KR20190010036A - Novel Lactobacillus plantarum K118 with Antioxidant and GABA-producing ability, composition comprising a fermented product of Dendropanax morbifera extract using the same and manufacturing method therof - Google Patents

Abstract

The present invention relates to a Lactobacillus plantarum K105 with antioxidant and GABA-producing ability, a composition comprising a fermented product of Dendropanax morbifera extract using the same and manufacturing method thereof and, more particularly, to a novel Lactobacillus plantarum strain having excellent activity for promoting antioxidation and GABA production, and fermenting a fermentation product by inoculating the novel Lactobacillus plantarum strain into extract of Dendropanax morbifera, thereby increasing the antioxidant and GABA content.

Description

Lactobacillus plantarum K118 strain having antioxidant activity and GABA production promoting activity, fermented product of Yellowlily chestnut tree using the same, and a method for producing the fermented product of Dillanthus chinensis extract using Dendropanax morbifera extract using the same. same and manufacturing method therof}

The present invention relates to a lactobacillus plantarum K118 strain having antioxidative and GABA production promoting activity, a fermented product of the same, and a method for producing the same. More particularly, the present invention relates to a novel fermented product of a novel fermented product Lactobacillus plantarum K118 strain, and a fermented product obtained by inoculating the fermented product with an extract of Fusarium oxysporum species, thereby increasing the antioxidant and GABA contents.

Gamma-amino butyric acid (GABA) is a nonprotein constituent amino acid that acts not only as a neurotransmitter in the brain but also as a brain stimulator, a mental stabilizer, a blood pressure lowering agent, a diuretic agent, , Alcohol metabolism promoting action, and deodorizing action. Especially, it is registered as a medicine and is used for treatment of headache, tinnitus and hypersensitivity due to stroke or cerebral artery sequelae.

GABA is known to be widely distributed in various vegetables, fruits, rice, beans, etc. However, its content is low and it has a disadvantage that it can not show the physiological function.

Dendropanax morbifera is a plant belonging to Araliaceae. It is a medicinal plant with excellent pharmacological efficacy. It has a scientific name Dendro-panax morbifera Nakai , meaning a panacea. In the past, it was used as a natural coating material because it is composed of 66.7% of nonvolatile matter, 10.8% of directional component, 8.1% of moisture, and 14.4% of solid content. However, recent studies have shown that diabetic, hypertensive, , Liver function improvement, antioxidation, osteoporosis, periodontal disease, immunity enhancement, nervous stability, antimicrobial, anticancer and so on.

In order to exert a pharmacological effect, such a method of extracting a useful component and taking it for a long time has been studied. However, the extract is not high in its pharmacological activity per unit weight, is slow in absorption into the human body, It is difficult to change the formulation.

As a part of this research, it has been known that fermented extracts of leafy leaves of Aspergillus oryzae were inoculated and cultured in the leaves of the leaves of Tochigi leaves, In order to increase the content of the present invention.

Therefore, the present inventors have made efforts to find a method of improving antioxidant and GABA content by using the extract of Hwangchujang. As a result, it has been found that a novel Lactobacillus plantarum strain is isolated, By confirming that the GABA content is improved, it is intended to provide a GABA high-content fermentation material and a production method thereof.

Patent Document 1. Korean Patent No. 10-1423433

SUMMARY OF THE INVENTION The present invention has been conceived to solve the above-mentioned problems, and an object of the present invention is to provide Lactobacillus plantarum strain K118 (accession number KCCM12015P) having antioxidative and GABA production promoting activity.

In addition, another object of the present invention is to provide a fermented yellowtail tree having increased antioxidative and GABA contents obtained by fermenting Lactobacillus plantarum K118 strain (accession number KCCM12015P) to a perennial tree.

It is still another object of the present invention to provide a fermented product of Fusarium oxysulfurum, which is obtained by inoculating Fusarium oxysporum with a strain of Lactobacillus plantarum K118 (Deposit No. KCCM 12015P).

In order to achieve the above object, the present invention provides a Lactobacillus plantarum K118 strain (Deposit No. KCCM 12015P) having antioxidant and GABA production promoting activity.

The strain is characterized in that it is for fermentation of yellow mulberry tree.

The strain is characterized in that it is cultured in the presence of an external carbon source in addition to Hwangchu.

The strain is characterized in that it has acid resistance and biliary cholesterol.

The strain may be selected from the group consisting of leucine arylamidase, valine arylamidase, N-acetyl-β-glucosaminidase, β-galactosidase (β -glucosidase, -galactosidase,? -glucosidase and? -glucosidase. However, it is not limited to β-glucuronidase beta-glucuronidase < / RTI > enzyme.

Wherein the strain is inhibited from growth by any one or more antibiotics selected from the group consisting of penicillin-G, ampicillin, rifampicin, novobiocin and chloramphenicol, and at least one selected from the group consisting of lincomycin, polymycin B and vancomycin And is resistant to antibiotics.

The strain is characterized in that it has antibacterial activity against food poisoning bacteria.

Wherein the food poisoning bacterium is any one or more selected from the group consisting of Escherichia coli , Salmonella Typhimurium , Staphylococcus aureus , and Listeria monocytogenes .

In order to accomplish the above-mentioned other objects, the present invention provides a fermented product of Fusarium oxysporum (Fusarium oxysporum) having increased antioxidative and GABA contents obtained by fermentation by inoculation with Lactobacillus plantarum strain K118 (accession number KCCM 12015P) .

The whitish wood is characterized by being a whitened wood pulverized product or an extract of Hokutogi tree.

 The extract is an extract of water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The extract is a hot-water extract.

According to another aspect of the present invention, there is provided a method for preparing a fermented Yellowlily of Korea, which comprises fermenting a Yellowlake tree by inoculating Lactobacillus plantarum strain K118 (Deposit No. KCCM12015P) .

The whitish wood is characterized by being a whitened wood pulverized product or an extract of Hokutogi tree.

 The extract is an extract of water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The extract is a hot-water extract.

The extract has a total solid content of 0.5 to 5% by weight, a pH of 2 to 7, and a sugar concentration of 1 to 5 Brrix.

The extract is a mixture of leaves and stems of Echinochloa crus-galli at 8 to 10: 1 weight ratio.

Wherein the fermentation is performed at 25 to 50 DEG C for 10 to 96 hours.

The fermentation is characterized in that it is cultured in the presence of an external carbon source in addition to Hwasungi.

The novel Lactobacillus plantarum K118 strain according to the present invention can increase the antioxidant and GABA content in the extract of Wuchulia chinensis using the Wuchu chrysanthemum extract as a fermentation substrate. This fermented fermented yellowtail has the advantage that various physiological activities are improved because it contains high concentration of GABA.

In addition, the fermented product according to the present invention can be used as safe medicines, food additives or cosmetic raw materials since it induces apoptosis of normal cells or does not show cytotoxicity, and thus has excellent stability to human body.

The novel Lactobacillus plantarum K118 strain of the present invention is isolated from kimchi, and the fermented product of Wuchulia japonica prepared by using the Lactobacillus plantarum K118 strain is a composition containing GABA at a high concentration And can reduce problems due to toxicity and side effects.

