiotics that can improve the anticancer and antioxidant activity of kimchi (Bong YJ, Jeong JK, Park KY. Fermentation properties and increased health functionality of kimchi by kimchi lactic acid bacteria starters. J Korean Soc Food Sci. 2013;42(11):1717-1726; doi: 10.3746/jkfn.2013.42.11.1717.). As such, there are many cases of developing probiotic lactic acid bacteria to manufacture healthy functional kimchi, but there are very few cases where they are actually used in kimchi and commercialized.

This is because there are important limitations when applying lactic acid bacteria developed as probiotics to kimchi. The first problem is that most of the 19 types of probiotics announced by the Ministry of Food and Drug Safety are developed based on milk-based milk-processed foods, and many of these types do well in the nutritional environment of vegetable-based kimchi. Most of the time, they don't grow. The second problem is that most probiotics are thermophilic lactic acid bacteria that grow above 37℃ and are not suitable for kimchi that is mainly fermented at low temperatures below 15℃. The third problem that arises is that even when probiotics are grown in kimchi, they often produce a large amount of lactic acid to lower the taste and shelf life, and create off-flavors that impair the flavor of kimchi. that there is

To overcome this, the existing method of culturing probiotic lactic acid bacteria and adding them after fermentation of kimchi has been proposed, but it is judged that the economic feasibility is lowered by increasing the price of kimchi. In addition, the genus Leuconostoc , which is currently widely used as a starter for kimchi, has weak resistance to acids and bile, and lacks the ability to adhere to human colon epithelial cells.

In Korean Patent Publication No. 1020180052032 (published on May 17, 2018), a novel lactic acid bacterium Lactobacillus plantarum DGK-17 (KACC 81028BP) derived from kimchi and kimchi comprising a fermented composition prepared therefrom and a method for manufacturing the same is described. In Korean Patent Publication No. 1020170080837 (published on July 11, 2017), a novel kimchi lactic acid bacterium Weissella cibaria MFST strain and a jujube seed composition using the same are described.

In the present invention, as a starter that can be used for fermentation of vegetables, fruits, grains, etc. represented by kimchi, a novel 'probiotic starter' that can enhance flavor and probiotic health functionality of the fermented product at the same time is discovered and provided want to

The present invention provides Lactobacillus fermentum EFEL 6800 (KACC 81106BP).

The Lactobacillus fermentum EFEL 6800 (KACC 81106BP) of the present invention is preferably characterized in that it has anti-inflammatory probiotic activity.

The present invention provides a fermented vegetable prepared by inoculating a vegetable with Lactobacillus fermentum EFEL 6800 (KACC 81106BP) as a seed and fermenting it. In this case, the fermented vegetable may be, for example, kimchi.

The present invention provides a fermented fruit prepared by inoculating a fruit with Lactobacillus fermentum EFEL 6800 (KACC 81106BP) as a seed and fermenting it.

The present invention provides a fermented grain produced by inoculating grains with Lactobacillus fermentum EFEL 6800 (KACC 81106BP) as a seed and fermenting it.

Lactobacillus fermentum EFEL6800 (KACC 81106BP) discovered in the present invention showed suitability as a fermentation seed and excellent probiotic properties. The strain of the present invention is suitable as a fermenting seed because it can grow even at a low temperature (15° C.), produces an appropriate amount of acid, and produces abundant aroma components and metabolites. In addition, it has probiotic properties with high acid bile tolerance and antioxidant activity. In addition, the strain of the present invention does not have a biogenic amine gene and is safe for use in food fermentation because it does not have hemolytic properties. In conclusion, Lactobacillus fermentum ( Lactobacillus fermentum ) EFEL6800 is suitable as a fermented strain of vegetables, fruits, grains, etc., and can be used as a healthy functional probiotic seed with anti-inflammatory activity.

1 is a safety test result of the present invention Lactobacillus fermentum EFEL6800.
Figure 2 shows the viability of Lactobacillus fermentum EFEL6800 in simulated gastrointestinal conditions.
Figure 3 shows the antioxidant activity of Lactobacillus fermentum EFEL6800.
Figure 4 shows the inhibitory effect of heat inactivated (death) strains on the expression of nitric oxide (NO) in LPS-induced RAW 264.7 cells.
Figure 5 shows the inhibitory effect of the heat inactivated (killed) strain on the expression of iNOS and COX-2 in LPS-induced RAW 264.7 cells.
6 is a measurement result of cytokine secretion of Lactobacillus fermentum EFEL6800.
Figure 7 is a measure of the growth rate of the present invention Lactobacillus fermentum EFEL6800 and control Leuconostoc mesenteroides DRC1506 (KCCM11712P) strain.
8 is a pH value measured during growth of Lactobacillus fermentum EFEL6800 and control DRC1506 strains of the present invention.
9 is a biplot result of principal components analysis (PCA) performed on the volatile fragrance component of nabak kimchi.
10 is a biplot result of principal components analysis (PCA) performed on the metabolites of kimchi.
11 is a sensory evaluation result of an optimally aged nabak kimchi sample fermented with a probiotic strain.

If the flavor and health functionality of fermented food represented by kimchi can be improved at the same time by developing 'probiotic spawn', the commercial value of kimchi will increase and it will have a desirable effect on the health of consumers.

To this end, in the present invention, the following four elements were determined as essential elements of a 'probiotic seed' to be developed. First, kimchi must be able to grow by using the nutrients well. Second, kimchi must be grown at a low temperature where fermentation takes place. Third, metabolites generated during fermentation should contribute to providing excellent flavor of kimchi. Fourth, it must possess health functionalities such as high acid-bile tolerance, intestinal adhesion, safety, and immunomodulatory ability.

In the present invention, a seed strain satisfying the above four requirements has been developed, and it is intended to provide Lactobacillus fermentum EFEL 6800. Lactobacillus fermentum EFEL 6800 discovered in the present invention is a seed that can be effectively applied to fermentation of kimchi, and at the same time has anti-inflammatory activity and suitability as a health functional probiotic.

