JP7766734B2 - A novel Lactobacillus fermentum strain that selectively decomposes acetaldehyde and a hangover-relieving composition containing the same as an active ingredient - Google Patents

Description

K.C.C.M. KCCM13264P

This application 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.

The present invention relates to novel lactic acid bacteria that selectively promote the activity of acetaldehyde-degrading enzymes that break down acetaldehyde, which causes hangover symptoms after alcohol consumption, thereby reducing blood acetaldehyde levels. More specifically, the present invention relates to the novel Lactobacillus fermentum strain HDB1098, which selectively degrades acetaldehyde, and a composition for relieving hangovers that contains the strain as an active ingredient.

Alcohol is a favorite food that humans have used since ancient times for purposes such as forming social relationships and relieving stress. It is still widely consumed in a variety of situations today, but after drinking alcohol, it can induce hangover symptoms such as nausea, vomiting, and headaches. For this reason, much research into hangovers has continued, and various products to relieve hangovers have been released.

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, and then converted into acetate by acetaldehyde dehydrogenase. Finally, acetate is completely broken down into carbon dioxide and water. Acetaldehyde, which is produced by the oxidation of ethanol, is a toxic substance known to cause hangover symptoms such as flushing, increased heart rate, and headaches.

However, 30-40% of Asians are unable to properly break down acetaldehyde. This is a genetic problem, and the presence of an acetaldehyde-degrading enzyme subtype (acetaldehyde dehydrogenase 2 mutation; ALDH2*2) plays a significant role. While this genetic factor does not pose a significant problem 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 but 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 both alcohol-degrading enzymes, may be more effective for people with acetaldehyde-degrading enzyme subtypes.

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 outdated literature, the Ministry of Food and Drug Safety of Korea (MFDS) has determined that there is insufficient evidence that commercially available products have hangover relief benefits. 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.

Meanwhile, the WHO has defined probiotics as "live microorganisms that, when ingested in adequate amounts, confer a beneficial effect on the host's health." However, through extensive research, the definition has expanded to include not only microorganisms but also their culture components, and "dead cells of lactic acid bacteria" are now included in the probiotics category.

In line with these changes, 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.

Meanwhile, Korean Patent Registration No. 10-2262646 discloses a composition for preventing or relieving hangovers, which contains 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, which contains 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 properties and fermented vegetables using the same. Korean Patent Publication No. 10-2022-0114993 discloses a novel Lactobacillus fermentum CJNU 1840 strain and its use, a novel microorganism resistant to coffee extract, and a bioconverted coffee extract using the same. Korean Patent Publication No. 10-2004-0104153 discloses lactic acid bacteria with alcohol-degrading properties, dairy products containing the same, functional health foods, and food additives, as well as live Lactobacillus fermentum bacteria and fermented products using the same. However, these documents do not disclose lactic acid bacteria that selectively decompose only acetaldehyde, the cause of hangovers.

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.

The primary object of the present invention is to provide a novel Lactobacillus fermentum strain HDB1098 that has selective acetaldehyde-degrading enzyme activity that is useful for relieving hangovers.

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 breakdown of acetaldehyde, reducing its blood concentration, and alleviating hangover symptoms.

In one aspect, the present invention provides 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.

In the present invention, the strain is a novel lactic acid bacterium isolated from raw makgeolli. It was identified as a Lactobacillus fermentum strain using an 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.

The present invention preferably provides Lactobacillus fermentum HDB1098, a bacterial strain 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, at least five times higher than that of other strains, was selected, isolated, and identified, and named Lactobacillus fermentum HDB1098.

The present invention preferably provides Lactobacillus fermentum HDB1098, a strain characterized by higher acetaldehyde-degrading enzyme activity in killed cells rather than in live cells. In an example of the present invention, to confirm that the acetaldehyde-degrading activity is derived from the 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 a metabolic product derived from the strain. In particular, the Lactobacillus fermentum HDB1098 strain of the present invention is unusual in that the killed cells have higher acetaldehyde-degrading enzyme activity than the live cells. Killed bacteria are easier to handle and more stable than live bacteria, which may be an advantage in the processing, storage, and distribution of hangover relief compositions made from them.