Figure 1 is a phylogenic tree of Lactobacillus plantarum strain K118.
2 is a result of measuring the number of viable cells (a) and pH (b) for 24 hours at 34 ° C, 37 ° C and 40 ° C using an MRS liquid medium for Lactobacillus plantarum K118 strain .
Fig. 3 shows the result of measuring the bile resistance of Lactobacillus plantarum K118 strain. A total of three experiments were performed at this time, and these data are expressed as mean ± standard deviation; * p < 0.05
FIG. 4 is a graph showing the acid resistance of Lactobacillus plantarum K118 obtained by culturing the strain K118 in various concentrations of HCl solution (pH 2.0, 3.0, 4.0 and 6.4) for 3 hours, and then counting viable cells. A total of three experiments were performed at this time, and these data are expressed as mean ± standard deviation; * p < 0.05, ** p < 0.01, *** p < 0.001
5 is a graph showing the adhesion ability of Lactobacillus plantarum strain K118 to HT29 cells. At this time, a total of three experiments were carried out and the data were expressed as mean ± standard deviation; ** p < 0.01

Accordingly, the inventors of the present invention have made extensive efforts to develop a novel strain for the fermentation of Fusarium oxysporum, which greatly improves the functionality and functionality of the Fusarium oxysporum extract. As a result, it has been found that the strain for fermentation of Fusarium oxysaccharus according to the present invention, And a method for producing the same, and have completed the present invention.

One aspect of the present invention relates to a Lactobacillus plantarum K118 strain (Deposit No. KCCM12015P) having antioxidant and GABA production promoting activity.

Lactobacillus strains belonging to the genus Lactobacillus , commonly referred to as bacilli, are part of lactic acid strains. Lactic acid bacteria are bacteria that produce lactic acid by decomposing saccharides such as glucose, and are often referred to as lactic acid bacteria.

The Lactobacillus plantarum K118 strain is isolated from kimchi and has a 16S rDNA sequence as shown in SEQ ID NO: 1, which is a 5 'to 3' direction sequence.

The Lactobacillus plantarum K118 strain is deposited at KCCM12015P.

The Lactobacillus plantarum K118 strain may have 99% or more homology with Lactobacillus plantarum .

The Lactobacillus plantarum strain K118 is fermented by inoculation on a wild ginseng tree to thereby produce a fermentation product containing a high concentration of GABA. In other words, the K118 strain is a strain having antioxidant activity and GABA production promoting activity through yellowtail millet fermentation, and it is possible to produce a fermented product of Ganoderma lucidum containing GABA at a higher concentration than Lactobacillus isolated from other methods or Lactobacillus plantarum In addition, the sensory and functional properties of the Fusarium oxysporum fungus can be greatly improved.

The above-mentioned strain is a lactic acid bacterium isolated from kimchi, and the kimchi may be one prepared by mixing with salt-seasoned vegetables and seasoning, but not particularly limited thereto. More preferably, It is preferable to use a product which has been aged for 10 days or more at a temperature.

The Lactobacillus plantarum strain K118 can be cultivated in the absence of an external carbon source in addition to Hwangchilmyeong. That is, even if it is used as a sole nutritional source, it grows well and has excellent antioxidant and GABA production activity. Therefore, It is preferable to contain Horticultura as a sole nutrient source to produce the fermented Horticulture.

In addition, it is advantageous in that it does not require additional external carbon sources and therefore does not require additional processing and costs for the fermentation substrate.

The strain may be fermented at 25 to 50 캜 for 10 to 96 hours, preferably at 34 to 45 캜 for 20 to 30 hours. And the pH of the finally produced fermentation product is 4.5 or less. The optimum fermentation conditions are not particularly limited to the above-mentioned conditions, but when the fermentation temperature is lower than 34 ° C, the growth rate is significantly lowered, and there is a problem that growth is not achieved at 15 ° C. However, since it is excellent in acid resistance, it can be grown under various pH conditions, so that the fermentation conditions are simple and the cost can be reduced.

Through the strain K118, the Fusarium oxysulfuric acid fermented product prepared from the Fusarium oxysporum fungi as a fermentation substrate has a pH of 4.5 or less, preferably pH 4-4.5, and an antioxidative activity by radical scavenging activity of 92% or more. , The GABA content was significantly higher than 1600 ㎍ / ㎖ in relation to the total weight of the perilla seedlings.

Therefore, if the pH, antioxidant ability and GABA content of the fermented Yellowlard seedlings are in the range of the upper limit value to the lower limit value, it is possible to obtain a fermented ferulic acid fermented product having significantly improved functionality compared to the extract of Yellowlake tree from other strains belonging to the genus Lactobacillus . Compared to Lactobacillus plantarum, which is a homologous strain, the growth activity was excellent and the GABA production activity was high even in the extreme conditions of the only nutrient source, and the sensory properties of the final fermented product 1.5 to 2 times better.

The Lactobacillus plantarum K118 strain is highly resistant to acid because it has bile resistance and stably maintains GABA production promoting activity even at pH 2, and its long-acting ability (adhesion ability) Have more than 5% higher values than the other strains of the parent strain.

The Lactobacillus plantarum strain K118 may also be selected from the group consisting of leucine arylamidase, valine arylamidase and N-acetyl-β-glucanamine -glucosaminidase, and then the enzymatic activity of β-galactosidase, α-glucosidase, and β-glucosidase Respectively. However, it does not have the activity of β-glucuronidase. Therefore, the strain K118 according to the present invention was found to be superior to the living organism.

The Lactobacillus plantarum K118 strain is resistant to penicillin-G, ampicillin, rifampicin, novobiocin and chloramphenicol, and is resistant to lincomycin, polymycin B and vancomycin antibiotics .

The Lactobacillus plantarum strain K118 has a high antimicrobial activity against food poisoning bacteria, specifically Escherichia coli , Salmonella Typhimurium , Staphylococcus aureus Staphylococcus aureus , and Listeria monocytogenes at least 18% and at most 50%.

Another aspect of the present invention relates to a Fagus crenata fermented product having an increased content of GABA obtained by inoculating L. bovis L. with Lactobacillus plantarum strain K118 (accession number KCCM 12015P).

The above-described Yellow Fungus tree fermented product is obtained by inoculating Lactobacillus plantarum K118 strain of the present invention into a perennial tree.

The perennial plant may be an extract of Echinochloa crataegus or an extract of Echinochloa crus-galli.

The Hwigaechuki extract includes not only the crude extract obtained by treating the extraction raw material with the extraction solvent but also the crude extract extract. For example, the extract may be prepared in powder form by an additional process such as vacuum distillation and lyophilization or spray drying. The extract also includes fractions obtained by further fractionating the crude extract.

The extract is an extract of water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, and the extraction method is not particularly limited.

When water is used as the solvent, hot water extraction is preferred. For example, at a temperature of 80 to 105 DEG C, preferably 90 to 100 DEG C for 0.5 to 24 hours, preferably 1 to 6 hours. Even if the hot water is not used as a solvent, the components of Hwangryu can be extracted from the diluted solution obtained by diluting the Hwangchu tree powder with cold water or room temperature water. When extracting the woody ingredients using cold water or room temperature water, it may be extracted separately from the fermentation, or may be extracted in the process of fermentation by inoculation with Lactobacillus plantarum K118 strain You may.