First, in order to investigate the characteristics of Lactobacillus fermentum EFEL 6800 of the present invention as a kimchi starter, metabolite analysis and sensory evaluation were performed using growth curves, GC/MS, and NMR in a kimchi imitation medium. The strain of the present invention can grow at both 37 ℃ and 15 ℃ in kimchi medium, and produced an appropriate amount of acid. In addition, when kimchi was prepared using the strain of the present invention as a seed, despite being a Lactobacillus genus strain, It was confirmed that kimchi with good flavor can be prepared, which contains aroma components and metabolites similar to Leuconostoc mesenteroides DRC1506 used as a positive control.

Next, in the present invention, the probiotic activity of the Lactobacillus fermentum EFEL 6800 strain was confirmed. It showed acid-bile tolerance and excellent intestinal stability, and health functionalities of high antioxidant activity and anti-inflammatory activity. That is, the heat inactivation sample of the strain of the present invention suppressed nitric oxide (NO) in macrophages stimulated by LPS, suppressed mRNA expression levels of inflammation-related genes iNOS and COX-2, suppressed pro-inflammatory cytokines, and -Induction of inflammatory cytokine showed high anti-inflammatory effect.

In addition, the Lactobacillus fermentum EFEL 6800 strain of the present invention does not possess the biogenic amine gene, and it is confirmed that it is safe for use in food fermentation because it does not have hemolytic properties.

Lactobacillus fermentum EFEL 6800 (KACC 81106BP) of the present invention having the above characteristics can be used as a fermented seed, it can be used as a fermented seed for vegetables, fruits, grains, and the like.

An example of a fermented vegetable prepared by using Lactobacillus fermentum EFEL 6800 (KACC 81106BP) as a fermented seed is kimchi, and Lactobacillus fermentum EFEL 6800 (KACC 81106BP) is used as a fermented seed. As an example of a fermented fruit produced using, there is atchara, and as an example of a fermented grain produced using Lactobacillus fermentum EFEL 6800 (KACC 81106BP) as a fermentation seed, there is oatmeal. .

On the other hand, in the production of fermented vegetables, fermented fruits, and fermented grains using Lactobacillus fermentum EFEL 6800 (KACC 81106BP) of the present invention as a fermented seed, there is no one in the art except for using the strain of the present invention as a seed. Well-known fermentation methods can be selectively applied and used. Therefore, a description of a specific fermentation method for each product will be omitted.

Hereinafter, the content of the present invention will be described in more detail through the following Examples and Experimental Examples. However, the scope of the present invention is not limited only to the following examples and experimental examples, and includes modifications of technical ideas equivalent thereto.

[Example 1: Lactobacillus fermentum EFEL6800 Isolation and Identification]

of the present inventionLactobacillus fermentum For separation, MRS+ bromophenol blue (BPB) medium supplemented with ribose was prepared. The human saliva of healthy adults was diluted with sterile saline (0.85% NaCl) and spread on a selective medium, and colonies were obtained by culturing at 37° C. under anaerobic conditions for 48 hours. Thereafter, genomic DNA was isolated from the isolated and selected colonies, the 16S rRNA gene sequence was analyzed, and it was identified by comparing the sequence of the type strain in NCBI and the Ribosomal Database Project (RDP).

The strain of the present invention isolated from human saliva through the above process was named EFEL6800, and the 16S rRNA gene sequence of the isolate was analyzed. As a result, it was identified as Lactobacillus fermentum with great similarity to the Lactobacillus fermentum type strain.

[Example 2: The present invention Lactobacillus fermentum Confirmation of biological amine gene and hemolytic activity of EFEL6800 strain]

In order for probiotics to be used in food, they must not form biogenic amines from dietary proteins that can adversely affect health, such as food poisoning. The present invention Lactobacillus fermentum To evaluate the stability of the EFEL6800 strain, the presence of genes related to biogenic amine formation was investigated by PCR.

Each gene 16s rRNA gene involved in biogenic amine production was detected using a multiplex PCR method. The hdc (histidine decarboxylase) and tyrdc (tyrosine decarboxylase) genes were amplified using the following primer pairs. hdc: HDC3 (5'-GATGGTATTGTTTCKTATGA-3') and HDC4 (5'-CAAACACCAGCATCTTC-3'). tyrdc: TD2 (5'-ACATAGTCAACCATRTTGAA-3') and TD5 (5'-CAAATGGAAGAAGAAGTAGG-3'). 16s rRNA genes: 27F and 1492R.

In multiplex PCR, each biogenic amine gene was amplified simultaneously with the 16s rRNA gene in a PCR tube by adding each corresponding primer set. The 16s rRNA gene was used as a positive control for the PCR reaction and template conditions. PCR conditions were as follows. After 32 cycles of 5 minutes at 95°C, 45 seconds at 95°C, 45 seconds at 58°C, and 75 seconds at 72°C, the final extension was performed at 72°C for 5 minutes. Genomic DNAs of L. reuteri ATCC 23272 and Enterococcus faecalis KCCM 11729 were used as positive controls for hdc and tyrdc genes, respectively. For hemolysis analysis, bacterial cells were inoculated on a BHI agar plate supplemented with 7% malpi (MB CELL, Seoul, Korea) by referring to Ryu and Chang's method. Listeria monocytogenes was used as a control and after anaerobic culture at 37° C. for 24 hours, red blood cell hemolysis was observed in the medium.

1 is a safety test result of the present invention Lactobacillus fermentum EFEL6800.

Figure 1 (A) is the detection result of the gene related to the biological amine production. Lane M is a 1 kb DNA marker, lane 1 (B) is a negative control without template DNA, lane 2 (P) is L with hdc (histidine decarboxylase, 440 bp) and tyrdc (tyrosine decarboxylase, 1100 bp) genes, respectively. .reuteri ATCC 23272 and Enterococcus faecalis KCCM 11729 genomic DNA as a positive control. The DNA of the 16S rRNA gene (1530bp) was also amplified. Lane 3 is the EFEL6800 strain. Figure 1 (B) is the result of analysis of hemolytic activity of L. fermentum EFEL6800. Hemolytic activity was measured in BHI broth containing 7% horse blood. The left is the EFEL6800 strain, and the right is the positive control Listeria monocytogenes , and a clear zone is seen around the cell drop.

