A strain of Bifidobacterium longum subsp. infantis TG010 with ethanol degradation function and its application

Abstract

The present invention discloses a strain of Bifidobacterium longum infantis subspecies TG010 with the function of degrading ethanol, and its deposit number is CCTCC NO: M20231775. The sequence of its 16SrDNA is shown in SEQ ID No. 1. The strain has the ability of high production of acetaldehyde dehydrogenase and alcohol dehydrogenase, can quickly decompose alcohol in the human body, reduce the harm of alcohol detoxification to the human body, and sober up faster.

Description

Technical Field

[0001] The invention relates to a strain of Bifidobacterium longum infantis subspecies TG010 with the function of degrading ethanol and application thereof, belonging to the technical field of microorganisms. Background Art

[0002] The main component of wine is alcohol, chemically called ethanol. Alcohol can diffuse directly into the blood and be distributed throughout the body without digestion in the human body. The process of alcohol absorption begins in the mouth. When it reaches the stomach, a small amount of alcohol can be directly absorbed by the stomach wall, which accounts for 20% of the total absorption. When it reaches the small intestine, the small intestine will quickly absorb the remaining 80%. The absorption rate of alcohol depends on whether there is food in the stomach, the functional state of the stomach wall, the amount of alcohol in the beverage, and drinking habits. Drinking on an empty stomach is the fastest, 50% is absorbed in 15 minutes, 60% to 90% is absorbed in half an hour, and it can be completely absorbed in 2 to 3 hours. After entering the blood, it flows to various organs with the blood circulation, mainly distributed in the liver and brain. A small part of the exudates from the blood vessels enters the interstitial fluid and is discharged through the skin, a small part is filtered out by the kidneys and discharged with urine, and a small part circulates to the lungs and evaporates and is discharged through breathing. These alcohols are not decomposed and are discharged in their original form, so if you drink too much, you will smell like alcohol all over your body. These are only a small part, accounting for only 10% of the alcohol that enters the body. 90% of the alcohol circulates to the liver and is then broken down by the liver. The liver relies on its own secretion, accumulation, and phagocytosis functions to process some of the alcohol, while most of it relies on the enzymes in the liver to undergo biochemical reactions with the alcohol.

[0003] When ethanol enters the human body, it can have many destructive effects. Excessive alcohol can cause intestinal flora disorder, which in turn damages the intestinal mucosa, destroys the barrier between the intestine and the blood, and increases the permeability of the intestinal mucosa, causing bacteria and metabolites to enter the body's circulatory system. Bacteria and their products promote the production of reactive oxygen molecules, pro-inflammatory cytokines, and chemokines, which will further aggravate liver cell damage and liver damage. At the same time, ethanol is an inhibitor of the central nervous system of the brain, which can act on GABA receptors and interfere with the autonomous regulation of brain nerves. As the concentration of ethanol in the blood circulation system continues to increase, it can inhibit the cerebral cortex, limbic system, reticular structure, and cerebellum, leading to corresponding clinical symptoms. Long-term alcoholism can cause atrophy and degeneration of the brain, damage and disappearance of neurons, and finally cause consequences such as neurological disorders, mania and personality changes.

[0004] Existing hangover products are mainly hangover tea, hangover soup or plant extracts, which are slow to sober up and take a long time. The hangover products on the market are mainly chemical synthetic drugs and natural Chinese herbal medicines. There are few hangover products with live probiotics as carriers. Their hangover sobering effect is relatively unsatisfactory and they fail to fundamentally reduce the body's absorption of ethanol and reduce the toxic effects of ethanol. Therefore, the development of an efficient and natural hangover sobering probiotic product has important theoretical value and application prospects. Summary of the invention

[0005] The present invention provides a strain of Bifidobacterium longum infantis subspecies TG010 having the function of degrading ethanol and application thereof, which can effectively solve the above problems.

[0006] The present invention is achieved in that:

[0007] The invention discloses a strain of Bifidobacterium longum infantis subspecies TG010 having the function of degrading ethanol, and its deposit number is CCTCC NO: M20231775.

[0008] In some embodiments, the sequence of the 16SrDNA of Bifidobacterium longum subsp. infantis having the function of degrading ethanol is shown as SEQ ID No.1.

