Lactobacillus plantarum capable of accelerating alcohol metabolism and improving hangover state and application thereof

Abstract

The application provides a lactobacillus plantarum capable of accelerating alcohol metabolism and improving hangover state and an application thereof. The strain has high safety, does not contain resistance genes and pathogenic virulence genes, and has strong tolerance to bile salts, artificial gastric juice and artificial intestinal juice. The strain has strong antioxidant capacity and strong scavenging capacity for ABTS, DPPH and hydroxyl radicals. The lactobacillus plantarum can significantly prolong the reaction time of mice from alcohol intake to drunkenness and shorten the sober-up time. Moreover, the lactobacillus plantarum can produce melatonin and convert NR into nicotinic acid.

Description

Technical Field

[0001] This invention relates to the field of probiotics technology, specifically to a plant-based Lactobacillus that can accelerate alcohol metabolism and improve hangover symptoms. Background Technology

[0002] During alcohol metabolism, a large amount of reactive oxygen species (ROS) and toxic intermediates such as acetaldehyde are produced, which in turn induce oxidative stress in the liver, mitochondrial dysfunction, inflammatory cascade reactions, and gut microbiota dysbiosis, ultimately leading to hepatocellular damage, fatty degeneration, alcoholic hepatitis, liver fibrosis, and even cirrhosis. In this process, lipid metabolism disorders manifest as a decrease in serum high-density lipoprotein cholesterol (HDL-C) levels, while triglyceride (TG), low-density lipoprotein cholesterol (LDL), and liver enzymes (ALT, AST) levels are abnormally elevated, further exacerbating liver pathological damage.

[0003] Currently, drugs such as metadoxine, glucocorticoids, and polyene phosphatidylcholine are used clinically to prevent and treat alcoholic liver injury, and have played a certain therapeutic role. Meanwhile, probiotics, as a safe adjunctive intervention strategy, are gradually gaining attention due to their potential advantages in regulating the gut microbiota, enhancing intestinal barrier function, reducing endotoxemia, and regulating host immune and metabolic responses, and are expected to provide a beneficial supplement for the repair of alcoholic liver injury. The paper "Multi-Strain Probiotics Alleviate Alcoholic Liver Disease Via Microbiota-Metabolism Axis: A Randomized Controlled Study" points out that compound probiotics enhance the efficacy of traditional hepatoprotective drugs by regulating the gut microbiota and metabolic network. A systematic review and meta-analysis (Saadh et al., Frontiers in Nutrition, 2025) included 12 clinical trials published between 2008 and 2025. The results showed that probiotic intervention reduced ALT by an average of 10.10 U / L (95% CI: -15.34, -4.87) and AST by an average of 13.05 U / L (95% CI: -21.33, -4.78) in patients with alcoholic liver injury. In terms of gut microbiota regulation, probiotics can increase the abundance of beneficial bacteria such as Lactobacillus, Bifidobacterium, Faecalibacterium, and Prevotella, while reducing pathogenic bacteria such as Escherichia coli and Shigella.

[0004] Lactobacillus plantarum, as one of the probiotic lactic acid bacteria, has shown promising application prospects in the field of hangover relief and liver protection. This strain can directly participate in alcohol metabolism through its own alcohol dehydrogenase and aldehyde dehydrogenase, accelerating the conversion of ethanol to acetaldehyde and further to acetic acid, thereby reducing the concentration of alcohol and its intermediate metabolite acetaldehyde in the blood. Simultaneously, Lactobacillus plantarum can also exert a comprehensive liver-protective effect through multiple pathways, including regulating intestinal microecological balance, enhancing intestinal epithelial barrier function, reducing endotoxin translocation to the liver, alleviating oxidative stress damage, and inhibiting liver inflammation. Therefore, it is considered an ideal candidate strain for preparing functional foods or pharmaceutical preparations for hangover relief and liver protection. For example, patent CN106867930B indicates that Lactobacillus plantarum PLH1405 can increase the activity of alcohol dehydrogenase and aldehyde dehydrogenase in the body, accelerate the metabolism of ethanol and acetaldehyde, and simultaneously increase antioxidant enzyme activity and reduce endotoxin production, thus better alleviating the damage to the body caused by alcohol poisoning, making it suitable as a drug for treating acute alcohol poisoning. Patent CN107760624A indicates that *Lactobacillus plantarum* FCJX 102 can produce high levels of acetaldehyde dehydrogenase under ethanol induction, and has broad application prospects in alcohol-relieving and liver-protecting drugs and foods. While patent CN114196593A discloses a strain of *Lactobacillus plantarum* P101 with high antioxidant activity and verifies its ability to effectively alleviate alcoholic liver damage, it does not analyze other metabolites of *Lactobacillus plantarum*, such as melatonin production and NR conversion, nor does it detect related indicators of HDL-C, LDL, and TG; its findings are limited to antioxidant effects and related animal experiments.

[0005] *Lactobacillus plantarum* possesses an enzymatic basis for metabolizing small organic acids. Formic acid can be metabolized via the pyruvate-formate lyase (PFL) system, oxidizing to CO2 and producing NADH under the catalysis of formate dehydrogenase, thus participating in energy metabolism. The PFL reaction is reversible, allowing exogenous formic acid to be replenished and enter central carbon metabolism. *Lactobacillus plantarum* also possesses an acetyl-CoA synthase (ACS) that converts acetic acid to acetyl-CoA in the presence of ATP (acetic acid + ATP + CoA → acetyl-CoA + AMP + PPi). Acetyl-CoA then enters the TCA cycle to produce energy, is used for cellular synthesis, or is converted to acetyl phosphate via the reverse AK-PTA pathway. These metabolic mechanisms provide a theoretical basis for *Lactobacillus plantarum* to utilize non-food carbon sources (such as electrochemically generated one- or two-carbon substances).

