Application of lactobacillus mucilaginosus strain A21196 in preparation of product for promoting ethanol metabolism

Abstract

The invention provides application of a lactobacillus mucus fermentation strain A21196 in preparation of a product for promoting ethanol metabolism, and belongs to the technical field of microbial preparations. The lactobacillus mucus fermentation strain A21196 has the high-yield ethanol dehydrogenase and acetaldehyde dehydrogenase, and has the effects of reducing the content of ethanol and acetaldehyde in the blood of mice suffering from acute alcoholism and shortening the central nervous inhibition time in vivo. Therefore, the invention provides the application of the fermented lactobacillus mucus strain A21196 in preparation of a product for preventing and / or treating the alcoholic liver injury or the alcoholic liver disease.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial preparations, and particularly relates to application of a fermented Lactobacillus mucilaginosus strain A21196 in preparing a product that promotes ethanol metabolism. Background Art

[0002] It is well known that excessive drinking has significant harmful effects on human health. Although some natural product extracts, small molecule drugs, and colloid detoxifiers have been developed as alcohol detoxification agents, most treatment options can only provide supportive treatment (e.g., restoring consciousness and alleviating adverse reactions) rather than directly decomposing alcohol.

[0003] The metabolism of alcohol mainly depends on alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) in liver cells. These two enzymes work together continuously to convert ethanol into acetic acid while eliminating the toxic intermediate metabolite, acetaldehyde. Acetaldehyde is a toxic carcinogen that causes double-strand breaks in cell DNA and chromosome rearrangements, and permanently changes the DNA sequence, thereby increasing the risk of cancer. In addition, excessive drinking can also cause severe liver damage. Studies have shown that exogenous administration of ADH and ALDH can effectively alleviate alcoholic liver damage caused by excessive drinking. However, direct oral administration will cause most enzymes to be inactivated in the gastrointestinal tract. To solve this problem, a large amount of enzyme carriers need to be administered, but the loading efficiency of most enzyme carriers is low, resulting in insufficient activity of ADH and ALDH in vivo. Therefore, there is an urgent need for a safe, convenient and effective in vivo alcohol detoxification strategy.

[0004] In recent years, more and more evidence has shown that bacteria can regulate the host's metabolism by degrading potential toxic substances. Studies have shown that oral administration of genetically engineered bacteria that highly express ADH and ALDH can reduce blood ethanol content in alcohol poisoning mouse models and alleviate liver damage, which indicates that the method based on bacterial delivery of ADH and ALDH has great application potential in alcohol detoxification treatment. At present, the prior art reports that Bacillus coagulans obtained by induction has the ability to degrade alcohol (CN 116731896A), Lactobacillus plantarum has the ability to produce high acetaldehyde dehydrogenase (CN107760624A), and Lactobacillus casei has the ability to produce high acetaldehyde dehydrogenase, which can be used to prepare alcohol detoxification products (CN20230822A). However, there is currently no report on the fermentation of mucus lactobacillus with ethanol degradation function. Summary of the invention

[0005] In view of this, the object of the present invention is to provide an application of fermented Lactobacillus mucoides strain A21196 in the preparation of products that promote ethanol metabolism. Fermented Lactobacillus mucoides strain A21196 has the performance of simultaneously producing high levels of ethanol dehydrogenase and acetaldehyde dehydrogenase, and has the characteristic of degrading alcohol poisoning in the body, providing a new choice for the preparation of products for preventing or treating alcoholic liver damage or alcoholic fatty liver.

[0006] The invention provides application of fermented Lactobacillus mucilaginosus strain A21196 in preparing a product promoting ethanol metabolism.

[0007] The invention provides application of fermented Lactobacillus mucilaginosus strain A21196 or a probiotic agent thereof in producing alcohol dehydrogenase and / or acetaldehyde dehydrogenase.

