CN114426942A - Recombinant lactococcus lactis, microcapsule and application thereof - Google Patents

Abstract

The invention provides a recombinant lactococcus lactis, a microcapsule and application thereof, belonging to the technical field of genetic engineering. The PNZ8149 is used as a basic vector, a first fusion gene or a second fusion gene is inserted into the PNZ8149, the first fusion gene comprises a signal peptide SPusp45, probiotic LEISS, an enterokinase recognition site DDDDK and human alcohol dehydrogenase which are sequentially connected in series, and the second fusion gene comprises a signal peptide SPusp45, probiotic LEISS, an enterokinase recognition site DDDDK and human aldehyde dehydrogenase which are sequentially connected in series and are respectively fused. The target enzyme cuts and separates the signal peptide sequences SPusp45 and LEISS sequences from the fusion protein under the action of endogenous enterokinase in intestinal tracts, and simultaneously releases free alcohol dehydrogenase or acetaldehyde dehydrogenase, and the released alcohol dehydrogenase or acetaldehyde dehydrogenase can decompose alcohol.

Description

A kind of recombinant Lactococcus lactis, microcapsule and application thereof

technical field

The invention belongs to the technical field of the technical field of genetic engineering, and in particular relates to a recombinant Lactococcus lactis, microcapsules and applications thereof.

Background technique

Alcohol is a commonly used drink in daily life. While drinking brings joy to people, it also brings certain harm to people's health and social stability. After people drink excessively, the accumulated ethanol will be metabolized in the body to produce many harmful substances and the deposition of a large number of free radicals, which will cause different degrees of damage to the human body, especially it will easily lead to liver lesions. Reducing alcohol intake or absorption is a direct way to reduce harm.

At present, most of the anti-alcoholic and liver-protecting products sold at home and abroad are chemically synthesized diuretic drugs, traditional Chinese medicines for protecting the liver and nourishing the stomach, hangover peptide beverages or drugs that increase the activity of enzymes related to alcohol metabolism. Most of its efficacy is based on the fact that such products can affect ethanol metabolism or inhibit ethanol absorption after being taken by the human body, thereby reducing the damage to other organs such as the liver. Although many medicines for hangover and liver protection have certain effects of hangover and hangover, they will also be accompanied by different side effects, which are not good for human health. Especially people with different constitutions have different reactions to different hangover medicines. .

SUMMARY OF THE INVENTION

In view of this, the purpose of the present invention is to provide a recombinant Lactococcus lactis, microcapsules and applications thereof. After oral administration of the recombinant Lactococcus lactis of the present invention, hADH1B and ALDH2 enzymes can be secreted in the intestine to decompose alcohol.

The invention provides a recombinant Lactococcus lactis, including the first recombinant Lactococcus lactis and/or the second recombinant Lactococcus lactis;

The first recombinant Lactococcus lactis comprises the first recombinant expression vector; the second recombinant Lactococcus lactis comprises the second recombinant expression vector;

The backbone plasmids used for the construction of the first recombinant expression vector and the second recombinant expression vector are PNZ8149 respectively;

A first fusion gene is inserted into the first recombinant expression vector, and the first fusion gene includes serially connected signal peptide SPusp45, probiotic LEISS, enterokinase recognition site DDDDK and human alcohol dehydrogenase;

A second fusion gene is inserted into the second recombinant expression vector, and the second fusion gene includes serially connected signal peptide SPusp45, probiotic LEISS, enterokinase recognition site DDDDK and human acetaldehyde dehydrogenase.

Preferably, the nucleotide sequence of the first fusion gene is shown in SEQ ID NO: 1; the nucleotide sequence of the second fusion gene is shown in SEQ ID NO: 2.

Preferably, the first fusion gene and the second fusion gene are inserted between the SphI and XbaI restriction sites of PNZ8149, respectively.

Preferably, when the recombinant Lactococcus lactis comprises the first recombinant Lactococcus lactis and the second recombinant Lactococcus lactis, the ratio of the effective viable counts of the first recombinant Lactococcus lactis to the second recombinant Lactococcus lactis is ( 1 to 3): (1 to 3).

The present invention also provides a microcapsule comprising the recombinant Lactococcus lactis described in the above scheme.

