KR102642548B1 - Composition for improving relieving hangover and liver function comprising the Pediococcus acidilactici WiKim0170 - Google Patents

Abstract

Pediococcus esidilactici WiKim0170 according to the present invention is a lactic acid bacterium isolated from makgeolli, and has excellent effects in reducing blood alcohol and improving liver damage. Therefore, it is used for relieving hangovers and improving liver function containing lactic acid bacteria as an active ingredient. It can be used in various ways.

Description

Composition for improving hangover and liver function comprising Pediococcus acidilactici WiKim0170 {Composition for improving relieving hangover and liver function comprising the Pediococcus acidilactici WiKim0170}

The present invention relates to Pediococcus esidilactisi WiKim0170, a lactic acid bacterium isolated from makgeolli, and a composition containing the same for relieving hangovers and improving liver function.

According to the Ministry of Food and Drug Safety's recent survey on alcohol consumption and intake in 2020, the average amount of alcohol consumed per time and the frequency of drinking have decreased due to the impact of COVID-19, but 'honsul (alcohol consumed alone)' and 'home alcohol (drinked at home)' have decreased. It is reported that there has been a change in drinking culture, such as an increase. In particular, the overall amount of drinking and alcohol consumption has decreased due to reasons such as the COVID-19 incident and the 52-hour workweek system, but the demand for hangover relievers is increasing due to changes such as the increase in high-risk drinking, drinking alone, and people drinking at home. .

A hangover refers to symptoms such as headache, diarrhea, loss of appetite, nausea, vomiting, chills, and cold sweat that appear after drinking alcohol. Objective symptoms include decreased cognition and motor skills, hematological changes, and hormonal changes. It is known that the causes of hangovers are dehydration, toxicity of alcohol and alcohol metabolites (acetaldehyde, formaldehyde, acetone, etc.), and nutrient deficiencies (blood sugar, vitamin, and mineral deficiencies) due to malabsorption. The degree of hangover varies greatly depending on individual differences in genetics and environmental conditions (nutrition, exercise, dehydration, and health).

After drinking, alcohol is metabolized through three pathways: when the concentration of ethanol is low, by the action of alcohol dehydrogenase (Acetaldehyde dehydrogenase) present in the gastrointestinal tract or liver, and when the concentration of ethanol is high, by the microsomal ethanol oxidation system present in the endoplasmic reticulum. It is metabolized into acetaldehyde and acetic acid by (MEOS: Microsomal ethanol oxidizing system), and then converted into carbon dioxide (CO ₂ ) and water (H ₂ O) through the action of catalase present in peroxisomes. is finally decomposed. When an appropriate amount of alcohol is introduced, the metabolic system described above operates smoothly and various alcohol-related symptoms do not occur. However, when a large amount of alcohol is introduced, the balance of the metabolic system is destroyed, making it impossible to maintain biological homeostasis, which can cause headaches or headaches in the short term. It causes symptoms such as loss of concentration, decreased concentration, heartburn, and indigestion, and in the long term, liver dysfunction may occur.

Meanwhile, the snack snack market size has grown significantly to KRW 700 billion as of 2019, and the dry snack market size has increased sales by 19.7% as of September 2020. Accordingly, in addition to existing formulations of hangover-relieving functional lactic acid bacteria, there is a need for the development of new types of products that can be applied to snacks and dry snacks, and probiotics with hangover-relieving function that have alcohol and acetaldehyde decomposition activity using lactic acid bacteria. There is a need to develop food containing hangover relievers.

Accordingly, the present inventors studied lactic acid bacteria with alcohol and acetaldehyde decomposition activity and compositions containing the same, and as a result, it was found that the Pediococcus esidilactisi WiKim0170 strain and the composition containing the same have alcohol decomposing enzyme activity, blood alcohol reduction, and liver function. It was confirmed that it was effective in improving damage.

The purpose of the present invention is to provide a strain of Pediococcus acidilactici WiKim0170 ( Pediococcus acidilactici WiKim0170) with accession number KCCM13113P.

Another object of the present invention is to provide a food composition for relieving a hangover containing Pediococcus acidilactici WiKim0170 (Pediococcus acidilactici WiKim0170) of accession number KCCM13113P, its culture, its lysate, or its extract as an active ingredient.

Another object of the present invention is to provide a food composition for improving liver function containing Pediococcus acidilactici WiKim0170 (Pediococcus acidilactici WiKim0170) of accession number KCCM13113P, its culture, its lysate, or its extract as an active ingredient. It is for this purpose.

Another object of the present invention is to provide a health functional food composition for improving liver function containing Pediococcus acidilactici WiKim0170 (Pediococcus acidilactici WiKim0170) of accession number KCCM13113P as an active ingredient.