[0008] Preferably, the method for producing alcohol dehydrogenase and / or acetaldehyde dehydrogenase comprises culturing a bacterial suspension of Lactobacillus mucosa strain A21196 in a culture medium containing 4% to 6% ethanol by volume, separating the bacterial cells from the culture medium, crushing the cells, collecting the cell contents, and removing cell fragments and impurities to obtain alcohol dehydrogenase and / or acetaldehyde dehydrogenase.

[0009] The invention provides application of fermented Lactobacillus mucilaginosus strain A21196 or a probiotic agent thereof in preparing a product for sobering up and protecting the liver.

[0010] Preferably, the alcohol sobering up and protecting the liver includes shortening the sobering up time and / or reducing the concentration of ethanol and acetaldehyde in the serum of drunk people.

[0011] The present invention provides application of fermented Lactobacillus mucilaginosus strain A21196 or a probiotic agent thereof in preparing a medicine for preventing and / or treating alcoholic liver injury.

[0012] Preferably, the concentration of live bacteria in the probiotics fermented from Lactobacillus mucilaginosus strain A21196 is not less than 5×10 9 CFU / g or 5×10 9 CFU / mL.

[0013] Preferably, the probiotic preparation of the fermented Lactobacillus mucilaginosus strain A21196 comprises a lyophilized powder or an aqueous solution.

[0014] Preferably, the lyophilized powder is prepared by mixing the fermented Lactobacillus mucilaginosus strain A21196 with a lyophilization protectant and then freeze-drying the mixture.

[0015] Preferably, the lyophilization protective agent is a skim milk aqueous solution with a mass percentage of 10% to 14%.

[0016] The present invention provides the application of fermented mucus lactobacillus strain A21196 in the preparation of products promoting ethanol metabolism. The strain A21196 has the characteristics of high production of ethanol dehydrogenase and acetaldehyde dehydrogenase at the same time, and the secretion concentrations reach 1048.15U / ml and 19.83gprot / L respectively, which is significantly better than other fermented mucus lactobacillus strains. Animal experiments have shown that the strain A21196 effectively reduces the ethanol and acetaldehyde content in the blood of mice with acute alcohol poisoning, and greatly shortens the sobering time. It can be seen that the fermented mucus lactobacillus strain A21196 provides a new and effective way for the rapid sobering of drunk people and the prevention and treatment of alcoholic liver damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The colony morphology of strain A21196 (left) and Gram staining microscopy (right);

[0018] Figure 2 The results are the results of the ADH and ALDH production ability test of strain A21196 in MRS medium containing 5% ethanol;

[0019] Figure 3 This is a bar graph of the time required for mice in each group to recover their righting reflex;

[0020] Figure 4 This is the result of the determination of ethanol content in mouse serum;

[0021] Figure 5 The results are for the determination of acetaldehyde content in mouse serum. DETAILED DESCRIPTION

[0022] The invention provides application of fermented Lactobacillus mucilaginosus strain A21196 in preparing a product promoting ethanol degradation or metabolism.

[0023] In the present invention, the fermented mucus Lactobacillus strain A21196 was isolated from a fecal sample of a healthy centenarian and was deposited in the Guangdong Microbial Culture Collection Center on November 12, 2022, with a deposit number of GDMCC No: 62959. The fermented mucus Lactobacillus strain A21196 is disclosed in the patent publication number CN116004455.