The present invention also provides the application of the recombinant Lactococcus lactis described in the above scheme or the microcapsules in the preparation of foods, health products or medicines for preventing drunkenness; the recombinant Lactococcus lactis includes the first recombinant Lactococcus lactis, or , the recombinant Lactococcus lactis includes a first recombinant Lactococcus lactis and a second recombinant Lactococcus lactis.

Preferably, the prevention of intoxication includes reducing alcohol absorption and/or prolonging alcohol tolerance time.

The present invention also provides the application of the recombinant Lactococcus lactis or the microcapsules described in the above scheme in preparing food, health products or medicines for hangover.

Preferably, the hangover includes shortening the recovery time after drinking and/or reducing acute liver and/or intestinal damage caused by excessive drinking and/or relieving vomiting and headache after drinking.

Preferably, the dosage form of the medicament includes an oral formulation.

The present invention provides a recombinant Lactococcus lactis. The present invention uses PNZ8149 (a food-grade expression vector for lactic acid bacteria) as a base vector, and a first fusion gene or a second fusion gene is inserted into PNZ8149, and the first fusion gene comprises serially connected The signal peptide SPusp45, probiotic LEISS, enterokinase recognition site DDDDK and human alcohol dehydrogenase, the second fusion gene includes serially connected signal peptide SPusp45, probiotic LEISS, enterokinase recognition site DDDDK and human The aldehyde dehydrogenases were fused separately. The target enzyme cleaves the signal peptide sequence SPusp45 and LEISS sequence from the fusion protein under the action of endogenous enterokinase in the intestine, and releases free alcohol dehydrogenase or aldehyde dehydrogenase, and the released ethanol Dehydrogenase or aldehyde dehydrogenase can break down alcohol. The present invention provides a kind of highly active human ADH1B (hADH1B and/or acetaldehyde dehydrogenase (ALDH2) expressing probiotics, after oral administration secretes hADH1B enzyme and ALDH2 enzyme to decompose alcohol. The results show that oral probiotics expressing hADH1B reduce Absorbed alcohol, prolonged alcohol tolerance time. Oral expression hADH1B and or ALDH2 shortened the recovery time after drinking, and more importantly, liver and intestinal tract are protected from acute damage caused by excessive drinking. Therefore, the present invention The recombinant Lactococcus lactis has the potential to be prepared as an alcohol partner. At the same time, the present invention proposes for the first time that ectopically expressed hADH1B and ALDH2 are used to catalyze the decomposition of ethanol, which provides a practical basis for the subsequent high-efficiency delivery of a super-biological antidote of various enzymes.

Description of drawings

Fig. 1 is the construction and action schematic diagram of recombinant Lactococcus lactis of the present invention;

Fig. 2 is the result that utilizes anti-hadh antibody to detect recombinant Lactococcus lactis and expresses ADH enzyme;

Fig. 3 is the result that utilizes hALDH antibody to detect recombinant Lactococcus lactis to express ALDH enzyme;

Fig. 4 is the result that the culture plate counting method detects the number of viable bacteria;

Fig. 5 is the situation that oral recombinant Lactococcus lactis extends alcohol tolerance time;

Fig. 6 is the time of drunken mice falling into drunk;

Figure 7 is the statistics of the number of movements of the drunk mice after sobering up;

Figure 8 shows the oral administration of recombinant Lactococcus lactis to alleviate liver-intestinal damage caused by acute alcohol intake, where A is the alcohol residue in serum, B is the blood triglyceride concentration, C is the intestinal mucosal lesions, and D is the liver of mice lipid levels.

Detailed ways

The invention provides a recombinant Lactococcus lactis, including the first recombinant Lactococcus lactis and/or the second recombinant Lactococcus lactis;

The first recombinant Lactococcus lactis comprises the first recombinant expression vector; the second recombinant Lactococcus lactis comprises the second recombinant expression vector;

The backbone plasmids used for the construction of the first recombinant expression vector and the second recombinant expression vector are PNZ8149 respectively;

A first fusion gene is inserted into the first recombinant expression vector, and the first fusion gene includes serially connected signal peptide SPusp45, probiotic LEISS, enterokinase recognition site DDDDK and human alcohol dehydrogenase;

A second fusion gene is inserted into the second recombinant expression vector, and the second fusion gene includes serially connected signal peptide SPusp45, probiotic LEISS, enterokinase recognition site DDDDK and human acetaldehyde dehydrogenase.

