A novel Lactobacillus fermentum strain that selectively decomposes acetaldehyde and a hangover relief composition containing the strain as an active ingredient

Abstract

To provide a composition capable of removing or alleviating hangover symptoms after alcohol consumption.SOLUTION: The present invention relates to a novel lactic acid bacterium that selectively promotes the activity of an acetaldehyde decomposing enzyme that decomposes acetaldehyde, which causes hangover symptoms after alcohol consumption, to reduce the concentration of acetaldehyde in the blood and, more specifically, to a lactobacillus fermentum HDB1098 strain that selectively decomposes acetaldehyde and a composition for removing a hangover containing the same as an active ingredient.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention claims priority to Korean Patent Application No. 10-2023-0044425, filed with the Korean Intellectual Property Office on April 4, 2023, the disclosure of which is incorporated herein by reference.

[0002] The present invention relates to a novel lactic acid bacterium that selectively promotes the activity of acetaldehyde-degrading enzymes that break down acetaldehyde, which causes hangover symptoms after alcohol consumption, thereby reducing the concentration of acetaldehyde in the blood. More specifically, the present invention relates to a novel Lactobacillus fermentum strain HDB1098 that selectively decomposes acetaldehyde, and a composition for relieving hangovers that contains the strain as an active ingredient. [Background technology]

[0003] Alcohol is a favorite food that humans have used since ancient times for purposes such as social connection and stress relief. It is still widely consumed in various situations today, but after drinking alcohol, it can cause hangover symptoms such as nausea, vomiting, and headaches. For this reason, much research on hangovers has been conducted, and various products to alleviate hangovers have been released.

[0004] When ethanol, the main component of alcohol, enters the body, it is converted into acetaldehyde by alcohol dehydrogenase, cytochrome P450 2E1 (CYP2E1), and various catalases. It is then converted into acetate by acetaldehyde dehydrogenase. Finally, acetate is completely broken down into carbon dioxide and water. Acetaldehyde, produced by the oxidation of ethanol, is a toxic substance known to cause hangover symptoms such as flushing, increased heart rate, and headache.

[0005] However, 30-40% of Asians are unable to properly break down acetaldehyde. This is a genetic problem, and the presence of an acetaldehyde dehydrogenase subtype (acetaldehyde dehydrogenase 2 mutation; ALDH2*2) plays a significant role. While this genetic factor does not cause significant problems in daily life, when alcohol is ingested, it is converted to acetaldehyde by alcohol-degrading enzymes. However, the conversion to acetate is reduced, resulting in the accumulation of acetaldehyde in the liver and subsequent toxin formation. While small amounts of alcohol can be broken down with limited enzymes, excessive drinking, due to this genetic problem, prevents the acetaldehyde broken down by alcohol-degrading enzymes from being converted to acetate, resulting in further accumulation of toxins in the liver (Chen et al., Physiol Rev 94:1-34, 2014). In other words, low acetaldehyde-degrading enzyme activity combined with high alcohol-degrading enzyme activity can increase blood acetaldehyde levels, potentially worsening hangover symptoms caused by acetaldehyde toxin. Therefore, activating only acetaldehyde-degrading enzymes to rapidly remove toxins that accumulate in the liver, rather than simultaneously activating alcohol-degrading enzymes and acetaldehyde-degrading enzymes, may be more effective for people with acetaldehyde-degrading enzyme subtypes.

[0006] According to a 2015 report by the National Health Insurance Service of Korea, the socioeconomic losses caused by drinking alcohol amounted to 9,452.4 billion won per year, and are showing a tendency to increase every year. For this reason, various hangover relief products have been developed, but based on old literature, the Ministry of Food and Drug Safety of Korea (MFDS) has determined that there is insufficient evidence for hangover relief from currently available products. As a result, the Ministry of Food and Drug Safety (MFDS) has strengthened regulations and established guidelines to only allow the sale of hangover relief products that include scientific evidence of their hangover relief efficacy.

[0007] Meanwhile, the WHO has defined probiotics as "live microorganisms that, when ingested in adequate amounts, confer beneficial health effects on the host." However, through numerous studies, the definition has expanded to include not only microorganisms but also their culture components, and "dead cells using lactic acid bacteria" are now included in the probiotics category.

