KR101768921B1 - Pediococcus acidilactici SRCM 102024 starter strain for producing fermented grain with improved antimicrobial activity and uses thereof - Google Patents

Abstract

The present invention relates to a Pediococcus acidilactici SRCM 102024 strain for a starter for producing a fermented grain with excellent antibacterial activity and uses thereof. The Pediococcus acidilactici SRCM 102024 strain, which is a lactic acid bacterium, enables a user to manufacture a functional fermented grain having antibacterial activity against propionibacterium acnes and Staphylococcus epidermidis, which are skin dermatitis causing bacteria. Accordingly, fermented grain liquid, obtained by using the Pediococcus acidilactici SRCM 102024 strain, can be used as an agent for treating acne and atopic dermatitis and thus can be advantageously used as functional food and a cosmetic composition for related industries.

Description

Pediococcus acedilactici SRCM 102024 starter for producing fermented grain having excellent antimicrobial activity and its use {Pediococcus acidilactici SRCM 102024 starter strain for producing fermented grain with improved antimicrobial activity and uses thereof}

The present invention relates to a strain Pediococcus aceyltracis SRCM 102024 for starter and a use thereof for producing a fermented grain having excellent antibacterial activity.

The desire for beautiful skin demands healthy skin regardless of age or gender. Healthy and clean skin is popular with the desire for beauty. In order to maintain clean skin according to these cosmetic trends, many people are worried about environmental problems in modern society.

Acne bacteria ( Propionibacterium acnes is known to be a causative agent of acne. When sebaceae are left in the hair follicle due to excessive secretion of sebum and excess and scale of cell keratin, acne bacteria and skin staphylococci, which are the skin- Lipase, a lipase of bacteria, breaks down triglyceride into lipid constituting sebum and changes it into free fatty acid. Acne bacterium stimulates the cells around the hair follicle walls or pores of the acne causing inflammation, and as a result, it is overshot, and at the moment clogs the initial cotton is produced. Free fatty acids produce enzymes that can amplify the inflammation, which causes the white blood cells to break down and the contents of the ruptured follicles enter the dermis, causing inflammation and causing acne.

Atopic dermatitis refers to a chronic, recurrent, inflammatory skin condition that is pruritic and usually occurs in infants and children. The duration of the disease is 1 month to 44 years, with an average of 11.40 ± 8.64 years. The prevalence rate is 27% for the year and 53.9% for the 10 years. Clinical features of atopic dermatitis are classified according to the developmental stage. In early childhood, the first signs of atopic dermatitis are eczema, acute blisters and spotty lesions on the cheeks, eczematous lesions in the childhood, antecubital fossae, neck, cuff, heel) and back of neck, foot and hands. When adolescent and adult eczema lesions continue from childhood to adolescence and adulthood, or when the disease is newly started in adulthood, the head and neck as well as the bending part are associated with lichenified plaques and one of the main features of atopic dermatitis is epidermal This dryness has been suggested as a mechanism of skin hypofunction in atopic dermatitis, dry skin is a problem, and in winter it is more problematic. It is true.

On the other hand, lactic acid bacterium is a generic term for lactic acid producing bacteria using saccharides such as glucose as an energy source, and has a function of inhibiting the growth of harmful bacteria, improving the function of lactic acid bacteria, improving immunity, improving diarrhea and constipation, , Lactobacillus lactic acid bacteria, Lactobacillus sp. Lactic acid bacteria, and Bifidobacterium sp. Lactic acid bacteria. The purpose of using lactic acid bacteria in food processing is to improve the preservation of food, improve flavor of food, and improve functionality.

Currently, adult lifestyles such as diabetes, obesity and hypertension are increasing in modern people due to the lifestyle of avoiding whole grains. People suffering from these adult diseases try to eat grains, but the digestion rate is low and the effect is great not. Therefore, it is necessary to eat gravy to prevent adult disease and increase digestion rate. Interest in cereals is rising day by day in pursuit of simplicity of eating habits in the rapidly changing modern age. Using lactic acid bacteria that grow well in mixed cereals by using human cereal, which is an important energy source, it promotes the consumption of cereals by promoting the consumption of cereals, which not only helps to solve the economic problems of the farmers but also provides excellent fermented cereals I want to.

Korean Patent Laid-Open Publication No. 2017-0045050 discloses a fermented product of lactic acid bacteria and a method for producing the fermented product. In Korean Patent Laid-Open Publication No. 2017-0048229, a novel strain derived from a traditional fermented food having excellent enzyme production ability, A method for producing enzyme food 'has been disclosed. However, as in the present invention, there has never been known a strain of Pediococcus acephaliticus SRCM 102024 for starter for producing a fermented grain having excellent antibacterial activity and its use .

The present invention has been made in view of the above-mentioned needs. In the present invention, lactic acid bacteria are isolated from a fruit fermentation broth. The lactic acid bacteria are selected from the group consisting of Propionibacterium acnes, acnes) and Staphylococcus epidermidis (Staphylcoccus epidermidis) Phedi O Lactococcus her CD rakti when (Pediococcus with antibacterial activity against acidilactici ) SRCM 102024.

In addition, as a result of using the strain as a starter strain for fermentation to prepare a fermented cereal grains, dermatitis causative agent of propionic sludge tumefaciens arc Ness (Propionibacterium acnes) and Staphylococcus epidermidis (Staphylcoccus epidermidis ), and the Pediococcus ( Pediococcus) acidic lactic acid, increased the content of amino acid nitrogen, organic acid, organic sugar and free amino acid, increased the inhibitory activity of alpha -glucosidase and protease activity and increased the total number of lactic acid bacteria, compared with the control without the acidilactici SRCM 102024 strain The present invention has been completed.

In order to solve the above-mentioned problems, the present invention relates to a method for producing an antimicrobial agent having antimicrobial activity against a causative agent of dermatitis isolated from a fruit fermentation broth and having a deposit number of KCCM12019P ( Pediococcus acidilactici ) SRCM 102024 strain.

In addition, the present invention provides the O Phedi Lactococcus her CD rakti when (Pediococcus acidilactici) SRCM 102024 fermentation for producing grain starter (starter) composition containing the strain or culture thereof as an active ingredient.

