Beneficial microorganism containing alcohol-degrading enzyme gene and application thereof

Abstract

The present invention relates to the field of microbial technology, and discloses a beneficial microorganism containing a hangover enzyme gene and an application. The strain is Fructilactobacillus fructivorans_XJ, with a deposit number of CGMCC NO.30574, which was deposited on May 9, 2024 at the General Microbiological Center of China Committee for the Collection of Microbiological Cultures, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. The present invention proposes a beneficial microorganism containing a hangover enzyme gene and an application. The Fructilactobacillus fructivorans_XJ strain provided by the present invention is isolated from the "Xijiao Live Wine" liquor, is a dominant strain of live wine fermentation liquid, and is used for live wine fermentation. XJ has strong acid and ethanol resistance, and has alcohol dehydrogenase and acetaldehyde dehydrogenase to live wine, and has the potential to sober up.

Description

Technical Field

[0001] The invention relates to the field of microorganisms, and in particular to a beneficial microorganism containing a hangover detoxifying enzyme gene and an application thereof. Background Art

[0002] Rice wine, as a traditional drink, is loved by people for its unique taste and rich nutritional value. Rice wine is made from glutinous rice and fermented by adding koji. It is rich in nutrients such as vitamins, trace elements and amino acids, is easy to be absorbed by the human body, and has many functions such as appetizing, refreshing, invigorating and nourishing blood, nourishing yin and tonifying the kidney.

[0003] As a type of rice wine, "Xijiao Huojiu" has a unique fermentation process and the characteristic of continued fermentation in the bottle, which makes its color and flavor change as the brewing time increases. According to the different brewing times, "Xijiao Huojiu" can be divided into different categories such as "Jade Wine", "Gold Wine" and "Daughter Red", each with its own characteristics. In addition to having the excellent qualities of general rice wine, "Xijiao Huojiu" also has unique vitality because of the large amount of probiotics in its wine.

[0004] In recent years, with the deepening of microbial research, people have gradually realized the important role of probiotics in human health. The probiotics in "Xijiao Huojiu" not only give it a unique taste and flavor, but also may have a potential hangover-relieving effect. However, the current research on this drink is still insufficient, and its fermentation mechanism and the hangover-relieving mechanism of probiotics are still unclear.

[0005] In order to further study the microbial flora in "Xijiao Live Wine" and its potential application value, the inventors enriched the microbial flora in "Xijiao Live Wine", extracted total microbial DNA and performed metagenomic analysis, and found that the main flora in the live wine was Lactobacilaceae strains.

[0006] On this basis, the inventors further isolated and cultured the probiotic strains in live wine, and successfully isolated a new lactobacillus strain - Fructilactobacillus fructivorans_XJ.

[0007] In order to solve the above problems, the present application proposes a beneficial microorganism containing a hangover detoxifying enzyme gene and its application. Summary of the invention

[0008] 1. Purpose of the invention In order to solve the technical problems existing in the background technology, the present invention proposes a beneficial microorganism containing a hangover enzyme gene and its application. The fructogenic Lactobacillus cane strain XJ provided by the present invention is separated from the "Xijiao Huojiu" liquor, is a dominant strain of the live wine fermentation liquid, and is applied to live wine fermentation. XJ has strong acid and ethanol resistance, and has alcohol dehydrogenase and acetaldehyde dehydrogenase to activate wine, and has the potential to sober up.

[0009] (II) Technical solution To solve the above problems, the present invention provides a beneficial microorganism containing a hangover enzyme gene, the strain is Fructilactobacillus fructivorans_XJ, the preservation number is: CGMCC NO.30574, and it was deposited on May 9, 2024 in the General Microbiology Center of China Culture Collection Administration, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0010] The beneficial microorganisms containing the alcohol-degrading enzyme gene as mentioned above are applied to live wine fermentation.

[0011] The beneficial microorganisms containing the alcohol-degrading enzyme gene as mentioned above are used for detoxification.