[0024] In the present invention, the dosage form of the probiotic agent of the fermented mucus lactobacillus strain A21196 preferably includes a lyophilized powder or an aqueous solution. The lyophilized powder is preferably prepared by mixing the bacterial body of the fermented mucus lactobacillus strain A21196 and a lyophilization protective agent and then freeze-drying. The lyophilization protective agent is preferably a skim milk aqueous solution with a mass percentage of 10% to 14%, and can be a skim milk aqueous solution with a mass percentage of 12%. The preparation method of the bacterial body of the fermented mucus lactobacillus strain A21196 preferably involves seed culture of the fermented mucus lactobacillus strain A21196 twice, and the obtained seed liquid is inoculated into an MRS liquid culture medium for expansion culture to obtain a large number of bacterial bodies. The culture medium for the seed culture is an MRS liquid culture medium. The time for the seed culture is 11 to 13 hours, and can be 12 hours. The inoculation amount during the seed culture is preferably 3%. The seed culture is preferably anaerobic culture. The time for the expansion culture is preferably 12 to 16 hours, and more preferably 14 hours. After obtaining the bacterial body of the fermented Lactobacillus mucoides strain A21196, it is preferably placed in an ice-water mixture and allowed to stand for 30 minutes before collecting the bacterial body, which is beneficial to increase the number of live bacteria. Before mixing with the freeze-drying protective agent, the bacterial body of the fermented Lactobacillus mucoides strain A21196 is preferably washed twice with physiological saline to remove residual culture medium. The mass volume ratio of the bacterial body of the fermented Lactobacillus mucoides strain A21196 and the freeze-drying protective agent is 1g:1mL. The preparation method of the aqueous solution preferably mixes the expanded culture solution of the fermented Lactobacillus mucoides strain A21196 with glycerol. The volume percentage of glycerol in the aqueous solution is preferably 40% to 80%, and can be 50%, 60%, or 70%. In the present invention, the concentration of live bacteria in the probiotic agent of the fermented Lactobacillus mucoides strain A21196 is preferably not less than 5×10 9 CFU / g or 5×10 9 CFU / mL, can be (10~100)×10 9 CFU / gCFU / g or (10~100)×10 9 CFU / mL.

[0025] In the present invention, the promotion of ethanol degradation or metabolism refers to the degradation or metabolism of ethanol under the action of alcohol dehydrogenase and / or acetaldehyde dehydrogenase secreted by the fermentation Lactobacillus mucosa strain A21196, specifically degrading ethanol into acetic acid and acetaldehyde in sequence, thereby fundamentally reducing the damage of ethanol to the liver.

[0026] In the present invention, the product preferably includes medicine, health care product or health food. The present invention has no special limitation on the preparation method of the product, and the preparation method of the product known in the art can be used.

[0027] The invention provides application of fermented Lactobacillus mucilaginosus strain A21196 or a probiotic agent thereof in producing alcohol dehydrogenase and / or acetaldehyde dehydrogenase.

[0028] In the present invention, the method for producing alcohol dehydrogenase and / or acetaldehyde dehydrogenase preferably comprises culturing a suspension of Lactobacillus mucilaginosus strain A21196 in a culture medium containing 4% to 6% ethanol by volume, separating the cells from the culture medium, crushing them, collecting the cell contents, removing cell fragments and impurities, and obtaining alcohol dehydrogenase and / or acetaldehyde dehydrogenase. The volume percentage of ethanol in the culture medium is 5%. The type of the culture medium preferably includes MRS culture medium. The OD value of the suspension of Lactobacillus mucilaginosus strain A21196 is 0.0447 W / m. 600 The value is preferably 0.6. The culture time is preferably 22 to 26 hours, and can be 24 hours. The culture is conducive to inducing the expression of alcohol dehydrogenase and / or acetaldehyde dehydrogenase. The method for separating bacteria preferably includes centrifugation. The speed of the centrifugation is preferably 5000 to 7000 rpm, and can be 6000 rpm. The time of the centrifugation is preferably 1 to 3 minutes, and more preferably 2 minutes. Before the crushing, the bacteria are preferably washed with PBS solution to remove the residual culture medium. The crushing method preferably includes a cell crusher. The method for removing bacterial fragment impurities is preferably to centrifuge the cell contents after crushing, discard the precipitate, and collect the supernatant. The speed of the centrifugation is preferably 12000g. The time of the centrifugation is preferably 20 minutes. The content of alcohol dehydrogenase and / or acetaldehyde dehydrogenase in the supernatant was detected using commercial detection kits, and the results showed that the concentrations of alcohol dehydrogenase and acetaldehyde dehydrogenase in the supernatant reached 1048.15U / ml and 19.83gprot / L.