An extract of an aqueous solution of an alcohol having 1 to 4 carbon atoms can be used. For example, extraction is carried out with an alcohol aqueous solution such as ethanol, methanol or isopropanol, preferably 20 to 80% by weight aqueous alcohol solution, more preferably 50 to 70% by weight aqueous alcohol solution. When alcohol extracts are used, before inoculation of Lactobacillus plantarum K118 strain, first concentrate the alcohol or alcohol concentrate or alcohol extract which has lowered the alcohol content by evaporating the alcohol of the alcohol extract, and dry it after dissolving in water. In addition, in the present invention, the extract of Hwangchu-myeon is broadly composed of a diluted solution of Hwangchu-myeon powder in water.

The hwangchil wood lactic acid nutrient source, such as Lactobacillus Planta room (Lactobacillus plantarum) Lactobacillus Planta room (Lactobacillus plantarum) in order to promote the growth of the K118 strain protein source, a carbohydrate source, vitamins or minerals prior to inoculation the K118 strain Further mixing is possible. The nutrient source of the strain may be a commercially available medium or may be supplemented with only necessary nutrients.

However, the Lactobacillus plantarum strain K118 according to the present invention is active even in the presence of an external carbon source, that is, the only nutrient source, in addition to the wild ginseng tree, and the GABA production activity is excellent, so that a high concentration of GABA is contained It is preferable to contain Hwangchujang as a sole nutrient source.

In addition, by making the Hwangchu tree the sole nutritional source, it is possible to obtain an effect of reducing the additional process and cost for the fermentation substrate, since no additional external carbon sources are added.

In addition, the mixture of the above Eustachia japonica extract or a mixture of the Eustachian japonica extract and the above-mentioned strain nutrient can be sterilized by heating before the inoculation of Lactobacillus plantarum K118 strain.

The solid content of the extract is not particularly limited, but is 0.5 to 1% by weight when there is no other concentration step after extraction. The extract may be used directly, or may be concentrated, or may be concentrated or dried and then diluted . When the extract is concentrated and used, the solid content may be 1 to 10% by weight.

It is preferable that the extract is used in an amount of 0.5 to 5% by weight, a pH of 2 to 7, and a sugar concentration of 1 to 5 Brix to increase the growth activity and GABA production promoting activity of the strain. Most preferred.

The fermentation conditions are the same as those of general lactic acid bacteria fermentation, so that the fermentation conditions are not particularly limited, but they may be performed at 25 to 50 ° C for 10 to 96 hours, preferably 34 to 45 ° C for 20 to 30 hours.

The Fusarium oxysulfuric acid fermented product may be a fermented product containing cells or a fermented product from which cells have been removed. The fermented product from which the cells have been removed may be sterilized to include dead cells, or may be a filtrate or a centrifuged supernatant from which cells have been removed by filtration or centrifugation. Since the components contained in the extract of the present invention are bioactivated or produced by the Lactobacillus plantarum K118 strain, which is an easily available component in the living body, the various physiological activity effects Respectively. This is remarkably higher than the effect obtained by adding the simple lactic acid bacteria themselves. The fermented product can be used as a liquid fermented product itself, or may be dried and pulverized.

The GABA content of the perilla seedlings was more than 1,600 ㎍ / ㎖ of the total weight of the perilla seedlings, which was 1.5 times more than the GABA content of the perennial seedlings.

Yet another aspect of the present invention relates to a method for producing fermented Yellowlings, which comprises fermenting Yellowlake wood by inoculating Lactobacillus plantarum K118 strain (Deposit No. KCCM12015P).

The Lactobacillus plantarum strain K118 of the present invention comprises a crushed cell wall fraction of the strain, a live cell, a dead cell or a dried cell, wherein the culture liquid is a culture medium itself obtained by culturing the strain in a liquid medium, Filtration or centrifugation to remove the strain (centrifuged supernatant), and the like.

The culture solution may be dried (e.g., lyophilized) and then powdered.

The perennial plant may be an extract of Echinochloa crataegus or an extract of Echinochloa crus-galli.

The Hwigaechuki extract includes not only the crude extract obtained by treating the extraction raw material with the extraction solvent but also the crude extract extract. For example, the extract may be prepared in powder form by an additional process such as vacuum distillation and lyophilization or spray drying. The extract also includes fractions obtained by further fractionating the crude extract.

The extract is an extract of water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, and the extraction method is not particularly limited.

When water is used as the solvent, hot water extraction is preferred. For example, at a temperature of 80 to 105 DEG C, preferably 90 to 100 DEG C for 0.5 to 24 hours, preferably 1 to 6 hours. Even if the hot water is not used as a solvent, the components of Hwangryu can be extracted from the diluted solution obtained by diluting the Hwangchu tree powder with cold water or room temperature water. When extracting the woody ingredients using cold water or room temperature water, it may be extracted separately from the fermentation, or may be extracted in the process of fermentation by inoculation with Lactobacillus plantarum K118 strain You may.

An extract of an aqueous solution of an alcohol having 1 to 4 carbon atoms can be used. For example, extraction is carried out with an alcohol aqueous solution such as ethanol, methanol or isopropanol, preferably 20 to 80% by weight aqueous alcohol solution, more preferably 50 to 70% by weight aqueous alcohol solution. When alcohol extracts are used, before inoculation of Lactobacillus plantarum K118 strain, first concentrate the alcohol or alcohol concentrate or alcohol extract which has lowered the alcohol content by evaporating the alcohol of the alcohol extract, and dry it after dissolving in water. In addition, in the present invention, the extract of Hwangchu-myeon is broadly composed of a diluted solution of Hwangchu-myeon powder in water.

However, when the degree of mixture of impurities and the degree of expression of flavor are considered in the extract, the most preferred method is to use the extracted water as a fermentation substrate.

In order to enhance the growth of Lactobacillus plantarum K118 strain before the inoculation of Lactobacillus plantarum K118 strain to the extract, the lactic acid bacteria nutrient source such as protein source, carbohydrate source, Can be further mixed. The nutrient source of the strain may be a commercially available medium or may be supplemented with only necessary nutrients.

However, the Lactobacillus plantarum strain K118 according to the present invention is active even in the absence of an external carbon source, that is, the only nutrient source, and has excellent antioxidative and GABA production activity, so that a high concentration of GABA It is preferable that the nutrient source is contained as a sole nutrient source.

In addition, by making the Hwangchu tree the sole nutritional source, it is possible to obtain an effect of reducing the additional process and cost for the fermentation substrate, since no additional external carbon sources are added.

In addition, the mixture of the above Eustachia japonica extract or a mixture of the Eustachian japonica extract and the above-mentioned strain nutrient can be sterilized by heating before the inoculation of Lactobacillus plantarum K118 strain.

The solid content of the extract is not particularly limited, but is 0.5 to 1% by weight when there is no other concentration step after extraction. The extract may be used directly, or may be concentrated, or may be concentrated or dried and then diluted . When the extract is concentrated and used, the solid content may be 1 to 10% by weight.