[0009] An application of the Bifidobacterium longum subspecies infantis in producing acetaldehyde dehydrogenase or alcohol dehydrogenase.

[0010] In some embodiments, the above application is to activate Bifidobacterium longum subsp. infantis and inoculate it into BHIS liquid culture medium, ferment and culture it to obtain fermentation liquid, collect the bacteria by centrifugation, crush the bacteria, collect the supernatant by centrifugation to obtain the initial enzyme liquid, and then extract acetaldehyde dehydrogenase or alcohol dehydrogenase from the initial enzyme liquid.

[0011] In some embodiments, the inoculum amount of Bifidobacterium longum subsp. infantis is 1-10%.

[0012] In some embodiments, the fermentation culture temperature is 37-39° C. and the fermentation time is 34-38 h.

[0013] A hangover-relieving probiotic product comprises the above-mentioned Bifidobacterium longum subspecies infantis.

[0014] In some embodiments, the viable count of Bifidobacterium longum subspecies infantis is 1×10 7 -1×10 9 CFU / mL.

[0015] A use of the above-mentioned Bifidobacterium longum infantis subspecies in the preparation of medicines for treating diseases caused by excessive alcoholism.

[0016] In some embodiments, the disease caused by excessive drinking is one or more of nerve damage, liver damage, cardiovascular damage, digestive system damage, lung damage, immune system damage, and decreased fertility caused by excessive drinking.

[0017] The beneficial effects of the present invention are:

[0018] The infant subspecies of Bifidobacterium longum of the present invention has the ability of high production of acetaldehyde dehydrogenase and alcohol dehydrogenase, can quickly decompose alcohol in the human body, reduce the harm of alcohol detoxification to the human body, and sober up faster.

[0019] The Bifidobacterium longum infantis subspecies of the present invention has the advantages of low cost, fast growth rate, large volume, small impact, strong biological accessibility, high safety, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0021] Figure 1 This is the phylogenetic tree of Bifidobacterium longum subsp. infantis TG010.

[0022] Figure 2 This is the growth curve of Bifidobacterium longum subsp. infantis TG010.

[0023] Figure 3 This is the optimal growth temperature curve of Bifidobacterium longum subspecies infantis TG010.

[0024] Figure 4 The figure is a statistical graph of the acetaldehyde dehydrogenase activity of the strains in Example 2 and 3 comparative examples.

[0025] Figure 5 Statistical graph of alcohol dehydrogenase activity of Example 2 and 3 comparative example strains. DETAILED DESCRIPTION

[0026] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. Therefore, the following detailed description of the embodiments of the present invention provided in the drawings is not intended to limit the scope of the invention claimed for protection, but merely represents the selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0027] The embodiment of the present invention provides a strain of Bifidobacterium longum subspecies infantis TG010 having the function of degrading ethanol, and its deposit number is CCTCC NO: M20231775.

[0028] In some embodiments, the sequence of the 16SrDNA of Bifidobacterium longum subsp. infantis having the function of degrading ethanol is shown as SEQ ID No.1.

[0029] An application of the above-mentioned Bifidobacterium longum subsp. infantis in producing acetaldehyde dehydrogenase or alcohol dehydrogenase. Acetaldehyde dehydrogenase and alcohol dehydrogenase are two important enzymes that play a key role in the metabolic process of ethanol. By utilizing this Bifidobacterium longum subsp. infantis, the production of these two enzymes can be effectively increased, thereby promoting the efficient conversion and utilization of ethanol. This application not only helps to improve the efficiency of ethanol metabolism, but may also play an important role in biofuel production and the field of medicine.

[0030] In some embodiments, the above application is to activate Bifidobacterium longum subsp. infantis and inoculate it into BHIS liquid culture medium, ferment and culture it to obtain fermentation liquid, collect the bacteria by centrifugation, crush the bacteria, collect the supernatant by centrifugation to obtain the initial enzyme liquid, and then extract acetaldehyde dehydrogenase or alcohol dehydrogenase from the initial enzyme liquid.

[0031] In some embodiments, the inoculation amount of Bifidobacterium longum subsp. infantis is 1-10%. Such an inoculation amount can ensure that the strain can effectively grow and reproduce in the culture medium, and is also beneficial to the subsequent enzyme extraction and purification process.