The present invention also provides a fermented grain having antimicrobial activity against a causative organism of dermatitis produced by fermenting cereals using Pediococcus acidilactici strain SRCM 102024 or a culture thereof as a starter strain do.

Also, the present invention provides a health functional food composition for preventing or ameliorating acne or atopic dermatitis containing an effective ingredient of a fermented grain having an antibacterial activity against the causative agent of dermatitis.

In addition, the present invention provides a cosmetic composition for improving acne or atopic dermatitis containing, as an active ingredient, a fermented grain having an antibacterial activity against the causative agent of dermatitis.

The strain Pediococcus acidilactici SRCM 102024, which is a lactic acid bacterium isolated in the present invention, can be used for the production of functional fermented grains having antimicrobial activity against the dermatitis causing bacteria such as Propionibacterium acnes and Staphylococcus epidermidis .

Thus, the Pediococcus acidilactici ) SRCM 102024 can be used as a therapeutic agent for acne and atopic dermatitis, and thus can be usefully used in related industries as a functional food and cosmetic composition.

1 shows a method for isolating and culturing a strain used in the present invention.
FIG. 2 shows a process for producing fermented grains using the strains isolated in the present invention.
FIG. 3 shows a method for separating microorganisms from a cereal fermentation broth using a strain isolated in the present invention.
Figure 4 shows the primer information used for 16S rRNA sequencing of strains isolated in the present invention.
FIG. 5 shows phylogenetic analysis based on the 16S rRNA nucleotide sequence of the lactic acid bacteria isolated in the present invention. Neighbor joining method was used and compared with 10 strains nearest to the blast result.
6 is Phedi of the present invention O Lactococcus her CD rakti when (Pediococcus acidilactici ) SRCM 102024 strain. It was treated with isolated strains. No clear zone was formed in the fungi. Two holes under the left plate culture medium with clear zone were formed by Pediococcus (Lactobacillus acidophilus) acidilactici ) SRCM 102024 strain.
7 is Phedi of the present invention O Lactococcus her CD rakti when (Pediococcus acidilactici ) SRCM 102024 was used for the analysis of organic acids in cereal fermentation broth. 1: Blank, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 6: Reuconostomyces teneroides SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stock Methenseeroides SRCM 101677, 9: Pediococcus acidiflactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
8 is Phedi of the present invention O Lactococcus her CD rakti when (Pediococcus acidilactici ) SRCM 102024 strain. 1: Blank, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 6: Reuconostomyces teneroides SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
9 is trying Phedi O Lactococcus according to the present invention when CD rakti (Pediococcus acidilactici ) SRCM 102024 strain of the present invention. 1: Blank, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 6: Reuconostomyces teneroides SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677
10 is Phedi of the present invention O Lactococcus her CD rakti when (Pediococcus acidilactici ) SRCM 102024 was used for the analysis of antioxidant activity of cereal fermentation broth. 1: Blank, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 6: Reuconostomyces teneroides SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677

In order to accomplish the object of the present invention, the present invention provides a method for producing an antimicrobial agent having antimicrobial activity against a causative agent of dermatitis isolated from a fruit fermentation broth and having a deposit number of KCCM12019P ( Pediococcus acidilactici ) SRCM 102024 strain.

In the present invention, lactic acid bacteria were isolated from a fruit fermentation broth. The lactic acid bacteria were selected from Propionibacterium acnes, acnes) and Staphylococcus epidermidis (Staphylcoccus epidermidis) Phedi O Lactococcus her CD rakti when (Pediococcus with antibacterial activity against acidilactici ) SRCM 102024, and the strain was deposited at the Korean Microorganism Conservation Center (KCCM) on April 20, 2017 (Accession No .: KCCM12019P).

By "antimicrobial" is meant the ability to reduce, prevent, inhibit, or eliminate microbial growth or survival at any concentration.

In the strain according to one embodiment of the present invention, the causative agent of dermatitis is Propionibacterium acnes causing acne It is acnes) or atopic pathogen is Staphylococcus epidermidis (Staphylcoccus epidermidis), but is not limited thereto.

In addition, the present invention relates to the aforementioned Pediococcus acid starch composition for the production of fermented grains containing an acidilactici SRCM 102024 strain or a culture thereof as an active ingredient.

In the present invention, a starter for manufacturing fermented grain means a preparation or a composition containing microorganisms involved in fermentation for producing fermented grain. It is used to provide a microorganism capable of growing in fermented grains or a microorganism capable of growing into a dominant species by addition at the time of fermented grain production. When the fermented grain is produced using the starter for cereal fermentation, the quality of the fermented grain is controlled by the microorganisms contained in the starter for cereal fermentation, , The purpose of reducing, or the purpose of enhancing the flavor or sweetness imparted by the fermented grain. In the present invention, Pediococcus acidilactici SRCM 102024 or a culture thereof as a starter strain has antimicrobial activity against a causative agent of dermatitis and furthermore has an antibacterial activity against Pediococcus acidic , lactic acid, lactic acid bacteria, acidic lactic acid , SRCM 102024 strain, amino acid nitrogen, organic acid, organic sugar and free amino acid content, α-glucosidase inhibitory activity and protease activity were increased, Can be manufactured.

The method for culturing the strain of the present invention can be carried out according to a method commonly used in the art, and is not limited to a specific method.

The present invention also provides a fermented grain having antimicrobial activity against a causative organism of dermatitis produced by fermenting cereals using Pediococcus acidilactici strain SRCM 102024 or a culture thereof as a starter strain do.

In the fermented grain according to one embodiment of the present invention, the dermatitis causing bacteria are selected from the group consisting of Propionibacterium acnes causing acne acnes) or atopic pathogen is Staphylococcus epidermidis (Staphylcoccus epidermidis ). < / RTI >

In a fermented grain according to an embodiment of the present invention, the fermented grain is selected from the group consisting of Pediococcus acidilactici) SRCM increases the amino nitrogen, organic acids, organic sugars and free amino acid content than the control not treated 102 024 the strain and, α- glucosidase, and the increase in the inhibitory activity and protease activity, may be a lactic acid bacteria is increased, but the total number , But is not limited thereto.

Also, the present invention provides a health functional food composition for preventing or ameliorating acne or atopic dermatitis containing an effective ingredient of a fermented grain having an antibacterial activity against the causative agent of dermatitis.