[0008] In line with this change, the Ministry of Food and Drug Safety in Korea has also defined probiotics as above, and the 19 types of lactic acid bacteria announced by the Ministry of Food and Drug Safety include the genus Lactobacillus, which includes Lactobacillus acidophilus and Lactobacillus gasseri; the genus Lactococcus, which includes Lactococcus lactis; the genus Enterococcus, which includes Enterococcus faecalis and Enterococcus spasium; the genus Streptococcus, which includes Streptococcus thermophilus; and the genus Bifidobacterium, which includes Bifidobacterium bifidum and Bifidobacterium brebu.

[0009] Korean Patent Registration No. 10-2262646 discloses a composition for preventing or relieving hangovers containing killed cells of Lactobacillus plantarum V135 strain as an active ingredient, and Korean Patent Registration No. 10-2251295 discloses a composition for preventing or relieving hangovers containing killed cells of Lactobacillus salivarius V133 strain as an active ingredient. Korean Patent Publication No. 10-2022-0050257 discloses a novel Lactobacillus fermentum EFEL6800 fermentation starter with probiotic function and fermented vegetables using the same. Korean Patent Publication No. 10-2022-0114993 discloses a novel Lactobacillus fermentum CJNU 1840 strain and a novel microorganism resistant to coffee extract and a bioconverted coffee extract using the same. In addition, Korean Patent Publication No. 10-2004-0104153 discloses lactic acid bacteria with alcohol-degrading ability, dairy products containing the same, functional health foods, and live Lactobacillus fermentum bacteria as a food additive, and fermented products using the same. However, these documents do not disclose lactic acid bacteria that selectively decompose only acetaldehyde, which causes hangovers.

[0010] The present inventors isolated and identified a novel Lactobacillus fermentum strain HDB1098 that can selectively decompose only acetaldehyde, an alcohol-derived hangover-inducing substance, and confirmed that this strain can be used as a functional ingredient in compositions for preventing or relieving hangovers, thereby completing the present invention. Summary of the Invention [Problem to be solved by the invention]

[0011] The main objective of the present invention is to provide a novel Lactobacillus fermentum strain HDB1098 having selective acetaldehyde-degrading enzyme activity that is useful for relieving hangovers.

[0012] Another object of the present invention is to provide a food or pharmaceutical composition for hangover relief, which contains the novel Lactobacillus famentum strain HDB1098 and is useful for inducing the decomposition of acetaldehyde, reducing its blood concentration, and alleviating hangover symptoms. [Means for solving the problem]

[0013] According to one aspect of the present invention, there is provided Lactobacillus fermentum HDB1098 strain, which has been deposited at the Korea Center for Microorganisms (KCCM) under accession number KCCM13264P and is characterized by its effectiveness in alleviating and relieving hangover symptoms.

[0014] In the present invention, the strain is a novel lactic acid bacterium isolated from raw makgeolli. It was identified as a Lactobacillus fermentum strain through API kit and 16S rRNA sequencing, and was confirmed to have selective acetaldehyde-degrading enzyme activity. It was named Lactobacillus fermentum HDB1098 and deposited with the Korea Center for Microorganisms (KCCM) on November 9, 2022, and assigned the accession number KCCM13264P.

[0015] The present invention preferably provides Lactobacillus fermentum HDB1098, which is characterized by selectively promoting acetaldehyde-degrading enzyme activity over alcohol-degrading enzyme activity, thereby reducing blood acetaldehyde levels. In an embodiment of the present invention, a strain with significantly higher acetaldehyde-degrading enzyme activity than alcohol-degrading enzyme activity, more than five times higher than that of other strains, was selected, isolated, and identified, and named Lactobacillus fermentum HDB1098.