[0012] The above technical solution of the present invention has the following beneficial technical effects: The fructophilous Lactobacillus fructophilus strain XJ provided by the present invention is separated from the "Xijiao Huojiu" liquor, is a dominant strain of the Huojiu fermentation liquor, and is applied to Huojiu fermentation. XJ has strong acid and ethanol resistance, and has alcohol dehydrogenase and acetaldehyde dehydrogenase to activate the liquor, and has the potential to sober up. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 This is a graph showing the content analysis of Lactobacillus frugivorus in the live wines brewed at different brewing times in the present invention.

[0014] Figure 2 This is a colony morphology observation diagram of Lactobacillus frugivorus F (XJ), a beneficial microorganism containing an alcohol-degrading enzyme gene proposed in the present invention.

[0015] Figure 3 The present invention discloses a genome circle map of Lactobacillus frugivorus F (XJ), a beneficial microorganism containing a hangover detoxifying enzyme gene.

[0016] Figure 4 This is a phylogenetic tree analysis diagram of Lactobacillus frugivorus F (XJ), a beneficial microorganism containing an alcohol-degrading enzyme gene proposed in the present invention.

[0017] Figure 5 This is a pH value detection diagram of live wine at different brewing times in a beneficial microorganism containing a hangover enzyme gene proposed by the present invention.

[0018] Figure 6 This is a graph for detecting the acid resistance of Lactobacillus fructovoracus in the beneficial microorganism containing the alcohol detoxification enzyme gene proposed by the present invention.

[0019] Figure 7 This is a graph showing the ethanol resistance characteristics of Lactobacillus fructovoracus, a beneficial microorganism containing an alcohol-degrading enzyme gene proposed in the present invention.

[0020] Figure 8 This is a graph for detecting the activity of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) in the present invention. DETAILED DESCRIPTION

[0021] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with specific embodiments and with reference to the accompanying drawings. It should be understood that these descriptions are only exemplary and are not intended to limit the scope of the present invention. In addition, in the following description, the description of well-known structures and technologies is omitted to avoid unnecessary confusion of the concept of the present invention.

[0022] Example 1. Source, Isolation and Identification of Strain Strain XJ was isolated from "Xijiao Huojiu". By observing the morphological characteristics of strain XJ and combining its 16SrDNA sequence and homology analysis with the completed bacterial genome map, the strain was identified as a new strain of Fructilactobacillus fructivorans. Its acid and ethanol resistance, as well as the activities of alcohol dehydrogenase and acetaldehyde dehydrogenase were also tested.

[0023] (1) Analysis of the content of Lactobacillus fructificus in Xijiao live wine The live wines brewed for 1 year and 1.5 years were centrifuged at high speed, 10,000 g for 12 minutes to collect the bacteria, and their DNA was extracted and sent to BGI Sequencing Company for metagenomic sequencing. The strains were classified according to the metagenomic sequence information. After analysis, we found that the live wines with different brewing times (1 year and 1.5 years) were mainly composed of Lactobacillus fructosus of the genus Lactobacillus fructosus of the Lactobacillaceae family. Among them, this species accounted for about 90% of the total number of species in the 1-year live wine, and 98% of the total number of species in the 1.5-year live wine, indicating that this species is likely to be the main effective species in the live wine ( Figure 1 A). We also tested the absolute content of this species in live wines of other brewing years and found that the concentration of this species in live wines is very high, with a concentration of up to 106 CFU / mL in jade wine brewed for 4-6 months, and a very high content in gin, about 104-105 CFU / mL ( Figure 1B). It further showed that Lactobacillus fructophilus is the main bacterial species of "Xijiao Huojiu".

[0024] (2) Isolation, culture and morphological observation of strain XJ We further isolated and cultured the fructophilous Lactobacillus, and the isolation and culture method was as follows: 100 μL of the 100-fold diluted "Xijiao Huojiu" liquor was applied to the MRS solid medium with pH = 4.8, and cultured at 30°C for 5-10 days. Milky white colonies were observed to grow on the surface of the medium. They were small, about 0.2 mm, round, convex, and moist. After Gram staining, the bacteria were reddish purple, and the bacteria were elongated, about 0.5 μm X 2-5 μm, and shaped like a matchstick ( Figure 3 ), we tentatively named this strain XJ.