The health functional food of the present invention is not particularly limited as long as it can be ingested to prevent or ameliorate acne or atopic dermatitis.

When the fermented grain of the present invention is used as a food additive, the fermented grain can be directly added or used together with other food or food ingredients, and can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, the fermented grains of the present invention are added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material, in the production of food or beverage. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .

There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including meats, sausages, breads, chocolates, candies, snacks, confectionery, pizza, ramen noodles, gums, ice cream, soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like.

In addition to the above, the extract of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention provides a cosmetic composition for improving acne or atopic dermatitis containing, as an active ingredient, a fermented grain having an antibacterial activity against the causative agent of dermatitis.

In the cosmetic composition for improving acne or atopic dermatitis according to the present invention, the cosmetic composition may be at least one selected from the group consisting of an ointment for external use, a cream, a softening agent, a nutrient lotion, a pack, an essence, a hair tonic, a shampoo, a rinse, a hair conditioner, Skin lotion, skin toner, astringent, lotion, milky lotion, moisturizing lotion, nutrition lotion, massage cream, nutritional cream, eye cream, moisturizing cream, hand cream, foundation, nutrition essence, sunscreen, soap, cleansing foam, But are not limited to, formulations selected from the group consisting of cleansing lotions, cleansing creams, body lotions and body cleansers. The composition of each of these formulations may contain various kinds of bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art.

When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier components .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Materials and methods

1. Isolation and Identification of Strain from Fruit Fermentation Liquor

(1) Microbial separation

Experimental materials were fruit fermented liquid prepared by various fruits at Cain farming in Sunchang - gun, Jeollabuk - do. The fermentation broth was fermented for 5 years and isolated microorganisms. For microbial separation, 10 g of each sample was diluted in 90 ml of sterilized physiological saline, homogenized, and sequentially diluted. The diluted solution was applied to NA (nutrient agar, Difco) and MRS medium and incubated at 37 ° C. for 24 hours, And cultured for 72 hours. At this time, specific colonies which are presumed to be bacteria and lactic acid bacteria were selected and purified. The method for isolating and culturing the strain was performed as shown in the schematic diagram of FIG. These isolates were identified and analyzed using 10% glycerol stock.

(2) Identification of microorganism-isolated strains

The isolated strains were subjected to 16S rRNA sequencing and BLAST test by Macrogen, and the identification results were obtained.

(3) Antibacterial activity of isolate strain

Agar well diffusion assay was used to confirm antimicrobial activity against the three microorganisms (Table 1). NA liquid medium, and the supernatant was centrifuged at 13,000 rpm for 30 minutes at 4 ° C to prepare 0.85% soft agar. 90 μl of the supernatant was dispensed into each well, and the supernatant was naturally dried in an aseptic state And then cultured at 37 ° C for 24 hours. After culturing, the diameter (Cm) of the inhibitory circle formed around the well was measured to confirm the presence of antibacterial activity.

Atopy and acne strains Strain name (scientific name) SRCM NO. One Starfilacocus epidermidis KCCM 35494 2 Starfilacocus epidermidis KCCM 40416 3 Propionibacterium acnes KCCM 41747

2. Analysis of general components of fermentation broth

(1) pH

5 g of the sample and 45 g of distilled water were stirred and a total of 50 g was measured at room temperature three times using an automatic titrator (Excellence Titrator T50M, Switzerland), and the average value was determined.

(2) pH

5 g of the sample and 45 g of distilled water were stirred and a total of 50 g was measured at room temperature three times using an automatic titrator (Excellence Titrator T50M, Switzerland), and the average value was determined.

(3) sugar content

1 g of the sample was diluted 10-fold and measured using a sugar meter (Master-2M, 1M ATAGO Co., LTD.

3. Grain fermentation experiment method

1) Experimental material

Germination barley, germinated brown rice, barley and brown rice were used for grain fermentation, and germinated barley, brown rice, germinated rice, barley, germinated barley and germinated brown rice were prepared. The strain was isolated from a fermentation broth of a farming cain, Pediococcus acidilactici ) and lactobacillus strains belonging to the fermentative microorganism industry promotion institute ( Leuconostoc mesenteroides ) 2 species, Lactobacillus plantarum ( Lactobacillus plantarum ) was distributed and tested.

2) Grain Fermentation Condition

(1) Culture of lactic acid bacteria and fermentation conditions of cereals

The strains used in this experiment were five strains of lactic acid bacteria including the strains isolated from the inventors' isolates and the fermented microorganism Industry Promotion Agency. The pre-cultured strain was streaked on Lactobacillus MRS agar (Difco) medium, and then primary cultured at 35 ° C for 48 hours, followed by secondary culturing for 24 hours at 35 ° C in Lactobacillus MRS medium.

Lactic acid bacteria strains for fermentation grain production Strain name (scientific name) SRCM No. One Pedio O'Corkus SRCM 102024 2 Lactobacillus Florata Room SRCM 101502 3 Lactobacillus Florata Room SRCM 101503 4 Ryukono Stock Methenseeroides SRCM 101665 5 Ryukono Stock Methenseeroides SRCM 101677

(2) Selection of raw materials for cereal fermentation

The raw materials of cereals are selected from barley, germinated barley, brown rice and germinated brown rice, and the strains which have been sold as R (germinated barley + brown rice), G (germinated barley + germinated brown rice) and Y Or in combination with each other. The whole process is schematically shown in Fig.

Raw grain ratio Strain number / grain R (germinated barley + brown rice) G (germinated barley + germinated brown rice) Y (germinated brown rice + barley) One 25g + 25g 25g + 25g 25g + 25g 2 25g + 25g 25g + 25g 25g + 25g 3 25g + 25g 25g + 25g 25g + 25g 4 25g + 25g 25g + 25g 25g + 25g 5 25g + 25g 25g + 25g 25g + 25g

(3) Setting of grain fermentation water addition amount

Germinated barley and brown rice, germinated barley and germinated brown rice, germinated rice and barley were weighed 1: 1. Moisture contents of 20%, 30%, 40%, 50% and 60% (W / W) were added to the cultured cereal grains and the fermented broth was inoculated with 10 ⁷ CFU / ㎖ of fermentation conditions.

