Lactobacillus reuteri BYN and application thereof in preparation of anti-helicobacter pylori beverage

Abstract

The invention provides lactobacillus reuteri BYN and an application of the lactobacillus reuteri BYN in preparation of an anti-helicobacter pylori beverage. Belongs to the technical field of biology, and the lactobacillus reuteri BYN is classified and named as Limosilicobacterium reuteri, is preserved in the China General Microbiological Culture Collection Center (CGMCC), and has the preservation number of CGMCC NO.31737. The lactobacillus reuteri BYN has the advantages that the lactobacillus reuteri BYN is named as Limosilicobacterium reuteri; the lactobacillus reuteri BYN with helicobacter pylori resisting activity is separated and purified from abalone viscera, the strain is applied to production of the carrot fermented beverage, natural nutrient substances in carrot juice are converted into metabolites with helicobacter pylori resisting activity through the lactobacillus reuteri BYN, and the helicobacter pylori resisting activity is obtained. The anti-scotophoresis activity is improved; meanwhile, the flavor of the carrot juice is improved. A new zymophyte source is provided for production, development and utilization of the carrot juice fermented beverage, and the carrot juice fermented beverage has a wide market application prospect.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a strain of Lactobacillus reuteri BYN and application thereof in preparing an anti-Helicobacter pylori beverage. Background Art

[0002] Helicobacter pylori ( Helicobacter pylori , Hp infection is associated with a variety of digestive tract diseases, and long-term infection is the most important microbial risk factor for gastric cancer. In recent years, due to the emergence of bacterial resistance, the eradication rate of Hp has gradually declined. Long-term and high-dose use of antibiotics can cause adverse reactions such as intestinal dysbiosis and gastrointestinal dysfunction. For some patients with unstable gastrointestinal microorganisms, especially children and elderly patients, eradication of Hp should be more cautious. Studies have shown that the use of probiotics in eradication treatment can reduce adverse reactions and improve patient compliance.

[0003] Carrot is one of the most commonly consumed root vegetables in the world. It is rich in carotenoids, vitamins, polyphenols, calcium, phosphorus, iron, zinc, manganese, molybdenum and other trace elements. It has multiple biological activities such as anti-oxidation, anti-aging, vision protection, immunity enhancement, and blood sugar reduction. However, its earthy and pungent smell makes carrot juice taste poor and not easily accepted by consumers. Fermentation is a traditional preservation and processing technology used to extend the storage period of food and improve the nutritional and sensory quality of food. During the fermentation process, many microbial and enzyme reactions occur, resulting in changes in the composition and compounds of food raw materials. Lactic acid bacteria are common microorganisms in fermented foods. Their application in the fermentation of vegetables and fruits has attracted widespread attention because it can improve the sensory properties, nutritional quality and health benefits of fermented foods. my country has vast sea areas and long coastlines, which contain rich and diverse marine resources. Marine microorganisms produce many undiscovered molecules in extreme living environments with unprecedented structural and pharmacological activities. Discovering new potential anti-Helicobacter pylori functional marine lactobacillus resources and establishing a marine lactobacillus resource library can not only fill the gap in marine-derived probiotics, but also lay the foundation for the development and utilization of functional anti-Helicobacter pylori probiotics.

[0004] Therefore, marine lactic acid bacteria with anti-Hp activity were combined with carrot juice to study its anti-Hp activity and changes in metabolites after fermentation. Only a simple fermentation process is needed to improve the flavor of carrot juice while giving it higher nutritional value and anti-Hp activity, which has important research value and market prospects. Summary of the invention

[0005] The purpose of the present invention is to provide a strain of Lactobacillus reuteri BYN and its application in an anti-Helicobacter pylori beverage. The present invention applies Lactobacillus reuteri BYN to the preparation of a fermented carrot beverage, and the provided anti-Helicobacter pylori fermented carrot beverage is rich in small molecule anti-Helicobacter pylori active ingredients, so that it can directly act on Helicobacter pylori in vitro to inhibit its growth and urease activity.