[0032] In some embodiments, the fermentation culture temperature is 37-39° C. and the time is 34-38 hours. Such temperature and time conditions can provide a suitable growth environment for Bifidobacterium longum subsp. infantis, thereby ensuring that it can efficiently produce the target enzyme, thereby improving the efficiency and yield of the entire extraction process.

[0033] In some embodiments, the viable count of Bifidobacterium longum subspecies infantis is 1×10 7 -1×10 9 CFU / mL. This number of active bacteria ensures their effectiveness and functionality in the relevant product or application.

[0034] The above-mentioned specific Bifidobacterium longum infantis subspecies has important application value in the development and preparation of drugs for treating various diseases caused by excessive drinking. In many embodiments, these diseases caused by excessive drinking include but are not limited to damage to the nervous system, damage to the liver, damage to the cardiovascular system, adverse effects on the digestive system, damage to the lungs, dysfunction of the immune system, and reduced fertility. These diseases can occur alone or in combination, and this Bifidobacterium longum infantis subspecies shows significant potential and effect in alleviating and treating these diseases.

[0035] Example 1 Isolation, purification and identification of Bifidobacterium longum subsp. infantis

[0036] The feces of healthy and alcohol-tolerant individuals from Xiamen, Fujian were used as samples. The samples were first inoculated into ethanol liquid culture medium for anaerobically cultured for more than 24 hours. Subsequently, the culture was spread on a solid culture medium with ethanol as the sole carbon source and cultured in an anaerobic incubator at 37°C for 48 hours. Next, a single colony was selected from the culture medium and purified by three streaking on a BHI agar plate. The purified single colony was then inoculated into BHI liquid culture medium and cultured in a shaker at 37°C and 200rpm for 24 to 48 hours. Afterwards, genomic DNA was extracted using a bacterial genomic DNA rapid extraction kit.

[0037] The formula of the ethanol liquid culture medium includes: 5g / L of bacteriological peptone, 1g / L of L-cysteine ​​hydrochloride, after high-pressure sterilization at 121°C for 20 minutes, 20% edible alcohol is added at 55°C to adjust the pH to 3.

[0038] The formula of BHI liquid culture medium is: 37 g of BHI pre-made powder, 5 g of yeast extract, 0.5 g of L-cysteine, 0.1 mg of resazurin, 2.50 μg of hematin, 0.001 mg of vitamin K1, 1 ml of Tween 80, add water to make up to 1 L, stir evenly, adjust the pH to 6.8 with HCl or NaOH, and sterilize by high pressure at 121°C for 20 min.

[0039] The extracted genomic DNA was used as a template, and PCR amplification was performed using a 16SrDNA full-length primer pair (forward primer 27F: 5'-AGAGTTTGATCCTGGCTCAG-3' (SEQ ID No.2); reverse primer 1492R: (5'-TACGGCTACCTTGTTACGACTT-3' (SEQ ID No.3). The PCR product was sent for sequencing, and its 16SrDNA sequence is shown in SEQ ID No.1. Through the Blast alignment function of the EzBioCloud website (https: / / www.ezbiocloud.net / ), the sequence alignment results obtained by sequencing showed that the bacteria was Bifidobacterium longum subsp. infantis, and its 16SrDNA sequence is shown in SEQ ID NO:1.

[0040] After BLAST comparison of the strain on the NCBI website, it was found that the similarity with the known Bifidobacterium longum subsp. infantis ATCC15697 was as high as 99.19%. In order to further determine the position of the strain in the phylogenetic relationship, the neighbor-joining method (NJ) was used to construct a phylogenetic tree in combination with the existing Bifidobacterium longum subsp. infantis sequence, with Enterobacter cloacae (ATCC13047) as the outgroup. The phylogenetic tree is shown in Figure 1. Figure 1 As shown, it is shown that the strain belongs to Bifidobacterium longum subsp. infantis. The strain of Bifidobacterium longum subsp. infantis was named Bifidobacterium longum subsp. infantis TreatGutProbioTG010 (Bifidobacterium longumsubsp. infantis TreatGutProbioTG010), and was deposited in the China Center for Type Culture Collection on September 25, 2023, with a deposit number of CCTCC NO: M20231775, and the deposit location is located in the Wuhan University Collection Center.

[0041] SEQ ID No.1:

[0042] .