[0016] The present invention preferably provides Lactobacillus fermentum HDB1098, a strain characterized by higher acetaldehyde-degrading enzyme activity in killed cells than in live cells. In an example of the present invention, to confirm that the acetaldehyde-degrading activity is due to metabolic products of the lactic acid bacteria strain, a comparative experiment was conducted on the acetaldehyde-degrading enzyme activity of medium components, live cells, and killed cells. The results showed that the acetaldehyde-degrading enzyme activity of live and killed cells was higher than that of medium components containing metabolic products, confirming that the activation of the acetaldehyde-degrading enzyme is a characteristic of the Lactobacillus fermentum HDB1098 cells themselves, rather than due to metabolic products derived from the strain. In particular, the Lactobacillus fermentum HDB1098 strain of the present invention is unusual in that killed cells exhibit higher acetaldehyde-degrading enzyme activity than live cells. Since killed bacteria are easier to handle and more stable than live bacteria, this may be advantageous in the processing, storage and distribution of hangover relief compositions made from them.

[0017] In another aspect, the present invention provides a composition for preventing or relieving hangovers, comprising live or killed cells, preferably killed cells, of the novel Lactobacillus fermentum HDB1098 strain according to the present invention as an active ingredient. According to the present invention, the novel Lactobacillus fermentum HDB1098 strain selectively promotes the activity of acetaldehyde-degrading enzymes over alcohol-degrading enzymes, thereby reducing blood acetaldehyde levels and thereby relieving or alleviating hangover symptoms after alcohol consumption.

[0018] The present invention provides a composition for preventing or relieving hangovers, preferably characterized in that the killed cells of the strain are obtained by culturing Lactobacillus fermentum HDB1098, removing medium components, and then heat-treating the cells to kill them. The strain can be cultured under any conditions suitable for culturing lactic acid bacteria, preferably at 35-40°C and pH 6.0-6.5. The heat treatment can be carried out at any temperature sufficient to kill the cells, preferably at 70-125°C for 30-120 minutes, and more preferably at 110-121°C for 60-90 minutes. The killed cells of the strain can also be prepared and used in the form of a dried powder or concentrated liquid.

[0019] The present invention preferably provides a composition for preventing or relieving hangovers, which is administered to a subject with an acetaldehyde-degrading enzyme subtype (acetaldehyde dehydrogenase 2 mutation: ALDH2*2) that cannot decompose acetaldehyde due to a genetic problem. In the examples of the present invention, it was confirmed that the strain of the present invention itself has acetaldehyde-degrading enzyme activity, and therefore hangovers can be relieved even when administered to a subject with an acetaldehyde-degrading enzyme subtype (acetaldehyde dehydrogenase 2 mutation: ALDH2*2) that cannot properly decompose acetaldehyde due to a genetic problem.

[0020] The present invention preferably provides a composition for preventing or relieving hangovers, characterized in that the composition contains 0.001 to 100% by weight of killed cells of Lactobacillus fermentum HDB1098 strain.

[0021] The present invention provides a composition for preventing or relieving hangovers, wherein the composition is preferably a food composition or a pharmaceutical composition.

[0022] In the present invention, the food composition may be any product selected from the group consisting of food, food additives, beverages, beverage additives, fermented milk, and functional health foods. The food composition may be, for example, various foods, fermented milk, meat, drinking water, chocolate, snacks, confectionery, pizza, ramen, other cotton, chewing gum, ice cream, alcoholic beverages, vitamin complexes, alcoholic beverages, and other functional health foods, but is not limited thereto.

[0023] The food composition may also contain ingredients commonly added during food production, such as proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. Carbohydrates include conventional sugars such as monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, oligosaccharides, etc.), and polysaccharides (e.g., dextrin, cyclodextrin, etc.), as well as sugar alcohols such as xylitol, sorbitol, and erythritol. Flavoring agents include natural flavoring agents such as taumarine and stevia extract (e.g., rebaudioside A, glycylrhizin, etc.), and synthetic flavoring agents such as saccharin and aspartame.

[0024] For example, when the food composition of the present invention is prepared as a drink, it may further contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, plum extract, or citric acid extract.

[0025] The food composition of the present invention includes all natural material product forms such as food, functional food, nutritional supplement, health food, food additive, etc. This type of food composition can be produced in various forms according to conventional methods known in the art.