[0006] In order to achieve the above-mentioned invention object, the present invention adopts the following technical solutions: The present invention provides a strain of Lactobacillus reuteri BYN, wherein the strain of Lactobacillus reuteri BYN is classified as Lactobacillus reuteri Limosilactobacillus reuteri , deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC NO.31737.

[0007] Furthermore, the 16S rDNA nucleotide sequence of the Lactobacillus reuteri BYN is shown in SEQ ID NO.1.

[0008] The present invention also provides an anti-Hepatitis beverage fermentation agent, which contains the activated Lactobacillus reuteri BYN fermentation bacterial liquid and sterile physiological saline.

[0009] Furthermore, the bacterial content of Lactobacillus reuteri BYN in the starter is 1×10 8 cfu / mL~1×10 9 cfu / mL.

[0010] The present invention also provides the application of the Lactobacillus reuteri BYN in the preparation of anti-Helicobacter pylori beverage.

[0011] The application is to inoculate Lactobacillus reuteri BYN into carrot juice, and obtain the anti-Helicobacter pylori-resistant carrot fermented beverage through fermentation, thereby providing a new fermentation bacteria source for the production and development of carrot fermented beverage.

[0012] Furthermore, the steps of the application are: (1) Preparation of carrot juice: Take carrot juice and dilute it with drinking water until the soluble solid content is 10.0±1.0, and the pH after adjustment is 4.85±0.10; sterilize and cool for later use; (2) Preparation of starter culture: The BYN strain of Lactobacillus reuteri was activated with MRS liquid culture medium, and the activated fermentation liquid was washed with sterile saline to prepare a starter culture of Lactobacillus reuteri BYN with a bacterial content of 1×10 8 cfu / mL~1×10 9 cfu / mL of starter culture; (3) Fermentation: The fermentation agent is inoculated into the prepared carrot juice at an inoculation rate of 2% to 5% by volume, and the anti-Helicobacter pylori-resistant carrot fermented beverage is prepared by shaking fermentation.

[0013] Furthermore, the anti-Helicobacter pylori fermented carrot beverage can inhibit the growth and colonization of Helicobacter pylori, and has the effect of preventing and treating Helicobacter pylori infection.

[0014] Furthermore, the anti-Helicobacter pylori fermented carrot beverage can inhibit the growth and colonization of Helicobacter pylori in the stomach by inhibiting urease activity, thereby achieving the effect of preventing and treating Helicobacter pylori infection.

[0015] Furthermore, the Helicobacter pylori is Helicobacter pylori SS1. The fermented carrot beverage can inhibit the growth of Helicobacter pylori and has the effect of preventing and treating Helicobacter pylori infection.

[0016] Preferably, the fermented carrot beverage has significant differences in metabolites compared to the unfermented carrot juice raw material, with a total of 131 differential metabolites, 47 differential metabolites whose contents significantly increased after fermentation, and 84 differential metabolites whose contents decreased, among which the contents of potential anti-Hemorrhagic active ingredients, such as succinic acid, 3-hydroxybenzoic acid, indole-3-acetamide and the like, were significantly increased.

[0017] Compared with the prior art, the present invention has the following beneficial effects: The present invention separates and screens Lactobacillus reuteri BYN with anti-Helicobacter pylori activity from abalone viscera, and applies the bacteria to the production of anti-Helicobacter pylori carrot fermented beverages, using lactic acid bacteria metabolism to produce a large number of small molecule active ingredients, improving the flavor while increasing the antibacterial activity of carrot juice. The carrot fermented beverage provided by the present invention is a postbiotic probiotic liquid beverage that has been sterilized at high temperature. It has the advantages of improving product stability, safety and application range.