[0025] The genomic DNA of the isolated fructophilous Lactobacillus strain was extracted, and the extracted genomic DNA was used as a template to amplify 16S rDNA using bacterial 16S rDNA universal primers.

[0026] 27F 5'- AGAGTTTGATCCTGGCTCAG-3', SEQ ID No. 2; 1492R 5'-GGTTACCTTGTTACGACTT-3', SEQ ID No. 3.

[0027] The high-fidelity DNA polymerase used was Phanta Max Super-Fidelity DNA Polymerase from Vayme. The PCR system was 50 μL: ddH2O: 20 μL; 2 X Phanta Max buffer: 25 μL; dNTP Mix (10 mM): 1 μL; 27F (10 mM): 1 μL; 1492R (10 mM): 1 μL; Phanta Max Super-Fidelity DNA Polymerase: 1 μL; bacterial genomic DNA: 1 μL (0.1-10 ng). The PCR conditions were: 95°C pre-denaturation for 2 minutes, 95°C denaturation for 15 seconds, 57°C annealing for 15 seconds, 72°C extension for 1 minute, repeated denaturation to extension for a total of 32 cycles, and 72°C extension for 5 minutes.

[0028] The target fragment obtained by PCR amplification was sequenced and analyzed, and the specific sequence is shown in SEQ ID No. 1. By consulting the relevant international gene library at the National Center for Biotechnology Information (NCBI), the 16S rDNA gene sequence of the strain XJ of the present invention was compared with the 16S rDNA gene sequence of the relevant strains to analyze their homology. The results showed that the strain was 100% similar to the sequence of Fructilactobacillus fructivorans strain LF543, and its 16S rDNA sequence was shown in Sequence 1.

[0029] By resequencing the whole genome, we constructed a complete bacterial genome map and found that the strain genome consists of 1,554,791bp, consisting of a chromatin and 3 plasmids. There are 1,445 genes in total, and the total gene length accounts for 81.7% of the genome. We annotated 1,408 genes among them using GO, KEGG, NR and other databases, accounting for 97.43% of the total number of genes. See the genome circle map for details. Figure 3 The conservation of common genes was analyzed using the same and similar species to construct an evolutionary tree ( Figure 4 ). This strain is most closely related to Fructilactobacillus fructivorans strain LF543 in evolution. We tentatively named this strain Fructilactobacillus fructivorans_XJ.

[0030] The strain was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, CGMCC on May 9, 2024. The deposit number is CGMCC NO. 30574. The address of the deposit unit is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0031] Example 2: Identification of the acid resistance of Fructilactobacillus fructivorans_XJ.

[0032] Xijiao Live Wine is different from other wines in that it does not make you drunk, but can relieve the symptoms of drunkenness. We speculate that it contains strains that can sober you up. As the main strain of live wine, Lactobacillus frugivorus is speculated to have the function of sobering up. The pH value in the human stomach and intestines is between 1.5-3.5, and this acidic environment can inhibit the normal functions of most bacteria. In order to further test whether Lactobacillus frugivorus can adapt to the acidic environment, we tested the pH value of Xijiao Live Wine and found that the pH value of "Xijiao Live Wine" is between 3-4, especially the pH value of Jade Wine is relatively low, even reaching 3.2 ( Figure 5 As the main bacteria in live wine, we speculate that it has strong acid resistance.

[0033] To further verify the acid resistance of Lactobacillus fructificus, we prepared an MRS medium with an H+ concentration gradient, in which the lowest pH value was 4.7 and the highest pH value was 6.5. 10 μL of Xijiao live wine (100) and a 100-fold dilution of Xijiao live wine (10-2) were evenly spread on the MRS medium with an H+ concentration gradient, and placed in a 30°C incubator for 5-10 days to observe the bacterial growth status ( Figure 6 A left); At the same time, the isolated Fructilactobacillus fructivorans_XJ was cultured in liquid MRS medium for 7 days until the stable growth period, and then the original bacterial solution and the 100-fold diluted bacterial solution were evenly spread on the MRS medium with H+ concentration gradient, placed in a 30℃ incubator for 5-10 days, and the bacterial growth status was observed ( Figure 6 A right). The results showed that whether it was in live wine or isolated and cultured from live wine, Fructilactobacillus fructivorans_XJ grew faster on the side of the MRS medium with a high H+ concentration, i.e., a low pH value, and formed larger colonies with denser bacterial mosses. This indicates that Fructilactobacillus fructivorans is acidophilic.