As shown in (A) of FIG. 1 , the EFEL6800 strain did not have hdc and tyrdc genes that produce histamine and tyramine, respectively. On the other hand, PCR products for these genes were positive controls L. reuteri ATCC 23272 and Ec. It was detected in the genomic DNA of faecalis KCCM 11729.

On the other hand, another important property for use in food is the absence of hemolytic activity of microorganisms. To evaluate the hemolytic activity of the EFEL6800 strain, the cells were inoculated on horse blood agar medium, and the hemolytic ability was examined after incubation at 37° C. for 24 hours. EFEL6800 strain did not show a clear zone around the cell droplet, whereas Listeria monocytogenes showed a clear zone that could be interpreted as hemolytic activity (FIG. 1 (B)).

These results show that the EFEL6800 strain does not have hdc and tyrdc genes and has no hemolytic ability.

[Example 3: The present invention Lactobacillus fermentum Confirmation of acid and bile resistance of EFEL6800 strain]

Resistance to acidic conditions was tested with slight modifications to the method proposed by Conway. Bacterial strains were obtained by culturing in MRS medium for 12 hours and centrifuging at 6,000xg for 10 minutes. The strain was washed three times using PBS (phosphate-buffered saline) of pH 7.2 and then resuspended in the same volume of PBS adjusted to pH 3.0 and 2.5 using HCl. After 0, 90, 180 minutes of incubation at 37 ° C., the strain was plated on MRS agar medium, and the viable cells were counted after incubation at 37 ° C for 48 hours to evaluate acid resistance.

Resistance to bile salts was evaluated by suspending cells in PBS solution containing 0.3% (w/v) bile salts (Sigma, St. Louis, MO, USA) and culturing them at 37°C. Bile salt tolerance was measured in the same way as acid tolerance.

Figure 2 shows the viability of Lactobacillus fermentum EFEL6800 in simulated gastrointestinal conditions. Figure 2 (A) is an acidic condition (pH 3.0), Figure 2 (B) is an acidic condition (pH 2.5), Figure 2 (C) is the result under 0.3% bile salt conditions. Results are expressed as mean±standard deviation (n=3). Significant differences were found in Lactobacillus plantarum WCFS1 (##p <0.01, ### p <0.001) or Lactobacillus rhamnosus GG (*p <0.05, **p <0.01).

As shown in (A) and (B) of Figure 2, the probiotic strains Lactobacillus rhamnosus GG (LGG, KCTC 5033) and Lactobacillus plantarum WCFS1 (WCFS1, ATCC BAA-793) used as positive controls and Lactobacillus fermentum EFEL6800 of the present invention Viability was maintained at pH 3.0. Upon 180 min incubation at pH 2.5, EFEL6800 had a significant decrease in viability, but was statistically similar to LGG.

In addition, as shown in (C) of Figure 2, as a result of analysis of bile tolerance in 0.3% bile salt, EFEL6800 showed a higher viability compared to LGG, although the viability was decreased than that of WSFS1. As a result, WCFS1 strain had the best intestinal stability, and EFEL6800 was able to survive at a similar level to LGG. These results suggest that the EFEL6800 strain is resistant to the gastrointestinal environment.

[Example 4: The present invention Lactobacillus fermentum Antioxidant activity of strain EFEL6800]

The antioxidant activity of the Lactobacillus fermentum EFEL6800 strain was measured with a slight modification of the DPPH inhibition assay proposed by Das & Goyal. For the preparation of intact cells and cell free extract, pre-culture and main culture of Lactobacillus fermentum EFEL6800 were performed for 12 hours each, and the absorbance (optical density) was set to 1.0 at 600 nm. Lactobacillus fermentum EFEL6800 cells were washed twice and resuspended in 0.85% saline to make intact cells. In the case of cell free extract, after sonication treatment for 10 minutes using a sonicator (VP-050N; Taitec Corp., Saitama, Japan), cell debris is removed by centrifugation (10,000xg 4℃ 5 minutes) did An ethanolic DPPH solution (100 μL, 0.4 mmol/L) was strongly mixed with a bacterial sample or water (control) and incubated at 37°C in the dark for 30 minutes. The absorbance of this mixture was measured at 517 nm using a microplate reader.

Figure 3 shows the antioxidant activity of Lactobacillus fermentum EFEL6800. Antioxidant activity was measured by DPPH inhibition assay, and the results were expressed as mean±standard deviation (n=3). Different letters in the error bars indicate significant differences. Significant differences were found with L. plantarum WCFS1 (#p <0.05, ### p <0.001) or L. rhamnosus GG (*** p <0.01).

As shown in FIG. 3 , the Intact cells of EFEL6800 showed 22% DPPH removal activity, which was statistically higher than that of WCFS1 (10.6%) and LGG (17.1%). The antioxidant activity of the cell free extract of EFEL6800 was 18.2%, which was significantly higher than that of WCFS1 (1.6%) and LGG (8.0%).

Therefore, Lactobacillus fermentum EFEL6800 of the present invention was judged to have higher antioxidant activity than commercial probiotics Lactobacillus plantarum WCFS1 and Lactobacillus rhamnosus GG.

[Example 5: The present invention Lactobacillus fermentum Anti-inflammatory activity of strain EFEL6800]

(1) Nitric oxide (NO) inhibitory ability

To analyze the anti-inflammatory activity of Lactobacillus fermentum EFEL6800, RAW 264.7 macrophages were stimulated with E. coli-derived lipopolysaccharide (LPS, 1 ug/ml) to excessively produce nitric oxide (NO), an inflammatory mediator, and then heat fire of the strain The NO inhibitory activity of the activated sample was measured.

For the preparation of heat-inactivated strains, the absorbance of the strain was adjusted to 1.0 (OD 600nm), centrifuged at 10,000xg for 5 minutes, the cell pellet was rinsed twice with PBS and suspended in DMEM, and heat-killed at 90°C for 30 minutes. . Macrophage line RAW 264.7 cell line was obtained from KCLB and maintained in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin (37°C, 5% CO ₂ ). Cells were sub-cultured in plates to the 80-90% confluency stage. RAW 264.7 cells (5x10 ⁵ cells per ml in 96 well plate) were stimulated for 24 hours by adding LPS (1 μg/mL) and heat-inactivated strain. After 24 h, the supernatant from each well was mixed with an equal volume of Griess reagent and placed in the dark at room temperature for 10 min. Then, the absorbance was measured at 540 nm using a microplate spectrophotometer (BioTek, Winooski, VT, USA), and the NO content was calculated by creating a standard curve with sodium nitrate. Methyl arginine was used as a positive control.