[0018] The present invention has been verified through experiments that the anti-Helicobacter pylori fermented carrot beverage has a significant inhibitory effect on Helicobacter pylori, can inhibit the growth and colonization of Helicobacter pylori, and provides more choices for the daily diet of people infected with Helicobacter pylori. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a diagram showing the effect of Lactobacillus reuteri BYN in inhibiting Helicobacter pylori.

[0020] Figure 2 This is a colony morphology diagram of Lactobacillus reuteri BYN.

[0021] Figure 3 This is a Gram stain of Lactobacillus reuteri BYN.

[0022] Figure 4 It is the agarose gel electrophoresis diagram of Lactobacillus reuteri BYN.

[0023] Figure 5 is the phylogenetic tree of Lactobacillus reuteri BYN.

[0024] Figure 6 It is the genome circle map of Lactobacillus reuteri BYN; A is the chromosome sequence; B is the plasmid 1 sequence; C is the plasmid 2 sequence.

[0025] Figure 7 is the COG functional annotation of Lactobacillus reuteri BYN.

[0026] Figure 8 This is the KEGG functional annotation of Lactobacillus reuteri BYN.

[0027] Fig. 9 This is a diagram showing the effect of fermented carrot juice in inhibiting Helicobacter pylori.

[0028] Fig.10 This is a graph showing the changes in the growth curve of Helicobacter pylori after intervention with fermented carrot juice.

[0029] Fig.11 This is a graph showing the changes in Helicobacter pylori urease activity after intervention with fermented carrot juice.

[0030] Fig.12 This is the PLS-DA score graph of carrot juice metabolites before and after BYN fermentation.

[0031] Fig.13 This is a volcano plot of the difference in metabolites of carrot juice before and after BYN fermentation; A is a volcano plot of data acquisition in positive ion mode, and B is a volcano plot of data acquisition in negative ion mode. DETAILED DESCRIPTION

[0032] The technical solution of the present invention is further described in detail with reference to the following specific examples, but the scope of protection claimed by the present invention is not limited to the scope described in the examples.

[0033] Concentrated carrot juice: from Tsingtao Brewery Co., Ltd.

[0034] MRS culture medium, MRS broth culture medium, Columbia blood agar plate, Lactobacillus biochemical identification tube (Qingdao Haibo Biotechnology Co., Ltd.), Gram staining kit (Beijing Solebold Technology Co., Ltd.), urea, phenol red, etc. were all domestic analytical grade.

[0035] Example 1: Isolation, screening, identification and preservation of strains 1. Isolation and screening of lactobacilli In February 2024, in Qingdao, Shandong Province, China, the contents of abalone viscera were aseptically picked and inoculated into 30 mL of sterile MRS liquid culture medium and fermented at 37°C for 36-48 h. The fermentation liquid was dipped with a sterile inoculation loop and streaked on an MRS solid plate and cultured at 37°C for 36-48 h. Single colonies with different morphology, size, and color were picked and streaked again, and cultured at 37°C for 36-48 h. Repeat the streaking and separation 2 to 3 times, determine the single colony, and number it, perform Gram staining and microscopic examination. The isolated strain was stored in glycerol at -80°C.

[0036] The isolated strain was inoculated with sterile MRS liquid medium, and after activation for 3 generations, the fermentation broth and the fermentation supernatant after centrifugation (5000 r / min, 5 min) were taken for in vitro anti-Helicobacter pylori activity determination. The concentration of Hp SS1 bacterial suspension was adjusted to 10 by turbidimetry. 8 CFU / mL, take 100 μL of bacterial solution and inoculate it on Columbia blood agar plate, and spread it evenly with a coating rod. Place the sterilized Oxford cup on the evenly coated Columbia blood agar plate at equal distances, and add 200 μL of amoxicillin (0.01 mg / mL), clarithromycin (0.01 mg / mL), sterile MRS medium, lactobacillus fermentation liquid and lactobacillus fermentation supernatant to each well in turn. Place the culture dish in a three-gas incubator and culture at 37°C for 48-72 hours before taking it out.