[0034] At the same time, we used liquid MRS culture medium with different pH values ​​to inoculate Fructilactobacillus fructivorans_XJ, and cultured at 30℃ until the stable growth period (the period with the maximum OD value), and recorded the OD values ​​of Fructilactobacillus fructivorans_XJ in MRS liquid culture medium with different pH values. Using common strains of Escherichia coli DH5α and Bacillus sp. as controls, the OD values ​​of the control strains Escherichia coli DH5α and Bacillus sp. were measured under different pH conditions. It was found that the OD values ​​of the control strains Escherichia coli DH5α and Bacillus sp. decreased with the decrease of pH, and their optimal pH values ​​were relatively high and close to neutral. When the pH value was as low as about 5.5, Escherichia coli and Bacillus sp. could hardly grow. As the pH value decreases, the OD value of Fructilactobacillus fructivorans_XJ shows a trend of first increasing and then decreasing. Its optimal growth pH is 4.7. When the pH value is lower than 4.7 or even around 3.7, Fructilactobacillus fructivorans_XJ can still grow, showing strong acidophilic and acid-resistant characteristics ( Figure 6 B).

[0035] Example 3: Identification of ethanol resistance of Fructilactobacillus fructivorans_XJ.

[0036] Ethanol is a common bactericidal disinfectant that has a killing or inhibitory effect on most microorganisms. To explore whether Lactobacillus frugivorus has ethanol resistance, we prepared MRS solid culture medium (pH=4.8,) (left side of the dichotomous plate) containing different ethanol concentrations (0-35%) to culture Lactobacillus frugivorus. At the same time, the main bacteria in the process of vinegar and sauce making, Acetobacter pasteurianus (Ap) and Zygosaccharomyces rouxii (Zr), were used as control strains and spread on normal MRS culture medium (right side of the dichotomous plate, ethanol-free). After 10 days of cultivation, it was observed that although Acetobacter pasteurianus (Ap) and Zygosaccharomyces rouxii (Zr) have ethanol resistance and are cultured on a medium without ethanol, their growth can be completely inhibited when the ethanol concentration on the left side of the dichotomous plate rises to 25%. It is speculated that their growth is inhibited by the volatile ethanol in the medium on the left. Lactobacillus frugivorus can grow and form bacterial moss on a medium containing 0-35% ethanol (). This suggests that Lactobacillus frugivorus has a strong ethanol tolerance ( Figure 7 A).

[0037] In order to further detect the ethanol resistance of XJ, we inoculated the control strains Escherichia coli DH5α, Bacillus sp. and Fructilactobacillus fructivorans_XJ into liquid culture media containing different ethanol concentrations. Escherichia coli was cultured in LB medium, Bacillus sp. and Fructilactobacillus fructivorans_XJ were cultured in MRS medium. All three bacterial strains were cultured at their optimal pH and temperature until the stable growth period. By detecting their OD values, it was found that the growth of the control strain gradually decreased with the increase of ethanol concentration, and 5% ethanol was enough to inhibit the growth of Escherichia coli and Bacillus sp. Fructilactobacillus fructivorans_XJ had a higher tolerance to ethanol. It can still grow normally in MRS medium containing 10% ethanol concentration, showing a high alcohol resistance ( Figure 7 B).