Figure 4 shows the inhibitory effect of the heat inactivated (death) strain on the expression of nitric oxide (NO) in LPS-induced RAW 264.7 cells. MA is methyl arginine, NO inhibition was measured by Griess reaction analysis, and the results are expressed as mean±standard deviation (n=3). Different letters in the error bars indicate significant differences. Statistical analysis was performed using an independent T-test compared to the LPS-treated group. (*p <0.05, **p <0.01, ***p <0.001).

As shown in FIG. 4 , NO production was significantly increased when treated with LPS compared to the control group not treated with LPS. At this time, treatment with methyl arginine, an NO synthetase inhibitor, inhibited NO production in a dose-dependent manner compared to the LPS-treated group (p <0.001). On the other hand, for all heat-inactivated strains tested, they showed a significant inhibitory effect on NO production compared to the LPS-treated group, and in particular, EFEL6800 showed a higher NO inhibitory ability than WCFS1 and LGG.

(2) Confirmation of inhibition of iNOS and COX-2 expression using RT-qPCR

The expression activity of inflammation-related mRNA of Lactobacillus fermentum EFEL6800 was measured by cDNA synthesis and RT-qPCR after RNA extraction in LPS-induced RAW 264.7 cells treated with heat-inactivated strains. RAW 264.7 cells (1x10 ⁶ cells/well) were cultured in a 6 well plate and stimulated with LPS (1 μg/mL) and heat-inactivated strain for 24 hours.

RNA was extracted from each well using Trizol reagent, and cDNA was synthesized using a cDNA synthesis kit (LeGene Express 1 ^st Standard cDNA Synthesis System). GADPH, iNOS, and COX-2 genes were amplified by RT-qPCR using the following primer pairs. GADPH is Forward (5'-TTGTCTCCTGCGACTTCAACA-3') and Reverse (5'-GCTGTAGCCGTATTCATTGTCATA-3'), iNOS is Forward (5'-ACCATGGAGCATCCCAAGTA-3') and Reverse (5'-CCATGTACCAACCATTGAAGG-3'), COX-2 was amplified by Forward (5'-AGCATTCATTCCTCTACATAAGC-3') and Reverse (5'-GTAACAACACTCACATATTCATACAT-3'). PCR conditions were as follows. After 5 minutes at 95°C, 40 cycles of 15 seconds at 95°C, 30 seconds at 60°C, and 30 seconds at 60°C were performed.

Figure 5 shows the inhibitory effect of the heat inactivated (killed) strain on the expression of iNOS and COX-2 in LPS-induced RAW 264.7 cells. Cells were treated with heat killed strains and stimulated with LPS for 24 h. The expressed mRNA levels of iNOS and COX-2 were determined by real-time PCR and relative quantification by normalization using GAPDH. Different letters on the error bars indicate significant differences. Statistical analysis was performed using an independent T-test compared to the LPS-treated group. (*p <0.05, **p <0.01, ***p <0.001).

As shown in FIG. 5, the mRNA expression of COX-2 (FIG. 5(B)) and iNOS (FIG. 5(A)) was hardly confirmed in RAW 264.7 cells in a stable state without LPS stimulation, but LPS stimulation The expression level was significantly increased. In contrast, the heat-inactivated samples of all tested strains significantly inhibited the mRNA expression of COX-2 and iNOS. In particular, EFEL6800 showed significant anti-inflammatory activity with WCFS1.

(3) Measurement of secretion of inflammatory cytokines (IL-12) and anti-inflammatory cytokines (IL-10) using ELISA

The immunomodulatory activity of Lactobacillus fermentum EFEL6800 was evaluated by measuring cytokine production in LPS-induced mouse macrophages using an ELISA kit. The animal protocol was approved by the Kyung Hee University Medical Center Institutional Animal Care and Use Committee (KHUASP(GC)-19-005). Peritoneal macrophages were collected from 7-week-old male Balb/c mice through peritoneal washing with DMEM, and cells (4x10 ⁵ cells/ml) were stimulated with LPS (1 μg/mL) and heat-inactivated strains for 24 hours. The supernatant was collected from each well, and IL-10 and IL-12 secreted into the culture medium were quantified using an ELISA assay kit (R&D systems Mouse DuoSet ELISA IL-10 and IL-12, Minnesota, USA).

6 is a measurement result of cytokine secretion of Lactobacillus fermentum EFEL6800. FIG. 6(A) is IL-12, FIG. 6(B) is IL-10, FIG. 6(C) is IL-10/IL-12. Results are expressed as mean±standard deviation (n=6). Different letters in the error bars indicate significant differences. Statistical analysis was performed using an independent T-test compared to the LPS-treated group. (*p <0.05, **p <0.01, ***p <0.001).

As a result of the experiment, LPS strongly induced the release of IL-12, an inflammatory cytokine (FIG. 6 (A)), whereas IL-10, an anti-inflammatory cytokine, was not affected (FIG. 6 (B)). All heat-inactivated strains tested completely inhibited IL-12 (FIG. 6(A)) and strongly induced IL-10 (FIG. 6(B)). In particular, the heat-inactivated strain of Lactobacillus fermentum EFEL6800 is a strong inducer of IL-10, and as an inhibitor of IL-12, it has a high anti-inflammatory index (IL-10/IL-12 ratio) at a level similar to that of LGG, a commercial probiotic (Fig. 6(c)).

Therefore, Lactobacillus fermentum EFEL6800 decreased IL-12 in LPS-stimulated mouse peritoneal macrophages, induced IL-10, and could be evaluated as having immunomodulatory ability ex vivo.

[Example 6: Invention in Kimchi Mosa Juice (SKJ) Lactobacillus fermentum Measurement of growth rate of EFEL6800]

Simulated kimchi juice (SKJ) was developed to monitor starter growth and obtain reproducible results. The ingredients and composition of the kimchi simulation juice are presented in Table 1.