[0037] The inhibition zone experiment was repeated three times. According to the drug sensitivity judgment criteria in the literature: the inhibition zone diameter ≥ 15 mm is highly sensitive; 10 mm ≤ inhibition zone diameter < 15 mm is moderately sensitive; 6 mm ≤ inhibition zone diameter < 10 mm is lowly sensitive; inhibition zone diameter < 5 mm or no obvious inhibition zone is insensitive. Finally, a lactobacillus strain that can produce a highly sensitive inhibition zone diameter was selected, namely Lactobacillus reuteri BYN. Its antibacterial results are shown in the figure. Figure 1 shown.

[0038] II. Identification of strain BYN (1) Morphological identification Figure 2 This is the BYN colony morphology. The isolated colonies are milky white, round, convex, moist, and have neat edges. Gram staining results show that it is a Gram-positive bacterium ( Figure 3 ).

[0039] (2) Physiological and biochemical identification The strain BYN was subjected to hydrogen sulfide test, sugar fermentation test, gelatin liquefaction test, indole test, etc. to observe the biochemical characteristics of the strain. For specific test operations, refer to the instructions of the biochemical identification tube.

[0040] The results are shown in Table 1. The strain BYN was able to utilize maltose, sucrose, raffinose and lactose.

[0041] Table 1: Physiological and biochemical results of strain BYN

[0042] Note: + represents a positive test result, - represents a negative test result (3) 16S rDNA molecular identification The single colony DNA of the selected BYN strain was extracted and used as a template for 16S rDNA amplification. The universal primers were: 27F: 5'-AGAGTTTGATCCTGGCTCAG-3'; 1492R: 5'-GGTTACCTTGTTACGACTT-3'.

[0043] PCR amplification program: pre-denaturation at 94°C for 3 min; denaturation at 94°C for 30 s, annealing at 56°C for 30 s; extension at 72°C for 1.5 min; a total of 30 cycles, post-extension at 72°C for 20 min, and storage at 4°C.

[0044] After the PCR reaction is completed, the PCR product is subjected to 1.5% agarose gel electrophoresis for quality control. Figure 4 As shown, the target band size is about 1500 bp. The PCR products that passed the quality control were sent to Shanghai Sangon Biotechnology Co., Ltd. for sequence determination and phylogenetic tree construction. The 16S rDNA nucleotide sequence of BYN is shown in SEQ ID NO.1.

[0045] Reference Figure 5 , the Lactobacillus reuteri BYN strain and Lactobacillus reuteri ( Limosilactobacillus reuteri ) with a homology of more than 99%, and was identified as Lactobacillus reuteri and named Limosilactobacillus reuteri BYN.

[0046] The screened Lactobacillus reuteri BYN was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration (CGMCC); address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; deposit date: August 26, 2024; Lactobacillus reuteri BYN Limosilactobacillus reuteri The deposit number is CGMCC No.31737.

[0047] Reference Figure 6 ,The whole genome sequencing results showed the basic characteristics of Lactobacillus reuteri BYN, such as Figure 1As shown, BYN's genome consists of a chromosome with a length of 2223753 bp and two plasmids with lengths of 19058 bp and 7047 bp, respectively, with GC contents of 39.06%, 36.87% and 39.24%, respectively. There are 2169 coding genes with a total length of 1947051 bp. The total length of the coding region accounts for 86.54% of the entire genome, and the average length of the coding gene is 897.67 bp.