In another aspect, the present invention provides a composition for preventing or relieving hangovers, comprising live or killed cells, preferably killed cells, of the 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, thereby relieving or alleviating hangover symptoms after alcohol consumption.

The present invention provides a composition for preventing or relieving hangovers, preferably characterized in that the killed cells of the strain are Lactobacillus fermentum HDB1098 strain, which have been cultured, and then the culture medium components removed, and then heat-treated to kill the cells. 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 that can kill the cells, preferably at 70-125°C for 30-120 minutes, and more preferably at 110-121°C for 60-90 minutes. Furthermore, the killed cells of the strain can be dried and then produced in the form of a powder or a concentrated liquid for use.

The present invention preferably provides a composition for preventing or relieving hangovers, characterized in that the composition 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.

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.

The present invention provides a composition for preventing or relieving hangovers, preferably characterized in that the composition is a food composition or a pharmaceutical composition.

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 cottonseeds, chewing gum, ice cream, alcoholic beverages, vitamin complexes, alcoholic beverages, and other functional health foods, but is not limited to these.

The food composition may also contain ingredients that are typically 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 that can be used 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.

For example, when the food composition of the present invention is manufactured 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.

The food compositions of the present invention include all natural product forms, such as foods, functional foods, dietary supplements, health foods, and food additives. These types of food compositions can be produced in various forms according to conventional methods known in the art.

For example, as a health food, the killed cells of the HDB1098 strain can be produced in the form of tea, juice, or drink, or can be ingested in the form of granules, capsules, or powder. Furthermore, as a food, it 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 protein, retort foods, frozen foods, and various seasonings (e.g., miso, soy sauce, etc.).

Furthermore, to use the killed cells of the HDB1098 strain of the present invention as a food additive, they can be produced and used in the form of a powder or concentrated liquid.

The present invention provides a composition for preventing or relieving hangovers, preferably in the form of a formulation selected from the group consisting of liquid phase, powder, granules, tablets, and capsules.

The pharmaceutical composition of the present invention can preferably be administered orally. The pharmaceutically effective dosage of the composition can be adjusted appropriately depending on individual differences such as age and health condition, as well as the formulation and form. However, it is generally appropriate to administer 10 to 3,000 mg of dried killed cells of the HDB1098 strain per day to an adult.

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

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 eliminating or alleviating hangover symptoms after alcohol consumption. Furthermore, because the strain itself possesses acetaldehyde-degrading enzyme activity, it has the advantage of being able to contribute to the prevention and elimination of hangover symptoms when 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) that is unable to properly break down acetaldehyde.

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. The graph shows the results of examining changes in blood alcohol concentration in experimental animals to which killed cells of Lactobacillus fermentum HDB1098 strain were administered 30 minutes before alcohol administration. The results show the alcohol-degrading enzyme activity in the liver tissue of experimental animals to which killed cells of Lactobacillus fermentum HDB1098 strain were administered 30 minutes before alcohol administration. The graph shows the results of examining changes in blood acetaldehyde concentrations in experimental animals to which killed cells of Lactobacillus fermentum HDB1098 strain were administered 30 minutes before alcohol administration. The results show the results of confirming the acetaldehyde-decomposing enzyme activity in the liver tissue of experimental animals to which killed cells of Lactobacillus fermentum HDB1098 strain were administered 30 minutes before alcohol administration.

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 provided merely to more specifically illustrate the present invention, and that the scope of the present invention is not limited to these examples.

Example 1: Screening of strains with selective acetaldehyde degradation ability To isolate lactic acid bacteria capable of selectively degrading acetaldehyde, 1 g of various 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 incubated on BCP agar at 37°C for 24 hours. When the bacterial cells turned yellow on the BCP agar as a result of the incubation, 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.