In the present invention, the nucleotide sequence of the first fusion gene (Noc1-Usp45-LESS-EK-ADH1B) is shown in SEQ ID NO: 1, specifically:

ccatggtc atgaaaaaaaagattatctcagctattttaatgtctacagtgatactttctgctgcagcc ccgttgtcaggtgtttacgctgatactaattctgatttggaaatatcgtcgacttgtgatgctgacgatgacgataag .

In the present invention, the nucleotide sequence of the second fusion gene (Noc1-Usp45-LESS-EK-ALDH2) is shown in SEQ ID NO: 2, specifically:

ccatggtc atgaaaaaaaagattatctcagctattttaatgtctacagtgatactttctgctgcagcc ccgttgtcaggtgtttacgctgatactaattctgatttggaaatatcgtcgacttgtgatgctgacgatgacgataag .

In the present invention, the first fusion gene and the second fusion gene are preferably obtained by full-sequence chemical synthesis.

In the present invention, the first fusion gene and the second fusion gene are inserted between the SphI and XbaI restriction sites of PNZ8149, respectively. The present invention has no particular limitation on the method for inserting the first fusion gene and the second fusion gene into PNZ8149, and conventional methods in the art may be used.

In the present invention, the original strains of the first recombinant Lactococcus lactis and the second recombinant Lactococcus lactis are preferably Lactococcus lactis NZ3900.

In the present invention, the first recombinant expression vector and the second recombinant expression vector are respectively transferred into Lactococcus lactis NZ3900 by electroporation; the electroporation conditions are preferably: 2000V, 25μF and 200Ω .

The present invention is based on PNZ8149 (a food-grade expression vector of lactic acid bacteria), and a first fusion gene or a second fusion gene is inserted into PNZ8149, and the first fusion gene includes signal peptide SPusp45, probiotic LEISS, enterokinase connected in series in series Recognition site DDDDK and human alcohol dehydrogenase, the second fusion gene includes serially connected signal peptide SPusp45, probiotic LEISS, enterokinase recognition site DDDDK and human aldehyde dehydrogenase are respectively fused. The target enzyme cleaves the signal peptide sequence SPusp45 and LEISS sequence from the fusion protein under the action of endogenous enterokinase in the intestine, and releases free alcohol dehydrogenase or aldehyde dehydrogenase at the same time.

In the present invention, when the recombinant Lactococcus lactis includes the first recombinant Lactococcus lactis and the second recombinant Lactococcus lactis, the ratio of the effective viable counts of the first recombinant Lactococcus lactis and the second recombinant Lactococcus lactis It is (1-3):(1-3), Preferably it is 1:1.

The present invention also provides a microcapsule comprising the recombinant Lactococcus lactis described in the above scheme.

In the present invention, there is no special limitation on the preparation method of the microcapsules, and conventional methods in the art can be used. The first recombinant Lactococcus lactis or the second recombinant Lactococcus lactis are separately embedded; the effective viable count of the recombinant Lactococcus lactis in each microcapsule is preferably 1-5*10 ⁹ cfu.

The present invention also provides the application of the recombinant Lactococcus lactis described in the above scheme or the microcapsules in the preparation of foods, health products or medicines for preventing drunkenness; the recombinant Lactococcus lactis includes the first recombinant Lactococcus lactis, or , the recombinant Lactococcus lactis includes a first recombinant Lactococcus lactis and a second recombinant Lactococcus lactis.

In the present invention, preventing drunkenness includes reducing alcohol absorption and/or prolonging alcohol tolerance time.

The present invention also provides the application of the recombinant Lactococcus lactis or the microcapsules described in the above scheme in preparing food, health products or medicines for hangover.

In the present invention, the hangover includes shortening the recovery time after drinking alcohol and/or reducing acute damage to the liver and/or intestine caused by excessive drinking and/or relieving vomiting and headache after drinking.

In the present invention, the dosage form of the drug preferably includes an oral preparation; the oral preparation preferably includes a microcapsule.

The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention.

Example 1

Cloning and expression of recombinant adh and ·ALDH genes

Studies have shown that the 143rd amino acid of ADH1 gene is mutated from glycine to alanine, which will greatly increase the activity of alcohol dehydrogenase to decompose ethanol (Lee, S.L., Chau, G.Y., Yao, C.T., Wu, C.W., and Yin , S.J.Functional assessment of human alcohol dehydrogenase family in ethanolmetabolism:significance of first-pass metabolism[J].Alcohol Clin Exp Res,2006,30(7):1132-1142.). We therefore synthesized the mutant gene.