[0006] In existing technologies, the industrial fermentation of *Lactobacillus plantarum* generally relies on grain-derived carbon sources such as glucose, which leads to problems such as competition for land with grains and high costs. For example, patent CN 103525740A uses hydrolysate of biomass raw materials rich in cellulose and / or hemicellulose to replace glucose in the universal culture medium MRS for *Lactobacillus*. Although this technology achieves the substitution of non-grain carbon sources, its focus is on the production of γ-aminobutyric acid (GABA) rather than the maintenance or enhancement of the viable cell count of the bacteria themselves. In contrast, the strain of this invention showed no significant change in the number of viable cells when replacing 30% of glucose with a non-grain carbon source, but the number of viable cells was significantly higher than that of conventional glucose fermentation when an additional 30% non-grain carbon source was added, demonstrating a unique advantage in carbon source utilization. Summary of the Invention

[0007] The technical problem this invention aims to solve is to provide a strain of *Lactobacillus plantarum* LT0077 with antioxidant functions, capable of accelerating alcohol metabolism and improving hangover symptoms. This bacterium was isolated from salted plum vegetables grown in Huizhou City, Guangdong Province. It can grow normally in a culture medium containing 10% 53% vol alcohol. This probiotic exhibits high tolerance in simulated gastric juice, simulated intestinal juice, and bile salts, especially in simulated full-feeding gastric juice, where its 3-hour tolerance rate reaches 91.73%, indicating that the bacterium can effectively colonize the intestines and exert its effects. In terms of antioxidant properties, compared to other *Lactobacillus plantarum* strains, this bacterium demonstrates superior ABTS, DPPH, and hydroxyl radical scavenging abilities, exhibiting excellent antioxidant capacity. Mouse animal experiments verified the bacterium's ability to detoxify and protect the liver. Experimental results showed that *Lactobacillus plantarum* LT0077 can reduce indicators related to alcoholic liver disease in mice and alleviate its pathological signs. Furthermore, this bacterium also has the ability to produce melatonin and convert NR to niacin.

[0008] This invention provides a strain of *Lactobacillus plantarum* LT0077, screened from salted plum vegetables grown in Huizhou City, Guangdong Province. This strain was deposited on December 16, 2025, at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou City, with accession number GDMCC No. 67489. The classification and naming are as follows. Lactiplantibacillus plantarum .

[0009] Furthermore, the 16S nucleotide sequence of the *Lactobacillus plantarum* LT0077 is shown in SEQ ID NO.1.

[0010] The present invention also provides a lyophilized powder formulation comprising the aforementioned Lactobacillus plantarum LT0077.

[0011] This invention also provides a method for producing the above-mentioned *Lactobacillus plantarum* LT0077 by fermentation and freeze-drying, comprising: The above-mentioned *Lactobacillus plantarum* LT0077 was inoculated into the fermentation medium at an inoculation rate of 2%-5% and cultured at 37°C for 12-24 hours to obtain a culture solution. The culture solution was centrifuged and the wet bacterial precipitate was collected. The wet bacterial precipitate was resuspended and mixed with the lyophilization protectant solution and then freeze-dried to obtain a lyophilized powder preparation.

[0012] The fermentation medium consists of 10g yeast powder, 10g peptone, 30g glucose, 1g dipotassium hydrogen phosphate, 0.1g magnesium sulfate, 0.01g manganese sulfate, 1g Tween 80, 1000ml water, and pH 7.0 before sterilization.

[0013] The freeze-drying protectant consists of 150g trehalose, 50g sucrose, 1g ascorbic acid, 1g Tween 80, and 1000mL water.

[0014] The present invention also provides a microbial preparation comprising the above-mentioned *Lactobacillus plantarum* LT0077, wherein the viable count of the above-mentioned *Lactobacillus plantarum* LT0077 is not less than 1 × 10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

[0015] The microbial preparation may also contain other probiotics, such as Pediococcus pentosus and Lactobacillus rhamnosus, and may also contain other auxiliary ingredients, such as vitamins and minerals.

[0016] The present invention also provides a pharmaceutical product that accelerates the metabolism of alcohols, comprising the above-mentioned microbial preparation.

[0017] The preferred dosage form of the medicine is tablets, capsules, granules, oral liquid, etc.

[0018] The medicine also contains other nutrients, such as turmeric powder and kudzu root powder.

[0019] The drug also includes a carrier.

[0020] This invention offers the following beneficial technical effects. The *Lactobacillus plantarum* LT0077 provided by this invention, possessing antioxidant functions, accelerating alcohol metabolism, and improving hangover symptoms, exhibits survival rates of 1.05±0.76, 90.37±3.51, and 91.73±4.24% in simulated gastric juice under simulated satiety, starvation, and standard conditions, respectively. In simulated intestinal juice containing 0.3% bile salts, the survival rate remains at 67.46% after 24 hours. Regarding antioxidant properties, the scavenging capacity of this bacterium for ABTS free radicals, DPPH free radicals, and hydroxyl free radicals in cell-free contents is 28.75%, 42.44%, and 6.04%, respectively, demonstrating strong free radical scavenging ability. Animal experiments show that in an alcoholic liver disease model, *Lactobacillus plantarum* LT0077 significantly improves the reaction time from alcohol intake to intoxication and significantly reduces the sobering-up time in mice. Serum results show that in an alcoholic liver disease model, *Lactobacillus plantarum* LT0077 reduces serum ALT, AST, TG, and LDL levels in mice. Pathological results showed that the Lactobacillus plantarum LT0077 could repair alcohol-induced liver damage.