[0029] The present invention provides application of fermented Lactobacillus mucilaginosus strain A21196 or a probiotic agent thereof in preparing a medicine for preventing and / or treating alcoholic liver injury.

[0030] In the present invention, the concentration of live bacteria in the drug is preferably not less than 5×10 9 CFU / g or 5×10 9 CFU / mL, can be (10~100)×10 9 CFU / gCFU / g or (10~100)×10 9 CFU / mL. The dosage form of the drug preferably includes oral solution, tablet, capsule, powder, etc. The present invention has no special limitation on the preparation method of the drug, and the preparation method of the drug dosage form well known in the art can be used.

[0031] The invention provides application of fermented Lactobacillus mucilaginosus strain A21196 or a probiotic agent thereof in preparing a product for sobering up and protecting the liver.

[0032] In the present invention, the alcohol sobering up and protecting the liver preferably includes shortening the sobering up time and / or reducing the concentration of ethanol and acetaldehyde in the serum of drunk people.

[0033] In the embodiment of the present invention, the Gaobinge alcoholic liver injury animal model was used as an object to verify the ability of fermented Lactobacillus mucosa strain A21196 to promote ethanol metabolism in vivo. The modeling method of the alcoholic liver injury animal model is simple and has good repeatability. The alcohol concentration in the serum is high, which can simulate the pathogenesis of human alcoholic liver disease. The experimental results show that fermented Lactobacillus mucosa strain A21196 can shorten the righting reflex time of mice and shorten the duration of drunkenness of mice. By detecting the ethanol and acetaldehyde content in the blood, the results show that fermented Lactobacillus mucosa strain A21196 has a significant effect in degrading ethanol and acetaldehyde.

[0034] The application of the fermented Lactobacillus mucilaginosus strain A21196 provided by the present invention in the preparation of products promoting ethanol metabolism is described in detail below in conjunction with the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0035] Example 1

[0036] 1. Screening and purification method of fermented Lactobacillus mucilaginosus strain A21196

[0037] The fermentative mucus Lactobacillus strain A21196 was isolated from the fecal sample of a healthy centenarian. The sample was diluted at different times and spread on MRS solid medium and cultured in a 37°C constant temperature incubator for 48 hours. The target single colony was picked and purified and cultured twice on MRS solid medium.

[0038] 2. Cryopreservation of strain A21196

[0039] Pick a single colony on the purification plate and culture it in a 37°C constant temperature incubator for 16 hours. Mix the bacterial solution with 40% glycerol at a ratio of 1:1 and freeze it at -80°C for later use, and name it A21196.

[0040] 3. Morphological identification of strains

[0041] The growth morphology of strain A21196 on MRS solid medium is as follows Figure 1 (Left) As shown. The colony is slightly white, about 3mm in diameter, convex, round, and smooth. A single colony was selected for Gram staining, and the microscopic examination was as follows Figure 1 (right) The strain is Gram-positive, non-spore-forming, and the cells are short rod-shaped, arranged singly or in pairs.

[0042] 4. 16S rDNA Identification of Strains

[0043] Sangon Biotech (Shanghai) Co., Ltd. was commissioned to perform 16S rDNA amplification and sequence determination on the A21196 strain. The 16S rDNA sequence of fermented mucus Lactobacillus A21196 was obtained and uploaded to the NCBI (National Center for Biotechnology Information) database for comparison. The results showed that the gene sequence of strain A21196 was most similar to that of fermented mucus Lactobacillus fermentum, with a homology of 99.86%. Therefore, strain A21196 was preliminarily identified as fermented mucus Lactobacillus fermentum. The strain was deposited in the Guangdong Microbial Culture Collection Center on November 12, 2022, with a deposit number of GDMCC No: 62959.