The plasmid pNZ8149 (Cat#VS-ELV00300-01) carrying a high-efficiency constitutive promoter (GapA promoter, patent CN111518801A) was selected as a vector (Fig. 1).

Human ADH1B gene and human ALDH2 gene were cloned into multiple cloning sites, and the N-terminus was fused with Usp45-LESS-EK sequence.

The present invention uses PNZ8149 (a food-grade expression vector of lactic acid bacteria) as a base vector, and a fusion gene is inserted into PNZ8149, and the fusion gene includes serially connected signal peptide SPusp45, probiotic LEISS and enterokinase recognition site DDDDK and human ethanol dehydration hydrogenase or acetaldehyde dehydrogenase, respectively. The full sequence is the full sequence chemical synthesis of SphI-SPusp45-LEISS-DDDDK-ADH1B-XbaI, and SphI-SPusp45-ALDH2-DDDDK-ADH1B-XbaI. The target enzyme cleaves the signal peptide sequence SPusp45 and LEISS sequence from the fusion protein under the action of endogenous enterokinase in the intestine, and releases free alcohol dehydrogenase or aldehyde dehydrogenase at the same time.

The expression construct was transformed into Lactococcus cremoris NZ3900 (Cat#VS-ELS03900-01) by electroporation, and the recombinant bacteria were screened on Elliker agar plates. The electroconversion conditions were 2000V, 25μF and 200Ω. Using forward (5'-catgccatggtcatgaaaaaaaagattatcagct-3', shown in SEQ ID NO: 3) and reverse (5'-gctctagatcaaaacgtcagacggtacg-3', shown in SEQ ID NO: 4) primers, the recombinant bacteria were identified and sequenced by PCR .

The recombinant Lactococcus lactis was resuspended in M17 liquid medium (1%), incubated at 30°C for 8h, centrifuged at 4°C (4000g for 10min), and the supernatant was collected. The supernatant protein was then precipitated by low temperature ethanol precipitation. Equal amounts of protein samples were separated by electrophoresis on 10% sds-polyacrylamide gels, proteins were transferred to PVDF membranes, incubated with blocking buffer (5% nonfat milk) for 1 h at room temperature, and treated with anti-hadh antibodies and hALDH antibodies (SantaCruz) Make a blot. The results of specific antibody detection showed that our recombinant bacteria could successfully express ADH enzyme (Fig. 2) and ALDH (Fig. 3).

Production of recombinant bacteria

The two recombinant bacteria were cultured for 6-8 hours and then collected by centrifugation (4000g, 4℃, 10min). Then microencapsulate the bacteria: the recombinant bacteria are suspended in 3% (w/v) sodium alginate solution. The suspension was dropped into soybean oil containing 0.2% Tween-80 at a ratio of 1:5 and then stirred with a magnetic stirrer at 600 rpm for 10 minutes. Then 0.05M CaCl2 solution was added slowly and stirred (600rpm, 10min). Microcapsules were collected by post-centrifugation (350 g at 4°C) for 10 minutes. Microcapsules were added to lyoprotectant (19.5% maltodextrin and 2.5% nonfat dry milk, respectively), resuspended and transferred to a petri dish, stored at -80°C until completely frozen, and then transferred to A vacuum freeze dryer was used to freeze-dry the recombinant Lactococcus lactis microcapsules for the two types of bacteria.

Recombinant Lactococcus lactis microcapsules were dissolved in artificial gastric acid (0.2% sodium chloride, pH 1.2), respectively, for 2 hours. Then add the bursting liquid (19:81 NaH ₂ PO ₄ 0.2 to 0.2 Na ₂ HPO ₄ ) and use a rotator at a speed of 200 rpm to break at 37 ° C for 30 minutes, the recombinant Lactococcus lactis was released, and the number of viable bacteria was detected by the culture plate counting method ( Figure 4).