Another object of the present invention is to provide a lactic acid bacteria starter containing Pediococcus acidilactici WiKim0170 (Pediococcus acidilactici WiKim0170) of accession number KCCM13113P, its culture, its lysate, or its extract as an active ingredient.

In addition, another object of the present invention is to provide a pharmaceutical composition for improving liver function containing Pediococcus acidilactici WiKim0170 (Pediococcus acidilactici WiKim0170) of accession number KCCM13113P, its culture, its lysate, or its extract as an active ingredient. It is provided.

Accordingly, the present inventors tried to find a lactic acid bacteria strain that shows excellent effects in alcohol decomposition enzyme activity, blood alcohol reduction, and liver damage improvement from traditional fermented foods, and as a result, Pediococcus strain of the new Pediococcus genus Lactobacillus strain, accession number KCCM13113P, was found. The present invention was completed by isolating and identifying the Pediococcus acidilactici WiKim0170 strain.

Hereinafter, the present invention will be described in more detail.

One aspect of the present invention is Pediococcus acidilactici WiKim0170 ( Pediococcus acidilactici WiKim0170) strain with accession number KCCM13113P.

In the present invention, Pediococcus esidilactici WiKim0170 is a Gram-positive bacterium, a facultative anaerobe capable of growing under both aerobic and anaerobic conditions, is non-motile, and its cells take the form of a coccus. Catalase and indole producing abilities are not observed.

In the present invention, the Pediococcus esidilactici strain WiKim0170 may have the 16S rDNA base sequence of SEQ ID NO: 1, but is not limited thereto.

In the present invention, the Pediococcus esidilactici strain WiKim0170 may exist as live or dead cells, and may also exist in dried or freeze-dried form. In addition, a culture of the Pediococcus esidilactisi WiKim0170 strain may be an active ingredient, and the culture may include a live cell culture medium or a dead cell supernatant. Forms and formulation methods of lactic acid bacteria suitable for inclusion in various compositions are well known to those skilled in the art. For example, the Pediococcus esidilactici WiKim0170 strain is a culture obtained by culturing the strain in a known liquid medium or solid medium, a fermentation product obtained by culturing the strain with additional components, or the strain is organically produced. It can be formulated in the form of an extract extracted with a solvent, a lysate (or lysate) obtained by dissolving the cell membrane of the strain, or crushed or homogenized, but is not limited thereto.

According to the following examples, it was confirmed that the Pediococcus esidilactici WiKim0170 strain is resistant to alcohol, is effective in reducing blood alcohol and improving hangovers, and has an alcohol decomposition effect.

In addition, Pediococcus esidilactisi WiKim0170 strain is effective in improving liver damage, reducing alanine aminotransferase (ALT) and aspartate aminotransferase (AST), which are indicators of liver damage in the blood. represents. In addition, it shows a high intestinal cell adhesion ability and a stable effect in the intestine.

Pediococcus esidilactisi WiKim0170 strain isolated and identified in the following examples shows acid resistance or bile resistance.

Another aspect of the present invention is a food composition for relieving a hangover containing Pediococcus acidilactici WiKim0170 (Pediococcus acidilactici WiKim0170) of accession number KCCM13113P, its culture, its lysate, or its extract as an active ingredient.

As used herein, the term "hangover" refers to temporary palpitations, headache, thirst, motion sickness, and gastrointestinal disorders caused by ethanol accumulated in the body without being decomposed into carbon dioxide and water due to excessive alcohol consumption, and acetaldehyde, a by-product of alcohol decomposition. , refers to phenomena such as general malaise, diarrhea, vomiting, abdominal bloating, indigestion, or vitamin deficiency.

In the present invention, the food composition for relieving hangovers includes milk thistle ( Silybum marianum ) extract, shiitake mushroom mycelium ( Lentinula edodes mycelium) extract, Ecklonia stolonifera extract, dogwood tree ( Lonicera caerulea ) fruit extract, bellflower root ( Platycodon grandiflorus ) It may further include one or more selected from the group consisting of extract, Rubus coreanus extract, lactic acid fermented kelp ( Saccharina japonica ) extract, lactic acid fermented garlic extract, and Hovenia dulcis fruit and fruit extract. However, it is not limited to this.

In the present invention, the food composition may include a beverage or bar.

In the present invention, the food composition can be used as a functional food or added to various foods. Foods that can be added to the composition of the present invention include, for example, beverages, vitamin complexes, and health supplements.

The food composition of the present invention may include ingredients commonly added during food production, and includes, for example, proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose, etc.; Disaccharides such as maltose, sucrose, oligosaccharides, etc.; and polysaccharides, such as common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is manufactured as a drink or beverage, citric acid, high fructose corn syrup, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be additionally included.