[0043] Example 2: Physiological characteristics of Bifidobacterium longum subsp. infantis TG010.

[0044] 1. Growth curve determination:

[0045] ① Activation of bacterial strains: The Bifidobacterium longum subsp. infantis TG010 purified in Example 1 was taken out from a -80°C refrigerator, activated by streaking on a BHI agar plate using a disposable inoculation loop, and cultured in an anaerobic incubator at 37°C until a single colony grew.

[0046] ② Inoculation liquid: Use a disposable inoculation loop to dip a single colony on the BHI agar plate, inoculate it into BHI liquid culture medium, shake it evenly, and culture it in a shaker at 37°C, 200 rpm for 24 hours.

[0047] ③Add culture medium: dilute the seed solution to OD 600 When the inoculation reaches the range of 0.8-1.0, the culture medium is inoculated with 1% of the inoculation amount into BHI liquid culture medium, and a total of 9 culture mediums are inoculated. After the inoculation is completed, the culture medium is placed in a shaking incubator at 37°C and 200 rpm for culture. A group is taken out at 2, 4, 6, 8, 10, 12, 14, 16, and 24 hours to measure the OD 600 , until the strain completes logarithmic growth and reaches the stationary phase.

[0048] The growth curve of Bifidobacterium longum subsp. infantis TG010 is as follows Figure 2 shown.

[0049] As can be seen from the figure, 2-8h is the logarithmic growth phase of the strain, and 14h enters the plateau phase.

[0050] 2. Determination of optimum temperature:

[0051] Inoculate the culture medium: dilute the seed solution in 1 to OD600 in the range of 0.8-1.0, inoculate it into BHI liquid culture medium at a 1% inoculation rate, and inoculate 3 groups of culture medium at each ambient temperature. After inoculation, place the culture medium in different temperature gradients (27, 32, 37, 42, 47°C) and culture it in a shaking incubator at 200 rpm. At the appropriate time point (when some groups of bacterial liquid become turbid), take out the culture medium and measure the OD 600 .

[0052] The optimal growth temperature of Bifidobacterium longum subsp. infantis TG010 is Figure 3 shown.

[0053] As can be seen from the figure, Bifidobacterium longum subspecies infantis TG010 grows best at an ambient temperature of 42°C.

[0054] Example 3

[0055] Acetaldehyde dehydrogenase activity assay steps: The purified Bifidobacterium longum infantis subsp. infantis of Example 1 was taken out from a -80°C refrigerator, streaked on a BHI agar plate for activation, cultured in an anaerobic incubator at 37°C until a single colony grew, inoculated into a BHIS liquid culture medium, and left to stand at 38°C for activation for 12 h. Then, the inoculum was inoculated at 2% in a 250 mL shake flask containing 50 mL of BHIS liquid culture medium and fermented at 38°C, 220 rpm, for 36 h to obtain a fermentation broth; the fermentation broth obtained above was centrifuged at 12000 r / min for 2 min, the supernatant was removed, the bacteria were collected, and the bacteria were diluted to 5×10 7After the elution was quantified, 500 μL of the diluted bacterial solution was taken for ultrasonic disruption (1000 W ultrasonic for 2 min), and the supernatant was obtained by centrifugation at 12000 r / min for 2 min to obtain the crude enzyme solution, and then the enzyme identification and enzyme activity were determined; Tris-HCl (pH 9.2), coenzyme NAD + , acetaldehyde, KCl, β-mercaptoethanol and ultrapure water were mixed in advance according to the addition amount in the ALDH activity assay system in the table below, and ALDH was added after warming at 35°C for 10 minutes. The change in absorbance at 340nm was quickly detected using an ELISA reader, and the absorbance was read once every 1 minute. The total reaction time was 5 minutes. The enzyme activity of ALDH was calculated based on the change in absorbance per minute. The ALDH activity assay system is shown in Table 1 below.

[0056] Table 1

[0057]

[0058] Definition of enzyme activity: At 35°C, a quantitative change of 0.001 in absorbance at 340 nm for 1 min is defined as 1 activity unit.