The components of the extract are selected from the group consisting of leaves, roots, roots, and fruits. However, the components of the extract are not required in the fermentation process of the extract. In order to prevent mixing, it is most preferable to use a mixture of the leaf and stem of the Hwigae-gil tree as a raw material.

Preferably, in the present invention, the extract is prepared from the mixture of leaves and stems of 8% to 10: 1 by weight, and when the leaves are out of the above range or other parts such as roots or fruit are included, Since the composition of the extract varies, the problem of lowering the GABA content in the fermented product may arise.

It is preferable that the extract is used in an amount of 0.5 to 5% by weight, a pH of 2 to 7, and a sugar concentration of 1 to 5 Brix to increase the growth activity and GABA production promoting activity of the strain. Most preferred.

In addition, the method may further include the step of inoculating the strain into a nutrient medium (peptone, yeast extract, glucose / distilled water) before inoculating the extract, and pre-culturing the strain at 20 to 30 DEG C for 18 to 28 hours, And then inoculating the pre-culture solution to the above-mentioned Hokutogi extract.

Preferably, the Lactobacillus plantarum K118 strain is added in an amount of 1 × 10 ⁴ to 1 × 10 ⁸ CFU per g of the extract, and more preferably the Lactobacillus planta / Room K118 The strain may be mixed with 1 x 10 ⁷ to 1 x 10 ⁸ CFU.

The fermentation conditions are the same as those of general lactic acid bacteria fermentation, so that the fermentation conditions are not particularly limited, but they may be performed at 25 to 50 ° C for 10 to 96 hours, preferably 34 to 45 ° C for 20 to 30 hours.

The Fusarium oxysulfuric acid fermented product may be a fermented product containing cells or a fermented product from which cells have been removed. The fermented product from which the cells have been removed may be sterilized to include dead cells, or may be a filtrate or a centrifuged supernatant from which cells have been removed by filtration or centrifugation. Since the components contained in the extract of the present invention are bioactivated or produced by the Lactobacillus plantarum K118 strain, which is an easily available component in the living body, the various physiological activity effects Respectively. This is remarkably higher than the effect obtained by adding the simple lactic acid bacteria themselves. The fermented product can be used as a liquid fermented product itself, or may be dried and pulverized.

The GABA content of the perennial fermented product produced by the above-mentioned process is 1,600 g / ml or more when compared with the total weight of the perilla seedlings. When GABA content is 1.5 times Or more.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Experimental Method

<Preparation of Yellowtail Tree Concentrate>

10 g of water was added to 1 g of the total solid weight of the mixture, and the mixture was heated at 100 ° C for 3 hours to obtain a hot-water extract. The total solid content contained in the hot water extract was 0.74% by weight and the pH was 5.1.

The concentrate was concentrated by using a vacuum decompressor (rotavator R-124, Buchi, Switzerland) at 100 rpm and 50 ° C to prepare an extract solution suitable for fermentation. The concentrate was 4.0 ㅀ Brix, pH 5.0 , And the total solid content contained in the concentrate was 4.2 wt%.

Experimental Example 1. Strain Collection and Separation from Raw Materials (Primary)

 1) Collecting and isolating strains in traditional fermented products

(Lactobacillus MRS agar, BD Difco, Sparks, MD, USA), which is a rich medium, is used as a nutrient-rich liquid medium for collecting and isolating strains from traditional fermentation products such as kimchi, And the ratio of the medium described in the manual was maintained.

Bromocresol purple and sodium azide were further added to the MRS solid medium, and the conventional fermented product was rubbed on each of the MRS solid medium, followed by incubation at 37 ° C. for 48 hours. Then, each of the yellow colonies Colonies were selected and purely isolated.

For pure isolation, the colonies were streaked on MRS solid medium, and the colonies were inoculated on a tryptic soy agar slant and incubated at 37 ° C for 18 hours. Then, Respectively. As a result, 170 kinds of conventional fermentation products were firstly selected.

2) Strain collection and separation in crude oil

Except for traditional fermented products, crude oil was used at each of the farms supported by the crude oil inspection centers (Gyeonggi and Gangwon provinces), Jeonbuk Livestock Research Institute, Gyeongbuk Livestock Hygiene Laboratory, and Cheju Provincial Livestock Promotion Agency in central Seoul, northern and western districts. 1.1) and the process of collecting and isolating strains were performed in the same manner. As a result, 100 kinds of crude oil isolates were firstly selected.

3) Collecting and isolating strains in feces

Experimental example 1.1) and strain collection and isolation were carried out in the same manner, except that feces instead of conventional fermentation products were used after 3 hours of bowel movement. As a result, 120 kinds of fecal isolates were firstly selected.

Experimental Example 2 Selection of Strains Growing in the Yellow Forest Tree Concentrate (Secondary)

In order to isolate strains with phagocytosis, acid production ability was required. Therefore, each strain was cultivated in an extract of Hwangchulchu and then a strain having a pH value of 4.5 or less was secondly selected.

The primary cultures (10 ⁹ CFU / ml) inoculated with MRS liquid medium and incubated at 37 ° C for 18 hours were inoculated 1% to the medium concentration and incubated at 37 ° C for 24 hours. A total of 13 kimchi isolates, 11 crude isolates, and 5 fecal isolates were screened by preparing a fermented tree and measuring the pH of the isolate. The strains selected in Table 1 are indicated as raw materials.

Raw material Strain name Kimchi isolate
(Traditional fermentation products) K4 K99 K112 K118 K266 KL69 KL71 KL72 KC3 KC13 KC19 KC20 KC23 Crude oil isolate Y48 Y51 Y106 Y122 Y140 Y142 Y158 Y161 R2 R24 R32 Fecal isolate BB7 BB21 BB32 JU4 JU9

Experimental Example 3. Selection of a strain having GABA production promoting activity

In the previous experiment, the strain was firstly isolated from the raw material, and the strain was inoculated with 1% of the extract in a concentration of yellowtail. After culturing for 24 hours, the pH was measured to secondarily select strains having a pH of 4.0 or less, The results were compared with those of the DHHP radical scavenging ability and the GABA productivity assay described in 1) and 2) below. As a result, the most active K118 strain was finally selected (Table 2).

At this time, the control group was a sample obtained by measuring the concentration of Hwangchil tree concentrate before the strain was inoculated.

1) Analysis method of DPPH radical scavenging ability

The sample was reacted with 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) to measure the radical scavenging activity of the sample. The DPPH radical scavenging activity was measured by using a spectrophotometer at 515 nm.

2) Analysis method of gamma-aminobutyric acid (GABA) content

Quantitative analysis of GABA was carried out by adjusting Zhang and Bown 's method to 96 well plate. 0.1 g of the sample and 400 μl of methanol were added to an eppendorf tube, mixed thoroughly, and thoroughly dried for about 30 minutes using a water bath preheated to 75 ° C. Then, 1 ml of 70 mM LaCl _{3 was} added, centrifuged at 13,000 ㅧ g for 5 minutes, 700 μl of the supernatant was taken, and the precipitate was discarded. 700 μl of the supernatant is taken up in an Eppendorf tube, and then 160 μl of 0.1 M KOH is added thereto. The mixture is stirred for 3-5 minutes, and further centrifuged at 13,000 × g for 5 minutes to obtain a supernatant. This was diluted and used for GABA content measurement. GABA content was determined by using an enzyme method using GABase and the amount of NADPH produced was measured at 340 nm using an ELISA reader.