Conditions for addition of grain fermented water Grain / water content 20% 30% 40% 50% 60% R (germinated barley + brown rice) 10 mL 15 mL 20 mL 25 mL 30 mL G (germinated barley + germinated brown rice) 10 mL 15 mL 20 mL 25 mL 30 mL Y (germinated brown rice + barley) 10 mL 15 mL 20 mL 25 mL 30 mL

3) Grain fermented  Characteristic evaluation experiment method

(1) Moisture

5 g of the sample was measured using an infrared water flow meter (Kette, 720).

(2) pH

5 g of sample, and 45 g of distilled water were stirred, and a total of 50 g was repeatedly measured three times using an automatic titrator (Excellence Titrator T50M, Switzerland) at room temperature, and the average value was determined.

(3) pH

5 g of sample, and 45 g of distilled water were stirred, and a total of 50 g was repeatedly measured three times using an automatic titrator (Excellence Titrator T50M, Switzerland) at room temperature, and the average value was determined.

(4) Amino nitrogen

Amino nitrogen content was measured by using an automatic titrator (Excellence Titrator T50M, Switzerland), taking 1 g of sample, adding 50 mL of distilled water, stirring for 1 hour, and titrating to pH 8.4 with 0.1 N NaOH solution. To this was added 20 mL of neutral formalin solution and titrated to pH 8.4 with 0.1 N NaOH solution.

(5) Organic acids and free sugars

To analyze the organic acids and free sugars, 45 mL of distilled water was added to 5 g of the sample, homogenized for 1 hour, passed through a 0.45 μm membrane filter, and simultaneously analyzed by HPLC under the analysis conditions shown in Table 5.

HPLC analysis conditions for organic acids and free sugars Instument Agilent Detector RID, DAD 210 nm Column Aminex column HPX-87P (300 x 7.8 mm) Temperature 50 ℃ Injection volume 20 쨉 l Flow rate 0.6 mL / min Mobile phase 5 mM H ₂ SO ₄

(6) free amino acids

For free amino acids, take 2 g of sample, add 30 mL of third-order distilled water, stir with 50 mL of distilled water for 20 minutes, centrifuge at 3,000 rpm for 10 minutes, add 2 mL of 5% TCA to 2 mL of supernatant After centrifugation (10,000 rpm, 10 min), the supernatant was diluted with 0.02 N HCl, passed through a 0.2 μm syringe filter, and analyzed with an amino acid analyzer under the analysis conditions of Table 6

Free amino acid analyzer analysis conditions Instument Amino acid analysis (Hitachi L-8900) Detector UV / Vis (440 nm - 570 nm) Column Hitachi 4.6 x 60 mm (speration)
Hitachi 4.6 × 40 mm (Ammonia filtering) Buffer flow rate 0.40 mL / min Ninhydrin flow rate 0.35 mL / min Temperature 50 ℃ Injection volume 20 쨉 l Mobile phase Buffer set (pH-SET KANTO)

(7) Measurement of DPPH radical scavenging ability

DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity was measured by the effect of electron donation abilities (EDA). That is, 150 μl of a 0.2 mM DPPH solution (dissolved in 99% MeOH) was added to 150 μl of a sample, and the mixture was allowed to stand at room temperature for 30 minutes. Then, the absorbance at 517 nm was measured using a spectrophotometer. The radical scavenging activity of each sample was expressed as a percentage of the absorbance difference between the sample addition and the no addition region by the following equation.

Electron donating ability (%) = (1-absorbance of sample added / absorbance of non-treated) x 100

(8)? -Glucosidase inhibitory activity

AGI (α-glucosidase inhibitory) inhibitory activity was measured by the following method. That is, 50 μl of 0.3 U / ml α-glucosidase enzyme solution (50 μl in 0.1 M PBS, pH 6.8) was mixed with 50 μl of the sample and pre-cultured at 37 ° C for 10 minutes. 3 mM ρ-NPG (ρ- nitro-phenyl-α-glucopyranoside, in 0.1 M PBS, pH 6.8) was added to 100 ㎕ After 10 min at 37 ℃ 0.1 M Na ₂ CO ₃ Was added to stop the reaction. The amount of ρ-nitrophenol produced at this time was measured at an absorbance of 405 nm using a spectrophotometer (ELISA reader, Infinite 200 TECAN). As a control, 0.1 M PBS (pH 6.8) was used instead of the culture, and 0.1 M PBS (pH 6.8) was used instead of the culture medium and enzyme solution as the blank.

Enzyme inhibitory activity (%) = (1-p-nitrophenol formation amount in reaction section / p-nitrophenol production amount in non-processing section) x 100

(9) Analysis of protein and starchase activity

In order to measure the activity of protein, fiber, and starch hydrolytic enzymes in the extracellular enzymes of the selective strains, a plate medium containing substrate components that specifically react with each enzyme was prepared. The activity of the protease was determined by adding 1.5% agar (Junsei Chemical Co. Ltd., Japan) to 2% skim milk (Difco ™, USA) A 5 mm hole was drilled in the medium. Was selected lactobacillus culture MRS broth ℃ 37, 24 hours with shaking at, screening strain culture solution is taken back by each supernatant was centrifuged for 10 minutes to 15,000x g in a filtering 0.45㎛ membrane filter (Sartorius, Germany) 100㎕ After extracellular enzymatic activity was measured, the cells were reacted at 37 DEG C for 24 hours, and the size of the circle around the wells was measured.

(10) Antimicrobial activity of grain fermented extract

Water activity, salting, and direct method are used to prevent food corruption. However, because of limited application range, artificial synthetic preservatives are used to improve storage stability. In order to increase the consumer 's health - oriented desire for atopy, we selected the microorganisms which cause atopy, and the three kinds of microorganisms were distributed in KCCM to test the antibacterial activity against them.

Atopy and acne strains Strain name (scientific name) KCCM No. One Starfilacocus epidermidis KCCM 35494 2 Starfilacocus epidermidis KCCM 40416 3 Propionibacterium acnes KCCM 41747

(11) Total bacteria count of lactic acid bacteria after cereal fermentation

25 g of the sample was aseptically taken, 225 mL of sterilized water was added, and homogenized with a bag mixer. Each sample was diluted by 10 dilution, 100 μL of the diluted solution was plated on a Lactobacillus MRS agar (Difo) medium and cultured at 35 ° C. for 48 hours The number of colonies was counted (CFU / mL).