[0048] Diamond blastp was used to compare the gene coding protein sequence of BYN with the COG database, and a total of 1819 genes were annotated. Among these genes, there are 24 secondary metabolite-related genes (Q), indicating that BYN has the ability to produce a variety of secondary metabolites. In addition, there are 73 genes with unknown functions (S), accounting for 23.25% of the total number of genes, indicating that BYN has the potential to produce new secondary metabolites ( Figure 7 ). Using the KEGG database, the KAAS automatic annotation system was used to annotate 1632 genes in the BYN genome. According to the function, the metabolic pathways provided by the KEGG database were divided into 6 categories: cellular processes, environmental information processing, genetic information processing, human diseases, metabolism, and body systems, with gene counts of 44, 133, 166, 81, 1173, and 35, respectively. In the metabolic category, carbohydrate metabolism genes, cofactor and vitamin metabolism genes, and amino acid metabolism genes accounted for the highest proportion. In the genetic information processing category, the number of translation genes was the largest. In the environmental information processing category, the number of signal transduction genes was the largest. The results showed that strain BYN has strong material metabolism ability, protein synthesis ability, and environmental adaptability ( Figure 8 ).

[0049] Example 2: Application of strain BYN in carrot fermented beverage (1) Preparation of carrot juice: Take commercially available concentrated carrot juice and dilute it with drinking water until the soluble solid content is 10.0±1.0, and adjust the pH to 4.85±0.10. Sterilize it in an autoclave at high temperature and high pressure for 20 min, then cool it for later use.

[0050] (2) Preparation of starter culture: The BYN strain of Lactobacillus reuteri was activated for three generations using MRS liquid medium at an inoculation rate of 3%. The activated fermentation liquid was washed three times with sterile saline and prepared into a culture medium with a bacterial count of 1×10 8 cfu / mL~1×10 9 cfu / mL of starter culture.

[0051] (3) Fermentation: The fermentation agent was inoculated into the prepared carrot juice at a volume ratio of 3%, and the fermentation was carried out in a shaking incubator at 37°C for 36 h. The pH of the fermented carrot juice was measured to be 3.65±0.10, and the number of viable bacteria was approximately 1.2×10 10 cfu / mL.

[0052] (4) Filling: Sterilize the fermented carrot juice in an autoclave at high temperature and high pressure for 20 min, fill it with sterile equipment, and refrigerate it at 0℃~4℃ for later use.

[0053] Example 3: Evaluation of Anti-Helicobacter pylori Activity of Fermented Carrot Juice 1: Determination of the inhibition of Helicobacter pylori growth by carrot juice before and after fermentation The concentration of Hp SS1 suspension was adjusted to 10 8 CFU / mL, take 100 μL of bacterial solution and inoculate it on Columbia blood agar plate, and spread it evenly with a spreading rod. Place the sterilized Oxford cup on the evenly coated plate at equal distances, and add 200 μL of unfermented carrot juice and BYN fermented carrot juice to each well in turn. Place the culture dish in a three-gas incubator and culture it at 37℃ for 48-72 hours before taking it out.

[0054] The results are as follows Fig. 9 As shown in the figure, there is no inhibition zone around the unfermented carrot juice, while there is an obvious inhibition zone around the carrot juice fermented by BYN, with an inhibition zone diameter of 10.45±0.15 mm. According to the drug sensitivity judgment standard in the reference literature: the inhibition zone diameter ≥ 15 mm is highly sensitive; 10 mm≤ inhibition zone diameter <15 mm; 6 mm≤ inhibition zone diameter <10 mm is low sensitivity; inhibition zone diameter <5 mm or no obvious inhibition zone is insensitive. BYN fermented carrot juice is moderately sensitive to Helicobacter pylori.

[0055] 2: Effects of carrot juice before and after fermentation on the growth curve of Helicobacter pylori Hp SS1 was diluted to 0.05 in BHI broth at an absorbance of 600 nm (OD600). It was mixed with unfermented carrot juice and BYN fermented carrot juice at a volume ratio of 3:1. Hp The BHI broth of SS1 was set as the control. The cells were cultured at 37°C in a three-gas incubator (5% O2, 10% CO2, 85% N2), and aliquots were drawn at 0, 12, 24, 36, and 48 h, and the △OD600 was measured using a microplate reader.

[0056] The results are as follows Fig.10 As shown, BYN-fermented carrot juice inhibited the growth of Helicobacter pylori by about two-thirds within 48 hours, and the inhibition rate was significantly higher than that of unfermented carrot juice.