Determination of intoxication scheme

Hongqiqi (Red Star) Erguotou (Fenweijiu, 56% vol) purchased from a supermarket was selected for the mouse intoxication experiment. In order to establish a safe and stable acute intoxication model, 9-week-old male C57BL/6J mice weighing 20-22 g were randomly divided into 3 groups (n=5 in each group), given 4 mg/g BW, 6 mg/g BW, 8 mg/g BW, respectively. g BW. Use the righting reflex as a criterion for determining the level of intoxication. Briefly, mice were placed on the ground with their backs and belly and limbs facing up. Mice were considered to have lost their normal righting reflex if they could not turn over within 30 seconds. The time point when the righting reflex disappeared was defined as the intoxication point, and the time point between the first drinking and intoxication was defined as the alcohol tolerance time. The alcohol tolerance time of the three groups of mice was less than 20 minutes. In order to exclude the influence of individual differences, we defined that the mice that did not lose the righting reflex within 1 hour were considered to be not acutely intoxicated. Considering the intoxication rate and safety, a dose of 6 mg/g BW was selected for subsequent experiments (Table 1).

Table 1. Establishment of intoxication model in mice Select 6mg/g BW alcohol as the therapeutic dose

Detection of alcohol tolerance time in different treatment groups

In the intestinal environment, the peak expression of recombinant Lactococcus lactis that secreted exogenous proteins appeared at about 1-2 hours. The anti-intoxication effect of oral recombinant L. lactis was tested by administration of hADH1B-expressing L. lactis (1X 109 CFU) (n=8). Lactobacillus lactis containing the pNZ construct served as a control (n=8). After 1 h, all mice were given 6 mg/g BW alcohol, and the alcohol tolerance time (time from drinking to loss of righting reflex) was recorded.

exercise recovery monitoring

After 1 h of drinking, the mice were placed in the motion detection system, and their motion was detected every 15 seconds. When the rat was drunk, the machine read 0. Mice were considered to have recovered exercise capacity when the number of movements at four consecutive recording time points was non-zero.

Histological analysis

Hematoxylin and eosin (H&E) staining: paraffin-embedded 4% paraformaldehyde-fixed tissue, 5 μm thick sections were stained, and observed under a 10X or 20X objective.

result

Prolonged alcohol tolerance time by recombinant oral recombinant Lactococcus lactis

The 3 groups of experimental mice were respectively administered with ADH, ALDH recombinant Lactococcus lactis and control bacteria (pNZ8149 empty-loaded Lactococcus lactis was transformed with pNZ instead), the results showed that the alcohol tolerance time of the hadh1b-expressing recombinant Lactococcus lactis mice (ie, the time from alcohol consumption to loss of righting reflex) was significantly prolonged, whereas ALDH probiotics had no significant effect (Table 2, Figure 5).

Table 2. Alcohol tolerance time of two groups of mice (that is, the time to disappearance of righting reflex, 8 mice in each group)

Effects of recombinant oral recombinant Lactococcus lactis on drunkenness rate of mice

The three groups of experimental mice were respectively given ADH, ALDH recombinant Lactococcus lactis and control bacteria (pNZ8149 empty-loaded Lactococcus lactis was transformed with pNZ instead), and the results showed that the drunken rate of mice expressing hadh1b recombinant Lactococcus lactis decreased significantly , while ALDH probiotics had no significant effect (Table 3).

Table 3. Effects of pNZ and hADH1B on drunkenness, recombinant hADH1B probiotics reduce the rate of drunkenness in mice

Recombinant oral probiotic shortens exercise recovery time in drunken mice

Drunk mice need 6 to 10 hours to recover. In order to accurately record the recovery time of mice in different treatment groups, mice were put into motion recorders after drinking alcohol for 1 h. The number of movements was recorded every 15 s (Figure 6). Mice were considered to have recovered their motor abilities when the recorder received 4 consecutive non-zero data. Statistically, we found that the exercise recovery time of the hADH1B-treated mice (5.5±0.41, n=6) was significantly shorter than that of the pNZ probiotic-treated group (6.4±0.41, n=7), the hADH1B-expressing probiotic-treated group One-quarter of the mice did not lose exercise capacity throughout the procedure (Figures 6, 7 and Table 4). These results suggest that probiotics expressing hADH1B can shorten recovery time in drunken mice.

Table 4. Effect of pNZ (n=8) and hADH1B (n=8) on exercise recovery time.