[0026] For example, as a health food, the killed cells of the HDB1098 strain can be prepared in the form of tea, juice, or drink, or can be ingested as granules, capsules, or powder. Furthermore, as a food, the killed cells can be added to beverages (including alcoholic beverages), fruits and processed foods (e.g., canned fruits, bottled fruits, jams, marmalade, etc.), fish, meat and processed foods (e.g., ham, sausage, corned beef, etc.), breads and noodles (e.g., udon, soba, ramen, spaghetti, macaroni, etc.), fruit juices, various drinks, cookies, candy, dairy products (e.g., yogurt, fermented milk, butter, cheese, etc.), edible vegetable oils and fats, macaroni, vegetable proteins, retort foods, frozen foods, and various seasonings (e.g., miso paste, soy sauce, etc.).

[0027] In addition, in order to use the killed cells of the HDB1098 strain of the present invention in the form of a food additive, they can be produced in the form of a powder or a concentrated liquid.

[0028] The present invention provides a composition for preventing or relieving hangovers, which is preferably in the form of any formulation selected from the group consisting of a liquid phase, a powder, a granule, a tablet, and a capsule.

[0029] The pharmaceutical composition of the present invention can be preferably administered orally. The pharmaceutically effective amount of the composition can be adjusted appropriately depending on individual differences such as age and health condition, as well as the formulation and form. Preferably, the composition is administered so that the weight of dried killed cells of the HDB1098 strain is generally 10 to 3,000 mg per day for an adult. [Effects of the Invention]

[0030] Lactobacillus fermentum HDB1098 of the present invention can selectively activate acetaldehyde-degrading enzymes to reduce blood acetaldehyde levels, thereby helping to alleviate or eliminate hangover symptoms caused by alcohol consumption. Lactobacillus fermentum HDB1098 of the present invention can reduce blood acetaldehyde levels by inducing the conversion of acetaldehyde to acetate in cases where acetaldehyde decomposition is difficult due to a lack of acetaldehyde-degrading enzymes or genetic inactivation.

[0031] According to the present invention, the novel lactic acid bacteria strain selectively promotes the activity of acetaldehyde-degrading enzymes over alcohol-degrading enzymes, thereby reducing blood acetaldehyde levels and thereby eliminating or alleviating hangover symptoms after alcohol consumption. Furthermore, because the strain itself possesses acetaldehyde-degrading enzyme activity, it can be advantageously administered to subjects who have difficulty relieving hangover symptoms, such as those with a genetically inherited acetaldehyde-degrading enzyme subtype (acetaldehyde dehydrogenase 2 mutation: ALDH2*2), which is unable to properly decompose acetaldehyde, thereby contributing to the prevention and elimination of hangover symptoms. [Brief explanation of the drawings]

[0032] [Figure 1] 1 is a graph showing a comparative experiment of the acetaldehyde decomposition activity of live cells, medium components, and dead cells of Lactobacillus fermentum HDB1098 strain. [Figure 2] These results show the change in blood alcohol concentration in experimental animals that were administered killed Lactobacillus fermentum HDB1098 cells 30 minutes before alcohol administration. [Figure 3] The results show that the activity of alcohol-degrading enzymes in the liver tissue of experimental animals was confirmed after administering killed cells of Lactobacillus fermentum HDB1098 strain 30 minutes before alcohol administration. [Figure 4]These results show the change in blood acetaldehyde concentration in experimental animals that were administered killed Lactobacillus fermentum HDB1098 cells 30 minutes before alcohol administration. [Figure 5] The results show that the acetaldehyde-degrading enzyme activity in the liver tissue of experimental animals was confirmed after administering killed Lactobacillus fermentum HDB1098 cells 30 minutes before alcohol administration. DETAILED DESCRIPTION OF THE INVENTION

[0033] The present invention will be described in more detail below using examples. It will be apparent to those skilled in the art that these examples are merely for the purpose of explaining the present invention in more detail, and that the scope of the present invention is not limited to these examples according to the gist of the present invention.

[0034] Example 1: Selection of strains with selective acetaldehyde degradation ability To isolate lactic acid bacteria capable of selectively decomposing acetaldehyde, 1 g of various types of raw makgeolli samples was suspended in 9 mL of sterile saline (0.85% (v / v) sodium chloride) and serially diluted 10-fold with water. The diluted solutions were then incubated on BCP agar at 37°C for 24 hours. When the bacterial cells turned yellow on the BCP agar, colonies were obtained and smeared on MRS agar. The smeared strains were serially subcultured on MRS agar to isolate pure strains so that they appeared as a single strain when observed under a microscope.