[0038] Alcohol dehydrogenase and acetaldehyde dehydrogenase play an important role in the ethanol metabolism process. Based on the genome annotation of Lactobacillus fructificans, we found a total of 15 genes involved in ethanol metabolism. The genes and annotations are shown in Table 1, and their gene sequences are shown in Sequence 2. We tested the activities of alcohol dehydrogenase and acetaldehyde dehydrogenase in Lactobacillus fructificans, and used Saccharomyces cerevisiae Y5 and Escherichia coli DH5a as controls. We found that Lactobacillus fructificans had strong alcohol dehydrogenase and acetaldehyde dehydrogenase activities, especially acetaldehyde dehydrogenase activity ( Figure 8 ).

[0039] Table 1 Genes related to ethanol metabolism in Lactobacillus fructovorax Gene_id KEGG:Ko_name KEGG:Ko_defi FGL000140 gd Glucose 1-dehydrogenase FGL000149 butA, budC meso-butanediol dehydrogenase / (S,S)-butanedioldehydrogenase / diacetyl reductase FGL000184 E1.1.1.51 3(or 17)beta-hydroxysteroid dehydrogenase FGL000200 qor, CRYZ NADPH:quinone reductase FGL000220 gldA Glycerol dehydrogenase FGL000249 adhP alcohol dehydrogenase, propanol-preferring FGL000262 fabG, AR1 3-oxoacyl-[acyl-carrier protein] reductase FGL000299 SORD,gutB L-iditol 2-dehydrogenase FGL000883 adhP alcohol dehydrogenase, propanol-preferring FGL001193 SORD, utB L-iditol 2-dehydrogenase FGL001240 adhE acetaldehyde dehydrogenase / alcohol dehydrogenase FGL001260 qor, CRYZ NADPH:quinone reductase FGL001324 E1.1.1.1, adh alcohol dehydrogenase FGL001327 SORD, gutB L-iditol 2-dehydrogenase FGL001340 E1.1.1.1, adh alcohol dehydrogenase Sequence 1: F. fructovorax Lactobacillus 16S rDNA sequence Sequence 2: Protein sequences of alcohol dehydrogenase and aldehyde dehydrogenase of Lactobacillus fructivorans (XJ): FGL000140 MYKDLEGKTAVITGGDSGIGAAIAERLGQEHMNVVINYHKNEEAANQVVEVVEKNGGHAVTKQGDIGDDAGAESLVKAAVDAFGDLDVWFNNAGMEKKTPTNEIPLEQWEAVVNVDLTGTFLGTKAALNYFLKKNKKGSIVQTSSVHQQIPWPTFASYAASKGGVKLFTETTAMEFAKQNIRVNSICPGAINTPINAKKFADPKTYESTVNMVPMHRIGKPSEVAAAAVWLASDQASYVTGVSLYVDGGMTLYPSFQNGNG FGL000149 MMDKKVALVTGAGQGIGAAIAKRLSKDGFSVALVGRHLDKVEEIAKEIQSDNGDAIAIKADVANRDEVFAAVEKAAKHFGDFNVIVNNAGIAPTTPIQTVTPEVLEQAEKINVGGTIWGIQAAAKKFKELGHSGKIINASSQAGMTGNENLTVYGSTKFAIRGITETTAKELADDGITVNAYCPGIVVTPMMNDIAQNVADEAGKPFEWGMEQFAKDIALKRLSKPEDVAGCVSYLAGPDSDYMTGQSLLIDGGMVFN FGL000184 