Raw material Strain DPPH (%) Relative value of DPPH versus control GABA generation
(ug / mL) Relative amount of GABA relative to control Control group 86.20 ± 0.20 100 966.11 + - 35.85 100 Kimchi isolate K4 88.90 ± 0.24 103.1 1244.23 + - 35.17 128.8 K99 91.20 ± 0.18 105.8 1244.23 ± 27.79 128.8 K112 89.48 ± 0.17 103.8 1127.12 + - 33.80 116.7 K118 90.55 + 0.24 105.0 1624.82 + - 33.80 168.2 K266 91.36 ± 0.14 106.0 1156.4 ± 36.90 119.7 KL69 87.32 + - 0.16 101.3 995.38 ± 25.28 103.0 KL71 87.70 ± 0.60 101.7 1010.02 + - 25.28 104.5 KL72 89.94 + 0.33 104.3 1214.95 ± 23.79 125.8 KC3 89.98 + 0.48 103.2 1097.85 ± 16.90 113.6 KC13 86.82 ± 0.45 100.7 936.83 + - 25.28 97.0 KC19 90.21 + - 0.41 104.5 995.38 +/- 13.79 103.1 KC20 86.90 ± 0.58 100.8 980.74 + - 66.90 101.5 KC23 88.40 +/- 0.39 102.6 1010.02 + - 25.28 104.5 Crude oil isolate Y48 88.47 ± 0.21 102.6 984.34 ± 22.28 101.9 Y51 87.40 ± 0.12 101.4 975.12 + - 17.45 100.9 Y106 88.85 ± 0.06 103.1 965.14 + 32.27 99.9 Y122 86.40 ± 0.21 100.2 954.84 + - 42.47 98.8 Y140 91.14 + 0.14 106.0 975.28 + 15.24 100.9 Y142 87.21 + - 0.45 101.2 992.14 + - 21.14 102.7 Y158 89.42 + 0.15 103.7 981.32 ± 30.14 101.6 Y161 86.78 ± 0.17 100.7 974.26 ± 14.28 100.8 R2 88.20 ± 0.11 102.3 962.74 + - 42.15 99.7 R24 87.08 + - 0.31 101.0 1012.02 + - 22.18 104.8 R32 88.12 + - 0.11 102.2 960.45 + - 39.96 99.4 Fecal isolate BB7 86.40 ± 0.21 100.2 978.40 ± 32.85 101.3 BB21 90.72 + 0.18 105.2 964.34 ± 15.73 99.8 BB32 88.30 ± 0.24 102.4 987.12 + - 24.80 102.2 JU4 87.21 + - 0.32 101.2 977.47 ± 21.52 101.2 JU9 90.02 + - 0.15 104.4 991.21 + 33.47 102.6

As shown in Table 2, it was confirmed that K118 strain had the best DPPH scavenging activity and GABA production activity.

Even though the same kimchi was separated into raw materials, K99 and K266 showed low DPPH scavenging ability and showed a difference of 400 ~ 700 ㎍ per 1 ㎖ of GABA production activity even if they were close to 90%.

This difference is only a difference of 1.5 to 2 times in the measured value based on ㎖, but it is a very significant difference in the process unit.

Experimental Example 4. Evaluation of functional (physiological) activity of strain K118

The final selected K118 strain is inoculated 1% in the Hwanyeongkuk concentrate and incubated at 37 ℃ for 24 hours to obtain a yellowtail tree fermented product. The fermented products were evaluated for functionality (bioactivity - antioxidant / cognitive function) before fermentation.

Among the isolates isolated from K118 isolates, the functionalities of K99 and K266 strains with excellent GABA production were also evaluated and compared with those of K118 (Table 3).

1) Analysis method of active oxygen (OH) scavenging activity

In order to measure the scavenging activity of the active oxygen (OH) in the yellowtail concentrate, 0.2 ml of the sample to be measured, 10 mM of FeSO _{4 ·} 7H ₂ O, 10 mM of EDTA, 2-deoxyribose (10 mM) 1.8 ml of a phosphate buffer solution (pH 7.4) and ₂ ml of H ₂ O ₂ (10 mM) were added thereto, and the cells were incubated at 37 ° C in an incubator. After completion of the incubation, 1 ml of TCA solution (2.8%) was added to the culture solution to stop the reaction. 1 ml of 1.0% TBA solution was added and the mixture was heated at 100 ° C for 10 minutes. After rapid cooling, Were measured.

2) Superoxide dismutase (SOD) activity assay method

3 ml of Tris-HCl buffer (50 mM Tris + 10 mM EDTA, pH 8.5) and 0.2 ml of 7.2 mM pyrogallol were added to 0.2 ml of the sample to be measured, and the mixture was incubated at 25 ° C for 10 minutes. Ml was added to stop the reaction, and the amount of oxidized pyrogallic acid present in the solution was measured at 420 nm. At this time, the SOD activity was expressed as% of absorbance reduction by comparing the addition of the sample with the addition of the sample.

pH SOD-like activity (%) H OH (%) Control group 4.5 16.86 + - 7.42 82.60 +/- 1.65 K99 3.81 17.27 + - 4.15 83.54 ± 1.28 K118 3.90 24.61 + - 2.63 87.71 + - 0.54 K266 3.85 18.64 + - 4.62 82.75 + - 0.55

As shown in Table 3, the finally selected K118 strain was found to be more functional than the other strains.

Experimental Example 5. Identification of the finally selected K118 strain

1) Physiological and biochemical characterization

First, in order to determine the genus and species of the finally selected strain K118, gran staining, microscopic observation, sporulation, aerobic and anaerobic growth, catalase production, growth at 15 ° C and 45 ° C, and ammonia production from arginine. The results of the sugar fermentation test were also shown in Table 4. The results of the fermentation experiments per API are shown in Table 5.

strains K118 Gram stain + Cell morphology rod Spore formation - Motility - Aerobic growth + Anaerobic growth + Catalase - Growth at 15 ℃ - Growth at 45 ° C + Gas from glucose - Ammonia from arginine -

As shown in Table 4, the K118 strain of the present invention was confirmed to be a Gram-positive group, and it was confirmed that the K118 strain was a rod-shaped bacterium. Also, aerobic and anaerobic growth were possible, and it was not possible to grow at 15 ℃, and it was confirmed that it could grow at 45 ℃.