[0021] The *Lactobacillus plantarum* LT0077 can increase the level of alcohol dehydrogenase in a mouse model of alcoholic liver disease, demonstrating significant application value in antioxidation, alcohol detoxification, and hangover prevention. Simultaneously, this bacterium can also produce melatonin and convert NR into niacin. Attached Figure Description

[0022] Figure 1 Growth inhibition rate of Lactobacillus plantarum LT0077 in this invention; Figure 2 The results of streak plate assay for Lactobacillus plantarum LT0077 in this invention; Figure 3 The liquid chromatogram of nicotinic acid detection in Lactobacillus plantarum LT0077 of this invention; Figure 4 The liquid chromatogram for melatonin detection by Lactobacillus plantarum LT0077 of this invention; Figure 5 Bar chart of serum ALT content in mice with *Lactobacillus plantarum* LT0077 according to this invention; Figure 6 Bar chart of serum AST content in mice with Lactobacillus plantarum LT0077 according to this invention; Figure 7 Bar chart of serum TG content in mice with Lactobacillus plantarum LT0077 according to this invention; Figure 8 Bar chart of LDL content in mouse serum of Lactobacillus plantarum LT0077 according to this invention; Figure 9 Bar chart of alcohol dehydrogenase content in mouse liver of Lactobacillus plantarum LT0077 according to the present invention; Figure 10 HE staining results of each group of mice in the Lactobacillus plantarum LT0077 animal experiment of this invention. Detailed Implementation

[0023] The following embodiments and accompanying drawings are used to describe in detail the implementation of the present invention, so that the process of how the present invention uses technical means to solve technical problems and achieve technical effects can be fully understood and implemented accordingly.

[0024] Example 1: Screening and Identification of Strains with Alcohol-Detoxifying Ability 1.1 Initial screening of strains Thirty samples of fresh milk, sauerkraut, and pickled vegetables were collected from various parts of the country. One mL of fresh milk or one mL of sauerkraut / pickled vegetable juice from each sample was placed in a test tube containing 9 mL of sterile physiological saline (0.85%, %w / v) and diluted thoroughly. The dilution gradient was recorded as 10. -1 Then, pipette 1 mL into a test tube containing 9 mL of sterile physiological saline and dilute and mix well. Record the dilution gradient as 10. -2 The samples were then serially diluted to 10⁻⁶. -6 The original sample solution and the above-mentioned dilution were spread onto MRS selection medium plates and incubated at 37°C in an anaerobic workstation for 48 h. Single colonies with different morphology, size, and color and obvious calcium dissolution zones were selected from the plates and streaked onto MRS plates for purification to obtain single colonies. The obtained single colony strains were subjected to Gram staining microscopy and catalase test. Gram-positive and catalase-negative strains were preliminarily identified as lactic acid bacteria, numbered, and stored for later use.

[0025] The MRS medium consisted of: 10 g peptone, 5 g yeast extract, 10 g beef extract, 20 g glucose, 5 g sodium acetate, 1.5 g diammonium citrate, 1 mL Tween 80, 0.5 g magnesium sulfate, 0.05 g manganese sulfate, and 2 g dipotassium hydrogen phosphate, brought to a final volume of 1 L and sterilized at 121°C for 15 min. MRS plate culture was prepared by adding 1.5-2% (w / v) agar powder to the MRS medium. MRS screening medium was prepared by adding 1.5% (w / v) CaCO3 and 1.5-2% (w / v) agar powder to the MRS medium.

[0026] 1.2 Rescreening of bacterial strains and identification of bacterial strains 1.2.1 Determination of the growth capacity of alcohol-containing MRS Commercially available 56-degree Hongxing Erguotou (a type of Chinese liquor) was added to sterilized MRS medium, resulting in an alcohol concentration of 5 degrees. The selected bacterial strain was inoculated into MRS medium and cultured overnight to obtain seed culture. The seed culture was then inoculated at a 3% inoculation rate into both alcohol-containing MRS and MRS medium, and cultured for 24 hours. The OD (octane rating) of the fermentation broth was measured. 600 Calculate the growth inhibition rate: Growth inhibition rate % = (M1 - M2) / M1 × 100% (M1: MRS medium). Results are as follows. Figure 1 As shown, strain A13 exhibited the lowest growth inhibition rate and a high tolerance to alcohol.

[0027] 1.2.2 Microscopic examination results and 16s identification of the strain A pure culture of A13 bacteria, which showed low MRS growth inhibition rate in the presence of Erguotou (a type of Chinese liquor), was selected and sent for 16S testing. After analysis using the NCBI BLAST database, the bacterium was confirmed as *Lactobacillus plantarum* and named LT0077. This bacterium was deposited on December 16, 2025, at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, with accession number GDMCC No. 67489.

[0028] SEQ ID NO.1:

[0029] Example 2: Safety Analysis of the Strains 2.1 Genome Analysis The complete genome sequence of *Lactobacillus plantarum* LT0077 was obtained through testing. The assembled genome sequence was then analyzed using the ABRicate tool to detect virulence and resistance genes. Comparisons were performed using locally installed databases: Megares, CARD, VFDB, PlasmidFinder, Ecoh, NCBI, ecoli_vf, ResFinder, and ARGANNOT. The comparison criteria were: similarity ≥70% and coverage ≥70%. The bacterium did not contain any anti-biotin resistance genes, but only contained stress-related survival, adhesion, and immune regulation genes related to environmental adaptation, indicating that the bacterium is a safe strain.

[0030] 2.2 Drug sensitivity and safety analysis The antimicrobial resistance of *Lactobacillus plantarum* LT0077 was determined using the micro-broth dilution method according to GB 31615.2-2025, National Food Safety Standard for Safety Evaluation of Microbial Strains for Food Use. *Lactobacillus plantarum* LT0077 was streaked from a glycerol tube to an MRS plate and anaerobically cultured at 36±1℃ for 20 h. Simultaneously, the quality control strain *Lactobacillus paracasei* ATCC334 was streaked to an MRS plate and anaerobically cultured for 20 h. Well-grown bacterial colonies were transferred from the plates to 15 mL centrifuge tubes containing 5 mL of sterile physiological saline, and the OD was measured using a spectrophotometer. 625 The bacterial concentration was adjusted with physiological saline to maintain an OD value between 0.16 and 0.2. *Lactobacillus plantarum* LT0077 and *Lactobacillus paracasei* ATCC334 were diluted 500-fold in LSM medium, and 50 μL of each solution was added to an antimicrobial microdilution plate. The plates were then anaerobically cultured at 28±1℃ for 48 h to obtain the MIC value of the antimicrobial agent for *Lactobacillus plantarum* LT0077. Each well contained 50 μL of antibiotic solution.