[0035] Fourteen strains of lactic acid bacteria were isolated using the above process. The isolated strains were cultured in MRS broth at 37°C for 24 hours and then used to evaluate alcohol-degrading enzyme activity and acetaldehyde-degrading enzyme activity. The ADH assay kit (Abcam, cat. AB102533, England) and ALDH assay kit (Abcam, cat. AB155893, England) were used for the evaluation, and the analytical method was performed according to the protocol included with the kit. After evaluation, the sum of alcohol-degrading enzyme and acetaldehyde-degrading enzyme activity was set at 100%, and one strain (HDB1098) with an acetaldehyde-degrading enzyme activity ratio of over 80% was selected as having selective acetaldehyde degradation ability. The selected strain, HDB1098, had significantly higher acetaldehyde-degrading enzyme activity than alcohol-degrading enzyme activity, more than five times higher. The activity (%) of each strain is shown in Table 1.

[0036] [Table 1]

[0037] Example 2: Identification of strains that selectively degrade acetaldehyde The strain (HDB1098) selected in Example 1, which selectively degrades acetaldehyde, was cultured in MRS liquid medium for one day, and then its carbon source utilization pattern was confirmed using the API 50 CHL medium kit for physicochemical identification. The purple medium turned yellow when the sugars in the kit were used, indicating a positive result (+), while a medium that did not change color was indicated as a negative result (-). The above results were confirmed via API Web to identify this strain as Lactobacillus famentum (99.6% identity). The results of physicochemical identification using the API kit are shown in Table 2.

[0038] [Table 2]

[0039] In addition, the isolated strain's genomic DNA was extracted and partial 16S rRNA sequencing was performed using the polymerase chain reaction (PCR) method to obtain its genetic information (the 16S rRNA sequence is set forth in SEQ ID NO: 1). The obtained genetic information was compared with the microbial genetic information program at the National Center for Biotechnology Information, USA, and the strain name was confirmed as Lactobacillus fermentum based on the strain similarity results. The strain was named Lactobacillus fermentum HDB1098 and deposited with the Korea Center for Microorganisms (KCCM) on November 9, 2022, and assigned the accession number KCCM13264P.

[0040] Example 3: Comparison of acetaldehyde-degrading enzyme activity of Lactobacillus fermentum strains The alcohol and acetaldehyde selective activity of Lactobacillus fermentum HDB1098 confirmed in Example 2 was compared with that of the same Lactobacillus fermentum strain, one Lactobacillus fermentum strain and two KCCM strains (KCCM35461 and KCCM35469) owned by the company. The experiment to measure the hydrolytic enzyme activity was carried out in the same manner as in Example 1. The results are shown in Table 3.

[0041] As shown in Table 3, even within the same Lactobacillus fermentum strain, the alcohol and acetaldehyde decomposition enzyme activities differed, and it was confirmed that the main enzymes activated by each strain were also different. Of these, the novel Lactobacillus fermentum strain HDB1098 had acetaldehyde decomposition enzyme as its main active enzyme, and had a high selective activity of over 80%, similar to the results in Example 1. However, KCCM35461 and KCCM35469 did not have any particular selective activity effect, and were shown to selectively act on the alcohol decomposition enzyme activity of HDB109.

[0042] [Table 3]

[0043] Example 4: Preparation of killed lactic acid bacteria that selectively decompose acetaldehyde Killed lactic acid bacteria were produced as follows. Lactobacillus fermentum HDB1098 strain was inoculated into sterilized MRS liquid medium and cultured at 37°C for 24 hours to prepare a seed culture. For the main culture, a basal medium was prepared using 1 L of water, 4% (w / v) glucose, 2% (w / v) yeast extract, 0.2% (w / v) potassium phosphate, 0.2% (w / v) ammonium sulfate, and 0.02% (w / v) magnesium sulfate. Each component was dissolved in water and sterilized at 121-123°C for 30 minutes. The Lactobacillus fermentum HDB1098 seed culture was inoculated into the basal medium in a sterilized fermenter and cultured at 35-40°C for 20 hours, maintaining the pH at 6.0-6.5 by adding sodium hydroxide solution dropwise at regular intervals.