Kimchi Mosa Juice Composition Ingredients Concentration Cabbage 700g Radish 200g Leek 50g Ginger 10g Garlic 20g Salt 3% Fish peptone 0.5%

Raw materials (cabbage, radish, garlic, ginger, leek) were purchased from the market. All the ingredients were cut using a blender, and salt was added for the amount of cabbage and salted overnight. Fish peptone (Bision Co., Seongnam, Korea) was added instead of salted fish, and SKJ was pasteurized at 70° C. for 30 minutes. After cooling to room temperature, the mixture was centrifuged at 7,000 rpm for 10 minutes to remove pulp, and only the supernatant was used for the experiment.

To investigate whether the isolate was suitable for kimchi fermentation, the fermentation properties of Lactobacillus fermentum EFEL6800 were monitored at SKJ. A commercial kimchi starter, Leuconostoc mesenteroides DRC1506, was used as a positive control. After pre-culturing each strain, 1% of SKJ was inoculated to reach 10 ⁷ CFU/mL and cultured at the optimal growth temperature (37°C for 24 hours, 15°C for 8 days) with a spectrophotometer (BioTek Instruments, Winooski, VT , USA) was used to measure optical density (OD600nm) and pH was measured using a pH meter (Orion Versa, Thermo, USA), that is, the optimum temperature 37 in SKJ to compare the adaptability of strains in the kimchi environment. The growth rate was measured at ℃ for 24 hours and at 15℃ for 8 days, and the pH value was measured to compare the effect on the taste of kimchi.

The experimental results were as shown in FIGS. 7 and 8 . Figure 7 is a measure of the growth rate of the present invention Lactobacillus fermentum EFEL6800 and the control Leuconostoc mesenteroides DRC1506 (KCCM11712P) strain, the optical density (OD) value is 37 ℃ (A) and 15 ℃ (B) SKJ inoculated with these strains was measured after Results are expressed as mean±standard deviation (n=3). Statistical analysis was performed using an independent T-test (* p <0.05, ** p <0.01, *** p <0.001). 8 is a pH value measured during the growth of Lactobacillus fermentum EFEL6800 and control DRC1506 strains of the present invention, measured after inoculating these strains into SKJ at 37° C. (A) and 15° C. (B). Results are expressed as mean±standard deviation (n=3). Statistical analysis was performed using an independent T-test (* p <0.05, ** p <0.01, *** p <0.001).

The Lactobacillus fermentum EFEL6800 strain of the present invention had a significantly higher growth rate than the positive control, Leuconostoc mesenteroides DRC1506, at the optimum temperature, and the pH after 24 hours of fermentation was statistically similar to that of DRC1506. As a result of culturing at 15° C., DRC1506 grew from the first day, whereas EFEL6800 grew from the fifth day of culture. These results indicate that EFEL6800 satisfies nutritional requirements for growth in plants, can grow at low temperatures, and produces an appropriate amount of acid during fermentation, making it suitable as a seed for kimchi.

[Example 7: The present invention Lactobacillus fermentum Analysis of fragrance components of Nabak Kimchi using EFEL6800]

To evaluate the effect of starter on kimchi flavor After fermenting nabak kimchi using different starters until it reached the proper season, volatile aroma components were analyzed through GC/MS. As starters, Lactobacillus fermentum EFEL6800, Lactobacillus brevis DRC301, Lactobacillus paracasei CBA3611, and Leuconostoc mesenteroides DRC1506 of the present invention were used as starters.

Nabak kimchi was prepared by the following method (the basic recipe for Nabak kimchi follows the nabak kimchi recipe manufactured by domestic company T). After cutting Chinese cabbage and radish, the main raw materials, to a certain size, it passed through a washing machine to select foreign substances to prepare raw materials. After washing according to the standards, the auxiliary ingredients such as chives, red pepper, water parsley, garlic, ginger, and carrots were pickled. Sub-materials that have been washed were prepared by cutting them to a prescribed size. Each raw material and auxiliary material was weighed according to the mixing ratio, mixed with the brine (salinity 1.8%) prepared in advance, and inoculated with the starter, respectively. After the mixing was completed, Nabakmul Kimchi was aged at low temperature (4℃) for 2 days, and the solid content was matched with the kimchi broth and packaged in a container.

2 mL of nabak kimchi prepared by the above method was placed in a headspace vial (20 mL, 22.5 mm×75.5 mm) and stirred at 51° C. for 20 minutes before analysis. The volatile aroma components of kimchi were extracted under the conditions shown in Table 2 using solid-phase microextraction (SPME) fibers (DVB/CAR/PDMS, 50/30 μm, Supelco, 57298-U).

SPME conditions Fiber ^DVB/CAR/PDMS Incubation 51℃ 20 min ^Adsorption 51℃ 30 min ^Desorption 250℃ 2 min

The extracted volatile fragrance components were analyzed under the conditions of Table 3 using a gas chromatography-mass spectrometer (7820A/5977E MSD, Agilent Technologies, USA).

GC/MS conditions Column DB-WAX (50m x 200um x 0.2um, Agilent Technologies, USA) ^{carrier gas} Helium ^{Oven temp.} 40℃(5min)
~150℃, 5℃/min (0min)
~200℃, 7℃/min (10min) ^{injector temp.} 250℃ ^{Ion source temp.} 250℃ ^{split ratio} splitless ^{Mass scan range} 35~350amu

The quantification of volatile aroma components of kimchi was calculated as the peak area of each compound relative to the peak area of methyl cinnamate (5 ppm), an internal standard.

The experimental results were shown in Table 4 and FIG. 9 .