Strains K118 K118 K118 Control - Inositol - D-Melezitose + Glycerol - D-Mannitol + D-Raffinose + Erythritol - D-Sorbitol + Amidon - D-Arabinose - Methyl-aD-Mannopyranoside - Glycogen - L-Arabinose + Methyl-aD-Glucopyranoside - Xylitol - D-Ribose + N-AcetylGlucosamine ± Gentiobiose ± D-Xylose - Amygdalin + D-Turanose + L-Xylose - Arbutin + D-Lyxose - D-Adonitol - Esculin ferric citrate + D-Tagatose - Methyl-β-D-Xylopyranoside - Salicin ± D-Fucose - D-Galactose + D-Celiobiose + L-Fucose - D-Glucose + D-Maltose + D-arabitol ± D-Fructose + D-Lactose + L-Arabitol - D-Mannose + D-Melibiose + potassium Gluconate ± L-Sorbose - D-Saccharose + potassium 2-KetoGluconate - L-Rhamnose ± D-Trehalose + potassium 5-KetoGluconate - Dulcitol - Inulin -

2) 16S rDNA sequencing

For the identification of the finally selected K118 strain, 16S rDNA sequencing was performed. For 16S rDNA sequencing, the selected K118 strain was cultured in MRS broth and DNA was extracted using genomic DNA extraction kit (iNtRON Biotechnology, Korea). The extracted DNA was confirmed using 1% agarose gel. The 16S rRNA gene was amplified by PCR using a universal primer.

The PCR conditions were 30 cycles of denaturation at 95 ° C for 1 min, annealing at 45 ° C for 1 min, and extension at 72 ° C for 1 min 30 sec.

The obtained PCR products were purified using a purification kit (iNtRON Biotechnology, Korea) for sequencing. The nucleotide sequence was determined using ABI PRISM 3799 DNA analyzer (Perkin Elmer, U.S.A), and the K118 strain was identified by comparing it with the GenBank database using NCBI's BLAST.

As a result of analysis of 16S rDNA sequence, K118 strain, which was finally isolated from kimchi, showed 99% homology with Lactobacillus plantarum , and it showed physiological characteristics such as antioxidant ability, It was confirmed that the active function was excellent. From the above results, strain K118 was named Lactobacillus plantarum K118 strain.

In the present invention, it was confirmed that 16S rDNA of Lactobacillus plantarum strain K118 was encoded by the nucleotide sequence of SEQ ID NO: 1, deposited with the Korean Microorganism Conservation Center (KCCM), and received KCCM12015P .

A phylogenic tree of the Lactobacillus plantarum K118 strain is shown in FIG.

Name of depository: Korea Microorganism Conservation Center (overseas)

Accession number: KCCM12015P

Checked on: 20170414

Experimental Example 6: Growth test of strain K118

Growth of Lactobacillus plantarum K118 strain was measured by measuring viable cell count and pH at various temperature conditions. The viable cell counts were obtained by inoculating 1 ml of lactic acid bacteria into 150 ml of MRS liquid medium, 1 ml of each pipette, and culturing at 34 ° C, 37 ° C, and 40 ° C for 3 hours at intervals of 24 hours. Each sample was diluted with 0.1% Plate count agar was poured on a plate, hardened, incubated at 35 ° C for 48 hours and counted by pH and temperature. The measurement results are shown in Figs.

2 is a result of measuring the number of viable cells (a) and pH (b) for 24 hours at 34 ° C, 37 ° C and 40 ° C using an MRS liquid medium for Lactobacillus plantarum K118 strain .

As can be seen from FIGS. 2A and 2B, the optimum temperature for growth of Lactobacillus plantarum strain K118 is 34 to 45 ° C, and it takes 13 hours to reach the optimum pH condition of the fermentation product, pH 4.3 to 4.4. Respectively.

Experimental Example 7. Bile tolerance test of strain K118

The bile tolerance test was performed according to the method of Gilliland and Walker (Gilliland & Walker, J. Dairy Sci., 73, 905, 1990), and the measurement results are shown in FIG.

Fig. 3 shows the result of measuring the bile resistance of Lactobacillus plantarum K118 strain. A total of three experiments were performed at this time, and these data are expressed as mean ± standard deviation; * p < 0.05

As can be seen in FIG. 3, after the 7-hour incubation, there was a slight difference between without oxgall and with oxgall, but it was basically insensitive to bile .

Experimental Example 7. Acid resistance test of strain K118

The pH tolerance test was carried out according to the method of Clark et al. (Clark et al., Cultured Dairy Products J., 28 (4), 11, 1993).

FIG. 4 is a graph showing the acid resistance of Lactobacillus plantarum K118 obtained by culturing the strain K118 in various concentrations of HCl solution (pH 2.0, 3.0, 4.0 and 6.4) for 3 hours, and then counting viable cells. A total of three experiments were performed at this time, and these data are expressed as mean ± standard deviation; * p < 0.05, ** p < 0.01, *** p < 0.001

As shown in FIG. 4, the resistance of Lactobacillus plantarum K118 after 3 hours in a hydrochloric acid solution was compared. As a result, the strain of the present invention showed almost no effect on growth even at pH 2, which is stronger than pH 6.4 It was confirmed that it has very strong acid resistance.

Experimental Example 8. Adhesion test of K118 strain to colon epithelial cells (HT cell)

The following experiment was conducted to determine the ability of the finally selected Lactobacillus plantarum strain K118 to coat the colon epithelial cells (HT29-MTX cell).

HT29-MTX cells were cultured in Dulbecco's modified Eagle's medium (DMEM) medium supplemented with 10% (v / v) heat-inactivated fetal bovine serum and antibiotic mixture. At this time, the antibiotics were finally made to be 50 mg / ml penicillin, 50 mg / ml streptomycin, 50 mg / ml gentamicin and 1.25 mg / ml amphotericin B, respectively.

HT29-MTX cells were seeded in a 30 mm culture dish to 1 x 10 ⁵ cells / ml and cultured at 37 ° C for 14 days in a 5% CO ₂ incubator (about 1 × 10 ⁷ cells / ml). All experiments were repeated three times. Lactobacillus plantarum K118 culture broth was collected by centrifugation, washed twice with phosphate buffer, and suspended in DMEM without antibiotics so that the bacterial: intestinal cells were 10: 1. To remove antibiotics, HT29-MTX monolayers were washed twice with Dulbecco's PBS and Lactobacillus plantarum strain K118 was added. After incubation at 37 ° C in a CO ₂ incubator for 1 hour, the supernatant was discarded and each well was gently washed three times with Dulbecco's PBS buffer to remove unattached bacteria. Cells were fixed with 4% fixative (35% formaldehyde 100 ml, Na ₂ HPO ₄ 16 g, NaH ₂ PO ₄ H ₂ O 4 g with distilled water to 1 L) and Lactobacillus plantarum K118 The strain was stained with Gram and observed with an optical microscope. Out to count the number of Lactobacillus Planta room (Lactobacillus plantarum) K118 strain attached to the cell before the cell and Lactobacillus Planta room (Lactobacillus plantarum) K118 strain by 0.1% TritonX-100 removed from the culture dish, and this serially diluted 100 [mu] l each were inoculated into the MRS solid medium by the spreading method. CFU was then counted for 2 days in anaerobic conditions. All of the experiments were repeated three times and the data were analyzed using SPSS 11.0 software for Windows (SPSS Inc., Chicago, IL, USA) (Gonzalez de los Reyes-Gavilan, Clara et al., 2011. Adhesion of even-adapted Bifidobacterium Kainulainena V., J. Reunanena, K. Hiippalaa, S. Guglielmettib, and S. Gullielmettib, J. Clin. Endocrinol. S. Vesterlundc, A. Palvaa and R. Satokaria, 2013. BopA does not have a major role in the adhesion of Bifidobacterium bifidum to intestinal epithelial cells, extracellular matrix proteins, and mucus. Appl. Environ Microbiol 79: 6989-6997 ).