[0044] Example 2

[0045] Determination of the ability of fermentative Lactobacillus mucilaginosus strain A21196 to secrete alcohol dehydrogenase and acetaldehyde dehydrogenase in vitro

[0046] 1. Preparation of MRS solid culture medium

[0047] Accurately weigh 10g of glucose, 5g of beef extract powder, 5g of peptone, 2.5g of yeast extract powder, 1g of dipotassium hydrogen phosphate, 1g of triammonium citrate, 2.5g of anhydrous sodium acetate, 40ml of Tween-80, 0.29g of magnesium sulfate, 0.125g of manganese sulfate, and 6g of agar, and dissolve them in 500ml of ultrapure water. Adjust the pH value to 6.2±0.2. Autoclave at 121℃ for 20min. When the temperature cools to about 70℃, take out and pour into a plate for later use.

[0048] 2. Activation of strains

[0049] The strains to be tested include the fermentative mucus lactobacillus strain A21196 and five strains of the same batch of fermentative mucus lactobacillus strains A20039-(10)-1, A20039-(11)-1, A20039-(12)-1, A20039-(13)-1, and A20039-(14)-1.

[0050] Take out the strain to be tested frozen at -80℃, pick a small amount of bacterial solution with an inoculation loop and streak it on the MRS solid culture medium plate. Incubate in a 37℃ constant temperature incubator for 48h. Then pick a single colony and activate it twice in 5ml MRS liquid culture medium with an inoculation volume of 1%. Incubate in a 37℃ constant temperature incubator for 16h to obtain fresh bacterial solution to be tested.

[0051] Determination of the ability of 3 strains to secrete alcohol dehydrogenase and acetaldehyde dehydrogenase in vitro

[0052] (1) Adjust the concentration of the fresh bacterial solution to OD 600 0.6, take 1 ml of bacterial suspension and add it into MRS liquid medium containing 5% ethanol and culture for 24 hours. 600 Adjust to 0.6, take 1 ml of bacterial suspension, centrifuge (6000rpm, 2min), discard the supernatant to obtain the bacteria. Wash twice with PBS, and then centrifuge to remove the supernatant. Add 800μl PBS and resuspend. Then use a cell disruptor to disrupt for 20min until the bacteria are completely lysed. After disruption, the bacteria are centrifuged at 12000g for 20min at 4°C, discard the precipitate and take the supernatant and freeze it in a -80°C refrigerator. Use a kit (ADH kit purchased from Nanjing Jiancheng Bioengineering Institute, item number: A083-2-1, ALDH kit purchased from Elabscience, item number: E-BC-K565-M), and determine the ADH and ALDH content in the supernatant according to the kit instructions.

[0053] The results are as follows Figure 2 As shown, after incubation in vitro with MRS liquid medium containing 5% ethanol, A21196 has the ability to produce high levels of ethanol dehydrogenase and acetaldehyde dehydrogenase, with concentrations reaching 1048.15 U / ml and 19.83 gprot / L. The ADH and ALDH production of strain A21196 was significantly higher than that of other control fermentation Lactobacillus mucoides strains.

[0054] Example 3

[0055] Preparation method of freeze-dried powder of fermented Lactobacillus mucilaginosus strain A21196

[0056] Use a coating rod to pick up a single colony of the fermented mucus lactobacillus strain A21196, activate and culture it twice in MRS solid culture medium, pick a single colony and inoculate it into 15ml MRS liquid culture medium, and culture it under anaerobic conditions for 12h to obtain seed liquid 1. Inoculate seed liquid 1 into a new MRS liquid culture medium with an inoculation amount of 3% and an inoculation volume of 150ml, and culture it under anaerobic conditions for 12h to obtain seed liquid 2. Inoculate seed liquid 2 into a new MRS liquid culture medium with an inoculation amount of 3% and an inoculation volume of 3.5L. Culture it under anaerobic conditions for 12 to 16h to obtain the amplified cultured bacterial liquid. Place the amplified cultured bacterial liquid in an ice-water mixture (4℃) for 30min, centrifuge it (4℃, 8000rpm, 6min), discard the supernatant, and collect the bacterial bodies. The collected bacterial bodies are washed twice with sterile saline (4℃, 6500rpm, 6min), and finally discard the saline to obtain A21196 bacterial bodies and weigh them. Mix the obtained bacterial cells in equal mass ratio with 12% skim milk (weigh 12g skim milk, dissolve it with ultrapure water and make up to 100ml. Sterilize at 115℃ for 20 minutes and set aside), stir evenly and dispense into 1.5ml EP tubes, place in a refrigerator and slowly freeze (-20℃, 30min, -80℃, 1h). Finally, place the frozen bacterial powder on a freeze dryer and freeze it to dry powder.