Aroma compounds Before fermentation After fermentation Nk 0day DRC1506 0day NK DRC1506 EFEL6800 CBA3611 DRC301 Total compounds 1188.19±182.49 1384.94±131.47 218.04±45.48 255.22±60.45 Sulfur-containing compounds 1019.85±146.83 1238.85±133.38 66.63±28.74 109.54±40.58 82.95±15.09 138.88±14.76 139.11±11.61 2-Thiophenecarboxylic acid, 5-(1,1-dimethylethoxy)- - 15.38±6.31 - - - - - N-Methyltaurine - 8.70±0.89 - - - - - Allyl methyl sulfide 218.41±30.02 230.57±28.76 - 21.36±1.25 20.20±0.38 18.72±3.87 15.18±1.73 Disulfide, dipropyl 4.03±0.64 5.31±0.19 - - - - - 1,5,2,4-Dioxadithiepane-2,2,4,4-tetraoxide - - - - - 3.07±0.65 2.52±0.09 1,2-Dithiolane 29.79±2.94 21.93±13.68 9.01±4.77 - - - - 1-Butene, 4-isothiocyanato- - 0.00±0.00 26.47±2.58 24.58±12.80 28.23±3.41 39.30±4.85 38.56±3.52 Diallyl disulfide 767.63±114.03 956.97±119.01 31.15±21.78 36.17±16.52 41.26±11.20 32.56±4.51 37.54±4.20 Cyclopentyl isothiocyanate - - - 27.43±13.65 - 45.23±4.35 45.30±3.08 Nitrile-containing compounds 0.98±0.09 7.95±2.86 34.50±5.87 - 40.35±4.58 5.02±2.51 1.24±0.14 2,6,6-Trimethyl-bicyclo[3.1.1]hept-3-ylamine - - - - 13.79±1.31 - - Hydroxyurea - - - - 1.16±0.60 - 1.24±0.14 2-t-Butyl-1-methyl-3-phenyl-imidazolidin-4-one - - - - - 3.85±1.72 - N-Methylcaprolactam - 7.95±2.86 34.50±5.87 - 30.00±2.08 - - R-(-)-Cyclohexylethylamine 0.98±0.09 - - - - - - N-Methoxy-1-ribofuranosyl-4-imidazolecarboxylic amide - - - - - 1.07±0.17 - N-Methoxy-1-ribofuranosyl-4-imidazolecarboxylic amide - - - - - 1.39±0.26 - Alcohols and Amino Alcohols 15.70±2.18 14.65±2.49 8.16±1.83 7.25±1.00 8.85±0.31 7.26±0.94 9.07±1.95 4-amino-1-pentanol 7.60±2.42 - 2.76±0.14 2.60±0.11 3.84±0.76 3.64±1.50 2.84±0.47 4-amino-1-pentanol 1.67±0.39 - 1.14±0.03 1.06±0.14 1.24±0.08 - 0.85±0.10 L-Valinol - 8.94±2.12 - - - - - 4-amino-1-pentanol 0.63±0.33 - - - - - 2.68±0.34 2-(2-Aminopropyl)phenol - 2.52±0.86 3.07±1.67 2.33±0.78 2.37±0.43 2.37±0.74 2.52±0.17 4-amino-1-pentanol - 1.22±0.35 - - - - - 1-Methylene-2b-hydroxymethyl-3,3-dimethyl-4b-(3-methylbut-2-enyl)-cyclohexane 2.12±0.22 1.98±0.20 1.20±0.11 1.26±0.13 1.41±0.21 1.25±0.18 1.08±0.06 2-Methyl-1-[3-methyl-6-(1-methylethylidene)-3-cyclohexen-1-yl]-3-buten-2-ol 3.89±1.40 - - - - - - Benzenes and benzene derivatives 9.62±3.10 6.24±0.54 13.35±2.54 27.01±6.33 26.71±4.00 24.63±2.97 35.49±21.58 Benzeneethanamine, N-[(4-hydroxy)hydrocinnamoyl]- - - - 9.98±0.15 12.10±2.15 - 8.13±0.18 Benzene, 1-methyl-3-(1-methylethyl)- 2.94±2.28 - - 10.11±1.08 - 13.16±1.97 - Benzene, 1,3-bis(1,1-dimethylethyl)- - - 3.35±1.67 2.37±0.16 4.51±3.37 - 20.35±25.24 Benzaldehyde, 3-(2,4,6-trichlorophenoxymethyl)-4-methoxy- - - 2.48±0.59 2.06±0.24 1.38±0.15 1.55±0.36 1.30±0.27 Benzaldehyde, 3-(2,4,6-trichlorophenoxymethyl)-4-methoxy- - - - - 1.17±0.42 3.21±0.64 1.34±0.41 Benzeneethanamine, 3-fluoro-β,5-dihydroxy-N-methyl- - - - - 0.90±0.13 - - Benzene, 1-(1,5-dimethyl-4-hexenyl)-4-methyl- 6.68±0.88 6.24±0.54 3.04±0.08 3.28±0.37 3.18±0.19 3.01±0.16 2.60±0.14 Benzene, (2-isothiocyanatoethyl)- - - 4.48±0.67 3.32±1.51 3.48±0.14 4.77±0.14 4.47±0.19 Acids 6.62±0.38 - 15.70±7.04 13.07±6.68 28.35±6.13 41.57±10.68 34.30±5.24 (Allythio)acetic acid - - - - 4.36±1.01 7.16±1.24 7.71±1.89 Oxaluric acid - - 15.70±7.04 13.07±6.68 23.98±5.93 34.40±9.50 26.59±3.37 2-Butoxyethyl acetate 6.62±0.38 - - - - - - Terpenes 14.81 ± 1.90 11.76±3.97 10.09±0.84 25.62±1.49 9.93±0.31 35.23±5.79 18.07±1.01 β-Terpinen - - - 15.34±0.16 - 23.60±5.40 9.98±0.59 β-Caryophyllene 14.81 ± 1.90 11.76±3.97 10.09±0.84 10.28±1.62 9.93±0.31 11.63±0.41 8.09±0.87 Aldehyde and ketone 1.72±0.47 12.22±2.75 1.32±0.54 1.44±0.54 1.33±0.55 1.43±0.62 1.28±0.47 Methyl salicylate 1.72±0.47 1.74±0.44 1.32±0.54 1.44±0.54 1.33±0.55 1.43±0.62 1.28±0.47 Dodecal - 8.63±2.35 - - - - - 1,1-Dodecanediol, diacetate - 1.85±0.67 - - - - - hydrocarbon 55.75±19.67 63.60±10.70 35.92±13.75 39.16±3.52 34.37±1.95 57.98±11.01 33.84±3.03 Limonene 55.75±19.67 63.60±10.70 35.92±13.75 36.00±2.81 34.37±1.95 57.98±11.01 33.84±3.03 propane - - - 0.73±0.25 - - - Miscellaneous 51.82±8.65 29.14±5.87 32.37±1.84 32.14±3.20 30.49±1.84 35.75±3.11 32.17±2.12 Penicillamine, tri-TMS 5.10±5.85 - 0.93±0.11 0.89±0.20 0.74±0.14 - 0.68±0.28 Silane, trichlorodocosyl- 10.67±5.04 5.05±2.96 5.71±0.28 5.55±0.17 6.16±0.99 5.48±0.22 5.10±0.36 Silane, trichlorodocosyl- - - - 3.22±1.66 5.00±0.92 - 3.96±0.50 2-Trifluoroacetoxydodecane 9.04±1.39 7.44±2.10 4.61±0.89 - - 4.36±0.76 - Silane, trimethyl(1-methyl-1-phenylethoxy)- - - - - - 3.45±0.04 - trans-(2-Chlorovinyl)dimethylethoxysilane 1.39±0.10 - - - - - - trans-(2-Chlorovinyl)dimethylethoxysilane 1.67±0.53 - 1.18±0.06 1.43±0.46 - - - Silanediol, dimethyl- 23.28±4.04 15.94±3.42 19.50±0.76 19.78±2.08 17.73±3.51 23.62±2.83 21.56±1.22 Metaraminol 0.67±0.09 - - - - - - Topotecan - 0.71±0.05 - 0.54±0.11 - - 0.86±0.02 Topotecan - - 1.15±0.20 0.90±0.51 0.86±0.09 - -