5 is a graph showing the adhesion ability of Lactobacillus plantarum strain K118 to HT29 cells. At this time, a total of three experiments were carried out and the data were expressed as mean ± standard deviation; ** p < 0.01

As shown in FIG. 5, the Lactobacillus plantarum strain K118 of the present invention was found to be well adhered to the HT29-MTX cell line.

Example 9. Enzyme activity test

The enzyme activity test was performed by diluting a Lactobacillus plantarum K118 strain cultured in an MRS liquid medium at 37 ° C. for 18 hours with physiological saline to prepare a sample of 10 ⁵ to 10 ⁶ cfu / (API bioMerieux, France) for 5 hours at 37 ° C, followed by enzymatic reaction. The enzyme activity was expressed as 0-5 in comparison with the standard color trademark, and the results are summarized in Table 6 below. Table 6 below shows the results of comparing enzyme activities of Lactobacillus plantarum strain K118.

Enzyme K118 Alkaline phosphatase 0 Esterase (C4) 0 Esterase Lipase (C8) 0 Lipase (C14) 0 Leucine arylamidase 5 Valine arylamidase 5 Cystinearylamidase 2 Trypsin 0 alpha-chymotrypsin 0 Acid phosphatase 3 Naphtol-AS-BI-phosphohydrolase 2 alpha -galactosidase 0 β-galactosidase 4 β-glucuronidase 0 α-glucosidase 4 β-glucosidase 4 N-acetyl-β-glucosaminidase 5 alpha-mannosidase 0 alpha-fucosidase 0

*: Standard color is specified in steps from 0 to 5, 0 is negative and 5 is maximum intensity. 1 shows 5 nano moles, 2 has 10 nano moles, 3 has 20 nano moles, 4 has 30 nano moles, and 5 has an enzyme activity of 40 nano moles or more.

As can be seen in Table 6 above, Lactobacillus plantarum K118 was found to inhibit the production of leucine arylamidase, valine arylamidase and N-acetyl-beta -glucosamine (N-acetyl-β-glucosaminidase).

The enzyme activity was then found to be high in β-galactosidase, α-glucosidase and β-glucosidase. In the case of β-glucuronidase, a carcinogenic enzyme that converts benzopyrene into a carcinogenic substance, it has been shown that it has no enzyme activity and thus has excellent safety.

Experimental Example 10. Antibiotic resistance test of strain K118

The antibiotic susceptibility test of Lactobacillus plantarum strain K118 was performed by the diffusion method of the National Committee for Clinical Laboratory Standards (NCCLS). Antibiotic discs (BBL Sensi-disc, Becton Dickinson Co., USA) used were amikacin (30 μg, AN), gentamicin (10 μg, GM), kanamycin (10 μg, AM), ampicillin (10 μg, S), neomycin (30 μg, N), streptomycin , 10 μg, AM), bacitracin (20 μg, AM), rifampicin (10 μg, AM), Novobiocin (10 μg, AM), lincomycin ), Polymyxin B (2 ㎍, L), chloramphenicol (2 ㎍, L) and vancomycin (5 ㎍, L) The final selected strain K118 was inoculated into Mueller Hinton Broth (MErck, Germany), incubated at 37 ° C for 5 hours, diluted with 0.5 MacFarland (BioMérieux), and spread on Muller-Hinton agar (Difco) After drying, 15 antibiotic discs were placed and incubated at 37 ° C for 1 day. The size of inhibitory ring for each antibiotic was measured and tolerance was determined by CLSI guideline.

R has antibiotic resistance (inhibitory ring size: 0 ㎜), IS has intermediate susceptibility (inhibitory ring size: 1-5 ㎜); S indicates susceptibility (inhibition ring size:> 5 mm).

Anti-microbial agents Antibiotic resistance K118 Aminoglycosides Amikacin IS (3 mm) Gentamycin IS (4 mm) Kanamycin IS (2 mm) Neomycin IS (4 mm) Streptomycin IS (3 mm) beta-lactams Penicillin-G S (6 mm) Oxacillin IS (2 mm) Ampicillin S (13 mm) Gram-positive spectrum Bacitracin IS (5 mm) Rifampicin S (6 mm) Novobiocin S (7 mM) Lincomycin R (0 mM) Gram-negative spectrum Polymyxin B R (0 mM) Broad spectrum Chloramphenicol S (9 mm) Vancomycin R (0 mM)

As shown in Table 7, the strain K118 according to the present invention was found to inhibit the growth of penicillin-G, ampicillin, rifampicin, novobiocin and chloramphenicol as antibiotics susceptibility, and the inhibition of growth of lincomycin, polymacin B, vancomycin And antibiotics of the family.

Experimental Example 11. Antibacterial Test of K118 Strain

The antimicrobial activity test was carried out according to Gilliland & Speck, J. Food Prot., 40 (12), 820, 1977. The measurement results are shown in Table 8 below. Table 8 below shows the results of measurement of antimicrobial activity of Lactobacillus plantarum K118 strain on MRS medium. The initial count of Lactobacillus plantarum K118 strain was 4.93 ± 0.25 × 10 ⁶ CFU / ml (*), and a was measured after incubation at 37 ° C. for 6 hours It means.

pathogens Growth pathogens ^a K118 + pathogens ^a Inhibition (%) CFU / mL CFU / mL Escherichia coli 3.23 ± 0.25 × 10 ⁶ 2.03 ± 0.53 × 10 ⁶ 37.11% Salmonella Typhimurium 6.46 ± 0.35 × 10 ⁶ 3.23 ± 0.49 × 10 ⁶ 50.00% Listeria monocytogenes 1.57 ± 0.20 × 10 ⁵ 1.29 ± 0.36 × 10 ⁵ 18.01% Staphylococcus aureus 3.46 ± 0.87 × 10 ⁶ 2.83 ± 0.61 × 10 ⁶ 18.27%

As can be seen in Table 8, the Lactobacillus plantarum K118 of the present invention showed an inhibitory effect on Escherichia coli of 37.11%, Salmonella Typhimurium , And 50%, respectively. And Staphylococcus aureus ( Staphylococcus aureus ) was 18.01%.

EXPERIMENTAL EXAMPLE 12. Confirmation of fermentation characteristics of fermented products according to kinds of fermentation medium.

10 parts by weight of water was added to 1 part by weight of the total solid content of these solid components, and the mixture was subjected to hot water extraction at 100 DEG C for 3 hours to prepare a hot water extract. The total solid content contained in the hot water extract was 0.74% by weight and the pH was 5.1.

The concentrate was concentrated using a vacuum decompressor (rotavator R-124, Buchi, Switzerland) at 100 rpm and 50 ° C to obtain an extract solution suitable for fermentation. Five kinds of Bulgogi concentrate having a weight ratio of 7: 1, a weight ratio of 11: 1, and a weight ratio of 15: 1, respectively, were used: .