[0044] The completed culture medium was centrifuged at 7,000 rpm for 20 minutes to remove medium components. After removing the medium components, the cells were suspended in sterilized water and centrifuged. The above procedure was repeated twice. After thoroughly removing the medium components by centrifugation, the lactic acid bacteria cells (live cells) were suspended in sterilized water and then heat-treated at 110-121°C for 60-90 minutes to kill the bacteria. After thorough sterilization, the cells were dried to obtain Lactobacillus fermentum HDB1098 killed cell powder (killed cells).

[0045] Example 5: Evaluation of acetaldehyde decomposition activity of live cells, medium components, and dead cells of selected strains The acetaldehyde decomposition activity was compared between live cells produced by culturing the lactic acid bacteria strain in Example 4 and then removing the medium components, medium components containing metabolic products, and the killed cells produced in Example 4. After drying, the mixture was diluted to the same concentration and the results are shown in Figure 1.

[0046] In an evaluation to confirm that the acetaldehyde-degrading activity was due to metabolic products of the lactic acid bacteria strain, the acetaldehyde-degrading enzyme activity of the medium components containing metabolic products was the lowest at 55.6 ± 13.9 nmol / min / g, while the viable cells showed higher results, and the dead cells showed the highest result at 235.13 ± 51.1 nmol / min / g. These results suggest that the activation of the acetaldehyde-degrading enzyme is a characteristic of the Lactobacillus famentum HDB1098 cells themselves, rather than a metabolic product derived from the strain.

[0047] Example 6: Evaluation of the selective ethanol and acetaldehyde degradation function of selected strains To evaluate the selective functionality of the Lactobacillus famentum HDB1098 culture powder prepared in Example 4 in relieving hangover symptoms, ethanol and acetaldehyde degradation and enzyme activity were evaluated in vivo.

[0048] For the study, 6-week-old male SD rats (Rats) were obtained from Orient Bio Co., Ltd. They were housed under specific conditions (temperature: 19-25°C, relative humidity: 30-70%, ventilation: 10-15 times / hour, lighting cycle: 12 hours / day, illuminance: 150-300 lux or more) and were fed solid food ad libitum. All animals were observed for general symptoms once daily during the quarantine and purification periods, and only animals showing no abnormal symptoms were used in the study.

[0049] The lactic acid bacteria culture powder was dissolved in sterilized distilled water and orally administered to rats at a low, medium, and high doses of 3.25, 6.5, and 13 mg / kg body weight / day, respectively, at a dose of 10 mL / kg.

[0050] After administration of the lactobacillus culture powder, rats were orally administered 10 mL / kg of 30% ethanol to induce a hangover. Blood samples were taken from the jugular vein of the blank group before administration of the lactobacillus, and from the negative control group (ethanol-administered group) and each test group at 1, 3, and 5 hours after ethanol administration. The blood was centrifuged to obtain serum, which was used to measure blood ethanol and acetaldehyde levels. After the 5-hour blood collection, the rats were euthanized with carbon dioxide and their livers were removed. The livers were frozen in liquid nitrogen and stored in a deep freezer at -80°C for use. The test methods and results used to evaluate the hangover-relief function are described below.

[0051] 6-1: Measurement of blood alcohol concentration of selected lactic acid bacteria culture powder Figure 2 shows the blood alcohol concentration in serum samples taken from the jugular vein. The negative control group, which was not treated with the selected lactobacillus sample, showed blood alcohol concentrations of 150±60 mg / dL before alcohol ingestion (5±6 mg / dL) at 1 hour, 194±68 mg / dL at 3 hours, and 99±9 mg / dL at 5 hours. The low-dose lactobacillus sample-treated group showed blood alcohol concentrations of 134±74 mg / dL, 190±48 mg / dL, and 114±25 mg / dL at each blood sampling time, showing no significant differences compared to the negative control group. The medium-dose test group showed blood alcohol concentrations of 128±82 mg / dL, 210±63 mg / dL, and 123±43 mg / dL, respectively. The high-dose group showed blood alcohol concentrations of 117±97 mg / dL, 226±41 mg / dL, and 127±20 mg / dL at each time point. Neither test group showed significant results when compared with the negative control group, as did the low-dose lactobacillus test group.