The above food composition may additionally contain food additives, and its suitability as a “food additive” is determined by the specifications for the item in accordance with the general provisions and general test methods of the food additive code approved by the Ministry of Food and Drug Safety, unless otherwise specified. and standards.

Items listed in the "Food Additives Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as subchromic pigment, licorice extract, crystalline cellulose, and guar gum, L- Examples include mixed preparations such as sodium glutamate preparations, noodle additive alkaline preparations, preservative preparations, and tar coloring preparations.

Foods containing the active ingredient of the present invention include confectionery such as bread, rice cake, dried fruit, candy, chocolate, chewing gum, and jam, ice cream products such as ice cream, frozen confectionery, and ice cream powder, milk, low-fat milk, lactose-digested milk, Dairy products such as processed milk, goat milk, fermented milk, butter milk, concentrated milk, milk cream, butter oil, natural cheese, processed cheese, powdered milk, and whey Processed meat products, processed egg products, meat products such as hamburgers Fish cakes, ham, Fish products such as processed fish products such as sausage and bacon, noodles such as ramen, dried noodles, fresh noodles, fried noodles, deluxe dried noodles, modified overnight noodles, frozen noodles, pasta, fruit drinks, vegetable drinks, carbonated drinks, soy milk, Lactobacillus beverages such as yogurt, beverages such as mixed drinks, seasonings such as soy sauce, soybean paste, red pepper paste, chunjang, cheonggukjang, mixed soybean paste, vinegar, sauces, tomato ketchup, curry, and dressing, but limited to margarine, shortening, and pizza. It doesn't work.

In addition to the above, the composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, It may include carbonating agents used in carbonated beverages. Additionally, the composition of the present invention may include pulp for the production of natural fruit juice, fruit juice beverages, and vegetable beverages. These ingredients can be used independently or in combination.

The beverage composition containing the active ingredient of the present invention has no particular limitations on other ingredients and may contain various flavoring agents or natural carbohydrates as additional ingredients like a conventional beverage. Examples of the above-mentioned natural carbohydrates include monosaccharides, (e.g., glucose, fructose, etc.); Disaccharides, (e.g. maltose, sucrose, etc.); and polysaccharides, common sugars such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. .

In the present invention, the amount of Pediococcus esidilactici WiKim0170 included in the composition may be about 10 ⁶ to 10 ¹² cfu/g, for example, 10 ⁷ to 10 ¹¹ cfu/g, 10 It may be ⁸ to 10 ¹⁰ cfu/g. When administering a strain, it is preferable to administer it in a live state, and it can be killed before ingestion or administered in an attenuated state. In addition, when manufacturing using culture supernatant, etc., it may additionally undergo a sterilization process through a heat treatment process. The amount of strain and daily intake required to have minimal efficacy may vary depending on the body or health condition of the ingestor, but may generally be about 10 ⁶ to 10 ¹² cfu/g, for example, 10 ⁷ to 10 ¹¹ cfu/g, may be 10 ⁸ to 10 ¹⁰ cfu/g.

Another aspect of the present invention is a food composition for improving liver function comprising Pediococcus acidilactici WiKim0170 (Pediococcus acidilactici WiKim0170) of accession number KCCM13113P, its culture, its lysate, or its extract as an active ingredient.

As used herein, the term “liver function improvement” means preventing, treating, or improving liver function decline or liver damage. Additionally, improving liver function may indicate that it supports or protects liver health.

In the present invention, improving liver function may mean protecting the liver from alcoholic damage, but is not limited thereto.

In the present invention, the food composition for improving liver function includes milk thistle ( Silybum marianum ) extract, shiitake mushroom mycelium ( Lentinula edodes mycelium) extract, Ecklonia stolonifera extract, dogwood ( Lonicera caerulea ) fruit extract, bellflower root ( Platycodon grandiflorus) ) extract, Rubus coreanus extract, lactic acid fermented kelp ( Saccharina japonica ) extract, lactic acid fermented garlic extract, and Hovenia dulcis fruit and fruit extract. However, it is not limited to this.

Another aspect of the present invention is a health functional food composition for improving liver function containing Pediococcus acidilactici WiKim0170 (Pediococcus acidilactici WiKim0170) with accession number KCCM13113P as an active ingredient.

In this specification, the term "health functional food" refers to food manufactured and processed using raw materials or ingredients with functionality useful to the human body in accordance with the Act on Health Functional Food (Article 3, Paragraph 1), and "functional" This means adjusting nutrients to the structure and function of the human body or obtaining useful effects for health purposes such as physiological effects (Article No. 2).