MTDRLKNKVAIITGGTLGIGYAIAEMFVKEGAKVAITGRNEDRGREAADAINKEYPNTAKFIKQDVSDPERWPEVFEETEKDFGQVTTLINNAGIAIGKSIEDTTTEEWKKVLSIDLDGVFYGMREGIKQMKNKNIGASIVNMSSIEGLVGDPDLGAYNASKGGVRMMSKSAALDCALKDYGIRVNSVHPGYIQTPMVKPDFVKAMSQRSKTPMGHLGKPKDVAYAAVYLASDESEFTTGSELVVDGGYTAQ FGL000200 MTKKMNAVGYTKYLPIDDPKSLFDFTTDVPESEPHDLLVKVEAVSVNPVDVFTRESQKVPLKDPKIIGWDAVGTVEKVGSKTNRFSVGDRVFYSGSHVRPGCDSEYQTVDERIVGIAPHSLDNAHAAAMPLTSLTAWEAMFEQMNIDFYDRDEQNHNKSILIINGAGGVGSIATQLAHRANLQVISTAGKDETKQWELDHGSNIVVDYHDDIIKQIHDKGFKYVDYILDRLNLDAYWDEICELIAPSGTIVSITGSGKSLDMSKLKEKRARFAWEYMFTKSEFHTPDMITQHEILDKISIMLDEGNLISTLTKQFGPINAENLRKATKLVESKQMIGKVVVANDEK FGL000220 MIEMVKLFASPSNYVQGPDVLMHSADRIGKFGKRVLLLSSPVLKIIGNDFAEYLKKNDFDVEVVNFTGESESEIDRVTKIGQTKGDNVVIGLGGGKVSDSAKAIADNLNTTVVIAPTIASTDAPCSRLSVIYTDNGQFDHYRFYKKNPDMVILDTKVISHAPVKFLIAGMADAMATNVEATDTRK SDGTNMLGGKSTLAGESIGQKCEQTLFKYGSLAVAANVVGSVTPELDNIVEANTLLSGIGFESGGLAAAHAIYNGFTALSGEIDQLSHGQKVAFGTLCELVLTGSDHRRFEKYLSFDLEFGLPTTLEDLKLGTASDKELMRAATQACDPSDTMVNLPVSVTPIDVMNAMKTVSEFSHEYQVKNGLGD FGL000249 MVKAAVVRDNSDGYVDVKDVKLRPIHEGEALVKVEYCGLCHTDLHVASGDFGEVPGRIIGHEGVGRVIQVADDVTDLKIGDRVSIAWFYSADGTCEYCVTGNETLCRNVKNSGFTVNGAMAEECIVDAKYAVKVPEGLDPVEATSLTCAGVTTYKALKIGDTKTGKWVEVVGAGGLGNLGVQLAHNVFGAPVVVVDGDPKKLEAAKKNGADVLINRHDGNVAEQIQDKVGGVYDSIVTAVNPDAFTQSVNALRPNGTLVAVALPKGDMALNIDKTVLDGIKVAGSLVGTRQDLRETFQFGAEGKVKPIVRTDKLENINNVIDDMKAGKIVGRVVFDFTGDEK FGL000262 MTTTNDKSTVLITGATCGIGLATAKRLAKNPNNLVIINSHRELNNDEVKDLKSQFDTEIEVLIGDVSKEEDAESMVNQILDEHDHLDLVNNAGITKDKLLTRMSEEDFKTVLDTNLVGTFNMTKYALKTMQKKRAGVIVNTSSISGLHGNMGQANYSASKSGIVGLTKTTAQEGSLRGVRCNAVAPGMINDTKLSDKIISKWEDTIPLHRFGNAEEVAETIEFLINNQYITGQVITVDGGLTM FGL000299 MTDVSIPNYDKNNMPKTTKSAVLEKVFDIDMKYTKLKEMGPTDVLIKVVAVGICGSDVHYYDQGHIGDFVVKKPLILGHESSGVIVAVGDKVTKFKRGDRVAMEPGVPCGHCEACRTGHYNLCPNMQFMATPPVNGDLTQFIVYPQDFVYPIPENVSYEEATLNEPLSVGVHATQKLGVDVGSSVLISGMGPIGLLAILAAKAHGADQIIVSDAEQSRLDVAKKLGATNAVNIKNADVLDTVKTLTNGVGVDYAIEASGTVPGEQTSLHALKRGGKVAYIGVPTTDQTPLDVPPFMTDHETTIMGIFRYCNNYQTGLKILAKNTKLVDNLLTNFYPLDQTKAALEKSRTDKSNSIKVIIYPNEQLRK FGL000883 MVEETKINIPKIMNAWEIVKPGPINGDKSPLKYVRKEVPTGDGEVLVKVLTCGVCHTDLHVTEGDLPVHKKNLTPGHEIIGKVVKNGPHTRRFKIGERIGIPWLRKTCGVCKYCRSGRENLCPYSKYTGWDHDGGYEEYTTVPEGFAYRIPERFNSLRAAPLLCAGIIGYRAFERANIPAGGTLGIYGFGGSAHITAQLAIDQGIEVHVFTRGEDAKKFALSLGAKSVNNTYDMSPVPLDSSIIFAPVGEAVPNALESLAPGGTLALAGIHMSDIPQMSYEDHIFHEKNLTSVESNTRSDGEEFLTLADRLHIEPDVTEFPLAKADEALKRVAKGDLKGACVLNVGLDSE FGL001193 MSKMKALVMTAKQHMEIEDRDVPTPKDDEVLVETAYAGICGTDHALYNGLPGSADATPPIVLGHENSGVVSAVGKNVKDFKKGDRVVVDPNIYCDKCFYCHTGRYELCENLSAVGVTRDGGLEDYFTAPEKVVYHIPDELSLQTACSMEPLSCAVHTMKITENITPYQHALIIGDGYMAKLFAQLLRIYGIQNIDMTGRHDDNLAWEKKELNLNKVINTTKEKITDKYDMVIECVGKPETQEQAVEATNPGAQVMMFGVGAPGQTFNMNTYEIFKKQLTIKGSMINPHSFEDSIALLSDGSVKVKPFQNNILTLEQVPGVLAGTDKVQGKSIVKVNGDLK