Example  1. Physicochemical analysis of fermentation broth

1) pH, acidity and sugar content

Table 8 shows the results of the analysis of the general components of raw materials that are fruit juices of Cain in Yeongdong corporation. As a result of the general composition analysis, the pH was 3.57 ± 0.02, the acidity was 7.44 ± 1.34, and the sugar content was 42.97 ± 0.05.

Standard solution Inspection items pH Acidity (%) Sugar (Briggs)
Raw material of fermentation liquid
3.57 + 0.02
7.44 ± 1.34
42.97 ± 0.05

2) Analysis of microorganisms in fermentation broth

The total number of bacteria in the fermentation broth was measured and the coliform group was analyzed. The results are summarized in Table 9. Microbial analysis results were negative for total bacteria, E. coli and B.C.

Microbiological review of raw materials of fermentation broth Inspection items Total number of bacteria Coliform group B.C Raw material of fermentation liquid voice voice voice

3) Isolation and identification of microorganisms from fermentation broth

The isolation of the strain was carried out as shown in Fig. 1, and the isolated strains were subjected to 16S rRNA sequencing and BLAST test by Macrogen, and the results were identified. The primers used in the analysis are shown in Fig. As a result of substituting the nucleotide sequence (SEQ ID NO: 1) obtained in the step of identification of the above strain into NCBI, 96% of the nucleotide sequences were read and 99% were identical to NR_042057.1 and Pediococcus acidilactici . The E-value indicating reliability was 0.0, confirming that the reliability was high. The results of the phylogenetic analysis of the newly isolated isolates were shown in Fig.

Pediococcus is a genus of gram-positive lactic acid bacteria belonging to Lactobacillaceae and family, which usually occur in pairs or pairs and maintain symmetry and are separated into two . These bacteria are bacteria that flow Stock Kono (Leuconostoc) and other lactic acid bacteria such as Lactobacillus for the cabbage such as (Lactobacillus) in fermented sauerkraut to change (Sauerkraut) bacteria that are known as the most likely cause.

4) Antimicrobial activity of fermentation broth

The results of the antimicrobial activity analysis of the fermentation broth are shown in Table 10 and FIG. In the antimicrobial activity, the isolates did not exhibit antimicrobial activity in the fungi, and lactic acid bacteria Staphylococcus epidermidis KCCM35494 showed high antimicrobial activity.

Antibacterial activity of fermentation broth (cm) Strain no. Identification Antimicrobial activity
(Halo size (cm), mean ± standard deviation) S. epidermidis
KCCM40416

S. epidermidis
KCCM40416

pH 7.0 S. epidermidis KCCM35494

S. epidermidis KCCM40416

pH 7.0 SREF008 Pedio O'Corkus w - 2.11 ± 0.01 - SREF009 Pedio O'Corkus w - 2.12 ± 0.01 -

5) Antibacterial activity of isolate of lactic acid bacteria

Table 11 shows the results of the antimicrobial activity analysis of the isolates of the lactic acid bacteria. The strain Pediococcus aceyltactii SRCM 102024 isolated and isolated from the fermentation broth provided by Cain, the Agricultural Cooperatives Act, is a strain of Staphylococcus epidermidis The halo size of KCCM35494 was 3.8cm, which showed higher antimicrobial activity than other strains.

Antibacterial activity of isolate of lactic acid bacteria (cm) Strain no. Identification Antimicrobial activity
(Halo size (cm), mean ± standard deviation) S. epidermidis KCCM40416
S. epidermidis KCCM35494
P. acnes KCCM41747
SRCM 102024 Pedio Caucus
Ashidi Laktishi 2.9 3.8 2.5 SRCM 101502 Lactobacillus
Flora Room 1.2 1.1 1.5 SRCM 101503 Lactobacillus
Flora Room 2.5 3.0 2.5 SRCM 101665 Ryukono Stock
Messeneroides 1.0 1.4 2.5 SRCM 101677 Ryukono Stock
Messeneroides 1.2 1.3 2.5

Example  2. Grain fermentation conditions

1) Selection of lactic acid bacteria

In this experiment, the fermented microorganism Industry Promotion Agency has leuconostoc mesenteroides ( Leuconostoc mesenteroides ) Two species Lactobacillus ( Lactobacillus) plantarum ), which was isolated from a fermentation broth of a farming corporation, and the strain Pediococcus acidilactici ) were added to each of the five cultivars.

Lactic acid bacteria strain for the production of grain fermentation Strain name (scientific name) SRCM NO. One Pedio O'Corkus SRCM 102024 2 Lactobacillus Florata Room SRCM 101502 3 Lactobacillus Florata Room SRCM 101503 4 Ryukono Stock Methenseeroides SRCM 101665 5 Ryukono Stock Methenseeroides SRCM 101677

2) Selection of raw materials for cereal fermentation

Germinated barley and brown rice, germinated barley and germinated brown rice, germinated rice and barley were prepared, respectively. Fermentation was carried out at 37 ℃ by inoculation with lactic acid bacteria strain (10 ⁷ CFU / ㎖). As a result, germinated barley and brown rice showed excellent results.

Example  3. Evaluation results of grain fermentation characteristics

(1) Moisture

Table 13 shows the results of moisture analysis of the fermented cereal fermented with different strains. In this study, the untreated control was 42.55% at day 0 and decreased to 39.10% at day 7. As the fermentation progressed from 42.56% on the 0th day, the number of grains inoculated with single strains tended to decrease to 38.65%, 39.07%, 39.51%, 41.25% and 40.48%, respectively. The fermentation rate decreased to 40.21%, 37.97% and 39.92%, respectively. There was no significant difference.

Grain fermentation water (%) Day One 2 3 4 5 6 7 8 9 0 42.55 42.56 42.56 42.56 42.56 42.56 42.56 42.56 42.56 3 39.80 40.05 39.91 42.15 41.55 40.64 34.91 39.36 40.16 5 39.30 39.77 39.62 41.24 41.35 40.37 39.92 38.05 40.00 7 39.10 38.65 39.07 39.51 41.25 40.48 40.21 37.97 39.92

1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677

(2) pH

Table 14 shows the pH analysis of the fermented cereal fermented with a single strain and a complex strain. In this study, the control value of cereal fermentation showed a value of 6.42 at the 0th day and decreased to 4.12 at the 7th day. As the fermentation progressed at 6.45, the fermentation progressed to 5.85, 4.81, 4.05, 4.71 and 4.60% at 0 day, and the fermentation progressed to 4.45 , 4.33, and 4.95, respectively. Among 9 strains including the control, Pediococcus acidilactic ) SRCM 102024 showed the lowest pH value. These results suggest that lactic acid bacteria and microorganisms formed during fermentation produce organic acids and pH is lowered.