Using the above process, 14 strains of lactic acid bacteria were isolated. 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 the ALDH assay kit (Abcam, cat. ab155893, England) were used for the evaluation, and the analysis 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 acetaldehyde-degrading enzyme activity superior to alcohol-degrading enzyme activity, exceeding 80%, was selected as a strain with selective acetaldehyde-degrading ability. The selected strain, HDB1098, had acetaldehyde-degrading enzyme activity that was significantly higher than alcohol-degrading enzyme activity by more than five times. The activity (%) results for each strain are shown in Table 1.

Example 2: Identification of a strain that selectively degrades acetaldehyde. The acetaldehyde-selective degrading strain (HDB1098) selected in Example 1 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 added, indicating a positive (+) result, while the medium that did not change color was indicated as a negative (-) result. The above results were confirmed via API Web as Lactobacillus famentum (99.6% identity). The physicochemical identification results using the API kit are shown in Table 2.

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 genetic information (the 16S rRNA base sequence is set forth as SEQ ID NO: 1). The obtained genetic information was compared with the genomic base sequences using the Microbial Gene Information Program of 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.

Example 3: Comparison of acetaldehyde decomposition enzyme activity of Lactobacillus fermentum strains The Lactobacillus fermentum HDB1098 confirmed in Example 2 was compared with the same Lactobacillus fermentum strains, which have one Lactobacillus fermentum strain and two KCCM strains (KCCM35461, KCCM35469) in-house owned.The experiment for measuring decomposition enzyme activity was carried out in the same manner as in Example 1.The results are shown in Table 3.

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

Example 4: Preparation of Killed Lactic Acid Bacteria Cells That Selectively Decompose Acetaldehyde Killed lactic acid bacteria cells were produced as follows. Lactobacillus fermentum HDB1098 strain was inoculated into a 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 by dissolving each component in water to a concentration of 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, based on 1 L of water, and sterilizing the mixture 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.

The completed culture medium was centrifuged at 7,000 rpm for 20 minutes to remove medium components. The cells from which the medium components had been removed were added to sterilized water, suspended, and then centrifuged. The above process was repeated twice. The lactic acid bacteria cells from which the medium components had been sufficiently removed by centrifugation (live cells) were added to sterilized water, suspended, and then heat-treated at 110-121°C for 60-90 minutes to kill the cells. After thorough sterilization, the cells were dried to obtain a killed Lactobacillus fermentum HDB1098 cell powder (killed cells).

Example 5: Evaluation of acetaldehyde decomposition activity of live cells, medium components, and killed cells of selected strains Live cells, prepared by culturing the lactic acid bacteria strain in Example 4 and then removing the medium components, and medium components containing metabolic products, and killed cells prepared in Example 4 were dried and diluted to the same concentration, and then the acetaldehyde decomposition activity was compared and evaluated. The results are shown in Figure 1.

In an evaluation to confirm that the acetaldehyde-degrading activity was derived from 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 bacteria showed higher results, and the dead bacteria showed the highest result at 235.13 ± 51.1 nmol/min/g. From these results, it appears that the activation of the acetaldehyde-degrading enzyme is a property of the Lactobacillus fementum HDB1098 bacteria itself, rather than a metabolic product derived from the strain.

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

For the study, 6-week-old male SD rats were obtained from Orient Bio Co., Ltd. They were kept under specified 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 for the study.

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

After administration of the lactic acid bacteria culture powder, 10 mL/kg of 30% ethanol was orally administered to induce a hangover. Blood was collected from the jugular vein of the blank group before administration of the lactic acid bacteria, 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 only, which was used to measure blood ethanol and acetaldehyde levels. After the 5-hour blood collection period, 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-relieving function are described below.

6-1: Measurement of blood alcohol concentration in serum from selected lactic acid bacteria culture powder. Figure 2 shows the blood alcohol concentration in serum from blood samples taken via the jugular vein. The negative control group, which was not treated with the selected lactic acid bacteria sample, showed blood ethanol concentrations of 150±60 mg/dL before alcohol intake (5±6 mg/dL) one hour later, 194±68 mg/dL three hours later, and 99±9 mg/dL five hours later. The low-dose lactic acid bacteria sample-treated group showed blood ethanol 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 group had blood alcohol levels of 128±82 mg/dL, 210±63 mg/dL, and 123±43 mg/dL, respectively, while the high-dose group had blood alcohol levels of 117±97 mg/dL, 226±41 mg/dL, and 127±20 mg/dL, respectively. Neither test group showed significant results compared with the negative control group, as did the low-dose lactobacillus test group.