In the present invention, the health functional food composition for improving liver function includes milk thistle ( Silybum marianum ) extract, shiitake mushroom mycelium ( Lentinula edodes mycelium) extract, Ecklonia stolonifera extract, dogwood tree ( Lonicera caerulea ) fruit extract, bellflower root ( Platycodon grandiflorus ) extract, bokbunja ( Rubus coreanus ) extract, lactic acid fermented kelp ( Saccharina japonica ) extract, lactic acid bacteria fermented garlic extract, and Hovenia dulcis fruit and bottle extract. It may be done, but it is not limited to this.

Another aspect of the present invention is a lactic acid bacteria starter containing Pediococcus acidilactici WiKim0170 (Pediococcus acidilactici WiKim0170) of accession number KCCM13113P, its culture, its lysate, or its extract as an active ingredient.

As used herein, the term "lactic acid bacteria starter" refers to the first start of fermentation in which lactic acid is produced through the decomposition of lactose in milk components by lactic acid bacteria. It means using after culturing and proliferating lactic acid bacteria stored as starter to manufacture an induced effect product. In the present invention, it means using after culturing and proliferating Pediococcus esidilactici WiKim0170.

In the present invention, the lactic acid bacteria starter can inhibit the growth of various bacteria by fermenting lactose into lactic acid and lowering the pH.

The lactic acid bacteria starter composition of the present invention can be used for promoting the health of humans and animals, that is, a composition for intestinal tract, a probiotic (probiotic) composition, or a lactic acid bacteria preparation, and the composition or preparation is a shredded cell wall fraction of the lactic acid bacteria of the present invention, live bacteria. , may contain dead cells, dried bacteria, or cultures as active ingredients, and may additionally include excipients or carriers. The culture includes the culture itself cultured in a liquid medium, a filtrate obtained by filtering or centrifuging the culture to remove strains (centrifuged supernatant), etc. The content of lactic acid bacteria starter in the composition may vary depending on the use and formulation of the composition. The enteral or probiotic composition according to the present invention can be prepared and administered in various formulations and methods, and can be prepared using Pediococcus esidilactisi WiKim0170 strain alone; its culture; Or Pediococcus esidilactisi WiKim0170 strain Lactobacillus acidophilus ( Lactobacillus acidophilus ) , Lactobacillus casei ( Lactobacillus casei ), Lactobacillus gasseri ( Lactobacillus gasseri ), Lactobacillus delbruecki ssp. Bulgaricus ( Lactobacillus delbrueckii ssp. bulgaricus ), Lactobacillus helveticus , Lactobacillus fermentum , Lactobacillus paracasei , Lactobacillus plantarum , Lactobacillus Lactobacillus reuteri , Lactobacillus rhamnosus , Lactobacillus salivarius , Lactococcus lactis , Enterococcus faecium , Enterococcus faecalis , Streptococcus thermophilus , Bifidobacterium bifidum , Bifidobacterium breve , Bifidobacterium longum and Bifidobacterium animalis ssp. A mixture containing one or more probiotics selected from Bifidobacterium animalis ssp. lactis is mixed with a carrier and flavoring agent commonly used in the pharmaceutical field to form tablets, troches, capsules, It can be manufactured and administered in the form of elixir, syrup, powder, suspension, or granule. The carrier may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, and suspending agents. The administration method may be oral, parenteral, or application, but oral administration is preferred. Additionally, the administered dose can be appropriately selected depending on the absorption, inactivation rate, and excretion rate of the active ingredient in the body, and the age, gender, and condition of the recipient.

In the present invention, the lactic acid bacteria starter includes milk thistle ( Silybum marianum ) extract, shiitake mushroom mycelium ( Lentinula edodes mycelium ) extract, Ecklonia stolonifera extract, dogwood ( Lonicera caerulea ) fruit extract, bellflower root ( Platycodon grandiflorus ) extract, and bokbunja. ( Rubus coreanus ) extract, lactic acid bacteria-fermented kelp ( Saccharina japonica ) extract, lactic acid bacteria-fermented garlic extract, and Hovenia dulcis ( Hovenia dulcis ) fruit and fruit extract may additionally be included. It is not limited.

In the present invention, the lactic acid bacteria starter can be used for adjuvant treatment of liver disease, but is not limited thereto.

Another aspect of the present invention is a pharmaceutical composition for improving liver function containing Pediococcus acidilactici WiKim0170 (Pediococcus acidilactici WiKim0170) of accession number KCCM13113P, its culture, its lysate, or its extract as an active ingredient.

According to the following examples, it was confirmed that the Pediococcus esidilactici WiKim0170 strain is resistant to alcohol, is effective in reducing blood alcohol and improving hangovers, and has an alcohol decomposition effect.