Grain fermentation pH Day One 2 3 4 5 6 7 8 9 0 6.42 6.45 6.45 6.45 6.45 6.45 6.45 6.45 6.45 3 5.96 6.20 4.19 4.45 5.53 4.28 4.22 4.46 4.08 5 4.69 5.90 4.88 4.10 4.80 4.11 4.20 4.23 4.15 7 4.12 5.85 4.81 4.05 4.71 4.60 4.77 4.33 4.95

1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677

(3) pH

Table 15 shows the titratable acidity of the fermented cereal fermented with different strains. In the present study, the control value of the unfermented grain was 0.28% at the 0th day and increased to 0.30% at the 7th day, but the difference was not significant. As the fermentation progressed from 0.27% on the 0th day, the number of grains increased to 1.59%, 1.16%, 2.55%, 1.20% and 1.40%, respectively. , Fermentation progressed by 1.50%, 1.70% and 1.90%, respectively. Of the nine strains, Pediococcus acidilactic ) SRCM 102024 showed the highest titratable acidity, indicating that lactic acid fermentation was better than other strains. No significant differences were found in other strains.

Grain fermentation titratable acidity (%) Day One 2 3 4 5 6 7 8 9 0 0.28 0.27 0.27 0.27 0.27 0.27 0.27 0.27 0.27 3 0.20 1.10 1.48 0.89 0.58 1.28 1.39 1.17 1.64 5 0.30 1.50 1.60 1.16 1.10 1.21 1.46 1.19 1.47 7 0.30 1.59 1.16 2.55 1.20 1.40 1.50 1.70 1.90

1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677

(4) Amino nitrogen

Amino nitrogen content of the fermented cereal fermented by different strains of the single strain and the complex strain is shown in Table 16. In the present study, 24.00 mg% of the control group without grain fermentation was observed at day 0, and decreased to 22.30 mg% at 7 days, but the difference was not significant. As the fermentation progressed from 24.10mg% on the 0th day, the grains inoculated with single strains tended to increase to 35.30mg%, 32.10mg%, 77.00mg%, 50.40mg% and 58.50mg%, respectively. In the day 0, the fermentation progressed from 24.10mg% to 45.30mg%, 49.50mg% and 37.66mg%, respectively. Of the nine strains, Pediococcus acidilactic ) SRCM 102024 showed the highest value of amino nitrogen, which is considered to be the best fermentation than other strains.

Grain fermentation amino nitrogen (mg%) Day One 2 3 4 5 6 7 8 9 0 24.00 24.10 24.10 24.10 24.10 24.10 24.10 24.10 24.10 3 25.20 58.50 26.20 31.00 33.50 40.60 38.10 39.80 37.30 5 38.60 31.40 53.60 69.50 40.10 49.00 42.40 48.30 35.60 7 22.30 35.30 32.10 77.00 50.40 58.50 45.30 49.50 37.66

1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677

(5) Organic acids and free sugars

After pretreatment according to the experimental method, samples were analyzed for oxalic acid, citric acid, malic acid, succinic acid, lactic acid, formic acid, acetic acid acid) were analyzed. Among them, citric acid was not detected, lactic acid was detected the most, and acetic acid, succinic acid, malic acid, formic acid and oxalic acid were detected in the order. FIG. 7 shows the total organic acid content and the highest concentration of organic acid was obtained in the fermentation of the combined strains of the 5-day fermentation conditions in which the three strains were fermented together. Glucose was analyzed by fructose, glucose, sucrose and maltose after HPLC pretreatment. Fructose, glucose and maltose were detected in maltose and maltose, respectively. FIG. 8 shows the total free sugar content of each sample and was highest in the combined fermentation conditions of three microorganisms under the 5-day fermentation condition.

(6) free amino acids

The free amino acids were analyzed by HPLC after fermentation by strain according to the number of days of fermentation. As a result, the fermentation conditions of Pediococcus acidilactic ) In SRCM 102024, the total amount of free amino acids was the highest. In addition, GABA in amino acid was increased during 5 days fermentation, which could be suggested as a characteristic of grain fermentation. FIG. 9 shows the results of the total free amino acids according to the experimental conditions.

(7) Measurement of DPPH radical scavenging ability

The DPPH radical scavenging reaction was tested for antioxidative activity and the results are shown in FIG. The water extracts showed little antioxidative effect and were tested with 70% ethanol extract. The changes in DPPH radical scavenging ability were not affected by fermentation conditions according to the fermentation conditions according to fermentation days 3 days / 5 days / 7 days.

Electron donating ability (%) = (1-absorbance of sample added / absorbance of non-treated) x 100

(8)? -Glucosidase inhibitory activity

AGI (α-glucosidase inhibitory) inhibitory activity was measured by using the supernatant of the strain used and the water extract of the fermentation of grain. The experimental results after the reaction were calculated by substituting the following equation.

Enzyme inhibitory activity (%) = (1-p-nitrophenol production amount in the reaction vessel / p-nitrophenol production amount in the untreated sphere) × 100

Single strain of Lactobacillus selected five kinds Planta Room (Lactobacillus plantarum) SRCM 101502, Lactobacillus Planta Room (Lactobacillus plantarum ) SRCM 101503, and Pediococcus acidilactic ) The highest activity of AGI activity was observed in SRCM 102024 and the results are shown in Table 17 below. Two kinds of Lactobacillus plantanum and Pediococcus acidilactici showed a high degree of AGI activity and showed almost no activity in Leuconostoc mesenteroides .