[0031] Experimental analysis results showed that the breakpoint values ​​of different antibiotics against *Lactobacillus plantarum* LT0077 were all equal to or less than the breakpoint values ​​of *Lactobacillus fermentum* required in Table E.6 of GB 31615.2-2025. This indicates that *Lactobacillus plantarum* LT0077 is not resistant to ampicillin, gentamicin, kanamycin, streptomycin, tetracycline, erythromycin, clindamycin, and chloramphenicol, demonstrating its good safety profile.

[0032] 2.3 Hemolysis Verification Lactobacillus plantarum LT0077 was streaked onto Columbia blood agar plates and cultured at 37°C for 48 h in an anaerobic workstation. The results are as follows: Figure 2The results indicate that *Lactobacillus plantarum* LT0077 is not hemolytic, while *Staphylococcus aureus* ATCC6538 exhibits β-hemolysis. As a positive control to verify the validity of the plate, *Staphylococcus aureus* ATCC6538 was purchased from the Guangdong Provincial Microbial Culture Collection Center.

[0033] Example 3: Fermentation Experiment Using Non-Grain Carbon Sources Single colonies of *Lactobacillus plantarum* LT0077, activated by streak plating, were inoculated into MRS medium supplemented with a non-grain mixed carbon source. The mass ratio of the non-grain carbon source components was ethanol:sodium formate:sodium acetate = 20:3:8. One group of MRS medium had 30% of the glucose replaced by the non-grain carbon source, while another group had an additional 30% carbon source added to the MRS medium. Specific formulations are shown in Table 1 below. The control group used MRS medium. After 16 h of anaerobic incubation at 37℃, the viable count of *Lactobacillus plantarum* LT0077 was measured. The results are shown in Table 2 below. After replacing 30% of the glucose in the medium with the non-grain carbon source, the viable count of *Lactobacillus plantarum* LT0077 was 4.53 ± 0.25 × 10⁻⁶. 9 CFU / mL, the effective viable count in the control group MRS medium was 4.68±0.21×10⁻⁶. 9 With an additional 30% non-grain carbon source, the viable cell count during fermentation was 6.16 ± 0.27 × 10⁻⁶ CFU / mL. 9 CFU / mL. The results show that replacing glucose with a non-grain carbon source did not significantly change the number of viable fermentation cells, but adding an additional non-grain carbon source significantly increased the number of viable fermentation cells.

[0034] Table 1. Culture medium composition table

[0035] Table 2. Count of viable bacteria in each group of Lactobacillus plantarum LT0077 fermentation

[0036] Note: 'a' indicates a comparison with the control group. P >0.05, n=3.

[0037] Example 4: High-density fermentation of bacterial strains and preparation of freeze-dried powder Lactobacillus plantarum LT0077 glycerol culture was streaked onto MRS plates and anaerobically cultured at 37°C for 24 h. Colonies were picked and inoculated into 12 ml shake tubes containing 5 ml of MRS medium and anaerobically cultured at 37°C for 20 h. 0.5 mL of the culture was then transferred to a 150 mL Erlenmeyer flask containing 30 mL of MRS medium and anaerobically cultured at 37°C for 10 h. The culture from the Erlenmeyer flask was then inoculated into a 5 L fermenter containing 3 L of fermentation medium and cultured at 37°C, 50 rpm, and aeration for 5 h to obtain the primary seed culture. This primary seed culture was then inoculated into a 100 L fermenter containing 70 L of fermentation medium and fermented at 37°C, 20 rpm, and aeration at a constant pH of 5.5 for 12 h. The pH was adjusted using 20% ​​sodium carbonate. The fermentation endpoint was determined by pH measurement; fermentation ended when the pH rose. At the end of fermentation, the viable cell count on MRS plates reached 14 billion CFU / mL. After fermentation, the fermentation broth was centrifuged at 8000 rpm and 4℃ for 15 min, the supernatant was discarded, and the bacterial sludge and freeze-drying protectant were mixed at a volume ratio of 1:2 to form an emulsion. The emulsion was poured into a stainless steel freeze-drying tray, with a thickness not exceeding 10 mm. The emulsion was pre-frozen in a freeze dryer at -45℃ for 4 hours. The freeze-drying curves were as follows: -40℃ for 2 hours, -30℃ for 2 hours, -20℃ for 10 hours, -10℃ for 14 hours, 0℃ for 8 hours, 10℃ for 6 hours, and 25℃ for 10 hours. The moisture content at the freeze-drying endpoint was less than 3%. 1.2 kg of *Lactobacillus plantarum* LT0077 freeze-dried powder was obtained, and the viable count of the freeze-dried powder reached 800 billion CFU / g.

[0038] Fermentation medium: 10g yeast powder, 10g peptone, 30g glucose, 1g dipotassium hydrogen phosphate, 0.1g magnesium sulfate, 0.01g manganese sulfate, 1g Tween 80, 1000ml water, pH 7.0 before sterilization, sterilize at 121℃ for 30min.

[0039] Freeze-drying protectant: 150g trehalose, 50g sucrose, 1g ascorbic acid, 1g Tween 80, 1000mL water.

[0040] Example 5: Detection of strain resistance 5.1 Simulated Gastric and Intestinal Fluid Tolerance Test Sterile simulated gastric fluid was purchased from Shanghai Yuanye Biotechnology Co., Ltd. Glycerol-coated bacterial cultures were streaked onto MRS plates and anaerobically cultured at 37°C for 24 h. Colonies were then picked and inoculated into 12 ml shake tubes containing 5 ml of MRS medium, and anaerobically cultured at 37°C for 20 h. 0.5 mL of the culture was then transferred to a 150 mL Erlenmeyer flask containing 30 mL of MRS medium and anaerobically cultured at 37°C for 16 h. 5 mL of the bacterial culture was centrifuged at 5000 rpm for 10 min, the supernatant was discarded, and the culture was resuspended in sterile physiological saline. The resuspended culture was mixed at a 1:1 ratio with simulated gastric fluid at pH 2.0, 3.0, and 4.0, and incubated in a water bath at 37°C. Counts were performed at serial dilutions at 0 h, 0.5 h, 2 h, and 3 h, with three replicates for each bacterial culture. The survival rate was calculated as (number of viable cells after culture / number of viable cells at 0 h) × 100%. pH 2.0 simulated fasting, pH 3.0 simulated normal conditions, and pH 4.0 simulated satiated conditions. Table 3 shows that Lactobacillus plantarum LT0077 had a high survival rate under all three simulated conditions, with a survival rate of over 90% under both standard simulated gastric juice and saturated simulated gastric juice conditions.