[0057] Example 4

[0058] Determination of the in vivo ethanol degradation ability of Lactobacillus mucilaginosus A21196 with high ADH and ALDH production.

[0059] 1 Animal experiments

[0060] After 21 6-week-old male C57BL / 6J mice were adapted to feeding for one week, the experiment was carried out according to the following steps:

[0061] (1) Random grouping: Twenty-one C57BL / 6J mice were randomly divided into three groups according to body weight: a normal group, a model group, and an A21196 group, with seven mice in each group. Except for the normal group, alcoholic liver injury models were established in all other groups.

[0062] (2) Modeling method: After the adaptation period, mice in each group were fed with non-alcoholic liquid feed (purchased from Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd.) for 5 consecutive days to allow the mice to adapt to the liquid feed feeding mode. Starting from the 6th day, the normal group was fed with non-alcoholic liquid feed every day, and the model group and A21196 group were fed with 5% alcoholic liquid feed for 10 consecutive days to obtain an alcoholic liver injury model.

[0063] (3) After the adaptation period, intervention treatment is carried out according to the following plan:

[0064] Normal group: gavage with 0.2 ml of normal saline for 10 consecutive days;

[0065] Model group: gavage with 0.2 ml of normal saline for 10 consecutive days;

[0066] A21196 group: intragastrically administered with 0.2 ml containing 5×10 9 CFU / ml of A21196 bacterial solution.

[0067] On the 11th day, the normal group was given maltodextrin solution (20 μl / Kg) by gavage, the model group was given ethanol (20 μl / Kg) by gavage, and the A21196 group was first given 0.2 ml of 5×10 9 CFU / ml of A21196 bacterial solution, half an hour later, the mice were intragastrically injected with alcohol (20 μl / Kg). Nine hours after the intragastrically injected with alcohol, the mice were anesthetized, and blood was collected from the heart and then the mice were killed by dislocating the neck.

[0068] (4) Observation indicators and sample collection

[0069] After alcohol gavage, the disappearance time and recovery time of the righting reflex of mice were observed, and the mental state of mice after drinking was recorded. Serum was collected 9 hours after drinking to measure the blood ethanol content.

[0070] (5) Observation and statistical results of the effect of strain A21196 on the incubation period of intoxication in mice

[0071] Table 1 Statistics of disappearance and recovery time of righting reflex in mice

[0072] Group Righting reflex disappearance time Righting reflex recovery time Interval time (min) Model Group No. 1 9:35AM 11:04AM 94 Model Group No. 2 9:35AM 12:20AM 165 Model Group No. 3 9:35AM 12:22AM 167 Model Group No. 4 9:35AM 11:04AM 89 Model set No. 5 9:36AM 11:05AM 89 Model Group No. 6 9:35AM 11:13AM 98 Model Group No. 7 9:46AM 11:13AM 93 A21196 Group No. 1 8:10AM 8:14AM 4 A21196 Group No. 2 8:13AM 9:47AM 94 A21196 Group No. 3 8:19AM 10:20AM 121 A21196 Group No. 4 8:22AM 9:14AM 52 A21196 Group No. 5 8:22AM 8:32AM 10 A21196 Group No. 6 8:22AM 9:49AM 87 A21196 Group No. 7 8:55AM 9:57AM 62

[0073] Figure 3 The statistical results in Table 1 show that compared with the model group, the righting reflex recovery time of mice in the A21196 group was shortened by nearly 1 times. The experimental results show that A21196 can significantly shorten the duration of drunkenness in mice and has a significant effect on degrading ethanol in the body.