[0059]

[0060] Alcohol dehydrogenase activity assay steps: take out Bifidobacterium longum infantis subsp. infantis from a -80°C refrigerator, streak on a BHI agar plate for activation, and culture in an anaerobic incubator at 37°C until a single colony grows; inoculate with BHIS liquid culture medium, culture anaerobically at 37°C for 12 h, and then inoculate with 2% inoculum in a 250 mL shake flask containing 50 mL BHIS liquid culture medium at 37°C, 220 rpm, and ferment for 36 h to obtain a fermentation broth; dilute the bacterial broth to 5×10 7 CFU / mL, take 500 μL of fermentation broth and centrifuge at 9000 r / min for 6 min to collect all the bacteria, wash and resuspend the bacteria with 1 ml of ADH extract, and then perform ultrasonic disruption (on 3s, off 7s, working time 3min), centrifuge at 16000g / min and 4°C for 20min, and take the supernatant to determine the enzyme activity; the enzyme activity determination is added in sequence according to the following Table 2.

[0061] Table 2

[0062]

[0063] After rapid mixing, measure the change in absorbance at 340 nm, record the absorbance at 15 s as Atest 1, Ablank 1, and the absorbance at 75 s as Atest 2, Ablank 2; △A assay tube = Atest 1-Atest 2, △A blank tube = Ablank 1-Ablank 2; Definition of activity unit: 1 μmol NADH oxidized per milliliter of sample per minute at 25°C is 1 enzyme activity unit.

[0064] ADH (U / mL) = [(ΔA test tube – ΔA blank tube) ÷ ε ÷ d × V total × 10 6 ]÷Vsample÷T=2.68×(ΔAtest tube – ΔAblank tube) (multiplied by the dilution factor when diluting the bacterial solution)

[0065] ε: NADH molar extinction coefficient, 6.22×10 3 L / mol / cm; d: 96-well plate light path, 0.6 cm; V: total volume of the reaction system, 200 μL = 2 × 10 -4 L; 10 6 :Unit conversion factor, 1mol=1×10 6 μmol; Cpr: supernatant protein concentration, mg / mL; W: sample mass, g; Vsample: supernatant volume added to the reaction system, 20μL=0.02mL; Vsampletotal: extract volume, 1mL; T: reaction time, 1min; number of cells: in tens of thousands.

[0066] According to the steps of acetaldehyde dehydrogenase activity determination, the acetaldehyde dehydrogenase activity of Bifidobacterium longum infantis subspecies of Example 1 was determined, and the acetaldehyde dehydrogenase activity of the strain was determined to be 54.27 U·mL -1 .

[0067] According to the alcohol dehydrogenase activity assay procedure, the alcohol dehydrogenase activity of Bifidobacterium longum subsp. infantis of Example 1 was assayed, and the alcohol dehydrogenase activity of the strain was determined to be 16.44 U·mL -1 .

[0068] Comparative Example 1

[0069] A strain of Bifidobacterium adolescentis (BNCC185974) with broad efficacy was purchased from Beina Biotech (https: / / www.bncc.com / ) and the acetaldehyde dehydrogenase and alcohol dehydrogenase activities of the strain were determined;

[0070] The acetaldehyde dehydrogenase activity of the strain of Bifidobacterium adolescentis was determined according to the steps of the acetaldehyde dehydrogenase activity determination in Example 3. The acetaldehyde dehydrogenase activity of the strain was determined to be 3.96 U·mL -1 .

[0071] The alcohol dehydrogenase activity of the strain of Bifidobacterium adolescentis was determined according to the steps of alcohol dehydrogenase activity determination in Example 3. The alcohol dehydrogenase activity of the strain was determined to be 0.82 U·mL -1 .

[0072] Comparative Example 2

[0073] A strain of Lactobacillus paracasei (BNCC336440) with broad efficacy was purchased from Beina Biotech (https: / / www.bncc.com / ) and the aldehyde dehydrogenase and alcohol dehydrogenase activities of the strain were determined;

[0074] The acetaldehyde dehydrogenase activity of the strain of Lactobacillus paracasei was determined according to the steps of the acetaldehyde dehydrogenase activity determination in Example 3, and the acetaldehyde dehydrogenase activity of the strain was determined to be 3.68 U·mL -1 .

[0075] The ethanol dehydrogenase activity of the strain of Lactobacillus paracasei was determined according to the steps of the ethanol dehydrogenase activity determination in Example 3. The ethanol dehydrogenase activity of the strain was determined to be 0.5 U·mL -1 .