6-2: Alcohol-degrading enzyme activity after administration of selected lactic acid bacteria culture powder. 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 based on the protocol enclosed with the kit.

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

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 blood acetaldehyde level of 4.8 ± 0.75 mg/dL after 1 hour, but decreased to 1.88 ± 0.25 mg/dL after 3 hours and 0.61 ± 0.12 mg/dL after 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 3.1±0.77mg/dL, 1.23±0.43mg/dL, and 0.49±0.15mg/dL, while the high-dose lactic acid bacteria sample group had 1.68±0.76mg/dL, 0.9±0.92mg/dL, and 0.44±0.21mg/dL. The blood acetaldehyde concentration tended to decrease at each blood sample collection, showing a significant difference compared to the negative control group. Furthermore, the concentration also showed a concentration-dependent result, with the acetaldehyde concentration decreasing rapidly as the concentration increased. This explains why the selected Lactobacillus fermentum HDB1098 is effective in relieving hangovers by rapidly breaking down acetaldehyde produced from alcohol.

6-4: Acetaldehyde-degrading enzyme activity of selected lactic acid bacteria culture powder. Frozen SD rat liver was used to evaluate the acetaldehyde-degrading enzyme activity of Lactobacillus fermentum HDB1098 killed cell powder. The 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.

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 of Lactobacillus fermentum HDB1098 killed cell powder had a value of 0.4 mU/mg, and the medium-dose group had a value of 0.6 mU/mg, showing no significant difference when compared to the negative control group. The high-dose group of killed cells showed a value of 0.8 mU/mg, which was significantly different from the negative control group and close to the blank group's result of 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.

To summarize these results, Lactobacillus fementum HDB1098 does not directly affect the activity of alcohol-degrading enzymes when alcohol is consumed, thereby preventing its oxidation to acetaldehyde and preventing fatigue caused by toxic substances from building up in the liver. Furthermore, it activates acetaldehyde-degrading enzymes, rapidly converting acetaldehyde in the blood to acetate, thereby reducing its concentration and alleviating potential hangover symptoms.

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.

When the hangover relief composition of the present invention is a functional health food or pharmaceutical product, it can be manufactured using the following formulation. The following formulation examples are illustrative of the present invention and are not intended to limit the scope of the present invention.

Formulation Example 1: Preparation of tablets Tablets are prepared by mixing the following ingredients and compressing them according to a conventional tablet preparation method.

Lactobacillus fermentum HDB1098 Killed Bacteria Powder 200 mg
Lactose 100mg
starch 100mg
Appropriate amount of magnesium stearate Formulation Example 2: Preparation of a liquid preparation The following ingredients were mixed according to a conventional method for preparing a liquid preparation, and then filled into a brown bottle and sterilized to prepare a liquid preparation.

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

Formulation Example 3: Preparation of capsules The following ingredients are mixed according to a conventional capsule preparation method, and filled into a gelatin capsule to prepare a capsule.

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 (8)

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. The Lactobacillus fermentum HDB1098 strain according to claim 1, characterized in that the strain selectively promotes the activity of acetaldehyde-degrading enzyme over alcohol-degrading enzyme, thereby reducing blood acetaldehyde levels. The Lactobacillus fermentum HDB1098 strain according to claim 1, characterized in that the dead cells of the strain have higher acetaldehyde-degrading enzyme activity than the live cells. 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. 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. The composition for preventing or relieving hangovers according to claim 4, wherein the composition is administered to a subject who, due to a genetic problem, has an acetaldehyde-degrading enzyme subtype that is unable to decompose acetaldehyde. The composition for preventing or relieving hangovers according to claim 4, wherein the composition is a food composition or a pharmaceutical composition. The composition for preventing or relieving hangovers according to claim 4, wherein the composition is in the form of a formulation selected from the group consisting of a liquid phase, a powder, a granule, a tablet, and a capsule.