[0074] Effect of 2A21196 strain on ethanol content in serum of drunk mice

[0075] The collected blood from each group of mice was allowed to stand for 2 hours, then centrifuged at 3000 rpm for 15 minutes, and the supernatant was taken. The blood ethanol kit (purchased from Nanjing Jiancheng Bioengineering Institute, item number: E036-1-1) was used to measure the ethanol content in the serum, and the analysis and statistics were performed. The results are as follows:

[0076] like Figure 4As shown in the figure, compared with the normal group of mice, the ethanol content in the serum of the model group mice was significantly increased, and the difference was statistically significant. Compared with the model group, the ethanol content in the serum of the A21196 group mice was significantly reduced, and the ethanol content in the blood decreased by nearly 50%, and the difference was statistically significant. This experimental result is consistent with the results of the righting reflex observation experiment, proving that A21196 has an excellent effect on metabolizing ethanol in vivo.

[0077] Effect of 3A21196 strain on acetaldehyde content in serum of drunk mice

[0078] The collected blood from each group of mice was allowed to stand for 2 hours, and then centrifuged at 3000 rpm for 15 minutes. The supernatant was taken and the acetaldehyde content in the serum was measured using an acetaldehyde detection kit (manufacturer: Megazyme, catalog number: K-ACHYD) and analyzed and statistically analyzed. The results are as follows:

[0079] like Figure 5 As shown in the figure, compared with the normal group of mice, the acetaldehyde content in the serum of the model group mice was significantly increased, and the difference was statistically significant. Compared with the model group, the acetaldehyde content in the serum of the A21196 group mice was significantly decreased, which was the same as that of the normal group, and the difference was statistically significant. This experimental result is consistent with the results of the righting reflex observation experiment, proving that A21196 has an excellent effect of metabolizing acetaldehyde in vivo.

[0080] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Application of fermented Lactobacillus mucilaginosus strain A21196 or its probiotic agent in the preparation of products promoting ethanol degradation or metabolism.

2. Application of fermented Lactobacillus mucilaginosus strain A21196 or its probiotic agent in producing alcohol dehydrogenase and / or acetaldehyde dehydrogenase.

3. The application according to claim 1, characterized in that: The method for producing alcohol dehydrogenase and / or acetaldehyde dehydrogenase comprises culturing a bacterial suspension of a fermented Lactobacillus mucosa strain A21196 in a culture medium containing 4% to 6% ethanol by volume, separating bacterial cells from the culture medium, crushing the cells, collecting the cell contents, removing cell fragments and impurities, and obtaining alcohol dehydrogenase and / or acetaldehyde dehydrogenase.

4. Application of fermented Lactobacillus mucilaginosus strain A21196 or its probiotic agent in the preparation of alcohol sobering and liver protecting products.

5. The application according to claim 4, characterized in that: The alcohol sobering and liver protecting method comprises shortening the sobering time and / or reducing the concentration of ethanol and acetaldehyde in the blood serum of drunk people.

6. Use of fermented Lactobacillus mucilaginosus strain A21196 or its probiotic agent in the preparation of a medicament for preventing and / or treating alcoholic liver damage.

7. The use according to any one of claims 1 to 6, characterized in that: The concentration of live bacteria in the probiotics fermented from Lactobacillus mucilaginosus strain A21196 is not less than 5×10 9 CFU / g or 5×10 9 CFU / mL.

8. The use according to claim 7, characterized in that: The dosage form of the probiotics prepared by fermenting the Lactobacillus mucilaginosus strain A21196 includes freeze-dried powder or aqueous solution.

9. The use according to claim 8, characterized in that: The freeze-dried powder is prepared by mixing the bacterial bodies of the fermented Lactobacillus mucilaginosus strain A21196 and a freeze-drying protective agent and then freeze-drying.

10. The use according to claim 9, characterized in that: The freeze-drying protective agent is a skim milk aqueous solution with a mass percentage of 10% to 14%.

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