9 is a biplot result of principal components analysis (PCA) performed on the volatile fragrance component of nabak kimchi. (A) is the result of inclusion before fermentation, (B) is the result of exclusion before fermentation).

As shown in FIG. 9 , as a result of PCA analysis, there was a clear difference in non-volatile fragrance components before and after fermentation (FIG. 9 (A) and (B)). However, when PCA was applied only to nabak kimchi after fermentation (Fig. 9 (B)), natural fermentation (NK), DRC1506, EFEL6800 fermented nabak kimchi based on the PC1 axis were located on the right , DRC301, CBA3611 Fermented nabak kimchi was found to be on the left. Through this, it is judged that EFEL6800 contains a fragrance component similar to that of Leuconostoc mesenteroides DRC1506.

[Example 8: the present invention Lactobacillus fermentum Analysis of metabolites of kimchi using EFEL6800]

Metabolite analysis of kimchi was performed using a 500 MHz NMR spectrometer. Briefly, 3 ml of kimchi broth was adjusted to pH 6.0 and then centrifuged at 13,000 rpm for 5 minutes to obtain a supernatant. The supernatant was dissolved in 99.9% D ₂ O (deuterium oxide; Sigma-Aldrich, USA) containing 0.5 mM 2,2-dimethyl-2-silapentane-5-sulfonate sodium salt (DSS; Sigma-Aldrich, USA) 1: It was suspended in a ratio of 1 and transferred to a 5 mm NMR tube. After that, ¹ H NMR spectra were obtained using a Bruker 500-MHz NMR spectrometer (Bruker Magnetics, Faellanden, Switzerland), and the identification and quantification of individual metabolites were performed using 2,2-dimethyl-2-silapentane-5-sulfonate sodium salt ( DSS; Sigma-Aldrich, USA) was used as an internal standard, and Chenomx NMR Suite v. 6.1 (Chenomx, Canada).

The results are shown in Table 5.

Quantitative value of metabolites (mM) of optimally aged nabak kimchi samples fermented with probiotic strains Group Compound NK 0day DRC1506 0day NK DRC1506 EFEL6800 DRC301 CBA3611 Carbohydrate Glucose 7.85±0.22 7.64±0.12 6.06±0.36 6.39±0.24 6.64±0.38 5.13±0.18 6.34±0.24 Fructose 9.90±0.07 9.41±0.05 2.65±0.79 1.73±0.05 4.60±0.10 3.62±0.04 4.36±0.09 Sucrose 0.61±0.02 0.30±0.00 0.75±0.23 1.05±0.08 0.37±0.20 0.22±0.04 0.41±0.02 Alcohol Ethanol 4.14±0.68 3.23±0.00 10.52±0.50 11.85±0.11 12.86±0.35 6.58±0.21 7.98±0.08 Mannitol 1.72±0.22 1.43±0.08 12.92±0.32 13.08±0.60 12.24±0.87 8.01±0.33 10.80±0.17 organic acid 2-Aminobutyrate 0.08±0.00 0.08±0.01 0.16±0.06 0.19±0.09 0.04±0.03 0.27±0.02 0.30±0.01 4-Aminobutyrate 0.64±0.01 0.58±0.01 0.99±0.06 1.25±0.08 1.08±0.08 0.82±0.11 1.31±0.03 Acetate 0.23±0.00 0.58±0.00 9.68±0.31 10.46±0.06 11.05±0.05 9.36±0.49 10.84±0.11 Lactate 0.44±0.02 0.79±0.02 13.16±0.44 13.59±0.04 13.93±0.37 13.92±0.32 15.15±0.06 Succinate 0.07±0.00 0.09±0.00 0.29±0.16 0.45±0.03 0.52±0.01 0.39±0.01 0.08±0.00 Pyruvate 0.14±0.01 0.13±0.01 0.04±0.02 0.12±0.04 0.05±0.01 0.08±0.01 0.03±0.01 amino acid Alanine 0.48±0.02 0.51±0.03 1.06±0.05 0.94±0.07 0.96±0.03 0.81±0.11 1.08±0.03 Arginine 0.24±0.01 0.23±0.05 0.38±0.24 0.91±0.02 0.41±0.20 0.46±0.00 1.15±0.01 Asparagine 0.53±0.00 0.61±0.10 0.57±0.12 1.21±0.07 0.48±0.20 0.35±0.00 0.98±0.25 Aspartate 0.45±0.09 0.26±0.06 0.30±0.09 0.99±0.08 0.45±0.20 0.28±0.04 0.94±0.11 Cysteine 0.29±0.04 0.23±0.02 0.19±0.10 0.55±0.03 0.36±0.16 0.35±0.00 0.45±0.02 Glutamate 1.43±0.01 1.32±0.02 0.91±0.12 1.11±0.00 1.26±0.10 1.17±0.07 1.35±0.09 Glutamine 0.18±0.03 0.18±0.00 0.95±0.25 0.72±0.08 1.08±0.07 1.05±0.00 1.51±0.04 Glycine 0.36±0.13 0.07±0.01 0.39±0.17 0.86±0.04 0.46±0.28 0.28±0.05 0.24±0.04 Histidine 0.00±0.00 0.00±0.00 0.08±0.06 0.05±0.05 0.01±0.02 0.06±0.09 0.04±0.03 Isoleucine 0.10±0.02 0.15±0.01 0.07±0.03 0.17±0.05 0.12±0.05 0.13±0.00 0.17±0.01 Leucine 0.10±0.01 0.14±0.01 0.10±0.06 0.17±0.02 0.20±0.03 0.16±0.04 0.24±0.02 Lysine 0.03±0.00 0.08±0.02 0.04±0.02 0.09±0.01 0.08±0.04 0.18±0.16 0.07±0.05 Malonate 0.13±0.02 0.12±0.02 0.04±0.06 0.34±0.05 0.16±0.04 0.16±0.03 0.15±0.04 Methionine 0.11±0.02 0.11±0.01 0.10±0.01 0.19±0.01 0.06±0.02 0.10±0.00 0.09±0.04 Proline 0.13±0.05 0.38±0.10 0.51±0.24 0.40±0.11 0.34±0.04 0.45±0.00 0.85±0.07 Phenylalanine 0.03±0.04 0.00±0.00 0.09±0.03 0.05±0.04 0.03±0.04 0.13±0.10 0.14±0.04 Serine 1.19±0.06 0.86±0.03 1.61±0.18 1.19±0.30 0.58±0.09 0.70±0.17 0.86±0.00 Ornithine 0.07±0.03 0.02±0.00 0.13±0.03 0.16±0.05 0.05±0.04 0.07±0.00 0.03±0.03 trans-4-Hydroxy-L-proline 0.54±0.00 0.53±0.02 0.34±0.13 0.92±0.13 0.34±0.21 0.37±0.13 0.52±0.03 Threonine 0.07±0.00 0.07±0.00 0.47±0.24 0.35±0.13 0.32±0.02 0.46±0.13 0.46±0.01 Tryptophan 0.00±0.00 0.00±0.00 0.05±0.03 0.02±0.02 0.02±0.03 0.09±0.04 0.23±0.02 Tyrosine 0.00±0.00 0.01±0.00 0.20±0.10 0.07±0.02 0.10±0.03 0.10±0.00 0.22±0.06 Valine 0.06±0.01 0.10±0.00 0.17±0.03 0.19±0.03 0.20±0.07 0.12±0.01 0.07±0.01