[0041] Table 3. Count of viable Lactobacillus plantarum LT0077 in simulated gastric fluid

[0042] 5.2 Simulated artificial intestinal fluid bile salt tolerance test The resuspended bacterial solution was mixed with simulated artificial intestinal fluid at a ratio of 1:9, with 0.3% bovine bile salts added to the intestinal fluid. The mixture was incubated at 37°C, and samples were taken at 0 h, 3 h, and 24 h for serial dilution and counting. Each strain was counted in triplicate. The survival rate of the strain was calculated as (number of viable bacteria at sampling time / number of viable bacteria at 0 h) × 100%. As shown in Table 4, *Lactobacillus plantarum* LT0077 exhibited good tolerance to intestinal fluid and bile salts.

[0043] Table 4. Number of viable Lactobacillus plantarum LT0077 in simulated intestinal fluid

[0044] 5.3 Salinity tolerance MRS culture media with sodium chloride concentrations of 0‰, 20‰, 40‰, 60‰, 80‰, and 100‰ were prepared. 20 mL of MRS culture medium was placed in a 50 mL Erlenmeyer flask, and inoculated at a 2.5% inoculum. The culture was then incubated at 37℃ for 18 hours, and the viable count was determined.

[0045] The results are shown in Table 5 below. Lactobacillus plantarum LT0077 can grow normally in MRS with a sodium chloride concentration of 60‰, and can even survive in MRS with a sodium chloride concentration of 100‰, indicating that Lactobacillus plantarum LT0077 has a strong salt tolerance.

[0046] Table 5. Viable count of *Lactobacillus plantarum* LT0077 in MRS media with different sodium chloride concentrations.

[0047] 5.4 Oxidative stress tolerance Prepare physiological saline solutions containing 0.2 mM, 0.5 mM, 1 mM, 2 mM, 5 mM, and 10 mM hydrogen peroxide (the physiological saline was sterilized before adding hydrogen peroxide). *Lactobacillus plantarum* LT0077 cultured for 16 h was centrifuged at 8000 rpm for 5 min, resuspended in physiological saline, and the resuspended solution was added to the hydrogen peroxide-containing physiological saline solution at a 1:1 ratio. After incubation at 37℃ for 1 h, the viable cell count and OD600 were measured.

[0048] The results are shown in Table 6 below. The survival rate of Lactobacillus plantarum LT0077 after standing in physiological saline containing 10 mM hydrogen peroxide for 1 hour was still 49.93%, indicating that Lactobacillus plantarum LT0077 has high antioxidant activity and can still survive under high concentration of oxygen stimulation.

[0049] Table 6. Survival rate of Lactobacillus plantarum LT0077 in different concentrations of hydrogen peroxide.

[0050] 5.5 Determination of Acid Production Capacity The titration method was used to determine the acid value of *Lactobacillus plantarum* LT0077 after 16 hours of culture. 100 mL of pure water was added, along with 3-4 drops of phenolphthalein indicator. The solution was titrated with 0.1 M / L sodium hydroxide until it turned pink and the color did not fade after 30 seconds. The formula is: X = ((V-V0)×c×40) / m. Where X = acid value (mg / g); V = volume of standard titrant consumed in the sample solution (mL); V0 = volume of standard titrant consumed in the blank test (mL); c = concentration of the standard titrant used (mol / L); 40 = molar mass of sodium hydroxide (g / mol); and m = mass of the bacterial culture used in the test. The results indicate that the fermentation acid value of *Lactobacillus plantarum* LT0077 is approximately 8.74.

[0051] 5.6 Antibacterial test Staphylococcus aureus ATCC 6538 and Escherichia coli ATCC25922 were cultured overnight on NB medium, and the bacterial cultures were diluted to 1-5 × 10⁻⁵ with physiological saline. 8The diluted bacteria (cfu / mL) were added at 1% to NA medium at 45℃, shaken well, and 20 mL was accurately pipetted into agar plates. After the plates solidified, Oxford cups were placed on top, and 100 μL of the probiotics cultured for 16 h were added. The plates were then placed in a 4℃ refrigerator for 6 h, followed by overnight incubation at 36℃. The size of the inhibition zone was measured using calipers. Table 7 shows that *Lactobacillus plantarum* LT0077 possesses strong antibacterial activity.

[0052] Table 7. Inhibition diameter of Lactobacillus plantarum LT0077

[0053] 5.7 Self-aggregation and co-aggregation Lactobacillus plantarum LT0077, Staphylococcus aureus ATCC 6538 (purchased from Guangdong Provincial Microbial Culture Collection Center), and Escherichia coli ATCC25922 (purchased from Guangdong Provincial Microbial Culture Collection Center), cultured for 18 h, were centrifuged at 3000×g for 10 min, and the supernatant was discarded. An equal volume of sterile PBS buffer was added, and the mixture was resuspended and mixed. The mixture was centrifuged at 3000×g for 10 min, and this step was repeated twice. A small amount of PBS buffer was added and the mixture was resuspended and mixed. The bacterial suspension to be tested was mixed with PBS buffer at a certain ratio so that the OD value of the mixed solution at a wavelength of 600 nm was about 0.8, which was recorded as A0. 4 ml of the bacterial suspension after adjusting the concentration was taken and thoroughly mixed, and incubated at room temperature for 5 h. 1 ml of the supernatant bacterial suspension was taken, and the OD value was measured at 600 nm using PBS buffer as a control, which was recorded as At. The self-aggregation rate (%) was calculated as: 1 - At / A0 × 100%.