In addition, Pediococcus esidilactisi WiKim0170 strain is effective in improving liver damage, reducing alanine aminotransferase (ALT) and aspartate aminotransferase (AST), which are indicators of liver damage in the blood. represents. In addition, it shows high intestinal cell adhesion ability and has a stable effect in the intestine.

Pediococcus esidilactisi WiKim0170 strain isolated and identified in the following examples shows acid resistance or bile resistance.

As used herein, the term “liver function improvement” means preventing, treating, or improving liver function decline or liver damage. Additionally, improving liver function may indicate that it supports or protects liver health.

In the present invention, improving liver function may mean protecting the liver from alcoholic damage, but is not limited to this.

In the present invention, the pharmaceutical composition for improving liver function can be used as an adjuvant therapy that can resolve a patient's liver disease, an agent for improving chronic liver disease patients, or an agent for improving liver function in alcoholism patients. .

When used as a pharmaceutical composition in the present invention, the pharmaceutical composition of the present invention can be prepared using pharmaceutically suitable and physiologically acceptable adjuvants in addition to the Pediococcus esidilactici WiKim0170 strain, As auxiliaries, excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, lubricants, or flavoring agents can be used.

For administration, the pharmaceutical composition may be preferably formulated as a pharmaceutical composition containing one or more pharmaceutically acceptable carriers in addition to the active ingredients described above.

For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, etc. Additionally, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tracacance or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate. Includes sodium acetate, sodium chloride, etc. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum, etc. Acceptable pharmaceutical carriers for compositions formulated as liquid solutions include those that are sterile and biocompatible, such as saline solution, sterile water, Ringer's solution, buffered saline solution, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these ingredients can be mixed and used, and other common additives such as antioxidants, buffers, and bacteriostatic agents can be added as needed. In addition, diluents, dispersants, surfactants, binders, and lubricants can be additionally added to formulate injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

The advantages and features of the present invention and methods for achieving them will become clear with reference to the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below and will be implemented in various different forms. The present embodiments are merely provided to ensure that the disclosure of the present invention is complete and to provide common knowledge in the technical field to which the present invention pertains. It is provided to fully inform those who have the scope of the invention, and the present invention is only defined by the scope of the claims.

Pediococcus esidilactici WiKim0170 according to the present invention is a lactic acid bacterium isolated from makgeolli, and has excellent effects in reducing blood alcohol and improving liver damage. Therefore, it is used for relieving hangovers and improving liver function containing lactic acid bacteria as an active ingredient. It can be used in various ways.

Figure 1 is a graph showing the results of comparing the growth of Pediococcus acidilactici WiKim0170 ( Pediococcus acidilactici WiKim0170) by alcohol content according to an embodiment of the present invention.
Figure 2 is a graph showing the results of comparing the alcohol degrading enzyme (ADH) activity of Pediococcus esidilactisi WiKim0170 according to an embodiment of the present invention.
Figure 3 is a graph showing the results of comparing the acetaldehyde degrading enzyme (ALDH) activity of Pediococcus esidilactisi WiKim0170 according to an embodiment of the present invention.
Figure 4 is a graph showing the results of measuring blood alcohol concentration in an animal test model administered Pediococcus esidilactisi WiKim0170 according to an embodiment of the present invention.
Figure 5 is a graph showing the change in blood concentration of aspartate aminotransferase (AST), an indicator of liver damage, over time in the blood of an animal model according to an embodiment of the present invention.
Figure 6 is a graph showing the change in blood concentration of alanine aminotransferase (ALT), an indicator of liver damage, over time in an animal model according to an embodiment of the present invention.
Figure 7 is a graph showing the results of comparing and evaluating hepatocellular toxicity after treating Pediococcus esidilactisi WiKim0170 according to an embodiment of the present invention.
Figure 8 is a graph showing the acid resistance results of Pediococcus esidilactisi WiKim0170 according to an embodiment of the present invention.
Figure 9 is a graph showing the biliary resistance results of Pediococcus esidilactisi WiKim0170 according to an embodiment of the present invention.
Figure 10 is a graph showing the results of enterocyte adhesion ability of Pediococcus esidilactisi WiKim0170 according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail through examples. The following examples are merely illustrative of the present invention and the scope of the present invention is not limited to the following examples.