[0076] Comparative Example 3

[0077] A strain of Bifidobacterium animalis subsp. lactis (BNCC134318) with a high usage rate was purchased from Beina Biotech (https: / / www.bncc.com / ) and the acetaldehyde dehydrogenase and alcohol dehydrogenase activities of the strain were determined;

[0078] The acetaldehyde dehydrogenase activity of the strain of Bifidobacterium animalis subsp. lactis was determined according to the steps of the acetaldehyde dehydrogenase activity determination in Example 3. The acetaldehyde dehydrogenase activity of the strain was determined to be 14.73 U·mL -1 .

[0079] The alcohol dehydrogenase activity of the strain Bifidobacterium animalis subsp. lactis was determined according to the steps of alcohol dehydrogenase activity determination in Example 3. The alcohol dehydrogenase activity of the strain was determined to be 13.53 U·mL -1 .

[0080] In the process of measuring the acetaldehyde dehydrogenase activity of the four probiotic strains in Example 1 and Comparative Examples 1-3, we found that the strain involved in Example 2, namely Bifidobacterium longum subsp. infantis, showed the highest acetaldehyde dehydrogenase activity. Specifically, the acetaldehyde dehydrogenase activity of this strain reached an impressive 54.27 U·mL -1 This result is in Figure 4 The figure clearly depicts the acetaldehyde dehydrogenase activity levels of different strains, among which the activity value of Bifidobacterium longum subspecies infantis is significantly higher than that of other strains.

[0081] Similarly, in the process of measuring the alcohol dehydrogenase activity of these four probiotic strains, we again observed that Bifidobacterium longum subsp. infantis in Example 2 showed the highest activity. The specific value was 16.44 U·mL -1 , this result is in Figure 5 It has been clearly reflected in. Figure 5 The alcohol dehydrogenase activity of different strains is shown in detail, and the activity value of Bifidobacterium longum subsp. infantis is particularly prominent in the figure, significantly higher than other strains. This finding further confirms the superior performance of Bifidobacterium longum subsp. infantis TG010 in Example 2 in terms of alcohol dehydrogenase.

[0082] Example 4

[0083] A probiotic product for alcohol detoxification, containing Bifidobacterium longum infantis subspecies of Example 1, with a viable bacterial count of 1×10 7 -1×10 9 CFU / mL.

[0084] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A strain of Bifidobacterium longum subspecies infantis TG010 having the function of degrading ethanol, characterized in that: Its deposit number is CCTCC NO: M20231775.

2. The Bifidobacterium longum subspecies infantis TG010 having the function of degrading ethanol according to claim 1, characterized in that: The sequence of its 16S rDNA is shown in SEQ ID No.

1.

3. Use of Bifidobacterium longum subspecies infantis TG010 according to claim 1 or 2 in producing acetaldehyde dehydrogenase or alcohol dehydrogenase.

4. The use according to claim 3, characterized in that: The activated Bifidobacterium longum subsp. infantis TG010 is inoculated into a BHIS liquid culture medium for fermentation to obtain a fermentation liquid, the bacterial cells are collected by centrifugation, the bacterial cells are crushed, the supernatant is collected by centrifugation to obtain a primary enzyme liquid, and then acetaldehyde dehydrogenase or alcohol dehydrogenase is extracted from the primary enzyme liquid.

5. The use according to claim 4, characterized in that: The inoculation amount of Bifidobacterium longum subspecies infantis TG010 is 1-10%.

6. The use according to claim 4, characterized in that: The fermentation culture temperature is 37-39° C. and the fermentation time is 34-38 hours.

7. A probiotic product for sobering up, characterized in that: The invention comprises the Bifidobacterium longum subspecies infantis TG010 as described in claim 1 or 2.

8. The alcohol sobering probiotic product according to claim 7, characterized in that: The viable count of the Bifidobacterium longum subspecies infantis TG010 is 1×10 7 -1×10 9 CFU / mL.

9. Use of Bifidobacterium longum subsp. infantis TG010 according to claim 1 or 2 in the preparation of a medicament for treating diseases caused by excessive alcoholism.

10. The use according to claim 9, characterized in that: The diseases caused by excessive drinking are one or more of nerve damage, liver damage, cardiovascular damage, digestive system damage, lung damage, immune system damage, and decreased fertility caused by excessive drinking.

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