[0054] Equal volumes (2 ml) of *Lactobacillus plantarum* LT0077 suspension and *Staphylococcus aureus* ATCC 6538 or *Escherichia coli* ATCC25922 suspension were mixed and incubated at room temperature for 5 h. A control group containing 4 ml of single-bacterial suspension was cultured under the same growth conditions, and the OD value was measured at 600 nm. The experiment was repeated three times, and the co-aggregation rate (%) was calculated as follows: ∫[(Ax+Ay) / 2 - A(x+y)] / (Ax+Ay) × 100%. (Ax and Ay represent the absorbance of the two bacterial suspensions, and A(x+y) represents the absorbance of the mixed bacterial suspension.) The test results are shown in Table 8 below. The results indicate that Lactobacillus plantarum LT0077 can effectively aggregate Staphylococcus aureus and Escherichia coli.

[0055] Table 8. Determination of autoaggregation and coagulation of Lactobacillus plantarum LT0077

[0056] Comparative Example 1 Lactobacillus plantarum CICC25125 was used to replace Lactobacillus plantarum LT0077 in Example 3. The other detection methods were the same as in Example 3. Lactobacillus plantarum CICC25125 was purchased from the China Industrial Microbial Culture Collection Center. The detection results are shown in Table 9. Under the three conditions of fasting simulated gastric juice, standard simulated gastric juice, and satiated simulated gastric juice, the survival rate of Lactobacillus plantarum CICC25125 was lower than that of Lactobacillus plantarum LT0077. Lactobacillus plantarum LT0077 has a stronger gastric juice tolerance.

[0057] Table 9. Count of viable Lactobacillus plantarum CICC25125 in simulated gastric fluid

[0058] Example 6: Determination of Free Radical Scavenging Ability 6.1 Determination of ABTS free radical scavenging ability After activation on agar plates, the culture was incubated in shake flasks for 16 hours. The fermentation broth was then centrifuged at 8000 rpm for 10 minutes. The supernatant was collected and set aside. The precipitate was resuspended in PBS, and the cell concentration was adjusted to 10⁻⁶. 9 CFU / mL. The resuspended solution was sonicated in an ice bath at 300 W for 3 seconds, with a 7-second interval, for a total of 3 min. The supernatant was collected and placed on ice for testing. The ABTS free radical scavenging capacity of the fermentation broth supernatant and cell-free intracellular contents was simultaneously measured. 3.6 mL of ABTS solution and 0.4 mL of glutathione solution were added to tube 1 as the experimental group (As); 3.6 mL of ABTS solution and 0.4 mL of sample solvent solution were added to tube 2 as the blank group (Ab). After thorough mixing, the mixture was reacted at room temperature in the dark for 5 min, centrifuged at 1000 rpm for 5 min at room temperature, and the absorbance of the supernatant was measured using a UV spectrophotometer at a wavelength of 734 nm (sample solvent was used for zeroing calibration). The ABTS scavenging rate was calculated as follows: P = (Ab - As) / Ab × 100%. P: Scavenging rate; Ab: Absorbance of the mixture of ABTS solution and sample solvent solution; As: Absorbance of the mixture of test solution and ABTS solution.

[0059] 6.2 Determination of DPPH free radical scavenging ability After activation on agar plates, the culture was incubated in shake flasks for 16 hours. The fermentation broth was then centrifuged at 8000 rpm for 10 minutes. The supernatant was collected and set aside. The precipitate was resuspended in PBS, and the cell concentration was adjusted to 10⁻⁶. 9CFU / mL. The resuspension was sonicated in an ice bath at 300 W for 3 seconds, with a 7-second interval, for a total of 3 min. The supernatant was collected and placed on ice for testing. The DPPH free radical scavenging capacity of the fermentation broth supernatant and cell-free intracellular contents was simultaneously measured. Glutathione standard solutions were prepared at concentrations of 0.03125 mg / mL, 0.625 mg / mL, 1.25 mg / mL, 2.5 mg / mL, and 5.0 mg / mL. 3.0 mL of 50 μg / mL DPPH solution and 1.0 mL of the above glutathione standard solution were added to a test tube as the experimental group; 1.0 mL of glutathione solution and 3.0 mL of anhydrous ethanol solution were added to a test tube as the control group; and 3.0 mL of DPPH solution and 1.0 mL of sample solvent solution were added to a test tube as the blank group. After thorough mixing, the mixture was reacted at room temperature in the dark for 30 min, centrifuged at 1000 rpm for 5 min at room temperature, and the supernatant was collected. The absorbance was measured using a UV spectrophotometer at a wavelength of 517 nm (zeroing was performed using the sample solvent). The DPPH scavenging rate was calculated as follows: P = (1 - (AS - AC) / AB)) × 100%. P: scavenging rate; AS: absorbance of the mixture of the test solution and DPPH solution; AC: absorbance of the mixture of the test solution and anhydrous ethanol solution; AB: absorbance of the mixture of DPPH solution and sample solvent solution.

[0060] 6.3 Hydroxyl radical scavenging capacity determination The hydroxyl radical scavenging capacity was determined using a kit from Beijing Solarbio Science & Technology Co., Ltd. After activation of the bacterial culture on agar plates, single colonies were inoculated into MRS medium and incubated statically in shake flasks for 16 h. The fermentation broth was then aspirated and centrifuged at 8000 rpm for 10 min. The supernatant was collected as the test sample. The fermentation broth was centrifuged at 8000 rpm for 10 min, and the precipitate was resuspended in PBS to adjust the cell concentration to 10-1. 9 CFU / mL. The resuspended solution was sonicated in an ice bath at 200 W for 3 seconds, with 10-second intervals, for a total of 6.5 min. Then, it was centrifuged at 8000 rpm at 4℃ for 10 min. The supernatant was collected and placed on ice for testing (no cell contents). The hydroxyl radical scavenging capacity of the fermentation broth supernatant and cell-free intracellular contents was simultaneously measured. The spectrophotometer or microplate reader was preheated for at least 30 min, and the wavelength was adjusted to 536 nm. The spectrophotometer was zeroed with distilled water. The reagents were added sequentially to 1.5 mL EP tubes according to the instructions, vortexed, and reacted accurately in a 37℃ water bath for 60 min. Afterward, the sample was centrifuged at 10000 rpm at room temperature for 10 min. 200 μL of the sample was measured at 536 nm in a microcup. Hydroxyl radical scavenging rate (%) = (Ameasured - Acontrol) / (Ablank - Acontrol) × 100%.