In order to confirm the fermentation characteristics of the fermented product when the Yellowish chestnut concentrate prepared from different raw materials was used as a fermentation substrate, Lactobacillus plantarum K118 strain was inoculated to each of the above five yellowtail concentrates, The percentage of DPPH (%) and GABA in the fermented product at 24 hours is shown in Table 9. In this example, the K118 strain was inoculated with the Hwanyeongkuk concentrate (weight ratio of leaves and stem of 9: 1) in Experimental Example 3 to measure the fermented broccoli extract.

number Mixed ratio of weed leaf and stem weight DPPH (%) GABA generation
(ug / mL) Comparative Example 1 Uses only Hwangchilok 76.22 0.30 906.11 ± 36.85 Comparative Example 2 Use only yellow trunk trunks 87.42 + 0.11 970.47 ± 33.85 Comparative Example 3 7: 1 82.71 ± 0.28 904.35 ± 11.73 Comparative Example 4 11: 1 78.38 + - 0.34 910.11 + - 24.80 Comparative Example 5 15: 1 83.23 + - 0.12 922.46 ± 22.52 Example 1 9: 1 90.55 + 0.24 1624.82 + - 33.80

As shown in Table 9, when the concentrated fermented product of the present invention was used in a weight ratio of 8: 10: 1, the highest GABA production activity and DPPH (% ).

Since the components of the extract vary depending on each site, the difference in the raw materials causes the growth and activation of the strain, and thus the functionality and GABA content of the final fermentation product are different from each other .

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. It is natural.

<110> KOREA FOOD RESEARCH INSTITUTE <120> Novel Lactobacillus plantarum K118 with Antioxidant and          GABA-producing ability, composition comprising a fermented          product of Dendropanax morbifera extract using the same and          manufacturing method therof <130> HP7343 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 1051 <212> DNA <213> Unknown <220> <223> 16S rDNA of Lactobacillus plantarum K118 <400> 1 gacttcgatg atgctagtgt tggagggttt ccgcccttca gtgctgcagc taacgcatta 60 agcattccgc ctggggagta cggccgcaag gctgaaactc aaaggaattg acgggggccc 120 gcacaagcgg tggagcatgt ggtttaattc gaagctacgc gaagaacctt accaggtctt 180 gacatactat gcaaatctaa gagattagac gttcccttcg gggacatgga tacaggtggt 240 gcatggttgt cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac 300 ccttattatc agttgccagc attaagttgg gcactctggt gagactgccg gtgacaaacc 360 ggaggaaggt ggggatgacg tcaaatcatc atgcccctta tgacctgggc tacacacgtg 420 ctacaatgga tggtacaacg agttgcgaac tcgcgagagt aagctaatct cttaaagcca 480 ttctcagttc ggattgtagg ctgcaactcg cctacatgaa gtcggaatcg ctagtaatcg 540 cggatcagca tgccgcggtg aatacgttcc cgggccttgt acacaccgcc cgtcacacca 600 tgagagtttg taacacccaa agtcggtggg gtaacctttt aggaaccagc cgcctaaggt 660 gggacagatg attagggtga agtcgtcaaa ggggtaacca aaaaaagtgg caccagggga 720 tctacggggg ttcgcccccg ttatccagaa gttaacaacc ggcttacccg agggttttcc 780 ccctcgttat ctcgcccgtt tttccactag attccagagg gcgtttcctc ctgttcttgt 840 tcgagggaac ctgcttccct atataacgcg ctcaactttc ttctgacgga gaacaagact 900 gtccatcaaa aaatacttgc aaacaagcct gttcggcgct ctaaaattcc ccacccaaag 960 cttccggcac agctattttg gcccctacaa gtgttatctc aggcccctgt ggacagggaa 1020 ctaattcttt cggcggggcg tttggattgg a 1051

Claims (21)

Lactobacillus plantarum K118 strain (Accession No. KCCM12015P) having antioxidant and GABA production promoting activity. The method according to claim 1,
The strain Lactobacillus plantarum K118 strain is characterized in that it is used for fermentation of yellowtail. The method according to claim 1,
A strain of Lactobacillus plantarum K118, wherein the strain is cultured in the absence of an external carbon source in addition to the woodweed. The method according to claim 1,
Wherein the strain has acid-fastness and biliary-bility. Lactobacillus plantarum K118 strain. The method according to claim 1,
The strain may be selected from the group consisting of leucine arylamidase, valine arylamidase, N-acetyl-β-glucosaminidase, β-galactosidase (β -glucosidase, -galactosidase,? -glucosidase and? -glucosidase. However, it is not limited to β-glucuronidase Lactobacillus plantarum K118 strain characterized in that it has no activity against the enzyme β-glucuronidase. The method according to claim 1,
The strain is inhibited by any one or more antibiotics selected from the group consisting of penicillin-G, ampicillin, rifampicin, novobiocin and chloramphenicol,
Lactobacillus plantarum K118 strain characterized in that it is resistant to any one or more antibiotics selected from the group consisting of lincomycin, polymycin B and vancomycin. The method according to claim 1,
The strain Lactobacillus plantarum K118 strain is characterized in that it has antibacterial activity against food poisoning bacteria. 8. The method of claim 7,
Wherein the food poisoning bacterium is any one or more selected from the group consisting of Escherichia coli , Salmonella Typhimurium , Staphylococcus aureus , and Listeria monocytogenes Lactobacillus plantarum K118 strain characterized. A fermented product of Ganoderma lucidum with increased GABA content obtained by inoculation with Lactobacillus plantarum strain K118 (Accession No. KCCM12015P) on a perennial tree. 10. The method of claim 9,
Characterized in that the Hwigae-jinja is an Euglena crush or a Hwigae-jinja extract. 11. The method of claim 10,
Wherein the extract is an extract of water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. 11. The method of claim 10,
Wherein the extract is a hydrothermal extract. 10. The method of claim 9,
Wherein the GABA content of the perennial fermented product is in the range of 1600 to 2000 g / ml or more based on the total weight of the perennial fermented product. A method for producing a fermented Yellowlily of Korea, characterized in that the Hwangjil tree is inoculated with a Lactobacillus plantarum K118 strain (Deposit No. KCCM12015P) and fermented. 15. The method of claim 14,
Wherein the Hwigae-jinja is an Euglena crush or a Hwangchu-jaeng extract. 16. The method of claim 15,
Wherein the extract is an extract of water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof. 16. The method of claim 15,
Wherein the extract is a hot-water extract. 16. The method of claim 15,
Wherein the extract has a total solid content of 0.5 to 5% by weight, a pH of 2 to 7, and a sugar concentration of 1 to 5 Brrix. 15. The method of claim 14,
Wherein the extract is a mixture of leaves and stems of a perennial plant of 8 to 10: 1 by weight. 15. The method of claim 14,
Wherein the fermentation is carried out at 25 to 50 DEG C for 10 to 96 hours. 15. The method of claim 14,
Wherein the fermentation is carried out in the absence of an external carbon source in addition to the woody spruce.

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