Example 1. Isolation and culture of Pediococcus esidilactisi WiKim0170

Lactic acid bacteria were isolated from makgeolli, inoculated into MRS liquid medium containing 10% (w/v) alcohol, and cultured at 30°C for 48 hours. Next, a pool of alcohol-resistant lactic acid bacteria resistant to 10% (w/v) alcohol content medium was spread on MRS agar medium and cultured at 30°C for 48 hours, and then the formed colonies were selected. As a result of analyzing the base sequence of the 16S rRNA of the selected colony, it was identified as a microorganism of the present invention having a 16S rDNA base sequence of SEQ ID No. 1, which was identified as Pediococcus acidilactici WiKim0170 ), and was deposited with the Korea Microbial Conservation Center (overseas) on January 18, 2022 (Accession Number: KCCM13113P)

Example 2. Evaluation of bacterial growth by alcohol content

Growth of Pediococcus acidilactici WiKim0170 (Pediococcus acidilactici WiKim0170) strain was evaluated by alcohol content.

Alcohol was added to MRS liquid medium at 0% (w/v), 7.5% (w/v), 10% (w/v), 12.5% (w/v), 15% (w/v) and 25% (w). /v) content, inoculated into MRS liquid medium, and cultured at 30°C for 48 hours. Next, the cultured bacterial solution was inoculated into MRS agar medium, and the growth of the strain was observed for 48 hours, 72 hours, and 120 hours, and the OD value was measured at an absorbance of 600 nm, and the results are shown in Figure 1.

As can be seen in Figure 1, Pediococcus esidilactici WiKim0170 was observed to grow at an OD of 0.6 or more at 600 nm even at an alcohol content of 15% (w/v), and was confirmed to be an alcohol-resistant lactic acid bacterium.

Example 3. Evaluation of alcohol degrading enzyme activity of strains

Alcohol is mainly broken down by the action of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) present in the liver. If it is not broken down, it can accumulate in the liver and cause various types of hangovers.

To evaluate the alcohol degrading enzyme activity of the strain, Pediococcus esidilactici WiKim0170 was cultured at 30°C for 48 hours and then alcohol degrading enzyme (ADH) and acetaldehyde degrading enzyme (ALDH) activities were observed. Alcohol degrading enzyme (ADH) activity was measured by absorbance measuring the amount of NADH product formed at 450 nm by reacting Pediococcus esidilactisi WiKim0170 with alcohol as a substrate (Figure 2). In addition, acetaldehyde degrading enzyme (ALDH) activity was confirmed by measuring the change in absorbance at 535 nm due to NADH production after reacting Pediococcus esidilactici WiKim0170 with ALDH (Figure 3).

As can be seen in Figures 2 and 3, Pediococcus esidilactisi WiKim0170 was observed to have alcohol degrading enzyme (ADH) activity of 137.2% compared to 100% of the control group, and acetaldehyde degrading enzyme (ALDH) activity was observed to be 224.9% compared to the PBS control group. was observed. Through this, the alcohol decomposition effect of Pediococcus esidilactici WiKim0170, which can be used to reduce blood alcohol and improve hangovers, was confirmed.

Example 4. Evaluation of blood alcohol reduction effect and liver damage improvement

Using experimental animals, the effect of reducing blood alcohol and the degree of improvement in liver damage were confirmed.

4-1. Evaluation of blood alcohol reduction effect

Pediococcus esidilactisi WiKim0170 was cultured and Pediococcus esidilactisi WiKim0170 strain and ethanol at a concentration of 25% (w/v) were administered to an animal model (Sprague-Dawley (SD) Rat, 6 weeks old) for 30 days. After 1, 60, 180, and 300 minutes, blood was collected from the tail and serum was separated. The serum of the experimental group administered Pediococcus esidilactisi WiKim0170 was mixed with ethanol assay buffer and reaction buffer and reacted for 30 minutes at room temperature blocked from light, and then the absorbance was measured at 570 nm. The control group in this experiment was compared to a group administered only ethanol to an animal model, and the results of the experiment are shown in Figure 4.

As can be seen in Figure 4, the blood alcohol reduction effect of the animal model administered Pediococcus esidilactici WiKim0170 was 31.5% after 30 minutes, 11.5% after 60 minutes, and 180 minutes, respectively, compared to the animal model administered only ethanol. It showed an alcohol reduction effect of 52.5% and 31.5% after 300 minutes.

4-2. Evaluation of liver damage improvement

Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are indicators of liver damage in the blood, and the degree of improvement in liver damage can be confirmed by measuring these two enzymes. Increased AST enzyme activity is known to be relatively highly correlated with the degree of hepatocellular dysfunction and is important as a marker of liver damage, while increased ALT enzyme activity is known to be relatively highly correlated with the degree of hepatocellular dysfunction and is important as a marker of liver damage.

Pediococcus esidilactisi WiKim0170 was cultured at 30°C for 48 hours, and then aspartate aminotransferase (AST) and alanine aminotransferase (ALT), indicators of liver damage, were measured in the blood using the same animal testing method as in Example 4-1. Measured. Pediococcus esidilactisi WiKim0170 was administered using serum from the experimental group and measured using biochemical analysis equipment. The relative decrease in the control group was compared with the group administered only ethanol.