In addition, as a result of PCA analysis, there was a clear difference in metabolites before and after fermentation. 10 is a biplot result of principal components analysis (PCA) performed on metabolites of kimchi. (A) is the result of inclusion before fermentation, and (B) is the result of exclusion before fermentation.

As shown in FIG. 10, when PCA was applied only to Nabak Kimchi after fermentation ((B) in FIG. 10), Natural Fermentation (NK), DRC1506, EFEL6800 Fermented Nabak Kimchi based on the PC1 axis were located on the right , DRC301, CBA3611 Fermented nabak kimchi was found to be on the left.

Through this, EFEL6800 is considered to contain metabolites similar to Leuconostoc mesenteroides DRC1506. In particular, EFEL6800 fermented nabak kimchi was statistically similar to DRC1506 in mannitol content. LAB produces mannitol by reducing fructose, which has a refreshing and soft sweetness and gives a refreshing feeling in kimchi. In addition, it has been reported that it helps to improve the taste of kimchi by suppressing sour taste and suppressing the phenomenon of excessive sourness of kimchi. These results indicate that EFEL6800 contains metabolites most similar to DRC1506, and can improve the taste and flavor of kimchi with high mannitol content.

[Example 9: The present invention Lactobacillus fermentum Sensory evaluation of kimchi using EFEL6800]

Sensory evaluation was conducted by a total of 30 sensory personnel according to the 9-point scale method after a total of 5 kinds of Nabak Kimchi reached the optimum season (acidity 0.17~0.31%/pH 3.99). The evaluation contents are based on preference criteria: aroma, sour taste, sweet taste, carbonic acid-like taste, off-flavor, and total preference. In terms of the degree, the closer to 1, the lower the preference, and the closer to 9, the higher the preference.

The results are shown in FIG. 11 . 11 is a sensory evaluation result of an optimally aged nabak kimchi sample fermented with a probiotic strain. Results are expressed as mean±standard deviation (n=30). There were no significant differences between samples.

In all evaluation items, all nabak kimchi scored an average of 5 or more, indicating a preference above average. As a result of the comprehensive preference evaluation, DRC301 fermented nabak kimchi showed the highest preference, and natural fermented (NK) nabak kimchi showed the lowest preference, but there was no significant difference. Although no significant difference was found in this sensory evaluation result, overall preference was shown in all fermented nabak kimchi, which means that the addition of the Lactobacillus fermentum EFEL6800 starter of the present invention does not have a negative effect on the sensory of nabak kimchi.

Deposited organization name: Agricultural Biotechnology Research Institute

Accession number: KACC81106

Deposit date: 20191025

Claims (6)

Lactobacillus fermentum EFEL 6800 (KACC 81106BP)
According to claim 1,
The Lactobacillus fermentum ( Lactobacillus fermentum ) EFEL 6800 (KACC 81106BP) is,
It is characterized by having anti-inflammatory and probiotic activity.
A fermented vegetable produced by inoculating vegetables with Lactobacillus fermentum EFEL 6800 (KACC 81106BP) as a seed and fermenting it.
According to claim 1,
The fermented vegetables are
A fermented vegetable, characterized in that it is kimchi.
A fermented fruit produced by inoculating a fruit with Lactobacillus fermentum EFEL 6800 (KACC 81106BP) as a seed and fermenting it.
Fermented grains produced by inoculating grains with Lactobacillus fermentum EFEL 6800 (KACC 81106BP) as a seed and fermenting them.

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