FGL001240 Aldehyde-alcohol dehydrogenase 2 MLEAGNMTDAKQQPNVDEIEKNIDAKVQKARIAVDEMSNFDQDKVDKITHAMVLAGVNAHQKLAKMAYEETGRGVEEDKAIKNLFATEEIWHDIRNDKTVGIISEDSENDLIKVAEPLGVIAGITPVTNPTSTTLFKAIISMKTRNAIIFSLHPQALQSSIEAARIVRDAAVKAGAPKDAIQWIEKPSREATNALIQNPGVDITLATGGPGLVKAAYSAGKPALGVGPGNGPVYVEKTANIPRAVNDIVLSKTFDNGMICATENSAVIDADIYKDVKEQLELNRVYFVPENEKQKLTDAMFDSKRLSVKGPIAGATAQQIADMAKIKIPKDTQVLAVEMTGIGPKYPMSGEKLSPVLSVYKSKDHKDAFRICAELLNYGGLGHTAAIQTSDDQLAVDFGVAVKASRVIVNSPAGLAGVGNLYNNMTPSLTLGTGSWGKNSISHNVTDYDLLNIKTIAKRRNNMQWIKLPRIYFEKNSVRYLHDMPGIKKAFIVTSPSMVKYHYVDKVLDELNRRQDDVQYYIFDDIHGEPTTDMVNAGVTQMNAFKPDTIIALGGGSPLDAAKMMWLIYENPETDFFGAKQKFLDIRKRAYRYNKPEKSQMVAIPSTSGTGSEVTPFSVITDSKLHVKYPLADYALTPDVAIVDPQFVKTVPKGTIAASGLDALTHAIESYVSTMASDYTRPLSLQAIKMIFDNLTDSYNGNMEARDKMHTASTIAGMAFANAFLGITHSIAHKIGGQFGITHGLAIAITLPHVIRYNFKKPKKIATWPKYESFRADDDYATIARYLGLPGNSNEELKEALVHRFIELAHSVGVELDLKSQGVSKAELEAAADHLAELAYSDQCTTANPKEPLIADLKKMILDEYEGVGVETQTDKEV FGL001260 Zinc-type alcohol dehydrogenase-like protein MEKDMKAIGFTKYLSIKNPDSLFEFTTKVPTPNSHDLLVKVEAVSVNPVDVFTRRSQRKQLASPKIMGWDAYGTVVSTGSEVSFFKPGDKVFYAGSYKRPGSDSEYQLVDERIVGHAPKNISIEESAAMPLTSLTAWESLFEQMDIDIDDKENNKSKSILIINGSGGVGSVATQLAHLANLTVIATAGNETTKQWELHHGNDHVVDYHDSIVDQVHDLGFQHVNYILELVNLDAYWSTITRLIGPNGSIVSTTGSGRNLNFSSLKSRMVRFGWEWMYEKSWYETPAMITQHEILDKIGKLLDQGVLKSTLTKTITPISAKNLRKATSMVESHHMMGKVVVKR FGL001324 MINIPKKMLARLIKYRHPFVKDTIPVPPQINDNEVLVSIRASSLNPIDYTIQSGKVKIIMPYKLPITVGNDFSGTVVRVGKKVTKFKLDDQVYGRPDDFQSGTLTEFIAINENDIAKTPRNLSLTDASAVPLTGLTSYQALIEIGKLKKGQKVFIQAGSGGVGTMAIQI AKAVGAYVATTTSKKNFDLVRSLGADQVIDYHTTDFSEVLKNYDLAFDTQGGKTLSKTFKILKPGGKLVTVNGIPTFKFGAQHHLGLLKSILFGIASSPLKLLAKHYVEYTFFIMHPSGRQLDLLTNMIENHELKPIVDRVFPFSEVFEGFKLLESHHATGKVIIEMK FGL001327 MNEKMNALVMTKKQKMEMEKLDVPTPKDDEVLVETAYAGLCGTDNGLFNGLPGSTDATPPIVLGHENSGVVAAVGKNVKDFKKGDRVAVDPNIYCDKCFYCRTGRYELCDNLSAIGVTRNGGLEQYFTAPEKVVYKLPDSISLQAACSLEPISCVVHAMKITENITPYQK ALIIGDGFMAKLFAQILRAYGIQSIDMAGLNDQVLHQMKTDFKLNKVINTSKTPIEENYDFVVEAVGRPETQEEAAEHTNKGAQVMMFGVGKPDQTFSMNTYNVFKKQLTIKGSIINPHAFEDSIALLESGAVQTEPFQNNILKLDEVEGALNHTTPKVGKIVVAVNPDLK FGL001340 MKATEFIKPGDVELVDTPKPKIEKPTDAIIKVVRASVCGSDLWWYRGIDERKANTFVGHEAIGIVDSVGSDVKDVKPGDFVIAPFTHGCGHCAACLAGFDGNCMNQEKDANIGYQAQYFRYINANWGLVKIPGQPEDYSDEMLKSLVTLSDVMATGYHAAASAEVKDGDTVVVM GDGAVGLCGVIVAKLRGAKRIIAMSRHEDRQKLAKEFGATDIVPERGDEAVKHVMELTDGAGADAILECVGTEQSVDTATKVARPGAVVGRVGIPQKAEMNTNNLFFNNTGLRGGIAPVTTYDREVLLKAVLDGKINPGKVFTKEFDLDHIQDAYDAMDKRKAIKSLVIVDKD.