Aspartate aminotransferase (AST), an indicator of liver damage, in the blood of experimental animals administered Pediococcus esidilactisi WiKim0170 decreased by 86.7% after 30 minutes, 35.3% after 60 minutes, respectively, compared to the group of experimental animals administered only ethanol. An AST reduction effect of 169.5% after 180 minutes and 378.9% after 300 minutes was confirmed (Figure 5).

In addition, alanine aminotransferase (ALT), an indicator of liver damage, in the blood of experimental animals administered Pediococcus esidilactisi WiKim0170 was confirmed to have a 38.6% ALT reduction effect after 300 minutes compared to the experimental animal group administered only ethanol. (Figure 6).

Example 5. Hepatocellular toxicity and safety evaluation

5-1. Hepatocellular toxicity assessment

HepG2 liver cancer cells were cultured, treated with Pediococcus esidilactisi WiKim0170, and after 48 hours, liver cell killing ability was observed based on 100% of the PBS-treated control group. The results are shown in Figure 7.

As can be seen in Figure 7, no hepatocellular toxicity was observed through co-culture of hepatocytes and Pediococcus esidilactici WiKim0170, and a survival rate similar to the control group was confirmed at 101.3% compared to the PBS control group.

5-2. Acid resistance and bile resistance assessment

For acid resistance, pH 2.5 MRS liquid medium was prepared with 1N HCl solution, and Pediococcus esidilactisi WiKim0170, cultured at 30°C for 48 hours, was inoculated and plated on MRS agar medium after 3 hours. Next, the number of viable bacteria formed after culturing at 30°C for 24 hours was expressed as cfu/ml, and it was judged to be an acid-resistant strain.

For bile resistance, make MRS liquid medium with Oxgall (difco) 1% (w/v), inoculate Pediococcus esidilactisi WiKim0170 cultured at 30°C for 48 hours, and spread on MRS agar medium after 6 hours. The number of viable bacteria formed after culturing at 30°C for 24 hours was expressed as cfu/ml, and the strain was judged to have bile resistance.

As a result of acid resistance confirmation, Pediococcus esidilactici WiKim0170 inoculated in MRS liquid medium corrected to ^{pH 2.5} at 1 8).

In addition, as a result of confirming bile resistance, Pediococcus esidilactisi WiKim0170 inoculated with 1% Oxgall (difco) in MRS liquid medium at 1 x 10 ⁸ cfu/ml was 3 x 10 ⁹ cfu/ml after 6 hours. Excellent bile resistance was confirmed (Figure 9).

5-3. Intestinal stability evaluation

Intestinal stability was evaluated through observation of enterocyte adhesion ability.

HT29 colon cancer cells were cultured, and Pediococcus esidilactisi WiKim0170, cultured at 30°C for 48 hours, was co-cultured for 2 hours, and then the cells were recovered and plated on MRS agar medium. Next, the plated medium was cultured at 30°C for 24 hours, and the number of viable cells formed was expressed in cfu/ml to observe enterocyte adhesion ability, and the results are shown in Figure 12.

As a result ^of confirming the ^stability in the intestine, 1 Intestinal stability was confirmed (Figure 12).

Example 6. Evaluation of mycological characteristics

The general mycological characteristics of Pediococcus esidilactisi WiKim0170 are that it is a Gram-positive bacterium, is a facultative anaerobic that can grow under both aerobic and anaerobic conditions, does not form spores, is non-motile, and its cells are similar to those of cocci. It is taking shape. And catalase and indole production were not observed. The growth temperature is 15°C to 45°C, and the optimal growth temperature is 30°C. The growth pH is 4.0 to 8.5, and the optimal growth pH is 6.0 to 6.5. Additional characteristics were observed through experiments using the API 50 CH kit for identification of lactic acid bacteria from Biomerieux, and the results are shown in Table 1 below.

number reaction substrate result number reaction substrate result One glycerol - 26 salicin - 2 Erythritol - 27 cellobiose + 3 D Arabinose - 28 maltose - 4 L Arabinose + 29 lactose - 5 Ribose D-Ribose + 30 melibius - 6 D xylose D-xylose + 31 saccharose - 7 L-xylose L-xylose - 32 trehalose + 8 Adonitol D-Adonitol - 33 Inulin - 9 Methyl-BD-xylofuranocycle - 34 Melezitos - 10 D-galactose + 35 Raffinose - 11 D-Glucose + 36 amidon - 12 D-fructose + 37 glycogen - 13 D-mannose + 38 xylitol - 14 L-sorbose - 39 gentiobios + 15 rhamnose - 40 D Turanos - 16 tulcitol - 41 D license - 17 inositol - 42 D tagatose + 18 Mannitol - 43 D fucose - 19 Sorbitol - 44 L fucose - 20 a-methyl-D-mannoside - 45 D-arabitol - 21 a-methyl-D-glucoside - 46 L-arabitol - 22 N-Acetyl-Glucosamine + 47 Gluconate - 23 Amygdalin - 48 2-keto-gluconate - 24 arbutin - 49 5-keto-gluconate - 25 Esculine +