[0061] The test results are shown in Table 10. The experimental results show that compared with Lactobacillus plantarum CICC25125 (purchased from China Industrial Microbial Culture Collection Center) and Lactobacillus plantarum CICC10481 (purchased from China Industrial Microbial Culture Collection Center), Lactobacillus plantarum LT0077 has a better ability to scavenge ABTS free radicals, DPPH free radicals and hydroxyl free radicals, indicating that it has certain potential in anti-oxidation and liver repair.

[0062] Table 10 Free radical scavenging capacity of Lactobacillus plantarum LT0077

[0063] Example 7 Verification of nicotinic acid and nicotinamide production After activation of the bacterial culture on plates, colonies were picked and transferred to MRS precursor medium, and incubated statically at 37°C for 16 hours. The MRS precursor medium was prepared as follows: MRS medium was prepared, sterilized at 121°C for 15 minutes, cooled, and then 500 mg / L of NR malate was added via filtration sterilization. The cultured bacterial solution was centrifuged at 10,000 rpm for 10 minutes at 4°C. The supernatant was collected, filtered through a 0.22 μm filter, and analyzed by liquid chromatography. The detection method was as follows: Mobile phase A: 1 ml phosphoric acid, 150 ml acetonitrile, 850 ml water, 1.22 g sodium 1-decanesulfonate; 100% A was eluted isocratically; flow rate: 1.0 mL / min; injection: 10 µL; detection time: 40 min. Experimental results are as follows: Figure 3 As shown, the nicotinic acid yield of Lactobacillus plantarum LT0077 is 13.12 mg / L.

[0064] Example 8: Verification of Indole Derivative Production After activation of the bacterial culture on agar plates, colonies were picked and transferred to MRS precursor medium, and incubated statically at 37°C for 16 h. The cultured bacterial solution was then centrifuged at 10,000 rpm for 10 min at 4°C. The supernatant was collected, filtered through a 0.22 μm filter, and analyzed by liquid chromatography. The detection method was as follows: using a C18 4.6 × 250 mm, 5 µm (40°C) mobile phase A: water; B: acetonitrile, gradient elution: 0–1 min 10% B; 1–8 min 10–95% B; 8–9.5 min 95% B; 9.6–12 min 10% B; 12–15 min 10% B, flow rate: 1.0 mL / min, injection volume: 10 µL. Figure 4 As shown, liquid chromatography analysis revealed that the melatonin production of Lactobacillus plantarum LT0077 was 42.282 mg / L.

[0065] Example 9 Animal Experiment Used 4-6 week old SPF-grade male mice. Animals were housed in an environment of 21-25°C and 48-55% humidity, maintaining a 12-hour light / 12-hour dark cycle. Mice were randomly divided into 4 groups of 10 mice each: Negative control group (NC): administered only saline, no alcohol; Model control group (AD): administered only alcohol; Lactobacillus plantarum LT0077 group: administered alcohol and 10... 8 CFU / probiotic. Positive drug group: Metadoxine capsules - Xinlide 200mg / kg·d. After one week of acclimatization, the model group and control group were administered 0.2 mL of physiological saline by gavage at 9:30 AM daily, while the experimental group was administered 0.2 mL of probiotic solution by gavage for 19 days. Starting from day 15, the model group and experimental group were administered 53% ABV Erguotou (a type of Chinese liquor) by gavage 1 hour after the administration of probiotics and a positive control drug, at a dose of 8 mL / kg body weight, adjusted to 14 mL / kg body weight on the last day. The control group was administered the same volume of physiological saline by gavage to establish an acute alcohol poisoning model. After the experiment, the animals were fasted for 6 hours but allowed free access to water. Blood was collected under inhalation anesthesia to test ALT, AST, TG, LDL, and carbon dioxide levels. The animals were euthanized by asphyxiation, and their livers were harvested to test for alcohol dehydrogenase. The livers were then used for pathological hematologic staining.

[0066] After the final gavage, six mice were selected from each group (excluding the control group) for testing. The mice were placed back-down in their cages, and their position was adjusted every 5 seconds until they could no longer correct themselves immediately. If a mouse could not roll over on its own within 30 seconds, the reflex was considered lost (intoxicated); if the mouse could roll over again on its own, the reflex was considered restored (sober). The latency period of intoxication (time from alcohol intake to intoxication reaction) and the sobering time (time from loss of reflex to recovery) were recorded after the mice were given alcohol via gavage. like Figures 5-8 The results showed that AST, ALT, TG and LDL levels in the Lactobacillus plantarum LT0077 group decreased compared with those in the model group, indicating that Lactobacillus plantarum LT0077 has a certain repair ability in liver injury repair in mice and can alleviate liver damage caused by ethanol.

[0067] like Figure 9 As shown, compared with the model group, the alcohol dehydrogenase activity in the Lactobacillus plantarum LT0077 group increased, indicating that oral administration of Lactobacillus plantarum LT0077 can promote the activity of alcohol dehydrogenase in mice after alcohol stimulation and help mice improve their alcohol metabolism.

[0068] The results in Table 11 show that, compared with the model group, the alcohol intake to intoxication reaction time and sobering time were significantly reduced in the positive drug group and the Lactobacillus plantarum LT-0119 group.