[0052] 6-2: Alcohol decomposition enzyme activity when selected lactic acid bacteria culture powder is administered To confirm the alcohol-degrading enzyme activity after administration of the selected lactic acid bacteria powder, livers frozen in liquid nitrogen were crushed and used. To evaluate the alcohol-degrading enzyme activity, a kit (Alcohol dehydrogenase colorimetric assay kit, cat. no. K787-100, Biovision, USA) was used. The analytical method was described in accordance with the protocol enclosed with the kit.

[0053] The blank group showed an alcohol-degrading enzyme level of 8.0 ± 2.8 mU / mg, while the ethanol-administered negative control group showed an enzyme activity level of 5.4 ± 3.7 mU / mg. The low-concentration lactic acid bacteria culture powder test group showed an alcohol-degrading enzyme level of 5.5 ± 3.1 mU / mg, showing no significant difference compared to the negative control group. The medium-dose and high-dose test groups showed levels of 6.3 ± 3.5 mU / mg and 7.0 ± 3.8 mU / mg, respectively, showing no statistically significant results compared to the negative control group. In other words, the selected lactic acid bacteria, Lactobacillus fermentum, does not activate alcohol-degrading enzymes and therefore does not promote the breakdown of alcohol into acetaldehyde. The results of the enzyme activity evaluation are shown in Figure 3.

[0054] 6-3: Changes in blood acetaldehyde levels after administration of selected lactic acid bacteria culture powder Figure 4 shows the changes in blood acetaldehyde levels following administration of culture powder of the selected lactic acid bacteria, Lactobacillus fermentum HDB1098. In the negative control group, blood acetaldehyde levels increased from 0 (0.81 mg / dL) to 1 hour after ethanol administration, reaching 6.05 ± 1.13 mg / dL, 3 hours later at 2.54 ± 0.68 mg / dL, and 5 hours later at 0.75 ± 0.19 mg / dL. The low-dose lactic acid bacteria culture powder test group showed a level of 4.8 ± 0.75 mg / dL at 1 hour, but decreased to 1.88 ± 0.25 mg / dL at 3 hours and 0.61 ± 0.12 mg / dL at 5 hours, showing a tendency to decrease from 1 hour onward. Significant differences were observed compared to the negative control group at all measurement intervals. The medium-dose group had blood acetaldehyde levels of 3.1±0.77 mg / dL, 1.23±0.43 mg / dL, and 0.49±0.15 mg / dL, respectively, while the high-dose Lactobacillus sample-treated group had blood acetaldehyde levels of 1.68±0.76 mg / dL, 0.9±0.92 mg / dL, and 0.44±0.21 mg / dL. Blood acetaldehyde levels tended to decrease at each blood sample collection, demonstrating a significant difference compared to the negative control group. Furthermore, the results showed a concentration-dependent pattern, with acetaldehyde levels decreasing rapidly as the concentration increased. These results suggest that the selected Lactobacillus fermentum HDB1098 is effective in relieving hangovers by rapidly breaking down acetaldehyde produced from alcohol.

[0055] 6-4: Acetaldehyde-degrading enzyme activity of selected lactic acid bacteria culture powder To evaluate the acetaldehyde-degrading enzyme activity of Lactobacillus fermentum HDB1098 killed cell powder, frozen SD rat liver was used. Acetaldehyde-degrading enzyme activity was measured using an Aldehyde dehydrogenase activity colorimetric assay kit (cat No. K731-100, Biovision, USA) according to the enclosed protocol.

[0056] In Figure 5, the acetaldehyde-degrading enzyme activity in the negative control group was measured at 0.5 mU / mg. Meanwhile, the low-dose group and the medium-dose group of Lactobacillus fermentum HDB1098 killed cell powder showed 0.4 mU / mg and 0.6 mU / mg, respectively, showing no significant difference compared to the negative control group. The high-dose group showed a significant difference from the negative control group at 0.8 mU / mg, a result close to the blank group's 0.9 mU / mg. In other words, Lactobacillus fermentum HDB1098 killed cell powder contributed to normalization by activating acetaldehyde-degrading enzymes and removing acetaldehyde from the blood.