Korea Center for Microbial Conservation (KCCM) KCCM13113P 20220118

Claims (11)

Pediococcus acidilactici WiKim0170 with accession number KCCM13113P strain. A food composition for improving liver function comprising Pediococcus acidilactici WiKim0170 with accession number KCCM13113P, its culture, its lysate, or its extract as an active ingredient. The method of claim 2, wherein the food composition for improving liver function is milk thistle ( Silybum marianum ) extract, shiitake mushroom mycelium ( Lentinula edodes mycelium) extract, Ecklonia stolonifera extract, dogwood tree ( Lonicera caerulea ) fruit extract, and bellflower root. ( Platycodon grandiflorus ) extract, bokbunja ( Rubus coreanus ) extract, lactic acid bacteria-fermented kelp ( Saccharina japonica ) extract, lactic acid bacteria-fermented garlic extract, and Hovenia dulcis ( Hovenia dulcis ) fruit and bottle extract, in addition to one or more selected from the group consisting of A food composition for improving liver function, comprising: A health functional food composition for improving liver function containing Pediococcus acidilactici WiKim0170 with accession number KCCM13113P as an active ingredient. The method of claim 4, wherein the health functional food composition for improving liver function includes milk thistle ( Silybum marianum ) extract, shiitake mushroom mycelium ( Lentinula edodes mycelium) extract, Ecklonia stolonifera extract, and dogwood ( Lonicera caerulea ) fruit extract. , bellflower root ( Platycodon grandiflorus ) extract, bokbunja ( Rubus coreanus ) extract, lactic acid fermented kelp ( Saccharina japonica ) extract, lactic acid fermented garlic extract, and Hovenia dulcis fruit and bottle extract. Additionally, a health functional food composition for improving liver function. A food composition for relieving a hangover containing Pediococcus acidilactici WiKim0170 with accession number KCCM13113P, its culture, its lysate, or its extract as an active ingredient. The method of claim 6, wherein the food composition for relieving a hangover is milk thistle ( Silybum marianum ) extract, shiitake mushroom mycelium ( Lentinula edodes mycelium) extract, Ecklonia stolonifera extract, dogwood tree ( Lonicera caerulea ) fruit extract, bellflower root ( Platycodon grandiflorus ) extract, bokbunja ( Rubus coreanus ) extract, lactic acid fermented kelp ( Saccharina japonica ) extract, lactic acid bacteria fermented garlic extract, and Hovenia dulcis fruit and bottle extract. A food composition for relieving hangovers. A lactic acid bacteria starter containing Pediococcus acidilactici WiKim0170 with accession number KCCM13113P, its culture, its lysate, or its extract as an active ingredient. The method of claim 8, wherein the lactic acid bacteria starter is milk thistle ( Silybum marianum ) extract, shiitake mushroom mycelium ( Lentinula edodes mycelium) extract, Ecklonia stolonifera extract, dogwood tree ( Lonicera caerulea ) fruit extract, bellflower root ( Platycodon grandiflorus ) extract, bokbunja ( Rubus coreanus ) extract, lactic acid fermented kelp ( Saccharina japonica ) extract, lactic acid fermented garlic extract, and Hovenia dulcis fruit and bottle extract. , Lactobacillus starter. A pharmaceutical composition for improving liver function in patients with chronic liver disease or alcoholism, comprising Pediococcus acidilactici WiKim0170 with accession number KCCM13113P, its culture, lysate thereof, or extract thereof as an active ingredient. The method of claim 10, wherein the pharmaceutical composition for improving liver function includes milk thistle ( Silybum marianum ) extract, shiitake mushroom mycelium ( Lentinula edodes mycelium) extract, Ecklonia stolonifera extract, dogwood tree ( Lonicera caerulea ) fruit extract, Add one or more selected from the group consisting of bellflower root ( Platycodon grandiflorus ) extract, bokbunja ( Rubus coreanus ) extract, lactic acid bacteria-fermented kelp ( Saccharina japonica ) extract, lactic acid bacteria-fermented garlic extract, and Hovenia dulcis fruit and bottle extract. A pharmaceutical composition for improving liver function in patients with chronic liver disease or alcoholism.