[0069] Table 11 Comparison of alcohol intake time to intoxication reaction time and sobering time in different groups of mice

[0070] Note: 'a' indicates a comparison with the model group. P < 0.05, n = 6.

[0071] Based on HE staining results ( Figure 10 It was found that the liver histological structure of the control group sections basically maintained the normal lobular structure. Specific pathological features: the lobular structure was relatively clear, hepatocytes were arranged in cords, and the hepatic cords and sinusoidal structures were distinguishable. Central veins (or vascular structures) and portal structures were visible, including a large luminal structure (possibly an intrahepatic vessel or bile duct, with no obvious contents or only a blank area). The surrounding hepatocytes were generally normal in morphology, with round nuclei and normal nucleoplasm ratio. No obvious ballooning degeneration or fatty degeneration of hepatocytes was observed. Inflammatory cell infiltration was not obvious, and there were no obvious areas of hepatocyte necrosis. There was no obvious fibrosis in the lobules and portal areas.

[0072] Pathological results of liver tissue from the model group showed numerous vacuoles in the hepatocyte cytoplasm, appearing as round vacuoles of varying sizes, with some areas showing vacuolation, suggesting moderate to severe fatty degeneration. Hepatocyte arrangement was slightly disordered, and the hepatic lamina structure was locally blurred. Fatty degeneration was evident in hepatocytes around the central vein, and the sinusoids were narrowed due to hepatocyte swelling. Inflammatory cell infiltration within the hepatic lobules was predominantly lymphocytes and monocytes, with a small number of neutrophils. There was no significant large-scale hepatocyte necrosis, but localized apoptosis (nuclear pyknosis, increased eosinophilicity) of individual hepatocytes was observed. No significant fibrosis was observed in the portal areas, and the vascular structure showed no obvious abnormalities. The study presented with moderate to severe hepatocyte fatty degeneration accompanied by an inflammatory response.

[0073] In the positive drug group, the liver lobule structure was basically intact, and the arrangement of hepatocyte cords was not significantly disordered. A small number of small vacuoles were visible in the hepatocyte cytoplasm, which was significantly reduced compared with the model group. The nuclei were normal in morphology and the nucleocytoplasmic ratio was normal. There was no obvious inflammatory cell infiltration, no signs of hepatocyte necrosis, and no fibrosis in the liver lobules or portal areas. Mild fatty degeneration of hepatocytes and local vascular congestion were the main characteristics.

[0074] In the *Lactobacillus plantarum* LT0077 group, the liver lobule structure was discernible, with mild morphological changes in local hepatocytes and focal congestion. Specific pathological features: hepatocyte cords were relatively well-arranged; small vacuoles (mild fatty degeneration) were visible in the cytoplasm of some hepatocytes; focal erythrocyte aggregation (focal congestion) was visible in the lower left corner; no obvious inflammatory cell infiltration was observed in the hepatic sinusoids; there was no proliferation of connective tissue between liver lobules, and the portal area structure was normal. Severity assessment: Mild lesion, mainly characterized by focal congestion and mild fatty degeneration of hepatocytes. The lesions showed significant improvement compared to the model group.

[0075] All of the foregoing primary implementations of this intellectual property right do not limit other forms of implementation of this new product and / or new method. Those skilled in the art will utilize this important information to modify the foregoing to achieve similar implementations. However, all modifications or alterations based on this new product invention are reserved rights.

[0076] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention in any other way. Any person skilled in the art may make changes or modifications to the above-disclosed technical content to create equivalent embodiments. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the protection scope of the present invention.

Claims

1. *Lactobacillus plantarum* can accelerate alcohol metabolism and improve hangover symptoms. Lactiplantibacillus plantarum ), characterized by: The plant lactobacillus ( Lactiplantibacillus plantarum It is named LT0077, with accession number GDMCC No. 67489, and its 16S nucleotide sequence is shown in SEQ ID NO.

1.

2. A product comprising the *Lactobacillus plantarum* as described in claim 1 (… Lactiplantibacillus plantarum ) freeze-dried powder formulation.

3. The *Lactobacillus plantarum* as described in claim 1 ( Lactiplantibacillus plantarum The method for producing the fermented freeze-dried product is characterized by, include: The plant lactobacillus ( Lactiplantibacillus plantarum The culture medium was inoculated and cultured at 37°C for 12-24 hours to obtain a culture solution. The culture solution was centrifuged, and the wet bacterial precipitate was collected. The wet bacterial precipitate was resuspended and mixed with a lyophilization protectant solution, and then freeze-dried to obtain a lyophilized powder formulation.

4. The *Lactobacillus plantarum* as described in claim 3 ( Lactiplantibacillus plantarum The method for producing the fermented freeze-dried product is characterized by: The fermentation medium consists of 10g yeast powder, 10g peptone, 30g glucose, 1g dipotassium hydrogen phosphate, 0.1g magnesium sulfate, 0.01g manganese sulfate, 1g Tween 80, and 1000ml water. The pH of the fermentation medium before sterilization is 7.

0.

5. The *Lactobacillus plantarum* as described in claim 3 ( Lactiplantibacillus plantarum The method for producing the fermented freeze-dried product is characterized by: The freeze-drying protectant solution consists of 150g trehalose, 50g sucrose, 1g ascorbic acid, 1g Tween 80, and 1000mL water.

6. Containing the *Lactobacillus plantarum* as described in claim 1 ( Lactiplantibacillus plantarum The microbial preparation of ) is characterized by: The plant lactobacillus ( Lactiplantibacillus plantarum The viable count is not less than 1×10⁻⁶. 6 CFU / mL or 1×10 6 CFU / g.

7. The microbial preparation according to claim 6, characterized in that: It also contains other probiotics.

8. A drug that can accelerate alcohol metabolism and improve hangover symptoms, characterized in that: It includes the microbial preparation described in claim 6 or 7.

9. The *Lactobacillus plantarum* as described in claim 1 ( Lactiplantibacillus plantarum Its application in the preparation of drugs that can accelerate alcohol metabolism and improve hangover symptoms.

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