[0057] 3: Determination of the inhibition of Helicobacter pylori urease activity by carrot juice before and after fermentation Use the inoculation loop to scrape fresh activated two generations of Hp SS1, washed twice with sterile PBS, centrifuged (8000 g, 5 min) to obtain bacterial slurry, and then adjusted with BHI Hp The concentration of SS1 is 1×10 7 CFU / mL. In a 96-well plate, take 40 μL Hp SS1 and 10 μL of the sample to be tested (unfermented carrot juice and carrot juice fermented by BYN) were mixed and placed in a three-gas incubator for 48 h. The 96-well plate was taken out and 150 μL of urea-phenol red solution was added. After shaking evenly, the absorbance at 550 nm was measured using an ELISA reader.

[0058] The results are as follows Fig.11 As shown in the figure, the urease inhibition rate of carrot juice fermented by BYN reached 70%, which was about 46% higher than that of unfermented carrot juice, indicating that carrot juice fermented by BYN had significant inhibitory activity on the urease of Hp SS1. Urease is the main factor for pathogen colonization and virulence. Helicobacter pylori promotes the decomposition of urea into NH3 and CO2, and ammonia is used to neutralize gastric acid to ensure the colonization and survival of Helicobacter pylori in the gastric acid environment. Therefore, carrot juice fermented by BYN can destroy the favorable environment for the survival of Helicobacter pylori by inhibiting urease activity, thereby effectively preventing its colonization on the gastric epithelium or mucosa.

[0059] Example 4: Analysis of differential metabolites before and after carrot juice fermentation The changes in metabolites in BYN carrot juice before and after fermentation were determined by LC-MS / MS. The determination was commissioned to Shanghai Meiji Biopharmaceutical Technology Co., Ltd. The specific conditions are as follows.

[0060] Chromatographic conditions: Mobile phase A is 95% water + 5% acetonitrile (containing 0.1% formic acid), and mobile phase B is 47.5% acetonitrile + 47.5% isopropanol + 5% water (containing 0.1% formic acid). Separation gradient: 0-0.1 min, mobile phase B increases linearly from 0% to 5%; 0.1-2 min, mobile phase B increases linearly from 5% to 25%; 2-9 min, mobile phase B increases linearly from 25% to 100%; 9-13 min, mobile phase B maintains linearity at 100%; 13.0-13.1 min, mobile phase B decreases linearly from 100% to 0%; 13.1-16 min, mobile phase B maintains linearity at 0%. Flow rate is 0.40 mL / min, column temperature is 40°C. Mass spectrometry conditions: Sample mass spectrometry signal acquisition adopts positive and negative ion scanning mode, and the mass scanning range is m / z: 70-1050. Ion spray voltage, positive ion voltage 3500 V, negative ion voltage 2800 V, sheath gas 40 psi, auxiliary heating gas 10 psi, ion source heating temperature 400 °C, 20-40-60 V cycle collision energy, MS1 resolution 70000, MS 2 Resolution 17500.

[0061] After the computer was completed, the LC-MS raw data were imported into the metabolomics processing software Progenesis QI (Waters Corporation, Milford, USA) for baseline filtering, peak identification, integration, retention time correction, peak alignment, and mass spectrometry information was matched with the metabolic public database HMDB (http: / / www.hmdb.ca / ) and Metlin (https: / / metlin.scripps.edu / ) as well as the Meiji self-built library to obtain metabolite information. The pre-processed matrix file was subjected to differential analysis.

[0062] By PLS-DA analysis ( Fig.12 ) showed that there were significant differences in metabolites between BYN fermented carrot juice (BYN) and unfermented carrot juice (Car). There were 131 differential metabolites, of which 47 had significantly increased levels after BYN fermentation and 84 had decreased levels. Fig.13 As shown in Table 2, the BYN-fermented carrot juice significantly increased the contents of succinic acid, 3-hydroxybenzoic acid, indole-3-acetamide and other substances compared with the unfermented carrot juice.