AGI activity of the strain supernatant Active / strain 2 3 4 5 6 AGI activity (%) 86.8 90.3 82.0 6.8 10.9

2; Lactobacillus Florata Room SRCM 101502, 3; Lactobacillus Florata Room SRCM 101503, 4; Pedio O'Corkus SRCM 102024, 5; Ryukono Stock Methenseeroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677

(9) Proteolytic enzyme activity assay

The results of proteolytic enzyme activity of the fermented cereal fermented with different strains of the single strain and the complex strain are shown in Table 18. The protease activity was not detected in the control group of 0 day old, but the different results of the protease activity test by the strain and fermentation period were shown differently. Among them, Pediococcus acidilactici SRCM 102024 fermented with a single strain showed the highest protease activity at 2.0 cm. And the other strains did not show any significant difference.

Protease activity (cm) Day One 2 3 4 5 6 7 8 9 0 ND ND ND ND ND ND ND ND ND 3 ND ND 1.8 1.9 ND 1.6 1.7 1.7 1.8 5 ND ND 1.8 2.0 ND 1.8 1.8 1.7 1.8 7 ND ND 1.9 2.0 ND 1.8 1.8 1.8 1.9

1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677

(10) Antimicrobial activity against atopy and acne

Table 19 shows the results of the antimicrobial activity of the fermented cereal fermented with a single strain and a complex strain. The antimicrobial activity was not observed in the 0 day old control, but the results of antimicrobial activity were different according to the fermentation period of each strain. Among them, Propionibacterium acnes Five the date of entry into force of a single strain KCCM 41747 strain Lactococcus Pedy five kids CD rakti City (Pediococcus acidilactic ) SRCM 102024 showed the highest antibacterial activity at 1.7 cm.

Propionibacterium acnes Antibacterial activity of cereal fermentation on KCCM 41747 strain (cm) Day One 2 3 4 5 6 7 8 9 0 ND ND ND ND ND ND ND ND ND 3 ND 1.5 1.3 1.5 ND 1.3 1.3 1.4 1.3 5 ND ND 1.4 1.7 ND 1.3 1.3 1.4 1.4 7 ND ND 1.4 1.5 ND 1.3 1.4 1.3 1.3

1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677

Table 20 shows the results of the antimicrobial activity of the fermented cereal fermented with a single strain and a complex strain. The antimicrobial activity was not observed in the 0 day old control, but the results of antimicrobial activity were different according to the fermentation period of each strain. Among them, acne strain Staphylococcus Epidermidis epidermidis) was trying Pedy five caucuses in the fermentation of a single strain KCCM40416 strain Sidi rakti City (Pediococcus acidilactic ) SRCM 102024 showed the highest antimicrobial activity as 2.9 cm.

Antibacterial activity of cereal fermentation on Staphylococcus epidermidis strain KCCM40416 (cm) Day One 2 3 4 5 6 7 8 9 0 ND ND ND ND ND ND ND ND ND 3 ND ND 2.3 2.3 ND 2.5 2.9 2.5 2.6 5 ND ND 2.6 2.8 ND 2.6 2.8 2.6 2.7 7 ND ND 2.6 2.9 ND 2.7 2.6 2.5 2.4

1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677

Table 21 shows the results of the antimicrobial activity of the fermented cereal grains fermented with different strains. The antimicrobial activity was not observed in the 0 day old control, but the results of antimicrobial activity were different according to the fermentation period of each strain. As the star of acne strains Philo Caucus epidermidis (Staphylcoccus epidermidis) was trying Pedy five caucuses in the fermentation of a single strain KCCM35494 strain Sidi rakti City (Pediococcus acidilactici ) SRCM 102024 showed the highest antimicrobial activity at 3.8cm and showed the highest antimicrobial activity at the 5th day of fermentation. This may be due to the fact that Starfilacococcus epidermidis The antimicrobial activity of KCCM40416 strain was much higher than 2.8cm.

Antibacterial activity of cereal fermentation on Staphylococcus epidermidis strain KCCM35494 (cm) Day One 2 3 4 5 6 7 8 9 0 ND ND ND ND ND ND ND ND ND 3 ND ND 2.6 3.5 ND 2.6 3.6 2.8 2.6 5 ND ND 3.0 3.8 ND 3.5 3.6 3.7 2.8 7 ND ND 2.9 3.4 ND 3.2 3.6 3.0 2.8

1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stokes meencereroides SRCM 101677, 9: Pediococcus acedilactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677

(11) Total number of lactic acid bacteria after grain fermentation

Table 22 shows the number of lactic acid bacteria in the fermented cereal fermented with a single strain and a complex strain. In this study, the control value of the untreated control was ND at day 0 and ND at day 7. On the 0th day of fermentation, the fermentation progressed from 1.3 × 10 ⁷ CFU / ㎖ to 4.6 × 10 ⁷ CFU / ㎖, 2.1 × 10 ⁷ CFU / ㎖, 9.5 × 10 ⁷ CFU / ^{7 CFU / ㎖, 2.1 × 10} 7 showed a tendency to increase in CFU / ㎖, the more the grain advanced to the composite strain even zero primary fermentation in a 1.3 × 10 ⁷ CFU / ㎖ progress ^{2.5 × 10 7 CFU / ㎖,} 7.0 × 10 ⁷ CFU / ml, and 3.8 × 10 ⁷ CFU / ml, respectively. Of the 9 strains, Pediococcus aceyltracis SRCM 102024 showed the highest number of lactic acid bacteria. The fermentation time of fermented grains increased to 1.4 × 10 ⁸ CFU / ㎖ at the 3rd day of fermentation, then increased to 4.6 × 10 ⁸ CFU / ㎖ at 5th day and decreased to 9.5 × 10 ⁷ CFU / ㎖ at 7th day .