Recombinant oral probiotics attenuate acute alcohol intake-induced liver-intestinal damage

Alcohol is primarily absorbed in the gut and eventually transported to the liver, where it is broken down. Therefore, the gut-liver axis plays an important role in regulating ethanol metabolism, and the gut and liver are also the most directly damaged organs after drinking. In order to detect the acute poisoning of the two groups of mice, the intestinal mucosal lesions were observed. The goblet cells of pnz-treated intoxicated mice exhibited more hypertrophy compared to non-intoxicated mice, and treatment with hadh1b-expressing probiotics attenuated acute drinking-induced intestinal damage (Figure 8, C), indicating that Decreased alcohol absorption through the gut. The alcohol content in the blood of mice reached a peak after 2 to 3 hours of drinking. To test the effect of different probiotics on alcohol absorption, blood alcohol levels at different points were measured after drinking using an ethanol assay kit (BioAssay Systems, ECET-100). During the first h, there was no difference in blood alcohol content between the two groups of mice. 2h after drinking, the serum alcohol residue in the pNZ group continued to increase, while the serum alcohol residue in the hADH group showed a significant downward trend, and was significantly lower than that in the pNZ group (A in Figure 8). Further investigation found that hADH1B treatment decreased blood triglyceride concentrations (B in Figure 8), as well as lipid levels in the liver of hADH1B-treated mice (D in Figure 8). Taken together, treatment with hadh1b-expressing probiotics attenuated acute alcohol intake-induced intestinal damage and reduced liver and blood fat content.

Although the above embodiment has made a detailed description of the present invention, it is only a part of the embodiments of the present invention rather than all of the embodiments, and people can also obtain other embodiments according to the present embodiment without creativity, and these embodiments are all It belongs to the protection scope of the present invention.

sequence listing

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Claims (10)

1. A recombinant lactococcus lactis comprising a first recombinant lactococcus lactis and/or a second recombinant lactococcus lactis;

the first recombinant lactococcus lactis comprises a first recombinant expression vector; the second recombinant lactococcus lactis comprises a second recombinant expression vector;

the skeleton plasmids for constructing the first recombinant expression vector and the second recombinant expression vector are respectively PNZ 8149;

a first fusion gene is inserted into the first recombinant expression vector, and the first fusion gene comprises a signal peptide Spusp45, probiotic LEISS, an enterokinase recognition site DDDDDDK and a human-derived alcohol dehydrogenase which are sequentially connected in series;

and a second fusion gene is inserted into the second recombinant expression vector and comprises a signal peptide SPusp45, probiotic LEISS, an enterokinase recognition site DDDDK and a human-derived acetaldehyde dehydrogenase which are sequentially connected in series.

2. The recombinant lactococcus lactis bacterium according to claim 1, wherein the nucleotide sequence of the first fusion gene is as set forth in SEQ ID NO: 1 is shown in the specification; the nucleotide sequence of the second fusion gene is shown as SEQ ID NO: 2, respectively.

3. The recombinant lactococcus lactis bacterium according to claim 1, wherein the first fusion gene and the second fusion gene are inserted between SphI and XbaI restriction sites of PNZ8149, respectively.

4. The recombinant lactococcus lactis bacterium according to claim 1, wherein when the recombinant lactococcus lactis bacterium comprises a first recombinant lactococcus lactis bacterium and a second recombinant lactococcus lactis bacterium, the ratio of the effective viable count of the first recombinant lactococcus lactis bacterium to that of the second recombinant lactococcus lactis bacterium is (1-3): (1-3).

5. A microcapsule comprising the recombinant lactococcus lactis bacterium according to claim 1.

6. Use of the recombinant lactococcus lactis of any one of claims 1 to 4 or the microcapsule of claim 5 for producing a food, health product or pharmaceutical product for preventing drunkenness; the recombinant lactococcus lactis comprises first recombinant lactococcus lactis, or the recombinant lactococcus lactis comprises first recombinant lactococcus lactis and second recombinant lactococcus lactis.

7. The use of claim 6, wherein said prevention of intoxication comprises a reduction in alcohol absorption and/or an extension of alcohol tolerance time.

8. Use of the recombinant lactococcus lactis bacterium of any one of claims 1 to 4 or the microcapsule of claim 5 for the preparation of a food, a health product or a pharmaceutical product for alleviating hangover.

9. The use according to claim 8, wherein the anti-hangover effect comprises a reduction in recovery time after drinking and/or a reduction in acute liver and/or intestinal damage caused by excessive drinking and/or a relief of post-hangover vomiting headache.

10. The use according to any one of claims 6 to 9, wherein the medicament is in a dosage form comprising an oral formulation.

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