[0040] Peptone 10.0 Beef dip powder 5.0 Yeast extract powder 4.0 glucose 20.0 Triammonium citrate 2.0 Sodium acetate 5.0 Magnesium sulfate 0.2 Manganese sulfate 0.05 Agar 15.0 Twain 80 1.0 pH 6.2 ± 0.2 25℃ Dipotassium hydrogen phosphate 2.0 It should be understood that the above specific embodiments of the present invention are only used to illustrate or explain the principles of the present invention, and do not constitute a limitation of the present invention. Therefore, any modifications, equivalent substitutions, improvements, etc. made without departing from the spirit and scope of the present invention should be included in the protection scope of the present invention. In addition, the appended claims of the present invention are intended to cover all changes and modifications that fall within the scope and boundaries of the appended claims, or the equivalent forms of such scope and boundaries.

Claims

1. A beneficial microorganism containing a hangover detoxifying enzyme gene, characterized in that: The strain is Fructilactobacillus fructivorans_XJ, with the deposit number: CGMCC NO. 30574. It was deposited on May 9, 2024 at the General Microbiology Center of China Culture Collection Administration, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

2. A beneficial microorganism containing a hangover enzyme gene is used in live wine fermentation.

3. A beneficial microorganism containing an alcohol-degrading enzyme gene is used to detoxify alcohol.

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