[0063] Succinic acid and 3-hydroxybenzoic acid belong to organic acid compounds, which can affect the growth and urease activity of Helicobacter pylori by changing the pH of its living environment. In addition, BYN was found to have a complete metabolic pathway for synthesizing succinic acid in the KEGG annotation. In the tricarboxylic acid cycle, L-malic acid is synthesized into fumaric acid under the action of fumarase (E4.2.1.2), and then succinic acid is synthesized by fumarate reductase (cytochrome) (EC1.3.2.4) and succinate dehydrogenase (ubiquinone) yellow protein subunit (EC1.3.5.1). Indole-3-acetamide is an indole derivative and has important biological functions in lactic acid bacteria metabolites. Various biological activities of indole derivatives have been revealed, including anti-inflammatory, analgesic, antibacterial, antibiotic, anticancer and anti-ulcer effects. The production of these metabolites not only gives carrot juice a better taste and nutritional value, but also acts on anti-Helicobacter pylori activity.

[0064] Table 2: Differential metabolites with significantly increased levels of BYN vs Car

[0065] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for a person skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to replace some of the technical features therein by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions claimed to be protected by the present invention.

Claims

1. A strain of Lactobacillus reuteri BYN, characterized in that: The classification of the Lactobacillus reuteri BYN is named Lactobacillus reuteri Limosilactobacillus reuteri , deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC NO.31737.

2. The Lactobacillus reuteri BYN according to claim 1, characterized in that The 16S rDNA nucleotide sequence of the Lactobacillus reuteri BYN is shown in SEQ ID NO.

1.

3. A fermentation agent for anti-hepatitis beverage, characterized in that: The starter contains the activated Lactobacillus reuteri BYN fermentation liquid of claim 1 and sterile physiological saline.

4. The anti-hepatitis beverage fermentation agent according to claim 3, characterized in that: The bacterial content of Lactobacillus reuteri BYN in the fermentation agent is 1×10 8 cfu / mL~1×10 9 cfu / mL.

5. Use of the Lactobacillus reuteri BYN according to claim 1 in preparing an anti-Helicobacter pylori beverage.

6. The use according to claim 5, characterized in that: The application is to inoculate Lactobacillus reuteri BYN into carrot juice, and obtain the anti-Helicobacter pylori-resistant carrot fermented beverage through fermentation.

7. The use according to claim 5, characterized in that: The steps of the application are: (1) Preparation of carrot juice: Take carrot juice and dilute it with drinking water; sterilize it and cool it for later use; (2) Preparation of starter culture: The BYN strain of Lactobacillus reuteri was activated with MRS liquid culture medium, and the activated fermentation liquid was washed with sterile saline to prepare a starter culture of Lactobacillus reuteri BYN with a bacterial content of 1×10 8 cfu / mL~1×10 9 cfu / mL of starter culture; (3) Fermentation: The fermentation agent is inoculated into the prepared carrot juice at an inoculation rate of 2% to 5% by volume, and the anti-Helicobacter pylori-resistant carrot fermented beverage is prepared by shaking fermentation.

8. The use according to claim 7, characterized in that: The pH of the anti-Helicobacter pylori fermented carrot beverage is 3.65±0.10, and the viable bacterial count is 10 9 cfu / mL~10 10 cfu / mL.

9. An anti-Hepatitis carrot fermented beverage, characterized in that: The anti-Helicobacterium fermented carrot beverage is produced by inoculating the Lactobacillus reuteri BYN described in claim 1 into sterilized carrot juice for fermentation.

10. The anti-hepatic carrot fermented beverage according to claim 9, characterized in that: The anti-Helicobacter pylori fermented carrot beverage contains anti-Helicobacter pylori active ingredients, namely, succinic acid, 3-hydroxybenzoic acid and indole-3-acetamide.

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