[0057] In summary, Lactobacillus fementum HDB1098 does not directly affect the activity of alcohol-degrading enzymes when alcohol is consumed, thereby preventing the oxidation of alcohol to acetaldehyde and preventing the liver from becoming fatigued due to toxic substances. Furthermore, it activates acetaldehyde-degrading enzymes to rapidly convert blood acetaldehyde to acetate, reducing its concentration and thereby alleviating hangover symptoms.

[0058] These effects explain why Lactobacillus fermentum HDB1098 selectively breaks down only acetaldehyde, helping to remove toxic substances derived from alcohol, and is effective in relieving hangover symptoms in people who are unable to effectively remove acetaldehyde.

[0059] The composition for relieving hangovers of the present invention, in the case of a functional health food or a pharmaceutical product, can be prepared using the following formulations. The following formulation examples are merely illustrative of the present invention and are not intended to limit the scope of the present invention.

[0060] Formulation Example 1: Tablet production The following ingredients are mixed and compressed into tablets according to a conventional tablet manufacturing method.

[0061] Lactobacillus fermentum HDB1098 Killed Bacteria Powder 200 mg Lactose 100mg starch 100mg Magnesium stearate dosage Formulation Example 2: Preparation of liquid formulation The following ingredients were mixed according to a conventional method for producing a liquid preparation, and then the mixture was filled into a brown bottle and sterilized to produce a liquid preparation.

[0062] Lactobacillus fermentum HDB1098 Killed Bacteria Powder 1000 mg 20g sugar 20g of high fructose corn syrup Lemon flavor to taste Add purified water to bring the total volume to 1000ml.

[0063] Formulation Example 3: Capsule manufacturing The following ingredients are mixed according to a conventional capsule manufacturing method, and the mixture is filled into a gelatin capsule to produce a capsule.

[0064] Lactobacillus fermentum HDB1098 Killed Bacteria Powder 300 mg Crystalline cellulose 3mg Lactose 14.8 mg Magnesium stearate 0.2 mg Although the present invention has been described with reference to the above embodiments, these are merely illustrative, and those skilled in the art will recognize that various modifications and equivalent embodiments are possible. Therefore, the true technical scope of protection of the present invention should be determined by the technical details of the appended claims.

Claims

1. Lactobacillus fermentum HDB1098 strain, deposited at the Korea Center for Microorganisms (KCCM) under accession number KCCM13264P, is characterized by its effectiveness in alleviating and relieving hangover symptoms.

2. 2. The Lactobacillus fermentum HDB1098 strain according to claim 1, characterized in that the strain selectively promotes the activity of acetaldehyde decomposition enzyme over alcohol decomposition enzyme, thereby reducing the concentration of acetaldehyde in the blood.

3. 2. The Lactobacillus fermentum HDB1098 strain according to claim 1, wherein the dead cells of said strain have higher acetaldehyde decomposition enzyme activity than the live cells.

4. A composition for preventing or relieving hangovers, comprising killed cells of the Lactobacillus fermentum HDB1098 strain according to any one of claims 1 to 3 as an active ingredient.

5. 5. The composition for preventing or relieving hangovers according to claim 4, wherein the killed cells of the strain are obtained by culturing Lactobacillus fermentum HDB1098 strain, removing medium components, and then heat-treating the cells to kill them.

6. 5. The composition for preventing or relieving a hangover according to claim 4, wherein the composition is administered to a subject who has an acetaldehyde-degrading enzyme subtype that is unable to decompose acetaldehyde due to a genetic problem.

7. The composition for preventing or relieving a hangover according to claim 4, wherein the composition is a food composition or a pharmaceutical composition.

8. 5. The composition for preventing or relieving a hangover according to claim 4, wherein the composition is in the form of any formulation selected from the group consisting of a liquid phase, a powder, a granule, a tablet, and a capsule.

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