Total number of fermented lactic acid bacteria in grain (CFU / ml) Day One 2 3 4 5 6 7 8 9 0 ND 1.3 x 10 ⁷ 1.3 x 10 ⁷ 1.3 x 10 ⁷ 1.3 x 10 ⁷ 1.3 x 10 ⁷ 1.3 x 10 ⁷ 1.3 x 10 ⁷ 1.3 x 10 ⁷ 3 ND 2.0 x 10 ⁸ 2.2 x 10 ⁸ 1.4 x 10 ⁸ 1.5 × 10 ⁸ 2.4 × 10 ⁸ 1.4 x 10 ⁸ 1.4 x 10 ⁸ 1.8 x 10 ⁸ 5 ND 7.6 × 10 ⁷ 5.5 × 10 ⁷ 4.6 × 10 ⁸ 3.6 × 10 ⁷ 6.6 x 10 ⁷ 6.6 x 10 ⁷ 1.4 x 10 ⁸ 5.6 x 10 ⁷ 7 ND 4.6 × 10 ⁷ 2.1 × 10 ⁷ 9.5 × 10 ⁷ 3.3 × 10 ⁷ 2.1 × 10 ⁷ 2.5 x 10 ⁷ 7.0 × 10 ⁷ 3.8 × 10 ⁷

1: Blank, 2: Lactobacillus plantarum SRCM 101502, 3: Lactobacillus plantarum SRCM 101503, 4: Pediococcus asiatici SRCM 102024, 5: Ryukono Stokes meencereroides SRCM 101665, 6: Leuconostomensee terroidis SRCM 101677, 7: Pediococcus acidifactii SRCM 102024 + Lactobacillus plantarum SRCM 101503, 8: Pediococcus asiatici SRCM 102024 + Ryukono Stock Methenseeroides SRCM 101677, 9: Pediococcus acidiflactii SRCM 102024 + Lactobacillus plantarum SRCM 101503 + Ryukono Stock Methenseeroides SRCM 101677

Korea Microorganism Conservation Center (overseas) KCCM12019P 20170420

<110> Microbial Institute for Fermentation Industry          SHIN, Yong Sung          LEE, Im Sook <120> Pediococcus acidilactici SRCM 102024 starter strain for producing          fermented grain with improved antimicrobial activity and uses          the <130> PN17191 <160> 5 <170> Kopatentin 2.0 <210> 1 <211> 1552 <212> DNA <213> Pediococcus acidilactici <400> 1 ttttcggggt tcccctgtac gacttcaccc taatcatctg tcccacctta gacggctagc 60 tcctaaaagg ttaccccacc ggctttgggt gttacaaact ctcatggtgt gacgggcggt 120 gtgtacaagg cccgggaacg tattcaccgc ggcatgctga tccgcgatta ctagcgattc 180 cgacttcgtg taggcgagtt gcagcctaca gtccgaactg agaatggttt taagagatta 240 gctaaacctc gcggtttcgc aactcgttgt accatccatt gtagcacgtg tgtagcccag 300 gtcataaggg gcatgatgat ttgacgtcgt ccccaccttc ctccggtttg tcaccggcag 360 tctcactaga gtgcccaact gaatgctggc aactagtaat aagggttgcg ctcgttgcgg 420 gacttaaccc aacatctcac gacacgagct gacgacaacc atgcaccacc tgtcattctg 480 tccccgaagg gaacgcctaa tctcttaggt tggcagaaga tgtcaagacc tggtaaggtt 540 cttcgcgtag cttcgaatta aaccacatgc tccaccgctt gtgcgggccc ccgtcaattc 600 ttttgagttt caaccttgcg gtcgtactcc ccaggcggat tacttaatgc gttagctgca 660 gcactgaagg gcggaaaccc tccaacactt agtaatcatc gtttacggca tggactacca 720 gggtatctaa tcctgttcgc tacccatgct ttcgagcctc agcgtcagtt acagaccaga 780 cagccgccct tcgccactgg tgttcttcca tatatctacg catttcaccg ctacacatgg 840 agttccactg tcctcttctg cactcaagtc tcccagtttc caatgcactt cttcggttga 900 gccgaaggct ttcacattag acttaaaaga ccgcctgcgc tcgctttacg cccaataaat 960 ccggataacg cttgccacct acgtattacc gcggctgctg gcacgtagtt agccgtggct 1020 ttctggttaa ataccgtcac tgggtgaaca gttactctca cccacgttct tctttaacaa 1080 cagagcttta cgagccgaaa cccttcttca ctcacgcggc gttgctccat cagacttgcg 1140 tccattgtgg aagattccct actgctgcct cccgtaggag tctgggccgt gtctcagtcc 1200 caatgtggcc gattaccctc tcaggtcggc tacgcatcat cgccttggtg agccgttacc 1260 tcaccaacta gctaatgcgc cgcgggtcca tccagaagtg atagcagagc catcttttaa 1320 aagaaaacca ggcggttttc tctgttatac ggtattagca tctgtttcca ggtgttatcc 1380 cctgcttctg ggcaggttac ccacgtgtta ctcacccgtc cgccactcac ttcgtgttaa 1440 aatctcattc agtgcaagca cgtcatgatc aattaacgga agttcgttcg acttgcatgt 1500 attaggcacg ccgccagcgt tcatcctgag cagagtttta aaatctataa gg 1552 <210> 2 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 ggattagata ccctggta 18 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 agagtttgat cmtggctcag 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 ccgtcaattc mtttragttt 20 <210> 5 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 5 tacggytacc ttgttacgac tt 22

Claims (7)

A Pediococcus acidilactici SRCM having antibacterial activity against Propionibacterium acnes or Staphylococcus epidermidis and having a deposit number of KCCM12019P isolated from a fruit fermentation broth, 102024 strain. delete A starter composition for the production of a fermented grain consisting of germinated barley and brown rice containing the strain Pediococcus acidilactici SRCM 102024 of claim 1 or a culture thereof as an active ingredient. A method for producing Pediococcus acidilactici SRCM 102024 of claim 1 or Propionibacterium acnes or Star produced by fermenting germinated barley and brown rice using a culture thereof as a starter strain, A fermented grain consisting of germinated barley and brown rice having antimicrobial activity against Staphylococcus epidermidis . 5. The fermented grain according to claim 4, wherein the fermented grain consisting of germinated barley and brown rice has an amino nitrogen, an organic acid, an organic sugar, and a free amino acid content in comparison with a control group not treated with Pediococcus acidilactici SRCM 102024 , The inhibitory activity of alpha -glucosidase and the activity of protease are increased, and the total number of lactic acid bacteria is increased, the fermented grain comprising germinated barley and brown rice. Prevention of acne or atopic dermatitis containing active ingredients of germinated barley and brown rice having antimicrobial activity against Propionibacterium acnes or Staphylococcus epidermidis of claim 4, Or improving health functional food composition. Improvement of acne or atopic dermatitis comprising as an active ingredient fermented cereals comprising germinated barley and brown rice having antimicrobial activity against Propionibacterium acnes or Staphylococcus epidermidis of claim 4 / RTI &gt;

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